KR20210013036A - 신규 벡터 및 그 이용 - Google Patents
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Abstract
포유동물 세포 숙주에 외래 유전자 유래 단백질의 고레벨 생산능을 부여하는 포유동물 세포용 발현 벡터를 제공한다. 디히드로 엽산 환원효소 유전자 결손 숙주 세포 염색체 상의 전사 핫 스팟에 플라스미드 DNA가 조립되고, 히포크산틴 티미딘(이하, 'HT'라 기재함) 불포함 배지 생육주로서 선별되는 구조를 갖는 발현 벡터에 편재성 크로마틴 오프닝 엘리먼트(ubiquitously acting chromatin opening element, UCOE)를 도입함으로써 형질전환체에서의 목적 단백질의 생산량을 증가시켰다.
Description
본 발명은 신규 벡터 및 그 이용에 관한 것이며, 보다 상세하게는 포유동물 세포 숙주에 외래 유전자 유래 단백질의 고(高)레벨 생산능을 부여하는 포유동물 세포용 발현 벡터에 관한 것이다. 본 발명의 발현 벡터는 특히 대장균이나 효모를 숙주로 한 유전자 조환(組換)에서는 충분한 활성이 얻어지기 어렵고, 포유류 독특한 당쇄 수식이나 폴딩이 필요한 포유동물 단백질의 생산에 적합하다.
조환 단백질 생산용 벡터는 다수 개발되고, 대장균을 대표로 하는 세균, 효모를 대표로 하는 진핵 미생물 및 곤충 세포를 숙주로 하는 발현계에서는 단백질의 발현량도 많다. 그렇지만 포유류 독특한 단백질을 발현시키는 경우, 정상적인 입체 구조를 취하지 않거나, 당쇄 부가 등의 번역 후 수식에 문제가 있는 경우가 많다. 여기서 포유류 세포를 숙주로 한 발현계의 확립이 필요하지만, 일반적으로 발현량이 낮은 경우가 많다. 또, 곤충 세포보다도 고등한 동물 세포에 대해서는 조환 바이러스 벡터를 이용한 발현계도 사용되지만, 조환 바이러스 벡터를 발현 단백질로부터 제거하는 작업은 매우 번잡하고, 추가로 바이러스 벡터 자체의 위험성도 부정할 수 없다.
포유동물 세포를 숙주로 한 조환 단백질 생산의 사례는 조직성 플라스미노겐 액티베이터(특허문헌 1), 에리트로포이에틴(특허문헌 2, 비특허문헌 1-3), IFN-γ(비특허문헌 4), IFN-β(특허문헌 3, 비특허문헌 5) 등에서 볼 수 있다. 또, 단일 클론 항체의 유전자 조환 생산에 관해서도 다수의 보고가 있다(특허문헌 4-6, 비특허문헌 6-8). 추가로, 포유동물 세포용 고(高)발현 벡터로서 pNOW/CMV-AA(특허문헌 7)를 들 수 있다. 이 벡터를 이용한 콘글루티닌의 생산량은 배양 4일간에서 최대 11.8 ㎍/㎖였다. 그렇지만, 이들 사례에 있어서 조환 단백질의 충분한 생산량이 얻어지고 있다고는 생각하기 어렵다.
포유동물 세포, 특히 차이니즈 햄스터 난소 세포(이하, 'CHO 세포')를 이용한 의약품의 제조는 안전이 확인되어 있고, 현재 일반적인 수법이 되어 있다. 포유동물 세포를 이용한 조환 단백질 제조에 있어서, 그 생산성을 올리는 것은 비용의 삭감·의료비의 억제 등의 면에서 매우 중요하다. 그 때문에, 효율적인 유전자 도입에 의해 고레벨 생산능을 갖는 형질전환체를 제작하기 위한 발현 벡터의 개발이 필수적이다.
포유동물 세포에 있어서 조환 단백질의 고레벨 생산을 용이하게 행하기 위해서는 효율적인 유전자 도입이 필요하다. 효율적인 유전자 도입이란 클론의 선별이 용이한 것임에도 불구하고 고레벨 생산성 클론이 얻어지는 확률이 높은 것이다. 즉, 피형질전환 세포 전체에 대하여 약제 선택 후의 생존 세포 클론 수가 상대적으로 적고, 그것에 의해 고레벨 생산성 클론의 선별이 용이한 것, 추가로 목적으로 하는 단백질을 생산하는 세포의 수가 적음에도 불구하고 고레벨 생산성 클론이 출현할 기대치가 큰 것이다. 얻어지는 세포의 수가 많으면 그만큼 선별에 시간과 노력을 필요로 하게 되어 효율이 나쁘고, 또 잠재적으로 고레벨 생산능을 갖는 클론을 간과할 가능성도 높다.
고레벨 생산능이란 유전자 도입에 의해 얻어진 형질전환 세포 클론에서의 조환 단백질의 발현량이 많은 것이고, 이것은 주로 발현 벡터의 성질, 성능에 유래한다고 생각된다. 유전자 발현의 레벨은 염색체 상의 위치에 의해 현저하게 다른 것이 발견되어 있고(비특허문헌 9), 염색체 상의 전사 활성이 높은 영역(이하, '전사 핫 스팟')에 목적 유전자가 도입됨으로써 조환 단백질의 생산 레벨이 높아질 것으로 예상된다.
스즈키 등은 디히드로 엽산 환원효소 유전자 결손 숙주 세포 염색체 상의 전사 핫 스팟에 플라스미드 DNA가 조립되고, 히포크산틴 티미딘(이하, 'HT'라 기재함) 불포함 배지 생육주로서 선별되는 구조를 갖는 발현 벡터의 개발에 성공하였다(특허문헌 8). 디히드로 엽산 환원효소(DHFR)는 핵산 염기의 생합성에 필요하기 때문에, 유전 정보 물질로서 DNA를 이용하는 모든 생물에게 필수인 효소이다. 그 때문에 디히드로 엽산 환원효소 유전자 결손 숙주 세포는 핵산의 성분인 HT를 포함하지 않는 배지에서는 생육할 수 없다. 목적 단백질 유전자와 DHFR 유전자를 조립한 구조체(construct)를 디히드로 엽산 환원효소 유전자 결손 숙주 세포에 도입하고, HT 불포함 조건에서 배양함으로써 목적 단백질을 발현하는 세포를 선별할 수 있다. 이 방법은 목적 단백질 유전자와 네오마이신 포스포트랜스퍼라아제 유전자를 조립한 구조체를 도입하고, G418로 선별하는 방법에 비해, DHFR 저해제인 MTX로 유전자 증폭하는 것이 가능하기 때문에, 목적 단백질의 고생산주를 얻는데 보다 적합하다. 그 결과, 고레벨이고 안정적인 단백질의 생산을 가능하게 하는 발현 벡터를 구축할 수 있었다.
Fermentation Bioengineering, 4, 257페이지, 1989년
Proc. Natl. Acad. Sci. USA, 83, 6465페이지, 1986년
Biotechnology, 6, 67페이지, 1988년,
Proc. Natl. Acad. Sci. USA, 80, 4564페이지, 1983년
Cytotechnology, 4, 173페이지, 1990년
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Annu. Rev. Cell Biol., 6, 679페이지, 1990년
본 발명은 포유동물 세포 숙주에 외래 유전자 유래 단백질의 고레벨 생산능을 부여하는 포유동물 세포용 발현 벡터를 제공하는 것을 목적으로 한다. 또, 본 발명은 상기 벡터를 이용한 형질전환체의 제조 방법, 및 상기 벡터를 이용한 외래 유전자 유래 단백질의 생산 방법을 제공하는 것도 목적으로 한다.
본 발명자들은 스즈키 등이 개발한 디히드로 엽산 환원효소 유전자 결손 숙주 세포 염색체 상의 전사 핫 스팟에 플라스미드 DNA가 조립되고, 히포크산틴 티미딘(이하, 'HT'라 기재함) 불포함 배지 생육주로서 선별되는 구조를 갖는 발현 벡터에 보편적 크로마틴 오프닝 엘리먼트(ubiquitously acting chromatin opening element, UCOE)를 도입함으로써, 형질전환체에서의 목적 단백질의 생산량을 증가시키는 것과 함께, 발현 안정성을 높이는 것에 성공하였다.
본 발명의 요지는 이하와 같다.
(1) 하기 (a), (b) 및 (c)를 포함하는 발현 벡터:
(a) 코돈을 개변한 번역 장해성 디히드로 엽산 환원효소 유전자 카세트(번역 장해성 DHFR 유전자 카세트)로서, 번역 장해성 DHFR 유전자 카세트의 코돈이, 알라닌은 GCA, 아르기닌은 CGA, 아스파라긴은 AAU, 아스파라긴산은 GAU, 시스테인은 UGU, 글루타민은 CAA, 글루탐산은 GAA, 글리신은 GGU, 히스티딘은 CAU, 루이신은 UUA, 리신은 AAA, 프롤린은 CCA, 페닐알라닌은 UUU, 세린은 UCA, 트레오닌은 ACU, 티로신은 UAU, 및/또는 발린은 GUA로 개변되어 있고, 코돈의 개변 영역이 DHFR 유전자의 5' 말단으로부터 DHFR 유전자의 전장(全長)의 30% 이상인 번역 장해성 DHFR 유전자 카세트;
(b) 전사 활성 프로모터와 안정성 폴리아데닐화 신호의 사이에 외래 유전자 조립용 클로닝 사이트를 포함하는 유전자 카세트; 및
(c) 편재성 크로마틴 오프닝 엘리먼트(UCOE).
(2) (1)(a)에 기재된 번역 장해성 DHFR 유전자 카세트가 프로모터로서 비포유동물 세포의 유전자를 유래로 하는 프로모터, 또는 인핸서(enhancer) 부분을 제거한 프로모터를 사용하고 있는 것을 특징으로 하는 (1)에 기재된 발현 벡터.
(3) UCOE가 서열번호 1의 뉴클레오티드 서열을 포함하는 (1) 또는 (2)에 기재된 발현 벡터.
(4) (1)∼(3) 중 어느 하나에 기재된 발현 벡터에 외래 유전자를 조립하는 공정, 및 상기 발현 벡터에 의해 디히드로 엽산 환원효소 유전자 결손 숙주 세포를 형질전환시키는 공정을 포함하는 외래 유전자 유래 단백질을 생산하는 형질전환체의 제조 방법.
(5) 이하 (a)∼(d)의 공정을 포함하는 외래 유전자 유래 단백질의 생산 방법:
(a) (1)∼(3) 중 어느 하나에 기재된 발현 벡터에 외래 유전자를 조립하는 공정,
(b) 상기 발현 벡터에 의해 디히드로 엽산 환원효소 유전자 결손 숙주 세포를 형질전환시키는 공정,
(c) 상기 형질전환체를 히포크산틴 티미딘 불포함 배지에서 배양하는 공정,
(d) 배양된 형질전환체로부터 외래 유전자 유래 단백질을 회수하는 공정.
(6) (5)(c)의 공정에 있어서, 화학적 구명 배지(Chemically Defined medium, CD 배지) 혹은 CD 배지에 비동물성의 첨가물을 가한 배지에서 배양하는 것을 특징으로 하는 (5)에 기재된 방법.
(7) 이하 (a)∼(c)의 공정을 포함하는 외래 유전자 유래 단백질을 생산하는 형질전환체의 스크리닝 방법:
(a) (1)∼(3) 중 어느 하나에 기재된 발현 벡터에 외래 유전자를 조립하는 공정,
(b) 상기 발현 벡터에 의해 디히드로 엽산 환원효소 유전자 결손 숙주 세포를 형질전환시키는 공정,
(c) 상기 형질전환체를 히포크산틴 티미딘 불포함 배지에서 배양하는 공정.
(8) (1)∼(3) 중 어느 하나에 기재된 발현 벡터에 외래 유전자가 조립한 외래 유전자 발현 벡터.
(9) (8) 기재의 외래 유전자 발현 벡터에 의해 형질전환된 숙주 세포.
본 명세서는 본원의 우선권의 기초인 일본 특허 출원 특원2018-99704 및 특원2018-168591의 명세서 및/또는 도면에 기재되는 내용을 포함한다.
도 1은 pDC62c5-U533 구조체를 나타내는 도면이다. 각각, PCMV5: 인간 사이토메갈로바이러스 프로모터와 아데노바이러스 프로모터의 융합 프로모터인 CMV5 프로모터(Nucleic Acid Research, 30, 2페이지, 2002년), PABGH: 소 성장 호르몬 유전자 폴리아데닐화 신호, UCOE: 편재성 크로마틴 오프닝 엘리먼트, PdSV: 인핸서를 결실시킨 시미안 바이러스(Simian Virus) 40 프로모터, mDHFR: 번역 장해성 디히드로 엽산 환원효소 유전자, SV40pA: 시미안 바이러스 40 유래 폴리아데닐화 신호, AMP: 대장균 중에서의 선택 마커(β-락타마아제 유전자), ori: pUC 플라스미드 유래의 복제 기점을 나타낸다. 또, AscI, AsiSI, NotI, SbfI는 제한 효소 절단 부위를 나타낸다.
도 2는 pDC62c5-U533-OMLH 구조체를 나타내는 도면이다. 각각, PCMV5: 인간 사이토메갈로바이러스 프로모터와 아데노바이러스 프로모터의 융합 프로모터인 CMV5 프로모터(Nucleic Acid Research, 30, 2페이지, 2002년), IgL-OM: 오말리즈맙(Omalizmab) 항체 경쇄 cDNA, PABGH: 소 성장 호르몬 유전자 폴리아데닐화 신호, UCOE: 편재성 크로마틴 오프닝 엘리먼트, IgH-OM: 오말리즈맙 항체 중쇄 cDNA, PdSV: 인핸서를 결실시킨 시미안 바이러스 40 프로모터, mDHFR: 번역 장해성 디히드로 엽산 환원효소 유전자, SV40pA: 시미안 바이러스 40 폴리아데닐화 신호, AMP: 대장균 중에서의 선택 마커(β-락타마아제 유전자), ori: pUC 플라스미드 유래의 복제 기점을 나타낸다. 또, AscI, AsiSI, NotI, SbfI는 제한 효소 절단 부위를 나타낸다.
도 3은 pDC61-OMLH 구조체를 나타내는 도면이다. 각각, PCMV: 인간 사이토메갈로바이러스 프로모터(Nucleic Acid Research, 30, 2페이지, 2002년), INRBG: 토끼 β-글로빈 유전자 인트론 서열, IgL-OM: 오말리즈맙 항체 경쇄 cDNA, PABGH: 소 성장 호르몬 유전자 폴리아데닐화 신호, IgH-OM: 오말리즈맙 항체 중쇄 cDNA, PdSV: 인핸서를 결실시킨 시미안 바이러스 40 프로모터, mDHFR: 번역 장해성 디히드로 엽산 환원효소 유전자, SV40pA: 시미안 바이러스 40 폴리아데닐화 신호, AMP: 대장균 중에서의 선택 마커(β-락타마아제 유전자), ori: pUC 플라스미드 유래의 복제 기점을 나타낸다. 또, AscI, AsiSI, NotI, SbfI는 제한 효소 절단 부위를 나타낸다.
도 4는 pNC32c-U533-OMLH 구조체를 나타내는 도면이다. 각각, PCMV: 인간 사이토메갈로바이러스 프로모터(Nucleic Acid Research, 30, 2페이지, 2002년), IgL-OM: 오말리즈맙 항체 경쇄 cDNA, SV40pA: 시미안 바이러스 40 폴리아데닐화 신호, UCOE: 편재성 크로마틴 오프닝 엘리먼트, IgH-OM: 오말리즈맙 항체 중쇄 cDNA, PSV40: 시미안 바이러스 40 프로모터, NTP: 네오마이신 포스포트랜스퍼라아제 유전자, SV40pA: 시미안 바이러스 40 폴리아데닐화 신호, AMP: 대장균 중에서의 선택 마커(β-락타마아제 유전자), ori: pUC 플라스미드 유래의 복제 기점을 나타낸다. 또, AscI, AsiSI, NotI, SbfI는 제한 효소 절단 부위를 나타낸다.
도 5는 pUCOE-Hu-P2-OMLH 구조체를 나타내는 도면이다. 각각, UCOE: 편재성 크로마틴 오프닝 엘리먼트, PgpCMV: 모르모트 사이토메갈로바이러스 프로모터(Nucleic Acid Research, 30, 2페이지, 2002년), IgL-OM: 오말리즈맙 항체 경쇄 cDNA, SV40pA: 시미안 바이러스 40 폴리아데닐화 신호, IgH-OM: 오말리즈맙 항체 중쇄 cDNA, PPGK: 마우스 포스포글리세레이트 키나아제(kinase) 프로모터, PUR: 퓨로마이신 N-아세틸트랜스퍼라아제 유전자, PAC2: 보체 제2 인자 유전자 폴리아데닐화 신호, AMP: 대장균 중에서의 선택 마커(β-락타마아제 유전자), ori: pUC 플라스미드 유래의 복제 기점을 나타낸다. 또, AscI, AsiSI, NotI, SbfI는 제한 효소 절단 부위를 나타낸다.
도 6은 pDC62c5-U533-OMLH(본 발명의 발현 벡터), pDC61-OMLH(종래 벡터), pNC32c-U533-OMLH(컨트롤 벡터) 또는 pUCOE-Hu-P2/OMLH(Millipore사 벡터)를 트랜스펙션(transfection)한 CHO 세포에 의해 생산된 항체 발현량의 분포를 나타낸다(3일간 배양).
도 7은 pDC62c5-U533-OMLH(본 발명의 발현 벡터) 또는 pDC61-OMLH(종래 벡터)를 트랜스펙션한 CHO 세포에 의해 생산된 항체량의 변화를 나타낸다(배양 개시부터의 주수 1-12).
도 8은 외래 유전자 삽입 부위가 1개 있는 pDC62c5-U533 구조체를 나타내는 도면이다. 각각, PCMV5: 인간 사이토메갈로바이러스 프로모터와 아데노바이러스 프로모터의 융합 프로모터인 CMV5 프로모터(Nucleic Acid Research, 30, 2페이지, 2002년), PABGH: 소 성장 호르몬 유전자 폴리아데닐화 신호, UCOE: 편재성 크로마틴 오프닝 엘리먼트, PdSV: 인핸서를 결실시킨 시미안 바이러스 40 프로모터, mDHFR: 번역 장해성 디히드로 엽산 환원효소 유전자, SV40pA: 시미안 바이러스 40 유래 폴리아데닐화 신호, AMP: 대장균 중에서의 선택 마커(β-락타마아제 유전자), ori: pUC 플라스미드 유래의 복제 기점을 나타낸다. 또, AscI, AsiSI, ClaI는 제한 효소 절단 부위를 나타낸다.
도 9는 pDC62c5-U533 벡터에 의해 수립한 고발현 CHO 세포주에 의한 개(犬) CTLA-4-Ig의 발현을 나타낸다. 14일간의 배양에서의 각 일수에서의 개 CTLA-4-Ig 발현량과 생존 세포 수를 나타낸다.
도 10은 배양 상청으로부터 정제한 개 CTLA-4-Ig의 SDS-PAGE 상을 나타낸다. 환원 조건 및 비환원 조건 하에서 개 CTLA-4-Ig를 전기영동하고 CBB 염색에 의해 가시화하였다. 환원 조건에서는 45 kDa 부근에, 비환원 조건에서는 90 kDa 부근에 단일 밴드를 볼 수 있고, 목적 이외의 밴드는 거의 검출되지 않았다.
도 11은 pDC62c5-U533-TRLH 구조체를 나타내는 도면이다.
도 12는 pDC62c5-U533-TRLH(본 발명의 발현 벡터)를 도입한 CHO 세포에 의해 생산된 항체 발현량의 분포를 나타낸다(3일간 배양).
도 13은 pDC62c5-U533-TRLH(본 발명의 발현 벡터)를 트랜스펙션한 CHO 세포에 의해 생산된 항체량의 변화를 나타낸다(생산성 시험 배양 개시부터의 주수 0-10).
도 14는 pDC62c5-U533-OMLH(본 발명의 발현 벡터)를 트랜스펙션한 CHO 세포(Clone01-10)를 접종한 플레이트로부터 얻어진 클론 각각을 60 nM MTX 존재 하에 배양하여 얻어진 세포주에 의해 생산된 항체량을 나타낸다(3일간 배양).
도 2는 pDC62c5-U533-OMLH 구조체를 나타내는 도면이다. 각각, PCMV5: 인간 사이토메갈로바이러스 프로모터와 아데노바이러스 프로모터의 융합 프로모터인 CMV5 프로모터(Nucleic Acid Research, 30, 2페이지, 2002년), IgL-OM: 오말리즈맙(Omalizmab) 항체 경쇄 cDNA, PABGH: 소 성장 호르몬 유전자 폴리아데닐화 신호, UCOE: 편재성 크로마틴 오프닝 엘리먼트, IgH-OM: 오말리즈맙 항체 중쇄 cDNA, PdSV: 인핸서를 결실시킨 시미안 바이러스 40 프로모터, mDHFR: 번역 장해성 디히드로 엽산 환원효소 유전자, SV40pA: 시미안 바이러스 40 폴리아데닐화 신호, AMP: 대장균 중에서의 선택 마커(β-락타마아제 유전자), ori: pUC 플라스미드 유래의 복제 기점을 나타낸다. 또, AscI, AsiSI, NotI, SbfI는 제한 효소 절단 부위를 나타낸다.
도 3은 pDC61-OMLH 구조체를 나타내는 도면이다. 각각, PCMV: 인간 사이토메갈로바이러스 프로모터(Nucleic Acid Research, 30, 2페이지, 2002년), INRBG: 토끼 β-글로빈 유전자 인트론 서열, IgL-OM: 오말리즈맙 항체 경쇄 cDNA, PABGH: 소 성장 호르몬 유전자 폴리아데닐화 신호, IgH-OM: 오말리즈맙 항체 중쇄 cDNA, PdSV: 인핸서를 결실시킨 시미안 바이러스 40 프로모터, mDHFR: 번역 장해성 디히드로 엽산 환원효소 유전자, SV40pA: 시미안 바이러스 40 폴리아데닐화 신호, AMP: 대장균 중에서의 선택 마커(β-락타마아제 유전자), ori: pUC 플라스미드 유래의 복제 기점을 나타낸다. 또, AscI, AsiSI, NotI, SbfI는 제한 효소 절단 부위를 나타낸다.
도 4는 pNC32c-U533-OMLH 구조체를 나타내는 도면이다. 각각, PCMV: 인간 사이토메갈로바이러스 프로모터(Nucleic Acid Research, 30, 2페이지, 2002년), IgL-OM: 오말리즈맙 항체 경쇄 cDNA, SV40pA: 시미안 바이러스 40 폴리아데닐화 신호, UCOE: 편재성 크로마틴 오프닝 엘리먼트, IgH-OM: 오말리즈맙 항체 중쇄 cDNA, PSV40: 시미안 바이러스 40 프로모터, NTP: 네오마이신 포스포트랜스퍼라아제 유전자, SV40pA: 시미안 바이러스 40 폴리아데닐화 신호, AMP: 대장균 중에서의 선택 마커(β-락타마아제 유전자), ori: pUC 플라스미드 유래의 복제 기점을 나타낸다. 또, AscI, AsiSI, NotI, SbfI는 제한 효소 절단 부위를 나타낸다.
도 5는 pUCOE-Hu-P2-OMLH 구조체를 나타내는 도면이다. 각각, UCOE: 편재성 크로마틴 오프닝 엘리먼트, PgpCMV: 모르모트 사이토메갈로바이러스 프로모터(Nucleic Acid Research, 30, 2페이지, 2002년), IgL-OM: 오말리즈맙 항체 경쇄 cDNA, SV40pA: 시미안 바이러스 40 폴리아데닐화 신호, IgH-OM: 오말리즈맙 항체 중쇄 cDNA, PPGK: 마우스 포스포글리세레이트 키나아제(kinase) 프로모터, PUR: 퓨로마이신 N-아세틸트랜스퍼라아제 유전자, PAC2: 보체 제2 인자 유전자 폴리아데닐화 신호, AMP: 대장균 중에서의 선택 마커(β-락타마아제 유전자), ori: pUC 플라스미드 유래의 복제 기점을 나타낸다. 또, AscI, AsiSI, NotI, SbfI는 제한 효소 절단 부위를 나타낸다.
도 6은 pDC62c5-U533-OMLH(본 발명의 발현 벡터), pDC61-OMLH(종래 벡터), pNC32c-U533-OMLH(컨트롤 벡터) 또는 pUCOE-Hu-P2/OMLH(Millipore사 벡터)를 트랜스펙션(transfection)한 CHO 세포에 의해 생산된 항체 발현량의 분포를 나타낸다(3일간 배양).
도 7은 pDC62c5-U533-OMLH(본 발명의 발현 벡터) 또는 pDC61-OMLH(종래 벡터)를 트랜스펙션한 CHO 세포에 의해 생산된 항체량의 변화를 나타낸다(배양 개시부터의 주수 1-12).
도 8은 외래 유전자 삽입 부위가 1개 있는 pDC62c5-U533 구조체를 나타내는 도면이다. 각각, PCMV5: 인간 사이토메갈로바이러스 프로모터와 아데노바이러스 프로모터의 융합 프로모터인 CMV5 프로모터(Nucleic Acid Research, 30, 2페이지, 2002년), PABGH: 소 성장 호르몬 유전자 폴리아데닐화 신호, UCOE: 편재성 크로마틴 오프닝 엘리먼트, PdSV: 인핸서를 결실시킨 시미안 바이러스 40 프로모터, mDHFR: 번역 장해성 디히드로 엽산 환원효소 유전자, SV40pA: 시미안 바이러스 40 유래 폴리아데닐화 신호, AMP: 대장균 중에서의 선택 마커(β-락타마아제 유전자), ori: pUC 플라스미드 유래의 복제 기점을 나타낸다. 또, AscI, AsiSI, ClaI는 제한 효소 절단 부위를 나타낸다.
도 9는 pDC62c5-U533 벡터에 의해 수립한 고발현 CHO 세포주에 의한 개(犬) CTLA-4-Ig의 발현을 나타낸다. 14일간의 배양에서의 각 일수에서의 개 CTLA-4-Ig 발현량과 생존 세포 수를 나타낸다.
도 10은 배양 상청으로부터 정제한 개 CTLA-4-Ig의 SDS-PAGE 상을 나타낸다. 환원 조건 및 비환원 조건 하에서 개 CTLA-4-Ig를 전기영동하고 CBB 염색에 의해 가시화하였다. 환원 조건에서는 45 kDa 부근에, 비환원 조건에서는 90 kDa 부근에 단일 밴드를 볼 수 있고, 목적 이외의 밴드는 거의 검출되지 않았다.
도 11은 pDC62c5-U533-TRLH 구조체를 나타내는 도면이다.
도 12는 pDC62c5-U533-TRLH(본 발명의 발현 벡터)를 도입한 CHO 세포에 의해 생산된 항체 발현량의 분포를 나타낸다(3일간 배양).
도 13은 pDC62c5-U533-TRLH(본 발명의 발현 벡터)를 트랜스펙션한 CHO 세포에 의해 생산된 항체량의 변화를 나타낸다(생산성 시험 배양 개시부터의 주수 0-10).
도 14는 pDC62c5-U533-OMLH(본 발명의 발현 벡터)를 트랜스펙션한 CHO 세포(Clone01-10)를 접종한 플레이트로부터 얻어진 클론 각각을 60 nM MTX 존재 하에 배양하여 얻어진 세포주에 의해 생산된 항체량을 나타낸다(3일간 배양).
본 발명자들은 DHFR 유전자의 코돈을 포유동물에 있어서 사용 빈도가 가장 낮은 것으로 개변하여 DHFR의 발현성을 매우 감쇠시킨 것으로 함으로써, 조립한 플라스미드 유전자가 디히드로 엽산 환원효소 유전자 결손 숙주 세포 염색체 상의 매우 높은 발현성을 갖는 위치에 도입되지 않는 한, 형질전환체이기는 해도 HT가 포함되지 않은 배지에서의 선별에 대하여 생존이 곤란하도록 하였다. 또, 플라스미드 유전자에 UCOE(편재성 크로마틴 오프닝 엘리먼트)를 도입함으로써, 형질전환체에서의 목적 단백질의 생산량을 증가시키는 것과 함께, 발현 안정성을 높이도록 하였다.
즉, 본 발명은 포유동물 숙주 세포에 있어서 유전자 조환 단백질의 고레벨 생산을 유도하기 위한 발현 벡터를 제공한다.
본 발명의 발현 벡터는 백본(backbone) 벡터 상에 하기 (a), (b) 및 (c)를 포함하는 것에 의해 구축된다.
(a) 코돈을 포유동물에 있어서 사용 빈도가 가장 낮은 것으로 개변하여 발현을 미약화시킨 번역 장해성 디히드로 엽산 환원효소 유전자 카세트(번역 장해성 DHFR 유전자 카세트)
(b) 프로모터와 폴리아데닐화 신호의 사이에 외래 유전자 조립용 클로닝 사이트를 포함하는 유전자 카세트
(c) 편재성 크로마틴 오프닝 엘리먼트(UCOE).
본 발명은 DHFR 유전자 카세트(시스트론)의 구성으로서, DHFR 유전자의 코돈을 포유동물에 있어서 사용 빈도가 가장 낮은 코돈으로 개변하고 DHFR의 발현 유도성을 저하시킨 프로모터를 사용하여, 유전자 도입에 의해 형질전환된 숙주 세포에서의 DHFR의 발현 기구를 현저하게 장해시킨 것이다. 본 발명에 있어서, 「유전자 카세트」란 프로모터, 구조 유전자, 폴리아데닐화 신호(polyA)를 기본 구성으로 하는 전사·번역에 의해 단백질을 발현하는 단위를 의미하지만, 이들 중 어느 하나에 관련되거나 혹은 임의의 DNA 서열을 삽입 서열로서 포함하고 있어도 된다. 본 발명의 DHFR 유전자 카세트는 프로모터를 약화시킬 뿐인 것과는 달리, 전사 핫 스팟에 플라스미드 유전자가 도입된 HT 불포함 배지 생육주를 특이적으로 획득할 수 있도록 유도하는 것으로부터, 「번역 장해성 DHFR 유전자 카세트」라고 정의한다.
본 발명에 있어서, 「포유동물에 있어서 사용 빈도가 가장 낮은 코돈」이란 인간에 있어서 사용 빈도가 가장 낮은 코돈을 바람직한 예로서 들 수 있다. 인간에 있어서 사용 빈도가 가장 낮은 코돈으로는 Kim 등의 문헌(Gene, 199, 293페이지, 1997년)에 개시된 코돈을 들 수 있다. 보다 구체적인 코돈으로서, 알라닌은 GCA, 아르기닌은 CGA, 아스파라긴은 AAU, 아스파라긴산은 GAU, 시스테인은 UGU, 글루타민은 CAA, 글루탐산은 GAA, 글리신은 GGU, 히스티딘은 CAU, 루이신은 UUA, 리신은 AAA, 프롤린은 CCA, 페닐알라닌은 UUU, 세린은 UCA, 트레오닌은 ACU, 티로신은 UAU, 및/또는 발린은 GUA를 예시할 수 있지만, 코돈은 이들에 한정되는 것은 아니다.
본 발명에 있어서, 「발현을 미약화하는」이란 유전자의 발현이 전사 및/또는 번역의 단계에서 약화되어 있는 것을 나타내며, 구체적으로는 코돈을 상기 「포유동물에 있어서 사용 빈도가 가장 낮은 코돈」으로 개변함으로써 얻을 수 있다.
상기 「번역 장해성 DHFR 유전자 카세트」에 있어서, 코돈이 개변되는 영역은 특별히 제한되는 것은 아니지만, 유전자 카세트의 전장의 30% 이상(예를 들면, 40% 이상, 50% 이상, 60% 이상, 70% 이상, 80% 이상, 90% 이상, 95% 이상, 또는 100%)의 영역에서의 코돈이 개변되는 것이 바람직하다. 코돈의 개변 영역의 범위는 벡터의 다른 조건을 고려하여 임의로 결정할 수 있다.
상기 「번역 장해성 DHFR 유전자 카세트」의 프로모터로는 포유동물 세포에서는 통상 발현하기 어려운 단백질 유전자의 프로모터(예를 들면, 비포유동물 세포의 유전자를 유래로 하는 프로모터)를 유래로 하는 것, 또는 통상의 프로모터로부터 인핸서를 제거한 것을 사용할 수 있고, 보다 구체적으로는 SV40의 바이러스 항원 프로모터로부터 인핸서 영역을 제거한 것이거나(Mol. Cell Biol., 6, 2593페이지, 1986년), 또는 이것과 동등하게 발현성이 매우 낮은 프로모터를 사용하는 것이 바람직하다.
플라스미드 DNA의 디히드로 엽산 환원효소 유전자 결손 숙주 세포 염색체 상의 전사 핫 스팟으로의 조립은 DHFR 유전자 카세트가 갖는 특성으로부터 HT를 포함하지 않는 배지에서의 선별에 의해 결과적으로 달성되게 되지만, 염색체 상의 전사 핫 스팟에서의 외래 유전자 유래 단백질의 발현 자체는 강력하게 유도될 필요가 있다. 이 때문에, 단백질 유전자를 조립하는 클로닝 사이트(이하, 'CS'라 기재함)의 프로모터 및 폴리아데닐화 신호(이하, 'polyA'라 칭함)는 가장 강한 발현 유도성을 갖는 것 중에서 선택하면 된다. 프로모터로는 인간 사이토메갈로바이러스 즉시 초기(Immediate Early)(hCMV MIE: Cell, 41, 521페이지, 1985년) 프로모터, 인간 사이토메갈로바이러스 프로모터와 아데노바이러스 프로모터의 융합 프로모터인 CMV5 프로모터(Nucleic Acid Research, 30, 2페이지, 2002년), β-액틴 프로모터(Proc. Natl. Acad. Sci. USA, 84, 4831페이지, 1987년), polyA에는 소 성장 호르몬 유래의 polyA 서열(DNA, 5,115페이지, 1986년) 등을 들 수 있다. 클로닝 사이트는 제한 효소 절단 부위에 의해 구성된다. 제한 효소 절단 부위로는 Asc I, AsiS I, Acc65I, BamHI, BclI, BsaI, BsiWI, BstBI, BstEI, Bsu36I, DraIII, EagI, FseI, KpnI, MboI, Nhe I, Not I, PacI, RsrII, SalI, Sbf I, SexAI, SgfI, XcmI 등을 들 수 있다. 본 명세서 중에서는 이 목적으로 하는 단백질 유전자를 조립하는 클로닝 사이트를 갖는 DNA 단편을 「유전자 발현 카세트」라 칭한다.
UCOE란 편재적으로 고발현하고 있는 유전자에 근접하여 존재하는 DNA 엘리먼트이며, 오픈 크로마틴 환경을 형성함으로써, 그 근린에 도입된 유전자가 메신저 RNA로 전사될 가능성을 최대한으로 높이고, 나아가서는 도입된 유전자가 코드하고 있는 단백질의 발현량을 최대로 하는 능력을 갖는다. 또, UCOE는 프로모터 영역의 DNA 메틸화를 억제하는 것에 의한 안티사이런싱 활성을 갖고, 장기에 걸친 계대 배양에 의한 도입된 유전자의 전사 활성의 저하를 억제하고, 나아가서는 도입된 유전자가 코드하고 있는 단백질의 발현량의 저하를 억제할 수 있다. 그 때문에, 본 발명의 발현 벡터에 UCOE를 도입함으로써, 형질전환체에서의 목적 단백질의 생산량을 증가시키는 것과 함께, 발현 안정성을 높이는 것을 기대할 수 있다.
후술하는 실시예에서 발현 벡터에 도입한 UCOE의 뉴클레오티드 서열을 서열번호 1에 나타낸다. 본 발명의 목적을 달성하는 한, 서열번호 1의 뉴클레오티드 서열에는 뉴클레오티드 염기의 치환, 뉴클레오티드의 결실이나 삽입 등의 변이가 도입되어도 된다. 발현 벡터에 도입하는 UCOE의 개수는 1개 또는 그 이상이어도 된다. UCOE의 도입 부위는 발현하고자 하는 유전자 카세트를 끼운 형태로 존재하면 된다.
본 발명의 발현 벡터는 선택 마커로서 약제 내성 유전자(예를 들면, 암피실린 내성 유전자, 카나마이신 내성 유전자, 클로람페니콜 내성 유전자 등), 복제 기점(예를 들면, pUC 유래의 복제 기점, ColE1 유래의 복제 기점, p15A 유래의 복제 기점, pSC101 유래의 복제 기점 등)을 탑재하면 된다.
본 발명의 발현 벡터로는 실시예에서 구체적으로 기재한 발현 벡터(pDC62c5-U533)를 예시할 수 있지만, 이것에 제한되는 것은 아니다.
또, 본 발명은 상기 발현 벡터에 외래 유전자를 조립하는 공정, 및 상기 발현 벡터에 의해 디히드로 엽산 환원효소 유전자 결손 숙주 세포를 형질전환시키는 공정을 포함하는 외래 유전자 유래 단백질을 생산하는 형질전환체의 제조 방법을 제공한다. 상기 형질전환체는 외래 유전자 유래 단백질의 고레벨 생산능 및 HT를 포함하지 않는 배지에서의 생육능을 가질 수 있다.
구체적으로는, 발현시키고자 하는 단백질을 코드하는 외래 유전자를 본 발명의 발현 벡터의 클로닝 사이트(이하, 'CS'로 기재)에 조립한 후, 트랜스펙션법(여기서 말하는 트랜스펙션법으로는 리포펙틴법, 전기천공법, 인산칼슘법, 마이크로인젝션법 등 통상의 기술자가 주지인 방법을 들 수 있음)을 이용하여 상기 발현 벡터에 의해 디히드로 엽산 환원효소 유전자 결손 숙주 세포를 형질전환하고, HT를 포함하지 않는 배지에서의 내성으로 선별을 행하여, 단백질의 고생산성 형질전환체를 얻는 방법을 들 수 있다. 예를 들면, pDC62c5-U533의 경우, Asc I와 Sbf I의 사이에 외래 유전자 cDNA를 삽입하면 된다. 외래 유전자가 항체 유전자인 경우에는 pDC62c5-U533의 Asc I와 AsiS I의 사이에 항체 경쇄 유전자 cDNA를, Not I와 Sbf I의 사이에 항체 중쇄 유전자 cDNA를 삽입하면 된다. pDC62c5-U533에 있어서는 Asc I와 AsiS I, 및 Not I와 Sbf I가 클로닝 사이트가 된다. 예를 들면, 외래 유전자 삽입 부위가 1개 있는 pDC62c5-U533의 경우, Asc I와 AsiS I의 사이에 외래 유전자 cDNA를 삽입하면 된다. 외래 유전자 삽입 부위가 1개 있는 pDC62c5-U533에 있어서는 Asc I와 AsiS I가 클로닝 사이트가 된다. 외래 유전자에는 Kozak를 부가하고, 본 발명의 발현 벡터에 조립하면 된다. Kozak는 최적화되면 되고, 최적화된 Kozak의 서열을 서열번호 2에 나타낸다. Kozak는 외래 유전자 cDNA의 개시 코돈의 상류에 부가하면 된다.
본 발명은 본 발명의 발현 벡터에 외래 유전자가 조립된 외래 유전자 발현 벡터도 제공한다.
본 발명에 있어서, 숙주 세포로는 외래 유전자 유래 단백질의 발현에 적절한 세포이면 특별히 제한은 없지만, 바람직하게는 디히드로 엽산 환원효소 유전자 결손 포유동물 세포, 보다 바람직하게는 디히드로 엽산 환원효소 유전자를 결손한 차이니즈 햄스터 난소 세포(CHO 세포)를 들 수 있다.
HT 불포함 배지에서의 선별에서 생존한 형질전환 세포의 상당수는 이미 상대적으로 단백질의 높은 발현 레벨을 달성하고 있지만, 그 중에서 추가로 고레벨 생산능을 갖는 형질전환 세포를 선택하기 위하여 단백질의 발현 레벨을 측정해도 된다.
본 발명은 본 발명의 외래 유전자 발현 벡터에 의해 형질전환된 숙주 세포도 제공한다.
또, 본 발명은 이하 (a)∼(d)의 공정을 포함하는 외래 유전자 유래 단백질의 생산 방법을 제공한다:
(a) 본 발명의 발현 벡터에 외래 유전자를 조립하는 공정,
(b) 상기 발현 벡터에 의해 디히드로 엽산 환원효소 유전자 결손 숙주 세포를 형질전환시키는 공정,
(c) 상기 형질전환체를 HT 불포함 배지에서 배양하는 공정,
(d) 배양된 형질전환체로부터 외래 유전자 유래 단백질을 회수하는 공정.
본 발명에 있어서, 상기 (c)의 공정에 있어서는 HT 불포함 배지에서 배양을 행함으로써, 고효율의 단백질 발현을 나타내는 형질전환체(콜로니)를 선택할 수 있다. 선택된 형질전환체는 계속해서 동일한 배지에 있어서 배양을 계속해도 되고, 다른 배지, 예를 들면 대량 발현용의 배지로 옮겨서 배양을 행해도 된다.
본 발명에 있어서, 형질전환체를 배양 또는 순화시키는 배지는 특별히 제한되지 않지만, 바람직하게는 무혈청 배지, 보다 바람직하게는 CD 배지 혹은 CD 배지에 비동물성의 첨가물(예를 들면, 염류나 아미노산, 당, 비타민류, 조환 인슐린, 조환 트랜스페린)을 가한 배지를 예시할 수 있다.
본 발명에 있어서, 배양된 형질전환체로부터 외래 유전자 유래 단백질을 회수할 때에는 통상의 기술자에게 공지인 방법(필터레이션(여과), 원심분리, 및 컬럼 정제 등)으로 단백질을 정제해도 된다. 외래 유전자 유래 단백질은 정제를 용이하게 하는 등의 목적으로 다른 단백질과의 융합 단백질로서 발현시키는 것도 가능하다.
추가로, 본 발명은 이하 (a)∼(c)의 공정을 포함하는 외래 유전자 유래 단백질을 생산하는 형질전환체의 스크리닝 방법을 제공한다:
(a) 본 발명의 발현 벡터에 외래 유전자를 조립하는 공정,
(b) 상기 발현 벡터에 의해 디히드로 엽산 환원효소 유전자 결손 숙주 세포를 형질전환시키는 공정,
(c) 상기 형질전환체를 HT 불포함 배지에서 배양하는 공정.
또한, 본 명세서에 있어서, 인용된 모든 선행 기술 문헌은 참조로서 본 명세서에 조립된다.
[실시예]
이하, 본 발명의 실시예를 설명한다.
[실시예 1] pDC62c5-U533의 제작
pDC6(일본 특허 5704753)을 개변하여 본 발명의 벡터인 pDC62c5-U533를 구축하였다. 백본 벡터 pDC62c5-U533의 전체 염기 서열을 서열번호 3에 기재한다. pDC62c5-U533는 6067-6630의 영역에 DHFR의 염기 서열을 5' 말단에서 180 염기의 범위에서 코돈을 포유동물에 있어서 사용 빈도가 가장 낮은 것으로 개변한 번역 장해성 DHFR 유전자가 도입되어 있다(도 1). 또, 1339-2889, 4234-5784 및 9771-11321의 영역에 UCOE가 도입되어 있다. UCOE의 염기 서열을 서열번호 1에 기재한다.
[실시예 2] pDC62c5-U533-OMLH의 제작
pDC62c5-U533의 염기 서열 No1098-No1108를 서열번호 4에 기재한 인간 오말리즈맙 항체(OMLH) 경쇄를 코드하는 cDNA(개시 코돈의 상류에 최적화한 Kozak가 부가되어 있음)로 치환하고, 또, pDC62c5-U533의 염기 서열 No3993-No4004를 서열번호 5에 기재한 인간 오말리즈맙 항체(OMLH) 중쇄를 코드하는 cDNA(개시 코돈의 상류에 최적화한 Kozak가 부가되어 있음)로 치환하여, pDC62c5-U533-OMLH(도 2)를 구축하였다. 최적화한 Kozak의 서열을 서열번호 2에 기재한다.
유전자 도입 전에 제한 효소 ClaI를 이용하여 벡터를 선상화(線狀化)하였다.
[실시예 3] pDC62c5-U533-OMLH의 CHO 세포로의 도입, 선별, 생산성 시험 및 발현 안정성 시험
18 ㎍의 pDC62c5-U533-OMLH를 리포펙틴법(FreeStyle MAX Reagent, Life Technologies를 사용)을 이용하여 125 ㎖ 용(容)의 배양 플라스크(Erlenmeyer Flask, Baffled, 125 ㎖, Vent Cap, cat#431405, corning) 중의 15,000,000개의 CHO 세포(CHO DG44 세포)에 유전자 도입하였다.
도입 방법에 대해서는 제조업자의 사용 설명서에 따랐다. 유전자 도입 48시간 후에 세포 수를 계측한 후, 세포를 4 mM GlutaMAX-I(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies)에서 희석하였다. 96 웰 미세적정 플레이트(microtiter plate) 중에 4,000 세포/웰의 농도로 12,000 세포/웰의 비유전자 도입 세포와 혼합한 후, 10매(960 웰)에 접종하고, 8% 탄산 가스 존재 하에서 37℃, 약 3주간 배양하고, 생존한 세포로부터 임의로 35 주의 HT 불포함 배지 내성 클론을 선택하였다. 새롭게 96 웰 미세적정 플레이트 중에 16,000 세포/웰의 농도로 12,000 세포/웰의 비유전자 도입 세포와 혼합한 후, 10매(960 웰)에 접종하고, 8% 탄산 가스 존재 하에서 37℃, 약 3주간 배양하고, 생존한 세포로부터 임의로 71 주의 HT 불포함 배지 내성 클론을 선택하여 합계 106 주를 얻었다. 얻어진 HT 불포함 배지 내성 클론에 대해서는 4 mM GlutaMAX-I(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies)와 함께 24 웰 플레이트로 옮기고, 세포가 각 웰 바닥 면적의 1/3 이상을 차지할 때까지 배양하였다. 24 웰 플레이트에서 증식한 세포에 대해서는 4 mM GlutaMAX-I(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies)와 함께 6 웰 플레이트로 옮기고, 세포가 각 웰 바닥 면적의 1/3 이상을 차지할 때까지 배양하였다.
각 주 1 ㎖를 멸균 튜브에 취하고, 300×g, 7분간 원심하였다. 상청을 버리고, 세포를 0.55 ㎖의 새로운 배지(4 mM GlutaMAX-I(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies))에 현탁하고, 세포 수를 계측하였다. 생존 세포 밀도를 2×105 세포/㎖가 되도록 배지에서 희석한 후, 0.4 ㎖를 새로운 24 웰 플레이트로 옮기고, 8% 탄산 가스 존재 하에서 37℃, 72시간 회전 진탕 배양(125 rpm)하였다. 배양 후, 세포 수를 계측하고, 9300×g, 2분간 원심분리하여 상청을 회수하였다.
이어서 배양 상청 중의 IgG 농도를 ELISA법으로 측정하였다. 그 결과, 최대 생산 클론의 IgG 생산량은 28.5 ㎍/㎖/3일이며, 10 ㎍/㎖ 이상의 IgG를 생산한 주는 106 주 중 16 주(15.1%)였다(도 6).
다음에 IgG 생산량 상위 10 주를 선택하여 발현 안정성 시험을 행하였다. 발현 안정성 시험에서는 각 주의 동결 스톡(stock)으로부터 계대 배양을 개시하였다. 계대 배양에서는 세포를 2×105 세포/㎖가 되도록 희석한 후, 3 내지는 4일 회전 진탕 배양하고, 이것을 또 2×105 세포/㎖가 되도록 희석하는 것을 반복 실시하였다. 회전 진탕 배양 개시 0, 7, 14, 21, 28, 35, 42, 49, 56, 63, 72, 79, 86일째에 세포 밀도 2×105 세포/㎖, 0.4 ㎖로 세포를 조정하여 24 웰-플레이트 상에서 72시간 회전 진탕 배양(125 rpm)하고, 회수한 배양 상청의 IgG 생산량을 ELISA로 측정하였다. 10 주의 IgG 생산량은 0주에 28.5∼13.0 ㎎/L이고, 12주째에는 15.6∼3.8 ㎎/L였다. pDC62c5-U533-OMLH 도입 세포의 발현 안정성에 대해서는 8주째까지 동결 스톡 제작 전의 70% 이상의 IgG 생산능을 유지하고 있던 것은 10 주 중 7 주, 50% 이상 70% 미만이 2 주, 50% 미만이 1 주이며, 12주째의 시점에서는 70% 이상 유지 주는 2 주, 50% 이상 70% 미만이 5 주, 50% 미만이 3 주였다(도 7).
[실시예 4] pDC61의 제작
pDC6(일본 특허 5704753)의 염기 서열 No3182-No5843을 서열번호 7에 기재된 서열로 치환하여 pDC61를 구축하였다. 백본 벡터 pDC61의 전체 염기 서열을 서열번호 6에 기재한다.
[실시예 5] pDC61/OMLH의 제작
pDC61의 염기 서열 No1267-No1273을 서열번호 8에 기재된 인간 오말리즈맙 항체(OMLH) 경쇄를 코드하는 cDNA로 치환하고, 또, pDC61의 염기 서열 No2765-No2771을 서열번호 9에 기재된 인간 오말리즈맙 항체(OMLH) 중쇄를 코드하는 cDNA에 치환하여 pDC61/OMLH(도 3)를 구축하였다.
유전자 도입 전에 제한 효소 ClaI를 이용하여 벡터를 선상화하였다.
[실시예 6] pDC61/OMLH의 CHO 세포로의 도입, 선별, 생산성 시험 및 발현 안정성 시험
18 ㎍의 pDC61/OMLH를 리포펙틴법(FreeStyle MAX Reagent, Life Technologies를 사용)을 이용하여 125 ㎖ 용의 배양 플라스크(Erlenmeyer Flask, Baffled, 125 ㎖, Vent Cap, cat#431405, corning) 중의 15,000,000개의 CHO 세포(CHO DG44 세포)에 유전자 도입하였다. 도입 방법에 대해서는 제조업자의 사용 설명서에 따랐다. 유전자 도입 48시간 후에 세포 수를 계측한 후, 세포를 4 mM GlutaMAX-I(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies)에서 희석하였다. 96 웰 미세적정 플레이트 중에 40,000 세포/웰의 농도로 12,000 세포/웰의 비유전자 도입 세포와 혼합한 후, 10매(960 웰)에 접종하고, 8% 탄산 가스 존재 하에서 37℃, 약 3주간 배양하고, 생존한 세포로부터 임의로 118 주의 HT 불포함 배지 내성 클론을 선택하였다. 얻어진 HT 불포함 배지 내성 클론에 대해서는 4 mM GlutaMAX-I(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies)와 함께 24 웰 플레이트로 옮기고, 세포가 각 웰 바닥 면적의 1/3 이상을 차지할 때까지 배양하였다. 24 웰 플레이트에서 증식한 세포에 대해서는 4 mM GlutaMAX-I(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies)와 함께 6 웰 플레이트로 옮기고, 세포가 각 웰 바닥 면적의 1/3 이상을 차지할 때까지 배양하였다. 각 주 1 ㎖를 멸균 튜브에 취하고, 300×g, 7분간 원심하였다. 상청을 버리고, 세포를 0.55 ㎖의 새로운 배지(4 mM GlutaMAX-I(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies))에 현탁하고, 세포 수를 계측하였다. 생존 세포 밀도를 2×105 세포/㎖가 되도록 배지에서 희석한 후, 0.4 ㎖를 새로운 24 웰 플레이트로 옮기고, 8% 탄산 가스 존재 하에서 37℃, 72시간 회전 진탕 배양(125 rpm)하였다. 배양 후, 세포 수를 계측하고, 9300×g, 2분간 원심분리하여 상청을 회수하였다. 이어서 배양 상청 중의 IgG 농도를 ELISA법으로 측정하였다. 그 결과, 최대 생산 클론의 IgG 생산량은 16.4 ㎍/㎖/3일이며, 10 ㎍/㎖ 이상의 IgG를 생산한 주는 118 주 중 2 주(1.7%)였다(도 6).
다음에 IgG 생산량 상위 10 주를 선택하여 발현 안정성 시험을 행하였다. 발현 안정성 시험에서는 각 주의 동결 스톡으로부터 계대 배양을 개시하였다. 계대 배양에서는 세포를 2×105 세포/㎖가 되도록 희석한 후, 3 내지는 4일 회전 진탕 배양하고, 이것을 또 2×105 세포/㎖가 되도록 희석하는 것을 반복 실시하였다. 회전 진탕 배양 개시 0, 7, 14, 21, 28, 35, 42, 49, 56, 63, 72, 79, 86일째에 세포 밀도 2×105 세포/㎖, 0.4 ㎖로 세포를 조정하여 24 웰-플레이트 상에서 72시간 회전 진탕 배양(125 rpm)하고, 회수한 배양 상청의 IgG 생산량을 ELISA로 측정하였다. 10 주의 IgG 생산량은 0주에 16.4∼3.1 ㎎/L 이며, 12주째에는 6.3∼0.1 ㎎/L였다. pDC61/OMLH 도입 세포의 발현 안정성에 대해서는 8주째까지 동결 스톡 제작 전의 70% 이상의 IgG 생산능을 유지하고 있던 것은 10 주 중 3 주, 50% 이상 70% 미만이 2 주, 50% 미만이 5 주이며, 12주째의 시점에서는 70% 이상 유지 주는 0 주, 50% 이상 70% 미만이 5 주, 50% 미만이 5 주였다.
[실시예 7] pNC32c-U533의 제작
pDC61(실시예 4에서 제작)를 개변하여 본 발명의 벡터인 pNC32c-U533를 구축하였다. 백본 벡터 pNC32c-U533의 전체 염기 서열을 서열번호 10에 기재한다. pNC32c-U533은 염기 서열 No5196-No5990의 영역에 네오마이신 포스포트랜스퍼라아제의 유전자가 도입되어 있다. 또, 염기 서열 No867-No2417, No3291-No4841 및 No9130-No10680의 영역에 UCOE가 도입되어 있다. UCOE의 염기 서열을 서열번호 1에 기재한다.
[실시예 8] pNC32c-U533-OMLH의 제작
pNC32c-U533의 염기 서열 No 658-No 668을 서열번호 11에 기재된 인간 오말리즈맙 항체(OMLH) 경쇄를 코드하는 cDNA로 치환하고, 또, pNC32c-U533의 염기 서열 No 3081-No 3092를 서열번호 12에 기재된 인간 오말리즈맙 항체(OMLH) 중쇄를 코드하는 cDNA로 치환하여 pNC32c-U533-OMLH(도 4)를 구축하였다.
유전자 도입 전에 제한 효소 ClaI를 이용하여 벡터를 선상화하였다.
[실시예 9] pNC32c-U533-OMLH의 CHO 세포로의 도입, 선별, 생산성 시험
18 ㎍의 pNC32c-U533-OMLH를 리포펙틴법(FreeStyle MAX Reagent, Life Technologies를 사용)을 이용하여 125 ㎖ 용의 배양 플라스크(Erlenmeyer Flask, Baffled, 125 ㎖, Vent Cap, cat#431405, corning) 중의 15,000,000개의 CHO 세포(CHO DG44 세포)에 유전자 도입하였다. 도입 방법에 대해서는 제조업자의 사용 설명서에 따랐다. 유전자 도입 48시간 후에 세포 수를 계측한 후, 4 mM GlutaMAX-I(Life Technologies) 및 400 ㎍/㎖ G418 설페이트(Wako), 1×HT 보충제(Supplement)(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies)에서 세포를 희석하였다. 96 웰 미세적정 플레이트 중에 800 세포/웰의 농도로 12,000 세포/웰의 비유전자 도입 세포와 혼합한 후, 10매(960 웰)에 접종하고, 8% 탄산 가스 존재 하에서 37℃, 약 3주간 배양하고, 생존한 세포로부터 임의로 105 주의 G418 내성 클론을 선택하였다. 얻어진 G418 내성 클론에 대해서는 4 mM GlutaMAX-I(Life Technologies) 및 400 ㎍/㎖ G418 설페이트(Wako), 1×HT 보충제(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies)와 함께 24 웰 플레이트로 옮기고, 세포가 각 웰 바닥 면적의 1/3 이상을 차지할 때까지 배양하였다. 24 웰 플레이트에서 증식한 세포에 대해서는 4 mM GlutaMAX-I(Life Technologies) 및 400 ㎍/㎖ G418 설페이트(Wako), 1×HT 보충제(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies)와 함께 6 웰 플레이트로 옮기고, 세포가 각 웰 바닥 면적의 1/3 이상을 차지할 때까지 배양하였다. 각 주 1 ㎖를 멸균 튜브에 취하고, 300×g, 7분간 원심하였다. 상청을 버리고, 세포를 0.55 ㎖의 새로운 배지(4 mM GlutaMAX-I(Life Technologies) 및 400 ㎍/㎖ G418 설페이트(Wako), 1×HT 보충제(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies))에 현탁하고, 세포 수를 계측하였다. 생존 세포 밀도를 2×105 세포/㎖가 되도록 배지에서 희석한 후, 0.4 ㎖를 새로운 24 웰 플레이트로 옮기고, 8% 탄산 가스 존재 하에서 37℃, 72시간 회전 진탕 배양(125 rpm)하였다.
배양 후, 세포 수를 계측하고, 9300×g, 2분간 원심분리하여 상청을 회수하였다. 이어서 배양 상청 중의 IgG 농도를 ELISA법으로 측정하였다. 그 결과, 최대 생산 클론의 IgG 생산량은 2.6 ㎍/㎖/3일이며, 10 ㎍/㎖ 이상의 IgG를 생산한 주는 105 주 중 0 주였다.
[실시예 10] UCOE-Hu-P2/OMLH의 제작
시판되는 벡터인 UCOE(등록상표) 발현 벡터 - 인간 4kb Puro Set(머크사 cat# 5.04867.0001)의 염기 서열 No5309-No5311을 서열번호 13에 기재된 서열로 치환함으로써 제한 효소 BstBI 인식 사이트를 증설하였다(UCOE-Hu-P2).
UCOE-Hu-P2 상의 FseI 인식 사이트 및 BstBI 인식 사이트를 사용하여, 서열번호 14에 기재된 인간 오말리즈맙 항체 경쇄(OML)를 코드하는 cDNA 및 시미안 바이러스 40 폴리아데닐화 신호(SV40pA), 모르모트 사이토메갈로바이러스 프로모터(PgpCMV), 인간 오말리즈맙 항체 중쇄(OMH)를 코드하는 cDNA를 연결한 서열을 UCOE-Hu-P2에 삽입하여 UCOE-Hu-P2/OMLH(도 5)를 구축하였다.
유전자 도입 전에 제한 효소 HindIII를 이용하여 벡터를 선상화하였다.
[실시예 11] UCOE-Hu-P2/OMLH의 CHO 세포로의 도입, 선별, 생산성 시험
18 ㎍의 UCOE-Hu-P2/OMLH를 리포펙틴법(FreeStyle MAX Reagent, Life Technologies를 사용)을 이용하여 125 ㎖ 용의 배양 플라스크(Erlenmeyer Flask, Baffled, 125 ㎖, Vent Cap, cat#431405, corning) 중의 15,000,000개의 CHO 세포(CHO DG44 세포)에 유전자 도입하였다. 도입 방법에 대해서는 제조업자의 사용 설명서에 따랐다. 유전자 도입 48시간 후에 세포 수를 계측한 후, 4 mM GlutaMAX-I(Life Technologies) 및 100 ㎍/㎖ 퓨로마이신 디히드로클로라이드(Puromycin dihydrochloride)(Thermo Fisher Scientific), 1×HT 보충제(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies)에서 세포를 희석하였다.
96 웰 미세적정 플레이트 중에 4,000 세포/웰의 농도로 12,000 세포/웰의 비유전자 도입 세포와 혼합한 후, 10매(960 웰)에 접종하고, 8% 탄산 가스 존재 하에서 37℃, 약 3주간 배양하고, 생존한 세포로부터 임의로 33 주의 퓨로마이신 내성 클론을 선택하였다. 새롭게 96 웰 미세적정 플레이트 중에 16,000 세포/웰의 농도로 12,000 세포/웰의 비유전자 도입 세포와 혼합한 후, 10매(960 웰)에 접종하고, 8% 탄산 가스 존재 하에서 37℃, 약 3주간 배양하고, 생존한 세포로부터 임의로 84 주의 퓨로마이신 내성 클론을 선택하여 합계 117 주를 얻었다. 얻어진 퓨로마이신 내성 클론에 대해서는 4 mM GlutaMAX-I(Life Technologies) 및 100 ㎍/㎖ 퓨로마이신 디히드로클로라이드(Thermo Fisher Scientific), 1×HT 보충제(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies)와 함께 24 웰 플레이트로 옮기고, 세포가 각 웰 바닥 면적의 1/3 이상을 차지할 때까지 배양하였다. 24 웰 플레이트에서 증식한 세포에 대해서는 4 mM GlutaMAX-I(Life Technologies) 및 100 ㎍/㎖ 퓨로마이신 디히드로클로라이드(Thermo Fisher Scientific), 1×HT 보충제(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies)와 함께 6 웰 플레이트로 옮기고, 세포가 각 웰 바닥 면적의 1/3 이상을 차지할 때까지 배양하였다. 각 주 1 ㎖를 멸균 튜브에 취하고, 300×g, 7분간 원심하였다. 상청을 버리고, 세포를 0.55 ㎖의 새로운 배지(4 mM GlutaMAX-I(Life Technologies) 및 100 ㎍/㎖ 퓨로마이신 디히드로클로라이드(Thermo Fisher Scientific), 1×HT 보충제(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies))에 현탁하고, 세포 수를 계측하였다. 생존 세포 밀도를 2×105 세포/㎖가 되도록 배지에서 희석한 후, 0.4 ㎖를 새로운 24 웰 플레이트로 옮기고, 8% 탄산 가스 존재 하에서 37℃, 72시간 회전 진탕 배양(125 rpm)하였다. 배양 후, 세포 수를 계측하고, 9300×g, 2분간 원심분리하여 상청을 회수하였다. 이어서 배양 상청 중의 IgG 농도를 ELISA법으로 측정하였다. 그 결과, 최대 생산 클론의 IgG 생산량은 4.7 ㎍/㎖/3일이며, 10 ㎍/㎖ 이상의 IgG를 생산한 주는 117 주 중 0 주였다.
[실시예 12] pDC62c5-U533를 이용한 개 CTLA-4-Ig의 발현
실시예 1에서 제작한 pDC62c5-U533의 염기 서열 No2896-No5784(PCMV5, PABGH 및 UCOE의 염기 서열)의 영역을 결실한 pDC62c5-U533(외래 유전자 삽입 부위가 1개 있음(AscI와 AsisSI의 사이))을 통상의 기술자에게 주지인 방법으로 제작하였다.
개 CTLA-4 및 개 IgG-D의 유전자 서열은 NCBI(The National Center for Biotechnology Information)에 이미 등록되어 있다(GenBank accession number; NM_001003106.1 및 AF354267.1). 개 CTLA-4의 예상 세포외 영역과 개 IgG-D의 힌지(hinge)부, CH2 및 CH3 영역을 결합시킨 아미노산 서열을 제작하여 CHO 세포에 대한 코돈 최적화를 행하였다(서열번호 15(아미노산 서열) 및 서열번호 16(코돈 최적화 후 뉴클레오티드 서열)). 그 후, AscI 제한 효소 인식 서열, KOZAK 서열, 개 CTLA-4-Ig 서열, AsiSI 제한 효소 인식 서열을 상기의 순서로 배치하도록 유전자 합성을 행하였다. 합성한 유전자 사슬을 외래 유전자 삽입 부위가 1개 있는 pDC62c5-U533(서열번호 17; 도 8)의 클로닝 사이트(염기 서열 No1098-No1108; PRCMV5와 PABGH의 사이에 있는 AscI 및 AsiSI 제한 효소 인식 서열)에 제한 효소 인식 서열을 이용해 상기의 순서로 배치하도록 조립하여 개 CTLA-4-Ig발현 벡터를 구축하였다. 이 발현 벡터를 제한 효소 ClaI에 의해 선상화한 후, 디히드로 엽산 환원효소 결손 세포인 CHO DG44 세포(CHO-DG44(dhfr-/-))에 리포펙타민 LTX(Thermo Fisher Scientific사)를 이용해 트랜스펙션하였다. 유전자 도입 48시간 후에, 티미딘 및 히포크산틴을 포함하지 않는 Opti-CHO 배지(Life Technologies사, 4 mM GlutaMAX-I(Life Technologies사)를 포함함)로 배지 교환하고, 선택함으로써 안정 발현 세포주를 얻었다. 추가로, 한계 희석법에 의해 세포의 클로닝을 행하고, 고발현 클론을 닷 블롯법 및 3일간의 진탕 배양 시험에 의해 선발하였다. 수립된 개 CTLA-4-Ig 고발현 CHO-DG44 세포 클론을 4 mM GlutaMAX-I(Life Technologies사)를 포함하는 Dynamis 배지(Life Technologies사)로 순화시키고, 5×105 세포/㎖, 액량 30 ㎖, 125 ㎖ 용의 배양 플라스크(Corning사)에서 14일간의 진탕 배양(125 rpm, 37℃, 8% CO2)을 행하였다. 배양 3, 5, 7, 10일째에 3배 농도로 조제한 Efficient Feed B+(Life Technologies사)를 3.3% v/v, 추가로 배양 3, 5, 7일째에 45% 글루코오스 용액(칸토 화학사)을 각각 최종 농도 4, 4, 6 g/L가 되도록 배지에 첨가하였다. 이 결과, 14일째의 배양 상청 중에 대략 300 ㎎/L의 개 CTLA-4-Ig 조환 단백질을 발현하였다(도 9). 아울러, 배양 상청 중의 개 CTLA-4-Ig 농도는 염소 항-개 IgG1 폴리클로날 항체(Bethyl Laboratories사)를 이용한 샌드위치 ELISA법에 의해 정량하고, 생존 세포 수는 트립판 블루 염색에 의해 죽은 세포를 제외하여 카운트하였다.
생산한 개 CTLA-4-Ig는 상청으로부터 Ab-Capcher ExTra(단백질 A 변이체, ProteNova사)를 이용해 정제를 행하였다. 레진에 대한 결합은 오픈 컬럼법을 이용하고, 평형화 버퍼 및 세정 버퍼로서 인산 완충 생리 식염수(PBS; pH 7.4)를 사용하였다. 용출 버퍼에는 0.1 M 글리신-HCl을, 중화 버퍼에는 1M Tris-HCl를 사용하였다. PD-MidiTrap G-25(GE Healthcare사)를 이용해 버퍼를 PBS로 치환하였다. 0.2 μm의 필터를 통과하고, 실험에 제공할 때까지 4℃에서 보존하였다. 단백질의 농도는 Pierce BCA 단백질 분석 키트(Thermo Fisher Scientific사)에 의해 정량하고, 이후의 실험에 이용하였다.
정제한 개 CTLA-4-Ig의 순도를 확인하기 위하여, SDS-PAGE 및 CBB 염색에 의해 Ig단백질의 검출을 행하였다. SuperSep Ace 5-20% 구배(gradient) 겔(Wako사)을 이용하여, 개 CTLA-4-Ig를 환원 조건 하 및 비환원 조건 하에서 전기영동하였다. Quick-CBB 키트(와코 순약 공업사)에 의해 염색을 행한 후, 증류수 중에서 탈색을 행하였다. 환원 조건에서는 약 45 kDa, 비환원 조건에서는 약 90 kDa에 상당하는 분자량의 위치에 밴드를 볼 수 있었다. 아미노산 서열로부터 계산되는 개 CTLA-4-Ig의 분자량은 2량체로 약 79 kDa, 단량체로 약 39.5 kDa이며, 당쇄 수식 등의 영향에 의해 큰 분자량의 위치에 밴드가 나온 것이라고 생각할 수 있었다. 협잡 단백질이라 생각되는 밴드는 거의 시인되지 않았다(도 10).
[실시예 13] pDC62c5-U533-TRLH의 제작
pDC62c5-U533의 염기 서열 No1098-No1108를 서열번호 18에 기재된 인간 트라스투주맙(TRLH) 경쇄를 코드하는 cDNA(개시 코돈의 상류에 최적화한 Kozak가 부가되어 있음)로 치환하고, 또, pDC62c5-U533의 염기 서열 No3993-No4004를 서열번호 19에 기재된 인간 트라스투주맙(TRLH) 중쇄를 코드하는 cDNA(개시 코돈의 상류에 최적화한 Kozak가 부가되어 있음)로 치환하여 pDC62c5-U533-TRLH(도 11)를 구축하였다. 최적화한 Kozak의 서열을 서열번호 2에 기재한다.
유전자 도입 전에 제한 효소 ClaI를 이용하여 벡터를 선상화하였다.
[실시예 14] pDC62c5-U533-TRLH의 CHO 세포로의 도입, 선별, 생산성 시험 및 발현 안정성 시험
18 ㎍의 pDC62c5-U533-TRLH를 리포펙틴법(FreeStyle MAX Reagent, Life Technologies를 사용)을 이용하여 125 ㎖ 용의 배양 플라스크(Erlenmeyer Flask, Baffled, 125 ㎖, Vent Cap, cat#431405, Corning) 중의 15,000,000개의 CHO 세포(CHO DG44 세포)에 유전자 도입하였다.
도입 방법에 대해서는 제조업자의 사용 설명서에 따랐다. 유전자 도입 48시간 후에 세포 수를 계측한 후, 세포를 4 mM GlutaMAX-I(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies)에서 희석하였다. 96 웰 미세적정 플레이트 중에 400 세포/웰의 농도로 12,000 세포/웰의 비유전자 도입 세포와 혼합한 후, 12매(1,152 웰)에 접종하고, 8% 탄산 가스 존재 하에서 37℃, 약 3주간 배양하고, 생존한 세포로부터 임의로 188 주의 HT 불포함 배지 내성 클론을 선택하였다. 얻어진 HT 불포함 배지 내성 클론에 대해서는 4 mM GlutaMAX-I(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies)와 함께 24 웰 플레이트로 옮겨 배양하였다.
각 주 0.6 ㎖를 멸균 튜브에 취하고, 300×g, 7분간 원심하였다. 상청을 버리고, 세포를 0.3 ㎖의 새로운 배지(4 mM GlutaMAX-I(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies))에 현탁하고, 세포 수를 계측하였다. 생존 세포 밀도를 2×105 세포/㎖가 되도록 배지에서 희석한 후, 0.4 ㎖를 새로운 24 웰 플레이트로 옮기고, 8% 탄산 가스 존재 하에서 37℃, 72시간 회전 진탕 배양(125 rpm)하였다. 배양 후, 세포 수를 계측하고, 9300×g, 2분간 원심분리하여 상청을 회수하였다.
이어서 배양 상청 중의 IgG 농도를 ELISA법으로 측정하였다. 그 결과, 최대 생산 클론의 IgG 생산량은 47.8 ㎍/㎖/3일이며, 10 ㎍/㎖ 이상의 IgG를 생산한 주는 188 주중 81 주(43.1%)였다(도 12).
다음에 IgG 생산량 상위 10 주를 선택하여 발현 안정성 시험을 행하였다. 발현 안정성 시험에서는 각 주의 동결 스톡으로부터 계대 배양을 개시하였다. 순화 배양에는 4 mM GlutaMAX-I(Life Technologies)를 포함하는 CD OptiCHO 배지(Life Technologies) 및 4 mM GlutaMAX-I(Life Technologies)를 포함하는 Dynamis 배지(Life Technologies)를 사용하여, 75:25, 50:50, 25:75로 계대마다 후자의 비율을 높임으로써 세포의 순화를 행하였다. 순화 후에 발현 안정성 시험을 개시하였다. 시험 기간 중의 계대 배양에서는 4 mM GlutaMAX-I(Life Technologies)를 포함하는 Dynamis 배지(Life Technologies)를 사용하고, 세포를 2 또는 1.5×105 세포/㎖가 되도록 희석한 후, 3 내지는 4일간 회전 진탕 배양하고, 이것을 또 2 또는 1.5×105 세포/㎖가 되도록 희석하는 것을 반복 실시하였다. 시험 배양 개시 0, 7, 14, 21, 28, 35, 42, 49, 56, 63, 70일째에 세포 밀도 2×105 세포/㎖, 0.4 ㎖로 세포를 조정하여 24 웰-플레이트 상에서 72시간 회전 진탕 배양(125 rpm)하고, 회수한 배양 상청의 IgG 생산량을 ELISA로 측정하였다. 10 주의 IgG 생산량은 0주에 50.9∼16.5 ㎎/L이며, 10주째에는 37.7∼0.5 ㎎/L였다. pDC62c5-U533-TRLH 도입 세포의 발현 안정성에 대해서는 8주째까지 동결 스톡 제작 전의 70% 이상의 IgG 생산능을 유지하고 있는 것은 10 주중 8 주, 50% 이상 70% 미만이 1 주, 50% 미만이 1 주이며, 10주째의 시점에서는70% 이상 유지 주는 9 주, 50% 미만이 1 주였다(도 13).
[실시예 15] pDC62c5-U533-OMLH를 도입한 CHO 세포의 MTX 선별 및 생산성 시험
실시예 3에서 얻어진 IgG 생산량 상위 10 주(Clone01-10)에 대하여, 각 주의 동결 스톡으로부터 2, 3회 계대 배양한 후, 4 mM GlutaMAX-I(Life Technologies) 및 60 nM MTX (Wako)를 포함하는 CD OptiCHO 배지(Life Technologies)에서 세포를 희석하였다. 1 주마다 96 웰 미세적정 플레이트 중에 1, 3, 10, 30, 100, 300, 1,000, 3,000, 10,000 세포/웰의 농도로 12,000 세포/웰의 비유전자 도입 세포와 혼합한 후, 각 농도 2매씩 10매(960 웰)에 접종하고, 8% 탄산 가스 존재 하에서 37℃, 3, 4주간 배양하고, 생존한 세포로부터 임의로 MTX 내성 클론을 선택하였다. 얻어진 클론을 24 웰 플레이트로 옮기고, 세포가 각 웰 바닥 면적의 1/3 이상을 차지할 때까지 배양하였다. 24 웰 플레이트에서 증식한 세포에 대해서는 각 주 0.5 ㎖를 멸균 튜브에 취하고, 300×g, 7분간 원심하였다. 상청을 버리고, 세포를 0.3 ㎖의 새로운 배지(4 mM GlutaMAX-I(Life Technologies) 및 60 nM MTX (Wako)를 포함하는 CD OptiCHO 배지(Life Technologies))에 현탁하고, 세포 수를 계측하였다. 생존 세포 밀도를 2×105 세포/㎖가 되도록 배지에서 희석한 후, 0.4 ㎖를 새로운 24 웰 플레이트로 옮기고, 8% 탄산 가스 존재 하에서 37℃, 72시간 회전 진탕 배양(125 rpm)하였다. 배양 후, 세포 수를 계측하고, 9300×g, 2분간 원심분리하여 상청을 회수하였다. 이어서 배양 상청 중의 IgG 농도를 ELISA법으로 측정하였다(도 14).
Clone01에 대해서는 접종 29-32일 후에 1, 3, 10 세포/웰의 농도로 접종한 플레이트로부터 18 클론을 얻었다. 최대 생산 클론의 IgG 생산량은 58.0 ㎍/㎖/3일이며, 친주(親株)(28.5 ㎍/㎖/3일)의 2.0배가 되었다.
Clone02에 대해서는 접종 22-32일 후에 1, 3 세포/웰의 농도로 접종한 플레이트로부터 24 클론을 얻었다. 최대 생산 클론의 IgG 생산량은 105.5 ㎍/㎖/3일이며, 친주(25.8 ㎍/㎖/3일)의 4.1배가 되었다.
Clone03에 대해서는 접종 22-32일 후에 1, 3 세포/웰의 농도로 접종한 플레이트로부터 19 클론을 얻었다. 최대 생산 클론의 IgG 생산량은 78.7 ㎍/㎖/3일이며, 친주(23.0 ㎍/㎖/3일)의 3.4배가 되었다.
Clone04에 대해서는 접종 22-32일 후에 1 세포/웰의 농도로 접종한 플레이트로부터 33 클론을 얻었다. 최대 생산 클론의 IgG 생산량은 83.7 ㎍/㎖/3일이며, 친주(20.9 ㎍/㎖/3일)의 4.0배가 되었다.
Clone05에 대해서는 접종 27-39일 후에 1, 3 세포/웰의 농도로 접종한 플레이트로부터 23 클론을 얻었다. 최대 생산 클론의 IgG 생산량은 71.0 ㎍/㎖/3일이며, 친주(19.2 ㎍/㎖/3일)의 3.7배가 되었다.
Clone06에 대해서는 접종 26-39일 후에 3, 10, 30 세포/웰의 농도로 접종한 플레이트로부터 16 클론을 얻었다. 최대 생산 클론의 IgG 생산량은 56.2 ㎍/㎖/3일이며, 친주(18.4 ㎍/㎖/3일)의 3.0배가 되었다.
Clone07에 대해서는 접종 29-39일 후에 3, 10, 30 세포/웰의 농도로 접종한 플레이트로부터 8 클론을 얻었다. 최대 생산 클론의 IgG 생산량은 65.3 ㎍/㎖/3일이며, 친주(17.1 ㎍/㎖/3일)의 3.8배가 되었다.
Clone08에 대해서는 접종 27-41일 후에 100, 300, 1,000 세포/웰의 농도로 접종한 플레이트로부터 24 클론을 얻었다. 최대 생산 클론의 IgG 생산량은 47.9 ㎍/㎖/3일이며, 친주(15.1 ㎍/㎖/3일)의 3.1배가 되었다.
Clone09에 대해서는 접종 26-41일 후에 100, 1,000, 3,000, 10,000 세포/웰의 농도로 접종한 플레이트로부터 12 클론을 얻었다. 최대 생산 클론의 IgG 생산량은 21.9 ㎍/㎖/3일이며, 친주(14.3 ㎍/㎖/3일)의 1.5배가 되었다.
Clone10에 대해서는 접종 22-39일 후에 1, 3, 10 세포/웰의 농도로 접종한 플레이트로부터 18 클론을 얻었다. 최대 생산 클론의 IgG 생산량은 131.4 ㎍/㎖/3일이며, 친주(13.0 ㎍/㎖/3일)의 10.1배가 되었다.
본 명세서에서 인용한 모든 간행물, 특허 및 특허 출원을 그대로 참고로서 본 명세서에 포함되는 것으로 한다.
본 발명에 의해, 디히드로 엽산 환원효소 유전자 결손 포유동물 세포를 숙주로서 고레벨의 외래 유전자 유래 단백질 생산을 가능하게 하는 발현 벡터를 제공할 수 있다. 또, 포유동물 본래의 번역 후 수식 및 높은 생물 활성을 갖는 단백질을 생산할 수 있다. 그러므로, 바이오 의약품 등의 단백질성 유용 물질의 생산 원가를 대폭 인하할 수 있다.
추가로, 본 발명에 의한 단백질의 생산 방법은 바이러스나 미생물을 사용하지 않기 때문에, 안전성이 높은 단백질 생산이 가능하다.
<서열번호 1> UCOE의 뉴클레오티드 서열
GGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGA
<서열번호 2> 최적화된 Kozak 서열을 나타낸다. 이 서열 중에서, ATG는 개시 코돈이 된다.
CCGCCGCCACCATGG
<서열번호 3> pDC62c5-U533의 전체 서열
CGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACACCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAATAACCCCGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCCTCACTCTCTTCCGCATCGCTGTCTGCGAGGGCCAGCTGTTGGGCTCGCGGTTGAGGACAAACTCTTCGCGGTCTTTCCAGTACTCTTGGATCGGAAACCCGTCGGCCTCCGAACGGTACTCCGCCACCGAGGGACCTGAGCGAGTCCGCATCGACCGGATCGGAAAACCTCTCGAGAAAGGCGTCTAACCAGTCACAGTCGCAAGGTAGGCTGAGCACCGTGGCGGGCGGCAGCGGGTGGCGGTCGGGGTTGTTTCTGGCGGAGGTGCTGCTGATGATGTAATTAAAGTAGGCGGTCTTGAGACGGCGGATGGTCGAGGTGAGGTGTGGCAGGCTTGAGATCCAGCTGTTGGGGTGAGTACTCCCTCTCAAAAGCGGGCATGACTTCTGCGCTAAGATTGTCAGTTTCCAAAAACGAGGAGGATTTGATATTCACCTGGCCCGATCTGGCCATACACTTGAGTGACAATGACATCCACTTTGCCTTTCTCTCCACAGGTGTCCACTCCCAGGTCCAAGGCGCGCCGCGATCGCGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGAGGATCTCCGCGGGGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGAGAATTCCGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACACCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAATAACCCCGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCCTCACTCTCTTCCGCATCGCTGTCTGCGAGGGCCAGCTGTTGGGCTCGCGGTTGAGGACAAACTCTTCGCGGTCTTTCCAGTACTCTTGGATCGGAAACCCGTCGGCCTCCGAACGGTACTCCGCCACCGAGGGACCTGAGCGAGTCCGCATCGACCGGATCGGAAAACCTCTCGAGAAAGGCGTCTAACCAGTCACAGTCGCAAGGTAGGCTGAGCACCGTGGCGGGCGGCAGCGGGTGGCGGTCGGGGTTGTTTCTGGCGGAGGTGCTGCTGATGATGTAATTAAAGTAGGCGGTCTTGAGACGGCGGATGGTCGAGGTGAGGTGTGGCAGGCTTGAGATCCAGCTGTTGGGGTGAGTACTCCCTCTCAAAAGCGGGCATGACTTCTGCGCTAAGATTGTCAGTTTCCAAAAACGAGGAGGATTTGATATTCACCTGGCCCGATCTGGCCATACACTTGAGTGACAATGACATCCACTTTGCCTTTCTCTCCACAGGTGTCCACTCCCAGGTCCAAGCGGCCGCCCTGCAGGGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGAGGATCTCCGCGGGGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGAACCGGTTGTGGAATGTGTGTCAGTTAGGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCATAGTCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGCTTTTGCAAAAAAGCTGCAGATGGTACGACCATTAAATTGTATTGTAGCAGTATCACAAAATATGGGTATTGGTAAAAATGGTGATTTACCATGGCCACCATTACGAAATGAATTTAAATATTTTCAACGAATGACTACTACTTCATCAGTAGAAGGTAAACAAAATTTAGTAATTATGGGTCGAAAAACTTGGTTTTCAATTCCTGAGAAGAATCGACCTTTAAAGGACAGAATTAATATAGTTCTCAGTAGAGAACTCAAAGAACCACCACGAGGAGCTCATTTTCTTGCCAAAAGTTTGGATGATGCCTTAAGACTTATTGAACAACCGGAATTGGCAAGTAAAGTAGACATGGTTTGGATAGTCGGAGGCAGTTCTGTTTACCAGGAAGCCATGAATCAACCAGGCCACCTCAGACTCTTTGTGACAAGGATCATGCAGGAATTTGAAAGTGACACGTTTTTCCCAGAAATTGATTTGGGGAAATATAAACTTCTCCCAGAATACCCAGGCGTCCTCTCTGAGGTCCAGGAGGAAAAAGGCATCAAGTATAAGTTTGAAGTCTACGAGAAGAAAGACTAAAGATCCGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTAAGGTAAATATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTGTGTATTTTAGATTCCAACCTATGGAACTGATGAATGGGAGCAGTGGTGGAATGCCTTTAATGAGGAAAACCTGTTTTGCTCAGAAGAAATGCCATCTAGTGATGATGAGGCTACTGCTGACTCTCAACATTCTACTCCTCCAAAAAAGAAGAGAAAGGTAGAAGACCCCAAGGACTTTCCTTCAGAATTGCTAAGTTTTTTGAGTCATGCTGTGTTTAGTAATAGAACTCTTGCTTGCTTTGCTATTTACACCACAAAGGAAAAAGCTGCACTGCTATACAAGAAAATTATGGAAAAATATTCTGTAACCTTTATAAGTAGGCATAACAGTTATAATCATAACATACTGTTTTTTCTTACTCCACACAGGCATAGAGTGTCTGCTATTAATAACTATGCTCAAAAATTGTGTACCTTTAGCTTTTTAATTTGTAAAGGGGTTAATAAGGAATATTTGATGTATAGTGCCTTGACTAGAGATCATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGGCCCATCGATGCCGACGTAGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGAAAGCTT
<서열번호 4> 개시 코돈의 상류에 최적화한 Kozak가 부가되어 있는 항체(OMLH) 경쇄 유전자 cDNA의 뉴클레오티드 서열
CGCGCCCCGCCGCCACCATGGGTTGGTCTTGTATCATCTTATTTTTAGTTGCTACTGCTACTGGTGTTCATTCTGATATACAGCTCACCCAAAGCCCATCATCTCTGTCTGCAAGCGTCGGCGACAGGGTGACCATTACCTGTCGCGCAAGCCAAAGCGTTGACTACGACGGCGACAGCTACATGAACTGGTACCAGCAGAAGCCCGGCAAGGCTCCTAAGCTGCTGATCTATGCCGCCTCCTACCTTGAATCTGGAGTGCCTTCTCGTTTTTCCGGCTCAGGGTCCGGAACTGATTTTACCCTGACCATTTCCTCCCTCCAGCCCGAGGATTTTGCCACTTACTACTGTCAGCAGTCCCACGAGGACCCATATACATTCGGACAAGGTACAAAGGTAGAAATCAAGCGTACGGTGGCTGCCCCATCCGTGTTCATATTTCCTCCTAGCGACGAACAACTCAAGTCCGGTACCGCCAGCGTGGTCTGCCTGTTGAACAATTTTTATCCAAGAGAAGCTAAGGTCCAGTGGAAGGTTGACAACGCCCTTCAGTCCGGAAATAGCCAAGAGAGCGTCACCGAACAGGACTCCAAGGACAGTACATACTCACTGAGCTCTACACTGACCCTTTCTAAGGCCGACTACGAGAAGCACAAGGTCTACGCATGCGAAGTGACCCATCAGGGACTCAGTAGCCCTGTAACAAAGAGTTTTAATCGAGGCGAGTGCTAAGCGAT
<서열번호 5> 개시 코돈의 상류에 최적화한 Kozak가 부가되어 있는 항체(OMLH) 중쇄 유전자 cDNA의 뉴클레오티드 서열
GGCCGCCCGCCGCCACCATGGAATTTGGTTTATCTTGGGTTTTTTTAGTTGCTTTATTAAGAGGTGTTCAATGTGAGGTGCAGTTGGTCGAATCCGGCGGCGGACTCGTGCAACCAGGCGGAAGTTTGCGGCTGTCCTGCGCAGTGTCTGGTTACAGCATCACCTCCGGGTATAGCTGGAACTGGATCCGCCAGGCTCCTGGAAAGGGGCTTGAGTGGGTGGCTTCCATTACCTACGACGGCTCCACTAACTATAACCCGAGCGTCAAAGGCAGAATCACCATCTCTCGGGACGACTCAAAGAATACCTTCTACCTGCAGATGAACTCACTGAGGGCCGAAGATACCGCAGTTTACTACTGCGCCAGGGGGTCCCACTATTTCGGCCACTGGCACTTCGCCGTGTGGGGACAGGGCACACTCGTGACCGTTAGTAGCGCTAGCACCAAAGGCCCCTCTGTGTTCCCACTTGCTCCCTCCAGTAAATCTACCTCCGGAGGAACCGCAGCCCTCGGCTGCCTGGTGAAGGATTACTTCCCAGAGCCCGTCACCGTCTCTTGGAACTCCGGAGCCTTGACTAGCGGAGTGCACACTTTCCCTGCTGTATTGCAGTCCAGCGGCTTGTATTCACTGAGTAGCGTCGTCACCGTGCCTTCAAGCAGCCTCGGGACACAGACATACATATGTAATGTCAACCATAAGCCATCAAACACTAAAGTTGATAAAAAGGTGGAACCTAAGAGTTGCGATAAGACCCATACCTGTCCTCCTTGCCCTGCTCCTGAGCTGCTGGGAGGCCCTAGCGTGTTTCTGTTCCCCCCCAAGCCCAAAGATACACTGATGATTTCCCGCACACCTGAAGTAACATGTGTCGTGGTTGATGTGAGTCACGAGGATCCAGAGGTCAAGTTTAATTGGTACGtGGACGGAGTGGAGGTGCACAACGCTAAGACTAAGCCTCGGGAGGAACAGTACAACAGCACATACCGCGTGGTCAGCGTTTTGACTGTGCTGCATCAAGACTGGCTCAATGGAAAGGAATACAAGTGCAAGGTCTCTAATAAAGCCCTCCCCGCTCCTATTGAGAAGACTATTTCTAAAGCCAAGGGCCAGCCTCGCGAACCTCAGGTATATACTTTGCCACCCTCTCGCGAAGAAATGACAAAGAATCAGGTCTCACTCACTTGCCTCGTCAAAGGGTTTTACCCTTCTGACATCGCTGTCGAATGGGAAAGTAATGGTCAGCCAGAAAACAATTACAAGACTACTCCACCAGTGCTCGATTCTGATGGAAGTTTCTTTCTCTACAGTAAGCTCACTGTGGACAAATCTCGCTGGCAGCAGGGTAACGTATTCTCATGCTCCGTGATGCATGAAGCCCTCCACAACCATTACACCCAGAAGAGCCTGTCTCTGAGCCCAGGCAAGTAACCTGCA
<서열번호 6> pDC61의 전체 서열
CGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCTCTGGCTAACTAGAGAACCCACTGTTAACTGGCTTATCGAAATTGTCGAGGAGAACTTCAGGGTGAGTTTGGGGACCCTTGATTGTTCTTTCTTTTTCGCTATTGTAAAATTCATGTTATATGGAGGGGGCAAAGTTTTCAGGGTGTTGTTTAGAATGGGAAGATGTCCCTTGTATCACCATGGACCCTCATGATAATTTTGTTTCTTTCACTTTCTACTCTGTTGACAACCATTGTCTCCTCTTATTTTCTTTTCATTTTCTGTAACTTTTTCGTTAAACTTTAGCTTGCATTTGTAACGAATTTTTAAATTCACTTTTGTTTATTTGTCAGATTGTAAGTACTTTCTCTAATCACTTTTTTTTCAAGGCAATCAGGGTATATTATATTGTACTTCAGCACAGTTTTAGAGAACAATTGTTATAATTAAATGATAAGGTAGAATATTTCTGCATATAAATTCTGGCTGGCGTGGAAATATTCTTATTGGTAGAAACAACTACATCCTGGTCATCATCCTGCCTTTCTCTTTATGGTTACAATGATATACACTGTTTGAGATGAGGATAAAATACTCTGAGTCCAAACCGGGCCCCTCTGCTAACCATGTTCATGCCTTCTTCTTTTTCCTACAGCTCCTGGGCAACGTGCTGGCGGCCGCCCCGGGGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACTCGAGCGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCTCTGGCTAACTAGAGAACCCACTGTTAACTGGCTTATCGAAATTGTCGAGGAGAACTTCAGGGTGAGTTTGGGGACCCTTGATTGTTCTTTCTTTTTCGCTATTGTAAAATTCATGTTATATGGAGGGGGCAAAGTTTTCAGGGTGTTGTTTAGAATGGGAAGATGTCCCTTGTATCACCATGGACCCTCATGATAATTTTGTTTCTTTCACTTTCTACTCTGTTGACAACCATTGTCTCCTCTTATTTTCTTTTCATTTTCTGTAACTTTTTCGTTAAACTTTAGCTTGCATTTGTAACGAATTTTTAAATTCACTTTTGTTTATTTGTCAGATTGTAAGTACTTTCTCTAATCACTTTTTTTTCAAGGCAATCAGGGTATATTATATTGTACTTCAGCACAGTTTTAGAGAACAATTGTTATAATTAAATGATAAGGTAGAATATTTCTGCATATAAATTCTGGCTGGCGTGGAAATATTCTTATTGGTAGAAACAACTACATCCTGGTCATCATCCTGCCTTTCTCTTTATGGTTACAATGATATACACTGTTTGAGATGAGGATAAAATACTCTGAGTCCAAACCGGGCCCCTCTGCTAACCATGTTCATGCCTTCTTCTTTTTCCTACAGCTCCTGGGCAACGTGCTGGCGCGCCTCTAGAGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGAGGATCTCCGCGGTGTGGAATGTGTGTCAGTTAGGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCATAGTCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGCTTTTGCAAAAAAGCTGCAGATGGTACGACCATTAAATTGTATTGTAGCAGTATCACAAAATATGGGTATTGGTAAAAATGGTGATTTACCATGGCCACCATTACGAAATGAATTTAAATATTTTCAACGAATGACTACTACTTCATCAGTAGAAGGTAAACAAAATTTAGTAATTATGGGTCGAAAAACTTGGTTTTCAATTCCTGAGAAGAATCGACCTTTAAAGGACAGAATTAATATAGTTCTCAGTAGAGAACTCAAAGAACCACCACGAGGAGCTCATTTTCTTGCCAAAAGTTTGGATGATGCCTTAAGACTTATTGAACAACCGGAATTGGCAAGTAAAGTAGACATGGTTTGGATAGTCGGAGGCAGTTCTGTTTACCAGGAAGCCATGAATCAACCAGGCCACCTCAGACTCTTTGTGACAAGGATCATGCAGGAATTTGAAAGTGACACGTTTTTCCCAGAAATTGATTTGGGGAAATATAAACTTCTCCCAGAATACCCAGGCGTCCTCTCTGAGGTCCAGGAGGAAAAAGGCATCAAGTATAAGTTTGAAGTCTACGAGAAGAAAGACTAAAGATCCGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTAAGGTAAATATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTGTGTATTTTAGATTCCAACCTATGGAACTGATGAATGGGAGCAGTGGTGGAATGCCTTTAATGAGGAAAACCTGTTTTGCTCAGAAGAAATGCCATCTAGTGATGATGAGGCTACTGCTGACTCTCAACATTCTACTCCTCCAAAAAAGAAGAGAAAGGTAGAAGACCCCAAGGACTTTCCTTCAGAATTGCTAAGTTTTTTGAGTCATGCTGTGTTTAGTAATAGAACTCTTGCTTGCTTTGCTATTTACACCACAAAGGAAAAAGCTGCACTGCTATACAAGAAAATTATGGAAAAATATTCTGTAACCTTTATAAGTAGGCATAACAGTTATAATCATAACATACTGTTTTTTCTTACTCCACACAGGCATAGAGTGTCTGCTATTAATAACTATGCTCAAAAATTGTGTACCTTTAGCTTTTTAATTTGTAAAGGGGTTAATAAGGAATATTTGATGTATAGTGCCTTGACTAGAGATCATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGGCCCATCGATGAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTT
<서열번호 7> pDC6 내의 서열과 치환한 서열(pDC61)
AATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCA
<서열번호 8> pDC61의 염기 서열 No1267-No1273과 치환한 인간 오말리즈맙 항체(OMLH) 경쇄를 코드하는 cDNA
GGCCGCCACCATGGGTTGGTCTTGTATCATCTTATTTTTAGTTGCTACTGCTACTGGTGTTCATTCTGATATACAGCTCACCCAAAGCCCATCATCTCTGTCTGCAAGCGTCGGCGACAGGGTGACCATTACCTGTCGCGCAAGCCAAAGCGTTGACTACGACGGCGACAGCTACATGAACTGGTACCAGCAGAAGCCCGGCAAGGCTCCTAAGCTGCTGATCTATGCCGCCTCCTACCTTGAATCTGGAGTGCCTTCTCGTTTTTCCGGCTCAGGGTCCGGAACTGATTTTACCCTGACCATTTCCTCCCTCCAGCCCGAGGATTTTGCCACTTACTACTGTCAGCAGTCCCACGAGGACCCATATACATTCGGACAAGGTACAAAGGTAGAAATCAAGCGTACGGTGGCTGCCCCATCCGTGTTCATATTTCCTCCTAGCGACGAACAACTCAAGTCCGGTACCGCCAGCGTGGTCTGCCTGTTGAACAATTTTTATCCAAGAGAAGCTAAGGTCCAGTGGAAGGTTGACAACGCCCTTCAGTCCGGAAATAGCCAAGAGAGCGTCACCGAACAGGACTCCAAGGACAGTACATACTCACTGAGCTCTACACTGACCCTTTCTAAGGCCGACTACGAGAAGCACAAGGTCTACGCATGCGAAGTGACCCATCAGGGACTCAGTAGCCCTGTAACAAAGAGTTTTAATCGAGGCGAGTGCTAAC
<서열번호 9> pDC61의 염기 서열 No2765-No2771로 치환한 인간 오말리즈맙 항체(OMLH) 중쇄를 코드하는 cDNA
CGCGCCACCATGGAATTTGGTTTATCTTGGGTTTTTTTAGTTGCTTTATTAAGAGGTGTTCAATGTGAGGTGCAGTTGGTCGAATCCGGCGGCGGACTCGTGCAACCAGGCGGAAGTTTGCGGCTGTCCTGCGCAGTGTCTGGTTACAGCATCACCTCCGGGTATAGCTGGAACTGGATCCGCCAGGCTCCTGGAAAGGGGCTTGAGTGGGTGGCTTCCATTACCTACGACGGCTCCACTAACTATAACCCGAGCGTCAAAGGCAGAATCACCATCTCTCGGGACGACTCAAAGAATACCTTCTACCTGCAGATGAACTCACTGAGGGCCGAAGATACCGCAGTTTACTACTGCGCCAGGGGGTCCCACTATTTCGGCCACTGGCACTTCGCCGTGTGGGGACAGGGCACACTCGTGACCGTTAGTAGCGCTAGCACCAAAGGCCCCTCTGTGTTCCCACTTGCTCCCTCCAGTAAATCTACCTCCGGAGGAACCGCAGCCCTCGGCTGCCTGGTGAAGGATTACTTCCCAGAGCCCGTCACCGTCTCTTGGAACTCCGGAGCCTTGACTAGCGGAGTGCACACTTTCCCTGCTGTATTGCAGTCCAGCGGCTTGTATTCACTGAGTAGCGTCGTCACCGTGCCTTCAAGCAGCCTCGGGACACAGACATACATATGTAATGTCAACCATAAGCCATCAAACACTAAAGTTGATAAAAAGGTGGAACCTAAGAGTTGCGATAAGACCCATACCTGTCCTCCTTGCCCTGCTCCTGAGCTGCTGGGAGGCCCTAGCGTGTTTCTGTTCCCCCCCAAGCCCAAAGATACACTGATGATTTCCCGCACACCTGAAGTAACATGTGTCGTGGTTGATGTGAGTCACGAGGATCCAGAGGTCAAGTTTAATTGGTACGtGGACGGAGTGGAGGTGCACAACGCTAAGACTAAGCCTCGGGAGGAACAGTACAACAGCACATACCGCGTGGTCAGCGTTTTGACTGTGCTGCATCAAGACTGGCTCAATGGAAAGGAATACAAGTGCAAGGTCTCTAATAAAGCCCTCCCCGCTCCTATTGAGAAGACTATTTCTAAAGCCAAGGGCCAGCCTCGCGAACCTCAGGTATATACTTTGCCACCCTCTCGCGAAGAAATGACAAAGAATCAGGTCTCACTCACTTGCCTCGTCAAAGGGTTTTACCCTTCTGACATCGCTGTCGAATGGGAAAGTAATGGTCAGCCAGAAAACAATTACAAGACTACTCCACCAGTGCTCGATTCTGATGGAAGTTTCTTTCTCTACAGTAAGCTCACTGTGGACAAATCTCGCTGGCAGCAGGGTAACGTATTCTCATGCTCCGTGATGCATGAAGCCCTCCACAACCATTACACCCAGAAGAGCCTGTCTCTGAGCCCAGGCAAGTAAT
<서열번호 10> 백본 벡터 pNC32c-U533의 전체 염기 서열
CGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCTCTGGCTAACTAGAGAACCCACTGTTAACTGGCTTATCGAAATTGGCGCGCCGCGATCGCCCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGATCTCCGCGGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGAGAATTCCGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCTCTGGCTAACTAGAGAACCCACTGTTAACTGGCTTATCGAAATTGCGGCCGCCCTGCAGGCCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGACTCCGCGGGGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGAACCGGTTGTGGAATGTGTGTCAGTTAGGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCAGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCATAGTCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGCTTTTGCAAAAAAGCTGCAGATGATTGAACAAGATGGATTGCACGCAGGTTCTCCGGCCGCTTGGGTGGAGAGGCTATTCGGCTATGACTGGGCACAACAGACAATCGGCTGCTCTGATGCCGCCGTGTTCCGGCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGACCGACCTGTCCGGTGCCCTGAATGAACTGCAGGACGAGGCAGCGCGGCTATCGTGGCTGGCCACGACGGGCGTTCCTTGCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAGGGACTGGCTGCTATTGGGCGAAGTGCCGGGGCAGGATCTCCTGTCATCTCACCTTGCTCCTGCCGAGAAAGTATCCATCATGGCTGATGCAATGCGGCGGCTGCATACGCTTGATCCGGCTACCTGCCCATTCGACCACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCGGATGGAAGCCGGTCTTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCAGCCGAACTGTTCGCCAGGCTCAAGGCGCGCATGCCCGACGGCGAGGATCTCGTCGTGACCCATGGCGATGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTCTGGATTCATCGACTGTGGCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCGTTGGCTACCCGTGATATTGCTGAAGAGCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGCTTTACGGTATCGCCGCTCCCGATTCGCAGCGCATCGCCTTCTATCGCCTTCTTGACGAGTTCTTCTGAGATCCGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTAAGGTAAATATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTGTGTATTTTAGATTCCAACCTATGGAACTGATGAATGGGAGCAGTGGTGGAATGCCTTTAATGAGGAAAACCTGTTTTGCTCAGAAGAAATGCCATCTAGTGATGATGAGGCTACTGCTGACTCTCAACATTCTACTCCTCCAAAAAAGAAGAGAAAGGTAGAAGACCCCAAGGACTTTCCTTCAGAATTGCTAAGTTTTTTGAGTCATGCTGTGTTTAGTAATAGAACTCTTGCTTGCTTTGCTATTTACACCACAAAGGAAAAAGCTGCACTGCTATACAAGAAAATTATGGAAAAATATTCTGTAACCTTTATAAGTAGGCATAACAGTTATAATCATAACATACTGTTTTTTCTTACTCCACACAGGCATAGAGTGTCTGCTATTAATAACTATGCTCAAAAATTGTGTACCTTTAGCTTTTTAATTTGTAAAGGGGTTAATAAGGAATATTTGATGTATAGTGCCTTGACTAGAGATCATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGGCCCATCGATGCCGACGTAGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGAAAGCTT
<서열번호 11> pNC32c-U533의 염기 서열 No 658-No 668로 치환한 인간 오말리즈맙 항체(OMLH) 경쇄를 코드하는 cDNA
CGCGCCACCATGGGTTGGTCTTGTATCATCTTATTTTTAGTTGCTACTGCTACTGGTGTTCATTCTGATATACAGCTCACCCAAAGCCCATCATCTCTGTCTGCAAGCGTCGGCGACAGGGTGACCATTACCTGTCGCGCAAGCCAAAGCGTTGACTACGACGGCGACAGCTACATGAACTGGTACCAGCAGAAGCCCGGCAAGGCTCCTAAGCTGCTGATCTATGCCGCCTCCTACCTTGAATCTGGAGTGCCTTCTCGTTTTTCCGGCTCAGGGTCCGGAACTGATTTTACCCTGACCATTTCCTCCCTCCAGCCCGAGGATTTTGCCACTTACTACTGTCAGCAGTCCCACGAGGACCCATATACATTCGGACAAGGTACAAAGGTAGAAATCAAGCGTACGGTGGCTGCCCCATCCGTGTTCATATTTCCTCCTAGCGACGAACAACTCAAGTCCGGTACCGCCAGCGTGGTCTGCCTGTTGAACAATTTTTATCCAAGAGAAGCTAAGGTCCAGTGGAAGGTTGACAACGCCCTTCAGTCCGGAAATAGCCAAGAGAGCGTCACCGAACAGGACTCCAAGGACAGTACATACTCACTGAGCTCTACACTGACCCTTTCTAAGGCCGACTACGAGAAGCACAAGGTCTACGCATGCGAAGTGACCCATCAGGGACTCAGTAGCCCTGTAACAAAGAGTTTTAATCGAGGCGAGTGCTAAGCGAT
<서열번호 12> pNC32c-U533의 염기 서열 No 3081-No 3092로 치환한 인간 오말리즈맙 항체(OMLH) 중쇄를 코드하는 cDNA
GGCCGCCACCATGGAATTTGGTTTATCTTGGGTTTTTTTAGTTGCTTTATTAAGAGGTGTTCAATGTGAGGTGCAGTTGGTCGAATCCGGCGGCGGACTCGTGCAACCAGGCGGAAGTTTGCGGCTGTCCTGCGCAGTGTCTGGTTACAGCATCACCTCCGGGTATAGCTGGAACTGGATCCGCCAGGCTCCTGGAAAGGGGCTTGAGTGGGTGGCTTCCATTACCTACGACGGCTCCACTAACTATAACCCGAGCGTCAAAGGCAGAATCACCATCTCTCGGGACGACTCAAAGAATACCTTCTACCTGCAGATGAACTCACTGAGGGCCGAAGATACCGCAGTTTACTACTGCGCCAGGGGGTCCCACTATTTCGGCCACTGGCACTTCGCCGTGTGGGGACAGGGCACACTCGTGACCGTTAGTAGCGCTAGCACCAAAGGCCCCTCTGTGTTCCCACTTGCTCCCTCCAGTAAATCTACCTCCGGAGGAACCGCAGCCCTCGGCTGCCTGGTGAAGGATTACTTCCCAGAGCCCGTCACCGTCTCTTGGAACTCCGGAGCCTTGACTAGCGGAGTGCACACTTTCCCTGCTGTATTGCAGTCCAGCGGCTTGTATTCACTGAGTAGCGTCGTCACCGTGCCTTCAAGCAGCCTCGGGACACAGACATACATATGTAATGTCAACCATAAGCCATCAAACACTAAAGTTGATAAAAAGGTGGAACCTAAGAGTTGCGATAAGACCCATACCTGTCCTCCTTGCCCTGCTCCTGAGCTGCTGGGAGGCCCTAGCGTGTTTCTGTTCCCCCCCAAGCCCAAAGATACACTGATGATTTCCCGCACACCTGAAGTAACATGTGTCGTGGTTGATGTGAGTCACGAGGATCCAGAGGTCAAGTTTAATTGGTACGtGGACGGAGTGGAGGTGCACAACGCTAAGACTAAGCCTCGGGAGGAACAGTACAACAGCACATACCGCGTGGTCAGCGTTTTGACTGTGCTGCATCAAGACTGGCTCAATGGAAAGGAATACAAGTGCAAGGTCTCTAATAAAGCCCTCCCCGCTCCTATTGAGAAGACTATTTCTAAAGCCAAGGGCCAGCCTCGCGAACCTCAGGTATATACTTTGCCACCCTCTCGCGAAGAAATGACAAAGAATCAGGTCTCACTCACTTGCCTCGTCAAAGGGTTTTACCCTTCTGACATCGCTGTCGAATGGGAAAGTAATGGTCAGCCAGAAAACAATTACAAGACTACTCCACCAGTGCTCGATTCTGATGGAAGTTTCTTTCTCTACAGTAAGCTCACTGTGGACAAATCTCGCTGGCAGCAGGGTAACGTATTCTCATGCTCCGTGATGCATGAAGCCCTCCACAACCATTACACCCAGAAGAGCCTGTCTCTGAGCCCAGGCAAGTAACCTGCA
<서열번호 13> UCOE-Hu-P의 염기 서열 No5309-No5311로 치환한 BstBI 인식 사이트를 포함하는 서열
CCTAGTAGTAGTAGTAGTTCGAAG
<서열번호 14> 인간 오말리즈맙 항체 경쇄(OML)를 코드하는 cDNA 및 시미안 바이러스 40 폴리아데닐화 신호(SV40pA), 모르모트 사이토메갈로바이러스 프로모터(PgpCMV), 인간 오말리즈맙 항체 중쇄(OMH)를 코드하는 cDNA를 연결한 서열
CCCACCATGGGTTGGTCTTGTATCATCTTATTTTTAGTTGCTACTGCTACTGGTGTTCATTCTGATATACAGCTCACCCAAAGCCCATCATCTCTGTCTGCAAGCGTCGGCGACAGGGTGACCATTACCTGTCGCGCAAGCCAAAGCGTTGACTACGACGGCGACAGCTACATGAACTGGTACCAGCAGAAGCCCGGCAAGGCTCCTAAGCTGCTGATCTATGCCGCCTCCTACCTTGAATCTGGAGTGCCTTCTCGTTTTTCCGGCTCAGGGTCCGGAACTGATTTTACCCTGACCATTTCCTCCCTCCAGCCCGAGGATTTTGCCACTTACTACTGTCAGCAGTCCCACGAGGACCCATATACATTCGGACAAGGTACAAAGGTAGAAATCAAGCGTACGGTGGCTGCCCCATCCGTGTTCATATTTCCTCCTAGCGACGAACAACTCAAGTCCGGTACCGCCAGCGTGGTCTGCCTGTTGAACAATTTTTATCCAAGAGAAGCTAAGGTCCAGTGGAAGGTTGACAACGCCCTTCAGTCCGGAAATAGCCAAGAGAGCGTCACCGAACAGGACTCCAAGGACAGTACATACTCACTGAGCTCTACACTGACCCTTTCTAAGGCCGACTACGAGAAGCACAAGGTCTACGCATGCGAAGTGACCCATCAGGGACTCAGTAGCCCTGTAACAAAGAGTTTTAATCGAGGCGAGTGCTAAGCGATCGCGCTAGCGCGGCCACGTAGTCGACTACGTAGAGCTCGGTACCCGGGGATCCTCTAGAGTCGACCTGCAGGCATGCAAGCTGGCCGCGACTCTAGATCATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTAACCGGTTATGTTACTTGGCAGAGGCCGCATGGAAAGTCCCTGGACGTGGGACATCTGATTAATACGTGAGGAGGTCAGCCATGTTCTTTTTGGCAAAGGACTACGGTCATTGGACGTTTGATTGGCATGGGATAGGGTCAGCCAGAGTTAACAGTGTTCTTTTGGCAAAGGGATACGTGGAAAGTCCCGGGCCATTTACAGTAAACTGATACGGGGACAAAGCACAGCCATATTTAGTCATGTATTGCTTGGCAGAGGGTCTATGGAAAGTCCCTGGACGTGGGACGTCTGATTAATATGAAAGAAGGTCAGCCAGAGGTAGCTGTGTCCTTTTTGGCAAAGGGATACGGTTATGGGACGTTTGATTGGACTGGGATAGGGTCAGCCAGAGTTAACAGTGTTCTTTTGGCAAAGGAAACGTGGAAAGTCCCGGGCCATTTACAGTAAACTGATACTGGGACAAAGTACACCCATATTTAGTCATGTTCTTTTTGGCAAAGAGCATCTGGAAAGTCCCGGGCAGCATTATAGTCACTTGGCAGAGGGAAAGGGTCACTCAGAGTTAAGTACATCTTTCCAGGGCCAATATTCCAGTAAATTACACTTAGTTTTATGCAAATCAGCCACAAAGGGGATTTTCCCGGTCAATTATGACTTTTTCCTTAGTCATGCGGTATCCAATTACTGCCAAATTGGCAGTACATACTAGGTGATTCACTGACATTTGGCCGTCCTCTGGAAAGTCCCTGGAAACCGCTCAAGTACTGTATCATGGTGACTTTGCATTTTTGGAGAGCACGCCCCACTCCACCATTGGTCCACGTACCCTATGGGGGAGTGGTTTATGAGTATATAAGGGGCTCCGGTTTAGAAGCCGGGCAGAGCGGAATTCGAGCTCCCTGCAGGTTAGTTAAGTTAACGGCGCGCCACCATGGAATTTGGTTTATCTTGGGTTTTTTTAGTTGCTTTATTAAGAGGTGTTCAATGTGAGGTGCAGTTGGTCGAATCCGGCGGCGGACTCGTGCAACCAGGCGGAAGTTTGCGGCTGTCCTGCGCAGTGTCTGGTTACAGCATCACCTCCGGGTATAGCTGGAACTGGATCCGCCAGGCTCCTGGAAAGGGGCTTGAGTGGGTGGCTTCCATTACCTACGACGGCTCCACTAACTATAACCCGAGCGTCAAAGGCAGAATCACCATCTCTCGGGACGACTCAAAGAATACCTTCTACCTGCAGATGAACTCACTGAGGGCCGAAGATACCGCAGTTTACTACTGCGCCAGGGGGTCCCACTATTTCGGCCACTGGCACTTCGCCGTGTGGGGACAGGGCACACTCGTGACCGTTAGTAGCGCTAGCACCAAAGGCCCCTCTGTGTTCCCACTTGCTCCCTCCAGTAAATCTACCTCCGGAGGAACCGCAGCCCTCGGCTGCCTGGTGAAGGATTACTTCCCAGAGCCCGTCACCGTCTCTTGGAACTCCGGAGCCTTGACTAGCGGAGTGCACACTTTCCCTGCTGTATTGCAGTCCAGCGGCTTGTATTCACTGAGTAGCGTCGTCACCGTGCCTTCAAGCAGCCTCGGGACACAGACATACATATGTAATGTCAACCATAAGCCATCAAACACTAAAGTTGATAAAAAGGTGGAACCTAAGAGTTGCGATAAGACCCATACCTGTCCTCCTTGCCCTGCTCCTGAGCTGCTGGGAGGCCCTAGCGTGTTTCTGTTCCCCCCCAAGCCCAAAGATACACTGATGATTTCCCGCACACCTGAAGTAACATGTGTCGTGGTTGATGTGAGTCACGAGGATCCAGAGGTCAAGTTTAATTGGTACGtGGACGGAGTGGAGGTGCACAACGCTAAGACTAAGCCTCGGGAGGAACAGTACAACAGCACATACCGCGTGGTCAGCGTTTTGACTGTGCTGCATCAAGACTGGCTCAATGGAAAGGAATACAAGTGCAAGGTCTCTAATAAAGCCCTCCCCGCTCCTATTGAGAAGACTATTTCTAAAGCCAAGGGCCAGCCTCGCGAACCTCAGGTATATACTTTGCCACCCTCTCGCGAAGAAATGACAAAGAATCAGGTCTCACTCACTTGCCTCGTCAAAGGGTTTTACCCTTCTGACATCGCTGTCGAATGGGAAAGTAATGGTCAGCCAGAAAACAATTACAAGACTACTCCACCAGTGCTCGATTCTGATGGAAGTTTCTTTCTCTACAGTAAGCTCACTGTGGACAAATCTCGCTGGCAGCAGGGTAACGTATTCTCATGCTCCGTGATGCATGAAGCCCTCCACAACCATTACACCCAGAAGAGCCTGTCTCTGAGCCCAGGCAAGTAATCTAGATT
<서열번호 15> 개 CTLA-4-Ig의 아미노산 서열
MAGFGFRRHGAQPDLASRTWPCTALFSLLFIPVFSKGMHVAQPAVVLASSRGVASFVCEYGSSGNAAEVRVTVLRQAGSQMTEVCAATYTVEDELAFLDDSTCTGTSSGNKVNLTIQGLRAMDTGLYICKVELMYPPPYYVGMGNGTQIYVIDPEPCPDSDPKESTCKCISPCPVPESLGGPSVFIFPPKPKDILRITRTPEITCVVLDLGREDPEVQISWFVDGKEVHTAKTQPREQQFNSTYRVVSVLPIEHQDWLTGKEFKCRVNHIGLPSPIERTISKARGQAHQPSVYVLPPSPKELSSSDTVTLTCLIKDFFPPEIDVEWQSNGQPEPESKYHTTAPQLDEDGSYFLYSKLSVDKSRWQQGDTFTCAVMHEALQNHYTDLSLSHSPGK
<서열번호 16> 개 CTLA-4-Ig의 코돈 최적화 후 뉴클레오티드 서열
ATGGCTGGATTTGGATTCAGAAGGCACGGAGCCCAGCCCGACCTGGCATCTCGCACTTGGCCCTGTACCGCACTGTTTTCACTGCTGTTCATCCCAGTGTTCAGCAAGGGAATGCACGTGGCTCAGCCAGCTGTGGTGCTGGCTTCCAGCAGAGGCGTGGCTTCCTTCGTGTGCGAGTACGGCTCTTCCGGCAACGCCGCTGAGGTGAGAGTGACCGTGCTGAGGCAGGCTGGCTCCCAGATGACAGAGGTGTGCGCCGCTACCTATACAGTGGAGGACGAGCTGGCTTTCCTGGACGATAGCACCTGTACAGGCACCAGCTCTGGCAACAAGGTCAATCTGACCATCCAGGGCCTGCGCGCCATGGATACAGGCCTGTACATCTGTAAGGTGGAGCTGATGTATCCCCCTCCATACTATGTGGGCATGGGCAATGGCACCCAGATCTACGTGATCGACCCCGAGCCTTGCCCAGACTCTGATCCAAAGGAGTCGACATGCAAGTGTATCTCTCCATGTCCTGTGCCAGAGAGCCTGGGAGGACCTTCCGTGTTCATCTTTCCCCCTAAGCCAAAGGATATCCTGAGGATCACACGGACCCCTGAGATCACCTGCGTGGTGCTGGACCTGGGAAGGGAGGATCCAGAGGTGCAGATCTCCTGGTTCGTGGACGGCAAGGAGGTGCATACCGCTAAGACACAGCCCAGAGAGCAGCAGTTTAACTCCACCTATCGCGTGGTGAGCGTGCTGCCTATCGAGCACCAGGATTGGCTGACAGGCAAGGAGTTTAAGTGCCGGGTGAATCATATCGGCCTGCCCTCTCCTATCGAGAGGACCATCTCCAAGGCTAGGGGACAGGCTCACCAGCCAAGCGTGTACGTGCTGCCACCCTCTCCTAAGGAGCTGTCCAGCTCTGACACAGTGACCCTGACATGTCTGATCAAGGACTTCTTTCCTCCAGAGATCGACGTGGAGTGGCAGTCCAACGGCCAGCCAGAGCCCGAGAGCAAGTATCATACCACAGCCCCCCAGCTGGACGAGGATGGCTCTTACTTCCTGTATTCCAAGCTGAGCGTGGACAAGTCCAGGTGGCAGCAGGGCGATACCTTTACATGTGCTGTGATGCACGAAGCCCTGCAGAATCATTACACCGACCTGTCACTGTCCCACTCCCCTGGCAAATAA
<서열번호 17> 외래 유전자 삽입 부위가 1개 있는 pDC62c5-U533의 전체 염기 서열
CGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACACCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAATAACCCCGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCCTCACTCTCTTCCGCATCGCTGTCTGCGAGGGCCAGCTGTTGGGCTCGCGGTTGAGGACAAACTCTTCGCGGTCTTTCCAGTACTCTTGGATCGGAAACCCGTCGGCCTCCGAACGGTACTCCGCCACCGAGGGACCTGAGCGAGTCCGCATCGACCGGATCGGAAAACCTCTCGAGAAAGGCGTCTAACCAGTCACAGTCGCAAGGTAGGCTGAGCACCGTGGCGGGCGGCAGCGGGTGGCGGTCGGGGTTGTTTCTGGCGGAGGTGCTGCTGATGATGTAATTAAAGTAGGCGGTCTTGAGACGGCGGATGGTCGAGGTGAGGTGTGGCAGGCTTGAGATCCAGCTGTTGGGGTGAGTACTCCCTCTCAAAAGCGGGCATGACTTCTGCGCTAAGATTGTCAGTTTCCAAAAACGAGGAGGATTTGATATTCACCTGGCCCGATCTGGCCATACACTTGAGTGACAATGACATCCACTTTGCCTTTCTCTCCACAGGTGTCCACTCCCAGGTCCAAGGCGCGCCGCGATCGCGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGAGGATCTCCGCGGGGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGAGAATTCaaatgggACCGGTTGTGGAATGTGTGTCAGTTAGGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCATAGTCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGCTTTTGCAAAAAAGCTGCAGATGGTACGACCATTAAATTGTATTGTAGCAGTATCACAAAATATGGGTATTGGTAAAAATGGTGATTTACCATGGCCACCATTACGAAATGAATTTAAATATTTTCAACGAATGACTACTACTTCATCAGTAGAAGGTAAACAAAATTTAGTAATTATGGGTCGAAAAACTTGGTTTTCAATTCCTGAGAAGAATCGACCTTTAAAGGACAGAATTAATATAGTTCTCAGTAGAGAACTCAAAGAACCACCACGAGGAGCTCATTTTCTTGCCAAAAGTTTGGATGATGCCTTAAGACTTATTGAACAACCGGAATTGGCAAGTAAAGTAGACATGGTTTGGATAGTCGGAGGCAGTTCTGTTTACCAGGAAGCCATGAATCAACCAGGCCACCTCAGACTCTTTGTGACAAGGATCATGCAGGAATTTGAAAGTGACACGTTTTTCCCAGAAATTGATTTGGGGAAATATAAACTTCTCCCAGAATACCCAGGCGTCCTCTCTGAGGTCCAGGAGGAAAAAGGCATCAAGTATAAGTTTGAAGTCTACGAGAAGAAAGACTAAAGATCCGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTAAGGTAAATATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTGTGTATTTTAGATTCCAACCTATGGAACTGATGAATGGGAGCAGTGGTGGAATGCCTTTAATGAGGAAAACCTGTTTTGCTCAGAAGAAATGCCATCTAGTGATGATGAGGCTACTGCTGACTCTCAACATTCTACTCCTCCAAAAAAGAAGAGAAAGGTAGAAGACCCCAAGGACTTTCCTTCAGAATTGCTAAGTTTTTTGAGTCATGCTGTGTTTAGTAATAGAACTCTTGCTTGCTTTGCTATTTACACCACAAAGGAAAAAGCTGCACTGCTATACAAGAAAATTATGGAAAAATATTCTGTAACCTTTATAAGTAGGCATAACAGTTATAATCATAACATACTGTTTTTTCTTACTCCACACAGGCATAGAGTGTCTGCTATTAATAACTATGCTCAAAAATTGTGTACCTTTAGCTTTTTAATTTGTAAAGGGGTTAATAAGGAATATTTGATGTATAGTGCCTTGACTAGAGATCATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGGCCCATCGATGCCGACGTAGCGCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGAAAGCTT
<서열번호 18> 개시 코돈의 상류에 최적화한 Kozak가 부가되어 있는 항체(TRLH) 경쇄 유전자 cDNA의 뉴클레오티드 서열
CGCGCCCCGCCGCCACCATGGGTTGGTCTTGTATCATCTTATTTTTAGTTGCTACTGCTACTGGTGTTCATTCTGATATACAGATGACCCAAAGCCCATCATCTCTGTCTGCAAGCGTCGGCGACAGGGTGACCATTACCTGTCGCGCAAGCCAAGACGTTAATACAGCAGTGGCTTGGTACCAGCAGAAGCCCGGCAAGGCTCCTAAGCTGCTGATCTATAGCGCCTCCTTTCTTTATTCTGGAGTGCCTTCTCGTTTTTCCGGCTCAAGGTCCGGAACTGATTTTACCCTGACCATTTCCTCCCTCCAGCCCGAGGATTTTGCCACTTACTACTGTCAGCAGCACTATACCACACCACCTACATTCGGACAAGGTACAAAGGTAGAAATCAAGCGTACGGTGGCTGCCCCATCCGTGTTCATATTTCCTCCTAGCGACGAACAACTCAAGTCCGGTACCGCCAGCGTGGTCTGCCTGTTGAACAATTTTTATCCAAGAGAAGCTAAGGTCCAGTGGAAGGTTGACAACGCCCTTCAGTCCGGAAATAGCCAAGAGAGCGTCACCGAACAGGACTCCAAGGACAGTACATACTCACTGAGCTCTACACTGACCCTTTCTAAGGCCGACTACGAGAAGCACAAGGTCTACGCATGCGAAGTGACCCATCAGGGACTCAGTAGCCCTGTAACAAAGAGTTTTAATCGAGGCGAGTGCTAAGCGAT
<서열번호 19> 개시 코돈의 상류에 최적화한 Kozak가 부가되어 있는 항체(TRLH) 중쇄 유전자 cDNA의 뉴클레오티드 서열
GGCCGCCCGCCGCCACCATGGAATTTGGTTTATCTTGGGTTTTTTTAGTTGCTTTATTAAGAGGTGTTCAATGTGAGGTGCAGTTGGTCGAATCCGGCGGCGGACTCGTGCAACCAGGCGGAAGTTTGCGGCTGTCCTGCGCAGCCTCTGGTTTTAACATCAAAGATACCTATATTCATTGGGTACGCCAGGCTCCTGGAAAGGGGCTTGAGTGGGTGGCTCGAATTTACCCAACCAATGGCTACACTCGGTATGCCGACAGCGTCAAAGGCAGATTCACCATCTCTGCAGACACATCAAAGAATACCGCTTACCTGCAGATGAACTCACTGAGGGCCGAAGATACCGCAGTTTACTACTGCTCCAGGTGGGGGGGGGACGGCTTCTACGCCATGGATTACTGGGGACAGGGCACACTCGTGACCGTTAGTAGCGCTAGCACCAAAGGCCCCTCTGTGTTCCCACTTGCTCCCTCCAGTAAATCTACCTCCGGAGGAACCGCAGCCCTCGGCTGCCTGGTGAAGGATTACTTCCCAGAGCCCGTCACCGTCTCTTGGAACTCCGGAGCCTTGACTAGCGGAGTGCACACTTTCCCTGCTGTATTGCAGTCCAGCGGCTTGTATTCACTGAGTAGCGTCGTCACCGTGCCTTCAAGCAGCCTCGGGACACAGACATACATATGTAATGTCAACCATAAGCCATCAAACACTAAAGTTGATAAAAAGGTGGAACCTAAGAGTTGCGATAAGACCCATACCTGTCCTCCTTGCCCTGCTCCTGAGCTGCTGGGAGGCCCTAGCGTGTTTCTGTTCCCCCCCAAGCCCAAAGATACACTGATGATTTCCCGCACACCTGAAGTAACATGTGTCGTGGTTGATGTGAGTCACGAGGATCCAGAGGTCAAGTTTAATTGGTACGTGGACGGAGTGGAGGTGCACAACGCTAAGACTAAGCCTCGGGAGGAACAGTACAACAGCACATACCGCGTGGTCAGCGTTTTGACTGTGCTGCATCAAGACTGGCTCAATGGAAAGGAATACAAGTGCAAGGTCTCTAATAAAGCCCTCCCCGCTCCTATTGAGAAGACTATTTCTAAAGCCAAGGGCCAGCCTCGCGAACCTCAGGTATATACTTTGCCACCCTCTCGCGAAGAAATGACAAAGAATCAGGTCTCACTCACTTGCCTCGTCAAAGGGTTTTACCCTTCTGACATCGCTGTCGAATGGGAAAGTAATGGTCAGCCAGAAAACAATTACAAGACTACTCCACCAGTGCTCGATTCTGATGGAAGTTTCTTTCTCTACAGTAAGCTCACTGTGGACAAATCTCGCTGGCAGCAGGGTAACGTATTCTCATGCTCCGTGATGCATGAAGCCCTCCACAACCATTACACCCAGAAGAGCCTGTCTCTGAGCCCAGGCTAACCTGCA
GGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGA
<서열번호 2> 최적화된 Kozak 서열을 나타낸다. 이 서열 중에서, ATG는 개시 코돈이 된다.
CCGCCGCCACCATGG
<서열번호 3> pDC62c5-U533의 전체 서열
CGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACACCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAATAACCCCGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCCTCACTCTCTTCCGCATCGCTGTCTGCGAGGGCCAGCTGTTGGGCTCGCGGTTGAGGACAAACTCTTCGCGGTCTTTCCAGTACTCTTGGATCGGAAACCCGTCGGCCTCCGAACGGTACTCCGCCACCGAGGGACCTGAGCGAGTCCGCATCGACCGGATCGGAAAACCTCTCGAGAAAGGCGTCTAACCAGTCACAGTCGCAAGGTAGGCTGAGCACCGTGGCGGGCGGCAGCGGGTGGCGGTCGGGGTTGTTTCTGGCGGAGGTGCTGCTGATGATGTAATTAAAGTAGGCGGTCTTGAGACGGCGGATGGTCGAGGTGAGGTGTGGCAGGCTTGAGATCCAGCTGTTGGGGTGAGTACTCCCTCTCAAAAGCGGGCATGACTTCTGCGCTAAGATTGTCAGTTTCCAAAAACGAGGAGGATTTGATATTCACCTGGCCCGATCTGGCCATACACTTGAGTGACAATGACATCCACTTTGCCTTTCTCTCCACAGGTGTCCACTCCCAGGTCCAAGGCGCGCCGCGATCGCGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGAGGATCTCCGCGGGGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGAGAATTCCGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACACCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAATAACCCCGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCCTCACTCTCTTCCGCATCGCTGTCTGCGAGGGCCAGCTGTTGGGCTCGCGGTTGAGGACAAACTCTTCGCGGTCTTTCCAGTACTCTTGGATCGGAAACCCGTCGGCCTCCGAACGGTACTCCGCCACCGAGGGACCTGAGCGAGTCCGCATCGACCGGATCGGAAAACCTCTCGAGAAAGGCGTCTAACCAGTCACAGTCGCAAGGTAGGCTGAGCACCGTGGCGGGCGGCAGCGGGTGGCGGTCGGGGTTGTTTCTGGCGGAGGTGCTGCTGATGATGTAATTAAAGTAGGCGGTCTTGAGACGGCGGATGGTCGAGGTGAGGTGTGGCAGGCTTGAGATCCAGCTGTTGGGGTGAGTACTCCCTCTCAAAAGCGGGCATGACTTCTGCGCTAAGATTGTCAGTTTCCAAAAACGAGGAGGATTTGATATTCACCTGGCCCGATCTGGCCATACACTTGAGTGACAATGACATCCACTTTGCCTTTCTCTCCACAGGTGTCCACTCCCAGGTCCAAGCGGCCGCCCTGCAGGGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGAGGATCTCCGCGGGGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGAACCGGTTGTGGAATGTGTGTCAGTTAGGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCATAGTCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGCTTTTGCAAAAAAGCTGCAGATGGTACGACCATTAAATTGTATTGTAGCAGTATCACAAAATATGGGTATTGGTAAAAATGGTGATTTACCATGGCCACCATTACGAAATGAATTTAAATATTTTCAACGAATGACTACTACTTCATCAGTAGAAGGTAAACAAAATTTAGTAATTATGGGTCGAAAAACTTGGTTTTCAATTCCTGAGAAGAATCGACCTTTAAAGGACAGAATTAATATAGTTCTCAGTAGAGAACTCAAAGAACCACCACGAGGAGCTCATTTTCTTGCCAAAAGTTTGGATGATGCCTTAAGACTTATTGAACAACCGGAATTGGCAAGTAAAGTAGACATGGTTTGGATAGTCGGAGGCAGTTCTGTTTACCAGGAAGCCATGAATCAACCAGGCCACCTCAGACTCTTTGTGACAAGGATCATGCAGGAATTTGAAAGTGACACGTTTTTCCCAGAAATTGATTTGGGGAAATATAAACTTCTCCCAGAATACCCAGGCGTCCTCTCTGAGGTCCAGGAGGAAAAAGGCATCAAGTATAAGTTTGAAGTCTACGAGAAGAAAGACTAAAGATCCGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTAAGGTAAATATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTGTGTATTTTAGATTCCAACCTATGGAACTGATGAATGGGAGCAGTGGTGGAATGCCTTTAATGAGGAAAACCTGTTTTGCTCAGAAGAAATGCCATCTAGTGATGATGAGGCTACTGCTGACTCTCAACATTCTACTCCTCCAAAAAAGAAGAGAAAGGTAGAAGACCCCAAGGACTTTCCTTCAGAATTGCTAAGTTTTTTGAGTCATGCTGTGTTTAGTAATAGAACTCTTGCTTGCTTTGCTATTTACACCACAAAGGAAAAAGCTGCACTGCTATACAAGAAAATTATGGAAAAATATTCTGTAACCTTTATAAGTAGGCATAACAGTTATAATCATAACATACTGTTTTTTCTTACTCCACACAGGCATAGAGTGTCTGCTATTAATAACTATGCTCAAAAATTGTGTACCTTTAGCTTTTTAATTTGTAAAGGGGTTAATAAGGAATATTTGATGTATAGTGCCTTGACTAGAGATCATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGGCCCATCGATGCCGACGTAGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGAAAGCTT
<서열번호 4> 개시 코돈의 상류에 최적화한 Kozak가 부가되어 있는 항체(OMLH) 경쇄 유전자 cDNA의 뉴클레오티드 서열
CGCGCCCCGCCGCCACCATGGGTTGGTCTTGTATCATCTTATTTTTAGTTGCTACTGCTACTGGTGTTCATTCTGATATACAGCTCACCCAAAGCCCATCATCTCTGTCTGCAAGCGTCGGCGACAGGGTGACCATTACCTGTCGCGCAAGCCAAAGCGTTGACTACGACGGCGACAGCTACATGAACTGGTACCAGCAGAAGCCCGGCAAGGCTCCTAAGCTGCTGATCTATGCCGCCTCCTACCTTGAATCTGGAGTGCCTTCTCGTTTTTCCGGCTCAGGGTCCGGAACTGATTTTACCCTGACCATTTCCTCCCTCCAGCCCGAGGATTTTGCCACTTACTACTGTCAGCAGTCCCACGAGGACCCATATACATTCGGACAAGGTACAAAGGTAGAAATCAAGCGTACGGTGGCTGCCCCATCCGTGTTCATATTTCCTCCTAGCGACGAACAACTCAAGTCCGGTACCGCCAGCGTGGTCTGCCTGTTGAACAATTTTTATCCAAGAGAAGCTAAGGTCCAGTGGAAGGTTGACAACGCCCTTCAGTCCGGAAATAGCCAAGAGAGCGTCACCGAACAGGACTCCAAGGACAGTACATACTCACTGAGCTCTACACTGACCCTTTCTAAGGCCGACTACGAGAAGCACAAGGTCTACGCATGCGAAGTGACCCATCAGGGACTCAGTAGCCCTGTAACAAAGAGTTTTAATCGAGGCGAGTGCTAAGCGAT
<서열번호 5> 개시 코돈의 상류에 최적화한 Kozak가 부가되어 있는 항체(OMLH) 중쇄 유전자 cDNA의 뉴클레오티드 서열
GGCCGCCCGCCGCCACCATGGAATTTGGTTTATCTTGGGTTTTTTTAGTTGCTTTATTAAGAGGTGTTCAATGTGAGGTGCAGTTGGTCGAATCCGGCGGCGGACTCGTGCAACCAGGCGGAAGTTTGCGGCTGTCCTGCGCAGTGTCTGGTTACAGCATCACCTCCGGGTATAGCTGGAACTGGATCCGCCAGGCTCCTGGAAAGGGGCTTGAGTGGGTGGCTTCCATTACCTACGACGGCTCCACTAACTATAACCCGAGCGTCAAAGGCAGAATCACCATCTCTCGGGACGACTCAAAGAATACCTTCTACCTGCAGATGAACTCACTGAGGGCCGAAGATACCGCAGTTTACTACTGCGCCAGGGGGTCCCACTATTTCGGCCACTGGCACTTCGCCGTGTGGGGACAGGGCACACTCGTGACCGTTAGTAGCGCTAGCACCAAAGGCCCCTCTGTGTTCCCACTTGCTCCCTCCAGTAAATCTACCTCCGGAGGAACCGCAGCCCTCGGCTGCCTGGTGAAGGATTACTTCCCAGAGCCCGTCACCGTCTCTTGGAACTCCGGAGCCTTGACTAGCGGAGTGCACACTTTCCCTGCTGTATTGCAGTCCAGCGGCTTGTATTCACTGAGTAGCGTCGTCACCGTGCCTTCAAGCAGCCTCGGGACACAGACATACATATGTAATGTCAACCATAAGCCATCAAACACTAAAGTTGATAAAAAGGTGGAACCTAAGAGTTGCGATAAGACCCATACCTGTCCTCCTTGCCCTGCTCCTGAGCTGCTGGGAGGCCCTAGCGTGTTTCTGTTCCCCCCCAAGCCCAAAGATACACTGATGATTTCCCGCACACCTGAAGTAACATGTGTCGTGGTTGATGTGAGTCACGAGGATCCAGAGGTCAAGTTTAATTGGTACGtGGACGGAGTGGAGGTGCACAACGCTAAGACTAAGCCTCGGGAGGAACAGTACAACAGCACATACCGCGTGGTCAGCGTTTTGACTGTGCTGCATCAAGACTGGCTCAATGGAAAGGAATACAAGTGCAAGGTCTCTAATAAAGCCCTCCCCGCTCCTATTGAGAAGACTATTTCTAAAGCCAAGGGCCAGCCTCGCGAACCTCAGGTATATACTTTGCCACCCTCTCGCGAAGAAATGACAAAGAATCAGGTCTCACTCACTTGCCTCGTCAAAGGGTTTTACCCTTCTGACATCGCTGTCGAATGGGAAAGTAATGGTCAGCCAGAAAACAATTACAAGACTACTCCACCAGTGCTCGATTCTGATGGAAGTTTCTTTCTCTACAGTAAGCTCACTGTGGACAAATCTCGCTGGCAGCAGGGTAACGTATTCTCATGCTCCGTGATGCATGAAGCCCTCCACAACCATTACACCCAGAAGAGCCTGTCTCTGAGCCCAGGCAAGTAACCTGCA
<서열번호 6> pDC61의 전체 서열
CGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCTCTGGCTAACTAGAGAACCCACTGTTAACTGGCTTATCGAAATTGTCGAGGAGAACTTCAGGGTGAGTTTGGGGACCCTTGATTGTTCTTTCTTTTTCGCTATTGTAAAATTCATGTTATATGGAGGGGGCAAAGTTTTCAGGGTGTTGTTTAGAATGGGAAGATGTCCCTTGTATCACCATGGACCCTCATGATAATTTTGTTTCTTTCACTTTCTACTCTGTTGACAACCATTGTCTCCTCTTATTTTCTTTTCATTTTCTGTAACTTTTTCGTTAAACTTTAGCTTGCATTTGTAACGAATTTTTAAATTCACTTTTGTTTATTTGTCAGATTGTAAGTACTTTCTCTAATCACTTTTTTTTCAAGGCAATCAGGGTATATTATATTGTACTTCAGCACAGTTTTAGAGAACAATTGTTATAATTAAATGATAAGGTAGAATATTTCTGCATATAAATTCTGGCTGGCGTGGAAATATTCTTATTGGTAGAAACAACTACATCCTGGTCATCATCCTGCCTTTCTCTTTATGGTTACAATGATATACACTGTTTGAGATGAGGATAAAATACTCTGAGTCCAAACCGGGCCCCTCTGCTAACCATGTTCATGCCTTCTTCTTTTTCCTACAGCTCCTGGGCAACGTGCTGGCGGCCGCCCCGGGGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACTCGAGCGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCTCTGGCTAACTAGAGAACCCACTGTTAACTGGCTTATCGAAATTGTCGAGGAGAACTTCAGGGTGAGTTTGGGGACCCTTGATTGTTCTTTCTTTTTCGCTATTGTAAAATTCATGTTATATGGAGGGGGCAAAGTTTTCAGGGTGTTGTTTAGAATGGGAAGATGTCCCTTGTATCACCATGGACCCTCATGATAATTTTGTTTCTTTCACTTTCTACTCTGTTGACAACCATTGTCTCCTCTTATTTTCTTTTCATTTTCTGTAACTTTTTCGTTAAACTTTAGCTTGCATTTGTAACGAATTTTTAAATTCACTTTTGTTTATTTGTCAGATTGTAAGTACTTTCTCTAATCACTTTTTTTTCAAGGCAATCAGGGTATATTATATTGTACTTCAGCACAGTTTTAGAGAACAATTGTTATAATTAAATGATAAGGTAGAATATTTCTGCATATAAATTCTGGCTGGCGTGGAAATATTCTTATTGGTAGAAACAACTACATCCTGGTCATCATCCTGCCTTTCTCTTTATGGTTACAATGATATACACTGTTTGAGATGAGGATAAAATACTCTGAGTCCAAACCGGGCCCCTCTGCTAACCATGTTCATGCCTTCTTCTTTTTCCTACAGCTCCTGGGCAACGTGCTGGCGCGCCTCTAGAGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGAGGATCTCCGCGGTGTGGAATGTGTGTCAGTTAGGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCATAGTCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGCTTTTGCAAAAAAGCTGCAGATGGTACGACCATTAAATTGTATTGTAGCAGTATCACAAAATATGGGTATTGGTAAAAATGGTGATTTACCATGGCCACCATTACGAAATGAATTTAAATATTTTCAACGAATGACTACTACTTCATCAGTAGAAGGTAAACAAAATTTAGTAATTATGGGTCGAAAAACTTGGTTTTCAATTCCTGAGAAGAATCGACCTTTAAAGGACAGAATTAATATAGTTCTCAGTAGAGAACTCAAAGAACCACCACGAGGAGCTCATTTTCTTGCCAAAAGTTTGGATGATGCCTTAAGACTTATTGAACAACCGGAATTGGCAAGTAAAGTAGACATGGTTTGGATAGTCGGAGGCAGTTCTGTTTACCAGGAAGCCATGAATCAACCAGGCCACCTCAGACTCTTTGTGACAAGGATCATGCAGGAATTTGAAAGTGACACGTTTTTCCCAGAAATTGATTTGGGGAAATATAAACTTCTCCCAGAATACCCAGGCGTCCTCTCTGAGGTCCAGGAGGAAAAAGGCATCAAGTATAAGTTTGAAGTCTACGAGAAGAAAGACTAAAGATCCGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTAAGGTAAATATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTGTGTATTTTAGATTCCAACCTATGGAACTGATGAATGGGAGCAGTGGTGGAATGCCTTTAATGAGGAAAACCTGTTTTGCTCAGAAGAAATGCCATCTAGTGATGATGAGGCTACTGCTGACTCTCAACATTCTACTCCTCCAAAAAAGAAGAGAAAGGTAGAAGACCCCAAGGACTTTCCTTCAGAATTGCTAAGTTTTTTGAGTCATGCTGTGTTTAGTAATAGAACTCTTGCTTGCTTTGCTATTTACACCACAAAGGAAAAAGCTGCACTGCTATACAAGAAAATTATGGAAAAATATTCTGTAACCTTTATAAGTAGGCATAACAGTTATAATCATAACATACTGTTTTTTCTTACTCCACACAGGCATAGAGTGTCTGCTATTAATAACTATGCTCAAAAATTGTGTACCTTTAGCTTTTTAATTTGTAAAGGGGTTAATAAGGAATATTTGATGTATAGTGCCTTGACTAGAGATCATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGGCCCATCGATGAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTT
<서열번호 7> pDC6 내의 서열과 치환한 서열(pDC61)
AATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCA
<서열번호 8> pDC61의 염기 서열 No1267-No1273과 치환한 인간 오말리즈맙 항체(OMLH) 경쇄를 코드하는 cDNA
GGCCGCCACCATGGGTTGGTCTTGTATCATCTTATTTTTAGTTGCTACTGCTACTGGTGTTCATTCTGATATACAGCTCACCCAAAGCCCATCATCTCTGTCTGCAAGCGTCGGCGACAGGGTGACCATTACCTGTCGCGCAAGCCAAAGCGTTGACTACGACGGCGACAGCTACATGAACTGGTACCAGCAGAAGCCCGGCAAGGCTCCTAAGCTGCTGATCTATGCCGCCTCCTACCTTGAATCTGGAGTGCCTTCTCGTTTTTCCGGCTCAGGGTCCGGAACTGATTTTACCCTGACCATTTCCTCCCTCCAGCCCGAGGATTTTGCCACTTACTACTGTCAGCAGTCCCACGAGGACCCATATACATTCGGACAAGGTACAAAGGTAGAAATCAAGCGTACGGTGGCTGCCCCATCCGTGTTCATATTTCCTCCTAGCGACGAACAACTCAAGTCCGGTACCGCCAGCGTGGTCTGCCTGTTGAACAATTTTTATCCAAGAGAAGCTAAGGTCCAGTGGAAGGTTGACAACGCCCTTCAGTCCGGAAATAGCCAAGAGAGCGTCACCGAACAGGACTCCAAGGACAGTACATACTCACTGAGCTCTACACTGACCCTTTCTAAGGCCGACTACGAGAAGCACAAGGTCTACGCATGCGAAGTGACCCATCAGGGACTCAGTAGCCCTGTAACAAAGAGTTTTAATCGAGGCGAGTGCTAAC
<서열번호 9> pDC61의 염기 서열 No2765-No2771로 치환한 인간 오말리즈맙 항체(OMLH) 중쇄를 코드하는 cDNA
CGCGCCACCATGGAATTTGGTTTATCTTGGGTTTTTTTAGTTGCTTTATTAAGAGGTGTTCAATGTGAGGTGCAGTTGGTCGAATCCGGCGGCGGACTCGTGCAACCAGGCGGAAGTTTGCGGCTGTCCTGCGCAGTGTCTGGTTACAGCATCACCTCCGGGTATAGCTGGAACTGGATCCGCCAGGCTCCTGGAAAGGGGCTTGAGTGGGTGGCTTCCATTACCTACGACGGCTCCACTAACTATAACCCGAGCGTCAAAGGCAGAATCACCATCTCTCGGGACGACTCAAAGAATACCTTCTACCTGCAGATGAACTCACTGAGGGCCGAAGATACCGCAGTTTACTACTGCGCCAGGGGGTCCCACTATTTCGGCCACTGGCACTTCGCCGTGTGGGGACAGGGCACACTCGTGACCGTTAGTAGCGCTAGCACCAAAGGCCCCTCTGTGTTCCCACTTGCTCCCTCCAGTAAATCTACCTCCGGAGGAACCGCAGCCCTCGGCTGCCTGGTGAAGGATTACTTCCCAGAGCCCGTCACCGTCTCTTGGAACTCCGGAGCCTTGACTAGCGGAGTGCACACTTTCCCTGCTGTATTGCAGTCCAGCGGCTTGTATTCACTGAGTAGCGTCGTCACCGTGCCTTCAAGCAGCCTCGGGACACAGACATACATATGTAATGTCAACCATAAGCCATCAAACACTAAAGTTGATAAAAAGGTGGAACCTAAGAGTTGCGATAAGACCCATACCTGTCCTCCTTGCCCTGCTCCTGAGCTGCTGGGAGGCCCTAGCGTGTTTCTGTTCCCCCCCAAGCCCAAAGATACACTGATGATTTCCCGCACACCTGAAGTAACATGTGTCGTGGTTGATGTGAGTCACGAGGATCCAGAGGTCAAGTTTAATTGGTACGtGGACGGAGTGGAGGTGCACAACGCTAAGACTAAGCCTCGGGAGGAACAGTACAACAGCACATACCGCGTGGTCAGCGTTTTGACTGTGCTGCATCAAGACTGGCTCAATGGAAAGGAATACAAGTGCAAGGTCTCTAATAAAGCCCTCCCCGCTCCTATTGAGAAGACTATTTCTAAAGCCAAGGGCCAGCCTCGCGAACCTCAGGTATATACTTTGCCACCCTCTCGCGAAGAAATGACAAAGAATCAGGTCTCACTCACTTGCCTCGTCAAAGGGTTTTACCCTTCTGACATCGCTGTCGAATGGGAAAGTAATGGTCAGCCAGAAAACAATTACAAGACTACTCCACCAGTGCTCGATTCTGATGGAAGTTTCTTTCTCTACAGTAAGCTCACTGTGGACAAATCTCGCTGGCAGCAGGGTAACGTATTCTCATGCTCCGTGATGCATGAAGCCCTCCACAACCATTACACCCAGAAGAGCCTGTCTCTGAGCCCAGGCAAGTAAT
<서열번호 10> 백본 벡터 pNC32c-U533의 전체 염기 서열
CGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCTCTGGCTAACTAGAGAACCCACTGTTAACTGGCTTATCGAAATTGGCGCGCCGCGATCGCCCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGATCTCCGCGGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGAGAATTCCGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCTCTGGCTAACTAGAGAACCCACTGTTAACTGGCTTATCGAAATTGCGGCCGCCCTGCAGGCCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGACTCCGCGGGGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGAACCGGTTGTGGAATGTGTGTCAGTTAGGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCAGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCATAGTCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGCTTTTGCAAAAAAGCTGCAGATGATTGAACAAGATGGATTGCACGCAGGTTCTCCGGCCGCTTGGGTGGAGAGGCTATTCGGCTATGACTGGGCACAACAGACAATCGGCTGCTCTGATGCCGCCGTGTTCCGGCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGACCGACCTGTCCGGTGCCCTGAATGAACTGCAGGACGAGGCAGCGCGGCTATCGTGGCTGGCCACGACGGGCGTTCCTTGCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAGGGACTGGCTGCTATTGGGCGAAGTGCCGGGGCAGGATCTCCTGTCATCTCACCTTGCTCCTGCCGAGAAAGTATCCATCATGGCTGATGCAATGCGGCGGCTGCATACGCTTGATCCGGCTACCTGCCCATTCGACCACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCGGATGGAAGCCGGTCTTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCAGCCGAACTGTTCGCCAGGCTCAAGGCGCGCATGCCCGACGGCGAGGATCTCGTCGTGACCCATGGCGATGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTCTGGATTCATCGACTGTGGCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCGTTGGCTACCCGTGATATTGCTGAAGAGCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGCTTTACGGTATCGCCGCTCCCGATTCGCAGCGCATCGCCTTCTATCGCCTTCTTGACGAGTTCTTCTGAGATCCGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTAAGGTAAATATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTGTGTATTTTAGATTCCAACCTATGGAACTGATGAATGGGAGCAGTGGTGGAATGCCTTTAATGAGGAAAACCTGTTTTGCTCAGAAGAAATGCCATCTAGTGATGATGAGGCTACTGCTGACTCTCAACATTCTACTCCTCCAAAAAAGAAGAGAAAGGTAGAAGACCCCAAGGACTTTCCTTCAGAATTGCTAAGTTTTTTGAGTCATGCTGTGTTTAGTAATAGAACTCTTGCTTGCTTTGCTATTTACACCACAAAGGAAAAAGCTGCACTGCTATACAAGAAAATTATGGAAAAATATTCTGTAACCTTTATAAGTAGGCATAACAGTTATAATCATAACATACTGTTTTTTCTTACTCCACACAGGCATAGAGTGTCTGCTATTAATAACTATGCTCAAAAATTGTGTACCTTTAGCTTTTTAATTTGTAAAGGGGTTAATAAGGAATATTTGATGTATAGTGCCTTGACTAGAGATCATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGGCCCATCGATGCCGACGTAGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGAAAGCTT
<서열번호 11> pNC32c-U533의 염기 서열 No 658-No 668로 치환한 인간 오말리즈맙 항체(OMLH) 경쇄를 코드하는 cDNA
CGCGCCACCATGGGTTGGTCTTGTATCATCTTATTTTTAGTTGCTACTGCTACTGGTGTTCATTCTGATATACAGCTCACCCAAAGCCCATCATCTCTGTCTGCAAGCGTCGGCGACAGGGTGACCATTACCTGTCGCGCAAGCCAAAGCGTTGACTACGACGGCGACAGCTACATGAACTGGTACCAGCAGAAGCCCGGCAAGGCTCCTAAGCTGCTGATCTATGCCGCCTCCTACCTTGAATCTGGAGTGCCTTCTCGTTTTTCCGGCTCAGGGTCCGGAACTGATTTTACCCTGACCATTTCCTCCCTCCAGCCCGAGGATTTTGCCACTTACTACTGTCAGCAGTCCCACGAGGACCCATATACATTCGGACAAGGTACAAAGGTAGAAATCAAGCGTACGGTGGCTGCCCCATCCGTGTTCATATTTCCTCCTAGCGACGAACAACTCAAGTCCGGTACCGCCAGCGTGGTCTGCCTGTTGAACAATTTTTATCCAAGAGAAGCTAAGGTCCAGTGGAAGGTTGACAACGCCCTTCAGTCCGGAAATAGCCAAGAGAGCGTCACCGAACAGGACTCCAAGGACAGTACATACTCACTGAGCTCTACACTGACCCTTTCTAAGGCCGACTACGAGAAGCACAAGGTCTACGCATGCGAAGTGACCCATCAGGGACTCAGTAGCCCTGTAACAAAGAGTTTTAATCGAGGCGAGTGCTAAGCGAT
<서열번호 12> pNC32c-U533의 염기 서열 No 3081-No 3092로 치환한 인간 오말리즈맙 항체(OMLH) 중쇄를 코드하는 cDNA
GGCCGCCACCATGGAATTTGGTTTATCTTGGGTTTTTTTAGTTGCTTTATTAAGAGGTGTTCAATGTGAGGTGCAGTTGGTCGAATCCGGCGGCGGACTCGTGCAACCAGGCGGAAGTTTGCGGCTGTCCTGCGCAGTGTCTGGTTACAGCATCACCTCCGGGTATAGCTGGAACTGGATCCGCCAGGCTCCTGGAAAGGGGCTTGAGTGGGTGGCTTCCATTACCTACGACGGCTCCACTAACTATAACCCGAGCGTCAAAGGCAGAATCACCATCTCTCGGGACGACTCAAAGAATACCTTCTACCTGCAGATGAACTCACTGAGGGCCGAAGATACCGCAGTTTACTACTGCGCCAGGGGGTCCCACTATTTCGGCCACTGGCACTTCGCCGTGTGGGGACAGGGCACACTCGTGACCGTTAGTAGCGCTAGCACCAAAGGCCCCTCTGTGTTCCCACTTGCTCCCTCCAGTAAATCTACCTCCGGAGGAACCGCAGCCCTCGGCTGCCTGGTGAAGGATTACTTCCCAGAGCCCGTCACCGTCTCTTGGAACTCCGGAGCCTTGACTAGCGGAGTGCACACTTTCCCTGCTGTATTGCAGTCCAGCGGCTTGTATTCACTGAGTAGCGTCGTCACCGTGCCTTCAAGCAGCCTCGGGACACAGACATACATATGTAATGTCAACCATAAGCCATCAAACACTAAAGTTGATAAAAAGGTGGAACCTAAGAGTTGCGATAAGACCCATACCTGTCCTCCTTGCCCTGCTCCTGAGCTGCTGGGAGGCCCTAGCGTGTTTCTGTTCCCCCCCAAGCCCAAAGATACACTGATGATTTCCCGCACACCTGAAGTAACATGTGTCGTGGTTGATGTGAGTCACGAGGATCCAGAGGTCAAGTTTAATTGGTACGtGGACGGAGTGGAGGTGCACAACGCTAAGACTAAGCCTCGGGAGGAACAGTACAACAGCACATACCGCGTGGTCAGCGTTTTGACTGTGCTGCATCAAGACTGGCTCAATGGAAAGGAATACAAGTGCAAGGTCTCTAATAAAGCCCTCCCCGCTCCTATTGAGAAGACTATTTCTAAAGCCAAGGGCCAGCCTCGCGAACCTCAGGTATATACTTTGCCACCCTCTCGCGAAGAAATGACAAAGAATCAGGTCTCACTCACTTGCCTCGTCAAAGGGTTTTACCCTTCTGACATCGCTGTCGAATGGGAAAGTAATGGTCAGCCAGAAAACAATTACAAGACTACTCCACCAGTGCTCGATTCTGATGGAAGTTTCTTTCTCTACAGTAAGCTCACTGTGGACAAATCTCGCTGGCAGCAGGGTAACGTATTCTCATGCTCCGTGATGCATGAAGCCCTCCACAACCATTACACCCAGAAGAGCCTGTCTCTGAGCCCAGGCAAGTAACCTGCA
<서열번호 13> UCOE-Hu-P의 염기 서열 No5309-No5311로 치환한 BstBI 인식 사이트를 포함하는 서열
CCTAGTAGTAGTAGTAGTTCGAAG
<서열번호 14> 인간 오말리즈맙 항체 경쇄(OML)를 코드하는 cDNA 및 시미안 바이러스 40 폴리아데닐화 신호(SV40pA), 모르모트 사이토메갈로바이러스 프로모터(PgpCMV), 인간 오말리즈맙 항체 중쇄(OMH)를 코드하는 cDNA를 연결한 서열
CCCACCATGGGTTGGTCTTGTATCATCTTATTTTTAGTTGCTACTGCTACTGGTGTTCATTCTGATATACAGCTCACCCAAAGCCCATCATCTCTGTCTGCAAGCGTCGGCGACAGGGTGACCATTACCTGTCGCGCAAGCCAAAGCGTTGACTACGACGGCGACAGCTACATGAACTGGTACCAGCAGAAGCCCGGCAAGGCTCCTAAGCTGCTGATCTATGCCGCCTCCTACCTTGAATCTGGAGTGCCTTCTCGTTTTTCCGGCTCAGGGTCCGGAACTGATTTTACCCTGACCATTTCCTCCCTCCAGCCCGAGGATTTTGCCACTTACTACTGTCAGCAGTCCCACGAGGACCCATATACATTCGGACAAGGTACAAAGGTAGAAATCAAGCGTACGGTGGCTGCCCCATCCGTGTTCATATTTCCTCCTAGCGACGAACAACTCAAGTCCGGTACCGCCAGCGTGGTCTGCCTGTTGAACAATTTTTATCCAAGAGAAGCTAAGGTCCAGTGGAAGGTTGACAACGCCCTTCAGTCCGGAAATAGCCAAGAGAGCGTCACCGAACAGGACTCCAAGGACAGTACATACTCACTGAGCTCTACACTGACCCTTTCTAAGGCCGACTACGAGAAGCACAAGGTCTACGCATGCGAAGTGACCCATCAGGGACTCAGTAGCCCTGTAACAAAGAGTTTTAATCGAGGCGAGTGCTAAGCGATCGCGCTAGCGCGGCCACGTAGTCGACTACGTAGAGCTCGGTACCCGGGGATCCTCTAGAGTCGACCTGCAGGCATGCAAGCTGGCCGCGACTCTAGATCATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTAACCGGTTATGTTACTTGGCAGAGGCCGCATGGAAAGTCCCTGGACGTGGGACATCTGATTAATACGTGAGGAGGTCAGCCATGTTCTTTTTGGCAAAGGACTACGGTCATTGGACGTTTGATTGGCATGGGATAGGGTCAGCCAGAGTTAACAGTGTTCTTTTGGCAAAGGGATACGTGGAAAGTCCCGGGCCATTTACAGTAAACTGATACGGGGACAAAGCACAGCCATATTTAGTCATGTATTGCTTGGCAGAGGGTCTATGGAAAGTCCCTGGACGTGGGACGTCTGATTAATATGAAAGAAGGTCAGCCAGAGGTAGCTGTGTCCTTTTTGGCAAAGGGATACGGTTATGGGACGTTTGATTGGACTGGGATAGGGTCAGCCAGAGTTAACAGTGTTCTTTTGGCAAAGGAAACGTGGAAAGTCCCGGGCCATTTACAGTAAACTGATACTGGGACAAAGTACACCCATATTTAGTCATGTTCTTTTTGGCAAAGAGCATCTGGAAAGTCCCGGGCAGCATTATAGTCACTTGGCAGAGGGAAAGGGTCACTCAGAGTTAAGTACATCTTTCCAGGGCCAATATTCCAGTAAATTACACTTAGTTTTATGCAAATCAGCCACAAAGGGGATTTTCCCGGTCAATTATGACTTTTTCCTTAGTCATGCGGTATCCAATTACTGCCAAATTGGCAGTACATACTAGGTGATTCACTGACATTTGGCCGTCCTCTGGAAAGTCCCTGGAAACCGCTCAAGTACTGTATCATGGTGACTTTGCATTTTTGGAGAGCACGCCCCACTCCACCATTGGTCCACGTACCCTATGGGGGAGTGGTTTATGAGTATATAAGGGGCTCCGGTTTAGAAGCCGGGCAGAGCGGAATTCGAGCTCCCTGCAGGTTAGTTAAGTTAACGGCGCGCCACCATGGAATTTGGTTTATCTTGGGTTTTTTTAGTTGCTTTATTAAGAGGTGTTCAATGTGAGGTGCAGTTGGTCGAATCCGGCGGCGGACTCGTGCAACCAGGCGGAAGTTTGCGGCTGTCCTGCGCAGTGTCTGGTTACAGCATCACCTCCGGGTATAGCTGGAACTGGATCCGCCAGGCTCCTGGAAAGGGGCTTGAGTGGGTGGCTTCCATTACCTACGACGGCTCCACTAACTATAACCCGAGCGTCAAAGGCAGAATCACCATCTCTCGGGACGACTCAAAGAATACCTTCTACCTGCAGATGAACTCACTGAGGGCCGAAGATACCGCAGTTTACTACTGCGCCAGGGGGTCCCACTATTTCGGCCACTGGCACTTCGCCGTGTGGGGACAGGGCACACTCGTGACCGTTAGTAGCGCTAGCACCAAAGGCCCCTCTGTGTTCCCACTTGCTCCCTCCAGTAAATCTACCTCCGGAGGAACCGCAGCCCTCGGCTGCCTGGTGAAGGATTACTTCCCAGAGCCCGTCACCGTCTCTTGGAACTCCGGAGCCTTGACTAGCGGAGTGCACACTTTCCCTGCTGTATTGCAGTCCAGCGGCTTGTATTCACTGAGTAGCGTCGTCACCGTGCCTTCAAGCAGCCTCGGGACACAGACATACATATGTAATGTCAACCATAAGCCATCAAACACTAAAGTTGATAAAAAGGTGGAACCTAAGAGTTGCGATAAGACCCATACCTGTCCTCCTTGCCCTGCTCCTGAGCTGCTGGGAGGCCCTAGCGTGTTTCTGTTCCCCCCCAAGCCCAAAGATACACTGATGATTTCCCGCACACCTGAAGTAACATGTGTCGTGGTTGATGTGAGTCACGAGGATCCAGAGGTCAAGTTTAATTGGTACGtGGACGGAGTGGAGGTGCACAACGCTAAGACTAAGCCTCGGGAGGAACAGTACAACAGCACATACCGCGTGGTCAGCGTTTTGACTGTGCTGCATCAAGACTGGCTCAATGGAAAGGAATACAAGTGCAAGGTCTCTAATAAAGCCCTCCCCGCTCCTATTGAGAAGACTATTTCTAAAGCCAAGGGCCAGCCTCGCGAACCTCAGGTATATACTTTGCCACCCTCTCGCGAAGAAATGACAAAGAATCAGGTCTCACTCACTTGCCTCGTCAAAGGGTTTTACCCTTCTGACATCGCTGTCGAATGGGAAAGTAATGGTCAGCCAGAAAACAATTACAAGACTACTCCACCAGTGCTCGATTCTGATGGAAGTTTCTTTCTCTACAGTAAGCTCACTGTGGACAAATCTCGCTGGCAGCAGGGTAACGTATTCTCATGCTCCGTGATGCATGAAGCCCTCCACAACCATTACACCCAGAAGAGCCTGTCTCTGAGCCCAGGCAAGTAATCTAGATT
<서열번호 15> 개 CTLA-4-Ig의 아미노산 서열
MAGFGFRRHGAQPDLASRTWPCTALFSLLFIPVFSKGMHVAQPAVVLASSRGVASFVCEYGSSGNAAEVRVTVLRQAGSQMTEVCAATYTVEDELAFLDDSTCTGTSSGNKVNLTIQGLRAMDTGLYICKVELMYPPPYYVGMGNGTQIYVIDPEPCPDSDPKESTCKCISPCPVPESLGGPSVFIFPPKPKDILRITRTPEITCVVLDLGREDPEVQISWFVDGKEVHTAKTQPREQQFNSTYRVVSVLPIEHQDWLTGKEFKCRVNHIGLPSPIERTISKARGQAHQPSVYVLPPSPKELSSSDTVTLTCLIKDFFPPEIDVEWQSNGQPEPESKYHTTAPQLDEDGSYFLYSKLSVDKSRWQQGDTFTCAVMHEALQNHYTDLSLSHSPGK
<서열번호 16> 개 CTLA-4-Ig의 코돈 최적화 후 뉴클레오티드 서열
ATGGCTGGATTTGGATTCAGAAGGCACGGAGCCCAGCCCGACCTGGCATCTCGCACTTGGCCCTGTACCGCACTGTTTTCACTGCTGTTCATCCCAGTGTTCAGCAAGGGAATGCACGTGGCTCAGCCAGCTGTGGTGCTGGCTTCCAGCAGAGGCGTGGCTTCCTTCGTGTGCGAGTACGGCTCTTCCGGCAACGCCGCTGAGGTGAGAGTGACCGTGCTGAGGCAGGCTGGCTCCCAGATGACAGAGGTGTGCGCCGCTACCTATACAGTGGAGGACGAGCTGGCTTTCCTGGACGATAGCACCTGTACAGGCACCAGCTCTGGCAACAAGGTCAATCTGACCATCCAGGGCCTGCGCGCCATGGATACAGGCCTGTACATCTGTAAGGTGGAGCTGATGTATCCCCCTCCATACTATGTGGGCATGGGCAATGGCACCCAGATCTACGTGATCGACCCCGAGCCTTGCCCAGACTCTGATCCAAAGGAGTCGACATGCAAGTGTATCTCTCCATGTCCTGTGCCAGAGAGCCTGGGAGGACCTTCCGTGTTCATCTTTCCCCCTAAGCCAAAGGATATCCTGAGGATCACACGGACCCCTGAGATCACCTGCGTGGTGCTGGACCTGGGAAGGGAGGATCCAGAGGTGCAGATCTCCTGGTTCGTGGACGGCAAGGAGGTGCATACCGCTAAGACACAGCCCAGAGAGCAGCAGTTTAACTCCACCTATCGCGTGGTGAGCGTGCTGCCTATCGAGCACCAGGATTGGCTGACAGGCAAGGAGTTTAAGTGCCGGGTGAATCATATCGGCCTGCCCTCTCCTATCGAGAGGACCATCTCCAAGGCTAGGGGACAGGCTCACCAGCCAAGCGTGTACGTGCTGCCACCCTCTCCTAAGGAGCTGTCCAGCTCTGACACAGTGACCCTGACATGTCTGATCAAGGACTTCTTTCCTCCAGAGATCGACGTGGAGTGGCAGTCCAACGGCCAGCCAGAGCCCGAGAGCAAGTATCATACCACAGCCCCCCAGCTGGACGAGGATGGCTCTTACTTCCTGTATTCCAAGCTGAGCGTGGACAAGTCCAGGTGGCAGCAGGGCGATACCTTTACATGTGCTGTGATGCACGAAGCCCTGCAGAATCATTACACCGACCTGTCACTGTCCCACTCCCCTGGCAAATAA
<서열번호 17> 외래 유전자 삽입 부위가 1개 있는 pDC62c5-U533의 전체 염기 서열
CGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACACCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAATAACCCCGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCCTCACTCTCTTCCGCATCGCTGTCTGCGAGGGCCAGCTGTTGGGCTCGCGGTTGAGGACAAACTCTTCGCGGTCTTTCCAGTACTCTTGGATCGGAAACCCGTCGGCCTCCGAACGGTACTCCGCCACCGAGGGACCTGAGCGAGTCCGCATCGACCGGATCGGAAAACCTCTCGAGAAAGGCGTCTAACCAGTCACAGTCGCAAGGTAGGCTGAGCACCGTGGCGGGCGGCAGCGGGTGGCGGTCGGGGTTGTTTCTGGCGGAGGTGCTGCTGATGATGTAATTAAAGTAGGCGGTCTTGAGACGGCGGATGGTCGAGGTGAGGTGTGGCAGGCTTGAGATCCAGCTGTTGGGGTGAGTACTCCCTCTCAAAAGCGGGCATGACTTCTGCGCTAAGATTGTCAGTTTCCAAAAACGAGGAGGATTTGATATTCACCTGGCCCGATCTGGCCATACACTTGAGTGACAATGACATCCACTTTGCCTTTCTCTCCACAGGTGTCCACTCCCAGGTCCAAGGCGCGCCGCGATCGCGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGAGGATCTCCGCGGGGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGAGAATTCaaatgggACCGGTTGTGGAATGTGTGTCAGTTAGGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCATAGTCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGCTTTTGCAAAAAAGCTGCAGATGGTACGACCATTAAATTGTATTGTAGCAGTATCACAAAATATGGGTATTGGTAAAAATGGTGATTTACCATGGCCACCATTACGAAATGAATTTAAATATTTTCAACGAATGACTACTACTTCATCAGTAGAAGGTAAACAAAATTTAGTAATTATGGGTCGAAAAACTTGGTTTTCAATTCCTGAGAAGAATCGACCTTTAAAGGACAGAATTAATATAGTTCTCAGTAGAGAACTCAAAGAACCACCACGAGGAGCTCATTTTCTTGCCAAAAGTTTGGATGATGCCTTAAGACTTATTGAACAACCGGAATTGGCAAGTAAAGTAGACATGGTTTGGATAGTCGGAGGCAGTTCTGTTTACCAGGAAGCCATGAATCAACCAGGCCACCTCAGACTCTTTGTGACAAGGATCATGCAGGAATTTGAAAGTGACACGTTTTTCCCAGAAATTGATTTGGGGAAATATAAACTTCTCCCAGAATACCCAGGCGTCCTCTCTGAGGTCCAGGAGGAAAAAGGCATCAAGTATAAGTTTGAAGTCTACGAGAAGAAAGACTAAAGATCCGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTAAGGTAAATATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTGTGTATTTTAGATTCCAACCTATGGAACTGATGAATGGGAGCAGTGGTGGAATGCCTTTAATGAGGAAAACCTGTTTTGCTCAGAAGAAATGCCATCTAGTGATGATGAGGCTACTGCTGACTCTCAACATTCTACTCCTCCAAAAAAGAAGAGAAAGGTAGAAGACCCCAAGGACTTTCCTTCAGAATTGCTAAGTTTTTTGAGTCATGCTGTGTTTAGTAATAGAACTCTTGCTTGCTTTGCTATTTACACCACAAAGGAAAAAGCTGCACTGCTATACAAGAAAATTATGGAAAAATATTCTGTAACCTTTATAAGTAGGCATAACAGTTATAATCATAACATACTGTTTTTTCTTACTCCACACAGGCATAGAGTGTCTGCTATTAATAACTATGCTCAAAAATTGTGTACCTTTAGCTTTTTAATTTGTAAAGGGGTTAATAAGGAATATTTGATGTATAGTGCCTTGACTAGAGATCATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGGCCCATCGATGCCGACGTAGCGCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGGCCCTCCGCGCCTACAGCTCAAGCCACATCCGAAGGGGGAGGGAGCCGGGAGCTGCGCGCGGGGCCGCCGGGGGGAGGGGTGGCACCGCCCACGCCGGGCGGCCACGAAGGGCGGGGCAGCGGGCGCGCGCGCGGCGGGGGGAGGGGCCGGCGCCGCGCCCGCTGGGAATTGGGGCCCTAGGGGGAGGGCGGAGGCGCCGACGACCGCGGCACTTACCGTTCGCGGCGTGGCGCCCGGTGGTCCCCAAGGGGAGGGAAGGGGGAGGCGGGGCGAGGACAGTGACCGGAGTCTCCTCAGCGGTGGCTTTTCTGCTTGGCAGCCTCAGCGGCTGGCGCCAAAACCGGACTCCGCCCACTTCCTCGCCCGCCGGTGCGAGGGTGTGGAATCCTCCAGACGCTGGGGGAGGGGGAGTTGGGAGCTTAAAAACTAGTACCCCTTTGGGACCACTTTCAGCAGCGAACTCTCCTGTACACCAGGGGTCAGTTCCACAGACGCGGGCCAGGGGTGGGTCATTGCGGCGTGAACAATAATTTGACTAGAAGTTGATTCGGGTGTTTCCGGAAGGGGCCGAGTCAATCCGCCGAGTTGGGGCACGGAAAACAAAAAGGGAAGGCTACTAAGATTTTTCTGGCGGGGGTTATCATTGGCGTAACTGCAGGGACCACCTCCCGGGTTGAGGGGGCTGGATCTCCAGGCTGCGGATTAAGCCCCTCCCGTCGGCGTTAATTTCAAACTGCGCGACGTTTCTCACCTGCCTTCGCCAAGGCAGGGGCCGGGACCCTATTCCAAGAGGTAGTAACTAGCAGGACTCTAGCCTTCCGCAATTCATTGAGCGCATTTACGGAAGTAACGTCGGGTACTGTCTCTGGCCGCAAGGGTGGGAGGAGTACGCATTTGGCGTAAGGTGGGGCGTAGAGCCTTCCCGCCATTGGCGGCGGATAGGGCGTTTACGCGACGGCCTGACGTAGCGGAAGACGCCTTAGTGGGGGGGAAGGTTCTAGAAAAGCGGCGGCAGCGGCTCTAGCGGCAGTAGCAGCAGCGCCGGGTCCCGTGCGGAGGTGCTCCTCGCAGAGTTGTTTCTCCAGCAGCGGCAGTTCTCACTACAGCGCCAGGACGAGTCCGGTTCGTGTTCGTCCGCGGAGATCTCTCTCATCTCGCTCGGCTGCGGGAAATCGGGCTGAAGCGACTGAGTCCGCGATGGAGGTAACGGGTTTGAAATCAATGAGTTATTGAAAAGGGCATGGCGAGGCCGTTGGCGCCTCAGTGGAAGTCGGCCAGCCGCCTCCGTGGGAGAGAGGCAGGAAATCGGACCAATTCAGTAGCAGTGGGGCTTAAGGTTTATGAACGGGGTCTTGAGCGGAGGCCTGAGCGTACAAACAGCTTCCCCACCCTCAGCCTCCCGGCGCCATTTCCCTTCACTGGGGGTGGGGGATGGGGAGCTTTCACATGGCGGACGCTGCCCCGCTGGGGTGAAAGTGGGGCGCGGAGGCGGGACTTCTTATTCCCTTTCTAAAGCACGCTGCTTCGGGGGCCACGGCGTCTCCTCGGAAAGCTT
<서열번호 18> 개시 코돈의 상류에 최적화한 Kozak가 부가되어 있는 항체(TRLH) 경쇄 유전자 cDNA의 뉴클레오티드 서열
CGCGCCCCGCCGCCACCATGGGTTGGTCTTGTATCATCTTATTTTTAGTTGCTACTGCTACTGGTGTTCATTCTGATATACAGATGACCCAAAGCCCATCATCTCTGTCTGCAAGCGTCGGCGACAGGGTGACCATTACCTGTCGCGCAAGCCAAGACGTTAATACAGCAGTGGCTTGGTACCAGCAGAAGCCCGGCAAGGCTCCTAAGCTGCTGATCTATAGCGCCTCCTTTCTTTATTCTGGAGTGCCTTCTCGTTTTTCCGGCTCAAGGTCCGGAACTGATTTTACCCTGACCATTTCCTCCCTCCAGCCCGAGGATTTTGCCACTTACTACTGTCAGCAGCACTATACCACACCACCTACATTCGGACAAGGTACAAAGGTAGAAATCAAGCGTACGGTGGCTGCCCCATCCGTGTTCATATTTCCTCCTAGCGACGAACAACTCAAGTCCGGTACCGCCAGCGTGGTCTGCCTGTTGAACAATTTTTATCCAAGAGAAGCTAAGGTCCAGTGGAAGGTTGACAACGCCCTTCAGTCCGGAAATAGCCAAGAGAGCGTCACCGAACAGGACTCCAAGGACAGTACATACTCACTGAGCTCTACACTGACCCTTTCTAAGGCCGACTACGAGAAGCACAAGGTCTACGCATGCGAAGTGACCCATCAGGGACTCAGTAGCCCTGTAACAAAGAGTTTTAATCGAGGCGAGTGCTAAGCGAT
<서열번호 19> 개시 코돈의 상류에 최적화한 Kozak가 부가되어 있는 항체(TRLH) 중쇄 유전자 cDNA의 뉴클레오티드 서열
GGCCGCCCGCCGCCACCATGGAATTTGGTTTATCTTGGGTTTTTTTAGTTGCTTTATTAAGAGGTGTTCAATGTGAGGTGCAGTTGGTCGAATCCGGCGGCGGACTCGTGCAACCAGGCGGAAGTTTGCGGCTGTCCTGCGCAGCCTCTGGTTTTAACATCAAAGATACCTATATTCATTGGGTACGCCAGGCTCCTGGAAAGGGGCTTGAGTGGGTGGCTCGAATTTACCCAACCAATGGCTACACTCGGTATGCCGACAGCGTCAAAGGCAGATTCACCATCTCTGCAGACACATCAAAGAATACCGCTTACCTGCAGATGAACTCACTGAGGGCCGAAGATACCGCAGTTTACTACTGCTCCAGGTGGGGGGGGGACGGCTTCTACGCCATGGATTACTGGGGACAGGGCACACTCGTGACCGTTAGTAGCGCTAGCACCAAAGGCCCCTCTGTGTTCCCACTTGCTCCCTCCAGTAAATCTACCTCCGGAGGAACCGCAGCCCTCGGCTGCCTGGTGAAGGATTACTTCCCAGAGCCCGTCACCGTCTCTTGGAACTCCGGAGCCTTGACTAGCGGAGTGCACACTTTCCCTGCTGTATTGCAGTCCAGCGGCTTGTATTCACTGAGTAGCGTCGTCACCGTGCCTTCAAGCAGCCTCGGGACACAGACATACATATGTAATGTCAACCATAAGCCATCAAACACTAAAGTTGATAAAAAGGTGGAACCTAAGAGTTGCGATAAGACCCATACCTGTCCTCCTTGCCCTGCTCCTGAGCTGCTGGGAGGCCCTAGCGTGTTTCTGTTCCCCCCCAAGCCCAAAGATACACTGATGATTTCCCGCACACCTGAAGTAACATGTGTCGTGGTTGATGTGAGTCACGAGGATCCAGAGGTCAAGTTTAATTGGTACGTGGACGGAGTGGAGGTGCACAACGCTAAGACTAAGCCTCGGGAGGAACAGTACAACAGCACATACCGCGTGGTCAGCGTTTTGACTGTGCTGCATCAAGACTGGCTCAATGGAAAGGAATACAAGTGCAAGGTCTCTAATAAAGCCCTCCCCGCTCCTATTGAGAAGACTATTTCTAAAGCCAAGGGCCAGCCTCGCGAACCTCAGGTATATACTTTGCCACCCTCTCGCGAAGAAATGACAAAGAATCAGGTCTCACTCACTTGCCTCGTCAAAGGGTTTTACCCTTCTGACATCGCTGTCGAATGGGAAAGTAATGGTCAGCCAGAAAACAATTACAAGACTACTCCACCAGTGCTCGATTCTGATGGAAGTTTCTTTCTCTACAGTAAGCTCACTGTGGACAAATCTCGCTGGCAGCAGGGTAACGTATTCTCATGCTCCGTGATGCATGAAGCCCTCCACAACCATTACACCCAGAAGAGCCTGTCTCTGAGCCCAGGCTAACCTGCA
<110> NATIONAL UNIVERSITY CORPORATION HOKKAIDO UNIVERSITY
Fuso Pharmaceutical Industries, Ltd.
<120> Novel vectors and use thereof
<130> 2020-FPA-0118
<150> JP2018-99704
<151> 2018-05-24
<150> JP2018-168591
<151> 2018-09-10
<160> 19
<170> PatentIn version 3.5
<210> 1
<211> 1551
<212> DNA
<213> Homo sapiens
<400> 1
ggccctccgc gcctacagct caagccacat ccgaaggggg agggagccgg gagctgcgcg 60
cggggccgcc ggggggaggg gtggcaccgc ccacgccggg cggccacgaa gggcggggca 120
gcgggcgcgc gcgcggcggg gggaggggcc ggcgccgcgc ccgctgggaa ttggggccct 180
agggggaggg cggaggcgcc gacgaccgcg gcacttaccg ttcgcggcgt ggcgcccggt 240
ggtccccaag gggagggaag ggggaggcgg ggcgaggaca gtgaccggag tctcctcagc 300
ggtggctttt ctgcttggca gcctcagcgg ctggcgccaa aaccggactc cgcccacttc 360
ctcgcccgcc ggtgcgaggg tgtggaatcc tccagacgct gggggagggg gagttgggag 420
cttaaaaact agtacccctt tgggaccact ttcagcagcg aactctcctg tacaccaggg 480
gtcagttcca cagacgcggg ccaggggtgg gtcattgcgg cgtgaacaat aatttgacta 540
gaagttgatt cgggtgtttc cggaaggggc cgagtcaatc cgccgagttg gggcacggaa 600
aacaaaaagg gaaggctact aagatttttc tggcgggggt tatcattggc gtaactgcag 660
ggaccacctc ccgggttgag ggggctggat ctccaggctg cggattaagc ccctcccgtc 720
ggcgttaatt tcaaactgcg cgacgtttct cacctgcctt cgccaaggca ggggccggga 780
ccctattcca agaggtagta actagcagga ctctagcctt ccgcaattca ttgagcgcat 840
ttacggaagt aacgtcgggt actgtctctg gccgcaaggg tgggaggagt acgcatttgg 900
cgtaaggtgg ggcgtagagc cttcccgcca ttggcggcgg atagggcgtt tacgcgacgg 960
cctgacgtag cggaagacgc cttagtgggg gggaaggttc tagaaaagcg gcggcagcgg 1020
ctctagcggc agtagcagca gcgccgggtc ccgtgcggag gtgctcctcg cagagttgtt 1080
tctccagcag cggcagttct cactacagcg ccaggacgag tccggttcgt gttcgtccgc 1140
ggagatctct ctcatctcgc tcggctgcgg gaaatcgggc tgaagcgact gagtccgcga 1200
tggaggtaac gggtttgaaa tcaatgagtt attgaaaagg gcatggcgag gccgttggcg 1260
cctcagtgga agtcggccag ccgcctccgt gggagagagg caggaaatcg gaccaattca 1320
gtagcagtgg ggcttaaggt ttatgaacgg ggtcttgagc ggaggcctga gcgtacaaac 1380
agcttcccca ccctcagcct cccggcgcca tttcccttca ctgggggtgg gggatgggga 1440
gctttcacat ggcggacgct gccccgctgg ggtgaaagtg gggcgcggag gcgggacttc 1500
ttattccctt tctaaagcac gctgcttcgg gggccacggc gtctcctcgg a 1551
<210> 2
<211> 15
<212> DNA
<213> Artificial Sequence
<220>
<223> Optimized Kozak sequence
<400> 2
ccgccgccac catgg 15
<210> 3
<211> 11327
<212> DNA
<213> Artificial Sequence
<220>
<223> pDC62c5-U533
<400> 3
cgatgtacgg gccagatata cgcgttgaca ttgattattg actagtcgcg ttacataact 60
tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat 120
gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta 180
tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc 240
tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg 300
ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg 360
gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct 420
ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa 480
atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt 540
ctatataagc agagctcgtt tagtgaaccg tcagatcctc actctcttcc gcatcgctgt 600
ctgcgagggc cagctgttgg gctcgcggtt gaggacaaac tcttcgcggt ctttccagta 660
ctcttggatc ggaaacccgt cggcctccga acggtactcc gccaccgagg gacctgagcg 720
agtccgcatc gaccggatcg gaaaacctct cgagaaaggc gtctaaccag tcacagtcgc 780
aaggtaggct gagcaccgtg gcgggcggca gcgggtggcg gtcggggttg tttctggcgg 840
aggtgctgct gatgatgtaa ttaaagtagg cggtcttgag acggcggatg gtcgaggtga 900
ggtgtggcag gcttgagatc cagctgttgg ggtgagtact ccctctcaaa agcgggcatg 960
acttctgcgc taagattgtc agtttccaaa aacgaggagg atttgatatt cacctggccc 1020
gatctggcca tacacttgag tgacaatgac atccactttg cctttctctc cacaggtgtc 1080
cactcccagg tccaaggcgc gccgcgatcg cgcctcgact gtgccttcta gttgccagcc 1140
atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt 1200
cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct 1260
ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc 1320
tggggaggat ctccgcgggg ccctccgcgc ctacagctca agccacatcc gaagggggag 1380
ggagccggga gctgcgcgcg gggccgccgg ggggaggggt ggcaccgccc acgccgggcg 1440
gccacgaagg gcggggcagc gggcgcgcgc gcggcggggg gaggggccgg cgccgcgccc 1500
gctgggaatt ggggccctag ggggagggcg gaggcgccga cgaccgcggc acttaccgtt 1560
cgcggcgtgg cgcccggtgg tccccaaggg gagggaaggg ggaggcgggg cgaggacagt 1620
gaccggagtc tcctcagcgg tggcttttct gcttggcagc ctcagcggct ggcgccaaaa 1680
ccggactccg cccacttcct cgcccgccgg tgcgagggtg tggaatcctc cagacgctgg 1740
gggaggggga gttgggagct taaaaactag tacccctttg ggaccacttt cagcagcgaa 1800
ctctcctgta caccaggggt cagttccaca gacgcgggcc aggggtgggt cattgcggcg 1860
tgaacaataa tttgactaga agttgattcg ggtgtttccg gaaggggccg agtcaatccg 1920
ccgagttggg gcacggaaaa caaaaaggga aggctactaa gatttttctg gcgggggtta 1980
tcattggcgt aactgcaggg accacctccc gggttgaggg ggctggatct ccaggctgcg 2040
gattaagccc ctcccgtcgg cgttaatttc aaactgcgcg acgtttctca cctgccttcg 2100
ccaaggcagg ggccgggacc ctattccaag aggtagtaac tagcaggact ctagccttcc 2160
gcaattcatt gagcgcattt acggaagtaa cgtcgggtac tgtctctggc cgcaagggtg 2220
ggaggagtac gcatttggcg taaggtgggg cgtagagcct tcccgccatt ggcggcggat 2280
agggcgttta cgcgacggcc tgacgtagcg gaagacgcct tagtgggggg gaaggttcta 2340
gaaaagcggc ggcagcggct ctagcggcag tagcagcagc gccgggtccc gtgcggaggt 2400
gctcctcgca gagttgtttc tccagcagcg gcagttctca ctacagcgcc aggacgagtc 2460
cggttcgtgt tcgtccgcgg agatctctct catctcgctc ggctgcggga aatcgggctg 2520
aagcgactga gtccgcgatg gaggtaacgg gtttgaaatc aatgagttat tgaaaagggc 2580
atggcgaggc cgttggcgcc tcagtggaag tcggccagcc gcctccgtgg gagagaggca 2640
ggaaatcgga ccaattcagt agcagtgggg cttaaggttt atgaacgggg tcttgagcgg 2700
aggcctgagc gtacaaacag cttccccacc ctcagcctcc cggcgccatt tcccttcact 2760
gggggtgggg gatggggagc tttcacatgg cggacgctgc cccgctgggg tgaaagtggg 2820
gcgcggaggc gggacttctt attccctttc taaagcacgc tgcttcgggg gccacggcgt 2880
ctcctcggag aattccgatg tacgggccag atatacgcgt tgacattgat tattgactag 2940
tcgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc 3000
attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg 3060
tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat 3120
gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca 3180
gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat 3240
taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg 3300
gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca 3360
acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg 3420
tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcctcactct 3480
cttccgcatc gctgtctgcg agggccagct gttgggctcg cggttgagga caaactcttc 3540
gcggtctttc cagtactctt ggatcggaaa cccgtcggcc tccgaacggt actccgccac 3600
cgagggacct gagcgagtcc gcatcgaccg gatcggaaaa cctctcgaga aaggcgtcta 3660
accagtcaca gtcgcaaggt aggctgagca ccgtggcggg cggcagcggg tggcggtcgg 3720
ggttgtttct ggcggaggtg ctgctgatga tgtaattaaa gtaggcggtc ttgagacggc 3780
ggatggtcga ggtgaggtgt ggcaggcttg agatccagct gttggggtga gtactccctc 3840
tcaaaagcgg gcatgacttc tgcgctaaga ttgtcagttt ccaaaaacga ggaggatttg 3900
atattcacct ggcccgatct ggccatacac ttgagtgaca atgacatcca ctttgccttt 3960
ctctccacag gtgtccactc ccaggtccaa gcggccgccc tgcagggcct cgactgtgcc 4020
ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 4080
tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 4140
gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 4200
caatagcagg catgctgggg aggatctccg cggggccctc cgcgcctaca gctcaagcca 4260
catccgaagg gggagggagc cgggagctgc gcgcggggcc gccgggggga ggggtggcac 4320
cgcccacgcc gggcggccac gaagggcggg gcagcgggcg cgcgcgcggc ggggggaggg 4380
gccggcgccg cgcccgctgg gaattggggc cctaggggga gggcggaggc gccgacgacc 4440
gcggcactta ccgttcgcgg cgtggcgccc ggtggtcccc aaggggaggg aagggggagg 4500
cggggcgagg acagtgaccg gagtctcctc agcggtggct tttctgcttg gcagcctcag 4560
cggctggcgc caaaaccgga ctccgcccac ttcctcgccc gccggtgcga gggtgtggaa 4620
tcctccagac gctgggggag ggggagttgg gagcttaaaa actagtaccc ctttgggacc 4680
actttcagca gcgaactctc ctgtacacca ggggtcagtt ccacagacgc gggccagggg 4740
tgggtcattg cggcgtgaac aataatttga ctagaagttg attcgggtgt ttccggaagg 4800
ggccgagtca atccgccgag ttggggcacg gaaaacaaaa agggaaggct actaagattt 4860
ttctggcggg ggttatcatt ggcgtaactg cagggaccac ctcccgggtt gagggggctg 4920
gatctccagg ctgcggatta agcccctccc gtcggcgtta atttcaaact gcgcgacgtt 4980
tctcacctgc cttcgccaag gcaggggccg ggaccctatt ccaagaggta gtaactagca 5040
ggactctagc cttccgcaat tcattgagcg catttacgga agtaacgtcg ggtactgtct 5100
ctggccgcaa gggtgggagg agtacgcatt tggcgtaagg tggggcgtag agccttcccg 5160
ccattggcgg cggatagggc gtttacgcga cggcctgacg tagcggaaga cgccttagtg 5220
ggggggaagg ttctagaaaa gcggcggcag cggctctagc ggcagtagca gcagcgccgg 5280
gtcccgtgcg gaggtgctcc tcgcagagtt gtttctccag cagcggcagt tctcactaca 5340
gcgccaggac gagtccggtt cgtgttcgtc cgcggagatc tctctcatct cgctcggctg 5400
cgggaaatcg ggctgaagcg actgagtccg cgatggaggt aacgggtttg aaatcaatga 5460
gttattgaaa agggcatggc gaggccgttg gcgcctcagt ggaagtcggc cagccgcctc 5520
cgtgggagag aggcaggaaa tcggaccaat tcagtagcag tggggcttaa ggtttatgaa 5580
cggggtcttg agcggaggcc tgagcgtaca aacagcttcc ccaccctcag cctcccggcg 5640
ccatttccct tcactggggg tgggggatgg ggagctttca catggcggac gctgccccgc 5700
tggggtgaaa gtggggcgcg gaggcgggac ttcttattcc ctttctaaag cacgctgctt 5760
cgggggccac ggcgtctcct cggaaccggt tgtggaatgt gtgtcagtta gggtgtggaa 5820
agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 5880
ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 5940
ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 6000
ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaaag 6060
ctgcagatgg tacgaccatt aaattgtatt gtagcagtat cacaaaatat gggtattggt 6120
aaaaatggtg atttaccatg gccaccatta cgaaatgaat ttaaatattt tcaacgaatg 6180
actactactt catcagtaga aggtaaacaa aatttagtaa ttatgggtcg aaaaacttgg 6240
ttttcaattc ctgagaagaa tcgaccttta aaggacagaa ttaatatagt tctcagtaga 6300
gaactcaaag aaccaccacg aggagctcat tttcttgcca aaagtttgga tgatgcctta 6360
agacttattg aacaaccgga attggcaagt aaagtagaca tggtttggat agtcggaggc 6420
agttctgttt accaggaagc catgaatcaa ccaggccacc tcagactctt tgtgacaagg 6480
atcatgcagg aatttgaaag tgacacgttt ttcccagaaa ttgatttggg gaaatataaa 6540
cttctcccag aatacccagg cgtcctctct gaggtccagg aggaaaaagg catcaagtat 6600
aagtttgaag tctacgagaa gaaagactaa agatccgtga cataattgga caaactacct 6660
acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta 6720
ctgattctaa ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag 6780
tggtggaatg cctttaatga ggaaaacctg ttttgctcag aagaaatgcc atctagtgat 6840
gatgaggcta ctgctgactc tcaacattct actcctccaa aaaagaagag aaaggtagaa 6900
gaccccaagg actttccttc agaattgcta agttttttga gtcatgctgt gtttagtaat 6960
agaactcttg cttgctttgc tatttacacc acaaaggaaa aagctgcact gctatacaag 7020
aaaattatgg aaaaatattc tgtaaccttt ataagtaggc ataacagtta taatcataac 7080
atactgtttt ttcttactcc acacaggcat agagtgtctg ctattaataa ctatgctcaa 7140
aaattgtgta cctttagctt tttaatttgt aaaggggtta ataaggaata tttgatgtat 7200
agtgccttga ctagagatca taatcagcca taccacattt gtagaggttt tacttgcttt 7260
aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt 7320
taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 7380
aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 7440
ttatcatgtc tgggcccatc gatgccgacg tagcgcctga tgcggtattt tctccttacg 7500
catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc 7560
gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt 7620
ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag 7680
aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt 7740
ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga 7800
aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 7860
atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt 7920
caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct 7980
cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt 8040
tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt 8100
tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac 8160
gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac 8220
tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct 8280
gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg 8340
aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg 8400
gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca 8460
atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa 8520
caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt 8580
ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc 8640
attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg 8700
agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt 8760
aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt 8820
catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 8880
ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 8940
tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 9000
ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 9060
ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac 9120
ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 9180
gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 9240
aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 9300
acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 9360
gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 9420
gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 9480
cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 9540
aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct 9600
gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct 9660
cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca 9720
atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg ggccctccgc 9780
gcctacagct caagccacat ccgaaggggg agggagccgg gagctgcgcg cggggccgcc 9840
ggggggaggg gtggcaccgc ccacgccggg cggccacgaa gggcggggca gcgggcgcgc 9900
gcgcggcggg gggaggggcc ggcgccgcgc ccgctgggaa ttggggccct agggggaggg 9960
cggaggcgcc gacgaccgcg gcacttaccg ttcgcggcgt ggcgcccggt ggtccccaag 10020
gggagggaag ggggaggcgg ggcgaggaca gtgaccggag tctcctcagc ggtggctttt 10080
ctgcttggca gcctcagcgg ctggcgccaa aaccggactc cgcccacttc ctcgcccgcc 10140
ggtgcgaggg tgtggaatcc tccagacgct gggggagggg gagttgggag cttaaaaact 10200
agtacccctt tgggaccact ttcagcagcg aactctcctg tacaccaggg gtcagttcca 10260
cagacgcggg ccaggggtgg gtcattgcgg cgtgaacaat aatttgacta gaagttgatt 10320
cgggtgtttc cggaaggggc cgagtcaatc cgccgagttg gggcacggaa aacaaaaagg 10380
gaaggctact aagatttttc tggcgggggt tatcattggc gtaactgcag ggaccacctc 10440
ccgggttgag ggggctggat ctccaggctg cggattaagc ccctcccgtc ggcgttaatt 10500
tcaaactgcg cgacgtttct cacctgcctt cgccaaggca ggggccggga ccctattcca 10560
agaggtagta actagcagga ctctagcctt ccgcaattca ttgagcgcat ttacggaagt 10620
aacgtcgggt actgtctctg gccgcaaggg tgggaggagt acgcatttgg cgtaaggtgg 10680
ggcgtagagc cttcccgcca ttggcggcgg atagggcgtt tacgcgacgg cctgacgtag 10740
cggaagacgc cttagtgggg gggaaggttc tagaaaagcg gcggcagcgg ctctagcggc 10800
agtagcagca gcgccgggtc ccgtgcggag gtgctcctcg cagagttgtt tctccagcag 10860
cggcagttct cactacagcg ccaggacgag tccggttcgt gttcgtccgc ggagatctct 10920
ctcatctcgc tcggctgcgg gaaatcgggc tgaagcgact gagtccgcga tggaggtaac 10980
gggtttgaaa tcaatgagtt attgaaaagg gcatggcgag gccgttggcg cctcagtgga 11040
agtcggccag ccgcctccgt gggagagagg caggaaatcg gaccaattca gtagcagtgg 11100
ggcttaaggt ttatgaacgg ggtcttgagc ggaggcctga gcgtacaaac agcttcccca 11160
ccctcagcct cccggcgcca tttcccttca ctgggggtgg gggatgggga gctttcacat 11220
ggcggacgct gccccgctgg ggtgaaagtg gggcgcggag gcgggacttc ttattccctt 11280
tctaaagcac gctgcttcgg gggccacggc gtctcctcgg aaagctt 11327
<210> 4
<211> 736
<212> DNA
<213> Artificial Sequence
<220>
<223> Antibody (OMLH) light chain gene cDNA with optimized Kozak
sequence added upstream of the initiation codon
<400> 4
cgcgccccgc cgccaccatg ggttggtctt gtatcatctt atttttagtt gctactgcta 60
ctggtgttca ttctgatata cagctcaccc aaagcccatc atctctgtct gcaagcgtcg 120
gcgacagggt gaccattacc tgtcgcgcaa gccaaagcgt tgactacgac ggcgacagct 180
acatgaactg gtaccagcag aagcccggca aggctcctaa gctgctgatc tatgccgcct 240
cctaccttga atctggagtg ccttctcgtt tttccggctc agggtccgga actgatttta 300
ccctgaccat ttcctccctc cagcccgagg attttgccac ttactactgt cagcagtccc 360
acgaggaccc atatacattc ggacaaggta caaaggtaga aatcaagcgt acggtggctg 420
ccccatccgt gttcatattt cctcctagcg acgaacaact caagtccggt accgccagcg 480
tggtctgcct gttgaacaat ttttatccaa gagaagctaa ggtccagtgg aaggttgaca 540
acgcccttca gtccggaaat agccaagaga gcgtcaccga acaggactcc aaggacagta 600
catactcact gagctctaca ctgacccttt ctaaggccga ctacgagaag cacaaggtct 660
acgcatgcga agtgacccat cagggactca gtagccctgt aacaaagagt tttaatcgag 720
gcgagtgcta agcgat 736
<210> 5
<211> 1436
<212> DNA
<213> Artificial Sequence
<220>
<223> Antibody (OMLH) heavy chain gene cDNA with optimized Kozak
sequence added upstream of the initiation codon
<400> 5
ggccgcccgc cgccaccatg gaatttggtt tatcttgggt ttttttagtt gctttattaa 60
gaggtgttca atgtgaggtg cagttggtcg aatccggcgg cggactcgtg caaccaggcg 120
gaagtttgcg gctgtcctgc gcagtgtctg gttacagcat cacctccggg tatagctgga 180
actggatccg ccaggctcct ggaaaggggc ttgagtgggt ggcttccatt acctacgacg 240
gctccactaa ctataacccg agcgtcaaag gcagaatcac catctctcgg gacgactcaa 300
agaatacctt ctacctgcag atgaactcac tgagggccga agataccgca gtttactact 360
gcgccagggg gtcccactat ttcggccact ggcacttcgc cgtgtgggga cagggcacac 420
tcgtgaccgt tagtagcgct agcaccaaag gcccctctgt gttcccactt gctccctcca 480
gtaaatctac ctccggagga accgcagccc tcggctgcct ggtgaaggat tacttcccag 540
agcccgtcac cgtctcttgg aactccggag ccttgactag cggagtgcac actttccctg 600
ctgtattgca gtccagcggc ttgtattcac tgagtagcgt cgtcaccgtg ccttcaagca 660
gcctcgggac acagacatac atatgtaatg tcaaccataa gccatcaaac actaaagttg 720
ataaaaaggt ggaacctaag agttgcgata agacccatac ctgtcctcct tgccctgctc 780
ctgagctgct gggaggccct agcgtgtttc tgttcccccc caagcccaaa gatacactga 840
tgatttcccg cacacctgaa gtaacatgtg tcgtggttga tgtgagtcac gaggatccag 900
aggtcaagtt taattggtac gtggacggag tggaggtgca caacgctaag actaagcctc 960
gggaggaaca gtacaacagc acataccgcg tggtcagcgt tttgactgtg ctgcatcaag 1020
actggctcaa tggaaaggaa tacaagtgca aggtctctaa taaagccctc cccgctccta 1080
ttgagaagac tatttctaaa gccaagggcc agcctcgcga acctcaggta tatactttgc 1140
caccctctcg cgaagaaatg acaaagaatc aggtctcact cacttgcctc gtcaaagggt 1200
tttacccttc tgacatcgct gtcgaatggg aaagtaatgg tcagccagaa aacaattaca 1260
agactactcc accagtgctc gattctgatg gaagtttctt tctctacagt aagctcactg 1320
tggacaaatc tcgctggcag cagggtaacg tattctcatg ctccgtgatg catgaagccc 1380
tccacaacca ttacacccag aagagcctgt ctctgagccc aggcaagtaa cctgca 1436
<210> 6
<211> 7317
<212> DNA
<213> Artificial Sequence
<220>
<223> pDC61
<400> 6
cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc 60
aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt 120
aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta 180
tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg 240
gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga 300
cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt 360
tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg 420
gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc 480
cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg 540
taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat 600
aagcagagct ctctggctaa ctagagaacc cactgttaac tggcttatcg aaattgtcga 660
ggagaacttc agggtgagtt tggggaccct tgattgttct ttctttttcg ctattgtaaa 720
attcatgtta tatggagggg gcaaagtttt cagggtgttg tttagaatgg gaagatgtcc 780
cttgtatcac catggaccct catgataatt ttgtttcttt cactttctac tctgttgaca 840
accattgtct cctcttattt tcttttcatt ttctgtaact ttttcgttaa actttagctt 900
gcatttgtaa cgaattttta aattcacttt tgtttatttg tcagattgta agtactttct 960
ctaatcactt ttttttcaag gcaatcaggg tatattatat tgtacttcag cacagtttta 1020
gagaacaatt gttataatta aatgataagg tagaatattt ctgcatataa attctggctg 1080
gcgtggaaat attcttattg gtagaaacaa ctacatcctg gtcatcatcc tgcctttctc 1140
tttatggtta caatgatata cactgtttga gatgaggata aaatactctg agtccaaacc 1200
gggcccctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt 1260
gctggcggcc gccccggggc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc 1320
ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa 1380
aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg 1440
gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggactcgagc 1500
gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca 1560
attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta 1620
aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat 1680
gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg 1740
taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac 1800
gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt 1860
cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg 1920
cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc 1980
attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt 2040
aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata 2100
agcagagctc tctggctaac tagagaaccc actgttaact ggcttatcga aattgtcgag 2160
gagaacttca gggtgagttt ggggaccctt gattgttctt tctttttcgc tattgtaaaa 2220
ttcatgttat atggaggggg caaagttttc agggtgttgt ttagaatggg aagatgtccc 2280
ttgtatcacc atggaccctc atgataattt tgtttctttc actttctact ctgttgacaa 2340
ccattgtctc ctcttatttt cttttcattt tctgtaactt tttcgttaaa ctttagcttg 2400
catttgtaac gaatttttaa attcactttt gtttatttgt cagattgtaa gtactttctc 2460
taatcacttt tttttcaagg caatcagggt atattatatt gtacttcagc acagttttag 2520
agaacaattg ttataattaa atgataaggt agaatatttc tgcatataaa ttctggctgg 2580
cgtggaaata ttcttattgg tagaaacaac tacatcctgg tcatcatcct gcctttctct 2640
ttatggttac aatgatatac actgtttgag atgaggataa aatactctga gtccaaaccg 2700
ggcccctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg 2760
ctggcgcgcc tctagagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 2820
ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 2880
tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 2940
gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg aggatctccg 3000
cggtgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa 3060
gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca 3120
tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 3180
ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag 3240
gcttttttgg aggcctaggc ttttgcaaaa aagctgcaga tggtacgacc attaaattgt 3300
attgtagcag tatcacaaaa tatgggtatt ggtaaaaatg gtgatttacc atggccacca 3360
ttacgaaatg aatttaaata ttttcaacga atgactacta cttcatcagt agaaggtaaa 3420
caaaatttag taattatggg tcgaaaaact tggttttcaa ttcctgagaa gaatcgacct 3480
ttaaaggaca gaattaatat agttctcagt agagaactca aagaaccacc acgaggagct 3540
cattttcttg ccaaaagttt ggatgatgcc ttaagactta ttgaacaacc ggaattggca 3600
agtaaagtag acatggtttg gatagtcgga ggcagttctg tttaccagga agccatgaat 3660
caaccaggcc acctcagact ctttgtgaca aggatcatgc aggaatttga aagtgacacg 3720
tttttcccag aaattgattt ggggaaatat aaacttctcc cagaataccc aggcgtcctc 3780
tctgaggtcc aggaggaaaa aggcatcaag tataagtttg aagtctacga gaagaaagac 3840
taaagatccg tgacataatt ggacaaacta cctacagaga tttaaagctc taaggtaaat 3900
ataaaatttt taagtgtata atgtgttaaa ctactgattc taattgtttg tgtattttag 3960
attccaacct atggaactga tgaatgggag cagtggtgga atgcctttaa tgaggaaaac 4020
ctgttttgct cagaagaaat gccatctagt gatgatgagg ctactgctga ctctcaacat 4080
tctactcctc caaaaaagaa gagaaaggta gaagacccca aggactttcc ttcagaattg 4140
ctaagttttt tgagtcatgc tgtgtttagt aatagaactc ttgcttgctt tgctatttac 4200
accacaaagg aaaaagctgc actgctatac aagaaaatta tggaaaaata ttctgtaacc 4260
tttataagta ggcataacag ttataatcat aacatactgt tttttcttac tccacacagg 4320
catagagtgt ctgctattaa taactatgct caaaaattgt gtacctttag ctttttaatt 4380
tgtaaagggg ttaataagga atatttgatg tatagtgcct tgactagaga tcataatcag 4440
ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa 4500
cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg 4560
ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 4620
tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgggccc atcgatgaat 4680
tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat 4740
cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat 4800
cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc 4860
cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct 4920
gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg 4980
gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg 5040
tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc 5100
ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt 5160
cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 5220
ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 5280
agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 5340
tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 5400
gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 5460
gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 5520
attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 5580
gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 5640
agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 5700
ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 5760
cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 5820
gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 5880
cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 5940
gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 6000
ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 6060
acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 6120
ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 6180
aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 6240
aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 6300
ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 6360
ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 6420
actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 6480
caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 6540
gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 6600
ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 6660
cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 6720
cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 6780
acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 6840
ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 6900
gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 6960
tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat 7020
accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag 7080
cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac 7140
gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc 7200
actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt 7260
gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctt 7317
<210> 7
<211> 2635
<212> DNA
<213> Artificial Sequence
<220>
<223> Sequence in pDC61, which is substitued for the sequence from No.
3182 to No. 5843 in pDC6
<400> 7
aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt 60
aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc 120
gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt 180
ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc 240
tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga 300
cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc 360
atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata 420
cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact 480
tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 540
tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt 600
atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct 660
gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca 720
cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc 780
gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc 840
cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg 900
gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta 960
tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc 1020
ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt 1080
gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg 1140
cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct 1200
tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc 1260
tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct 1320
cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac 1380
acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc 1440
tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat 1500
ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 1560
accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 1620
aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 1680
ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 1740
gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta 1800
ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 1860
ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 1920
ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 1980
gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 2040
cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 2100
cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 2160
cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 2220
aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 2280
ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 2340
gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa 2400
gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg 2460
cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag 2520
ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga 2580
attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgcca 2635
<210> 8
<211> 725
<212> DNA
<213> Artificial Sequence
<220>
<223> cDNA encoding human omalizumab antibody light chain (OML) to be
substitued for the sequence from No. 1267 to No. 1273 in pDC61
<400> 8
ggccgccacc atgggttggt cttgtatcat cttattttta gttgctactg ctactggtgt 60
tcattctgat atacagctca cccaaagccc atcatctctg tctgcaagcg tcggcgacag 120
ggtgaccatt acctgtcgcg caagccaaag cgttgactac gacggcgaca gctacatgaa 180
ctggtaccag cagaagcccg gcaaggctcc taagctgctg atctatgccg cctcctacct 240
tgaatctgga gtgccttctc gtttttccgg ctcagggtcc ggaactgatt ttaccctgac 300
catttcctcc ctccagcccg aggattttgc cacttactac tgtcagcagt cccacgagga 360
cccatataca ttcggacaag gtacaaaggt agaaatcaag cgtacggtgg ctgccccatc 420
cgtgttcata tttcctccta gcgacgaaca actcaagtcc ggtaccgcca gcgtggtctg 480
cctgttgaac aatttttatc caagagaagc taaggtccag tggaaggttg acaacgccct 540
tcagtccgga aatagccaag agagcgtcac cgaacaggac tccaaggaca gtacatactc 600
actgagctct acactgaccc tttctaaggc cgactacgag aagcacaagg tctacgcatg 660
cgaagtgacc catcagggac tcagtagccc tgtaacaaag agttttaatc gaggcgagtg 720
ctaac 725
<210> 9
<211> 1423
<212> DNA
<213> Artificial Sequence
<220>
<223> cDNA encoding human omalizumab antibody heavy chain (OMH) to be
substitued for the sequence from No. 2765 to No. 2771 in pDC61
<400> 9
cgcgccacca tggaatttgg tttatcttgg gtttttttag ttgctttatt aagaggtgtt 60
caatgtgagg tgcagttggt cgaatccggc ggcggactcg tgcaaccagg cggaagtttg 120
cggctgtcct gcgcagtgtc tggttacagc atcacctccg ggtatagctg gaactggatc 180
cgccaggctc ctggaaaggg gcttgagtgg gtggcttcca ttacctacga cggctccact 240
aactataacc cgagcgtcaa aggcagaatc accatctctc gggacgactc aaagaatacc 300
ttctacctgc agatgaactc actgagggcc gaagataccg cagtttacta ctgcgccagg 360
gggtcccact atttcggcca ctggcacttc gccgtgtggg gacagggcac actcgtgacc 420
gttagtagcg ctagcaccaa aggcccctct gtgttcccac ttgctccctc cagtaaatct 480
acctccggag gaaccgcagc cctcggctgc ctggtgaagg attacttccc agagcccgtc 540
accgtctctt ggaactccgg agccttgact agcggagtgc acactttccc tgctgtattg 600
cagtccagcg gcttgtattc actgagtagc gtcgtcaccg tgccttcaag cagcctcggg 660
acacagacat acatatgtaa tgtcaaccat aagccatcaa acactaaagt tgataaaaag 720
gtggaaccta agagttgcga taagacccat acctgtcctc cttgccctgc tcctgagctg 780
ctgggaggcc ctagcgtgtt tctgttcccc cccaagccca aagatacact gatgatttcc 840
cgcacacctg aagtaacatg tgtcgtggtt gatgtgagtc acgaggatcc agaggtcaag 900
tttaattggt acgtggacgg agtggaggtg cacaacgcta agactaagcc tcgggaggaa 960
cagtacaaca gcacataccg cgtggtcagc gttttgactg tgctgcatca agactggctc 1020
aatggaaagg aatacaagtg caaggtctct aataaagccc tccccgctcc tattgagaag 1080
actatttcta aagccaaggg ccagcctcgc gaacctcagg tatatacttt gccaccctct 1140
cgcgaagaaa tgacaaagaa tcaggtctca ctcacttgcc tcgtcaaagg gttttaccct 1200
tctgacatcg ctgtcgaatg ggaaagtaat ggtcagccag aaaacaatta caagactact 1260
ccaccagtgc tcgattctga tggaagtttc tttctctaca gtaagctcac tgtggacaaa 1320
tctcgctggc agcagggtaa cgtattctca tgctccgtga tgcatgaagc cctccacaac 1380
cattacaccc agaagagcct gtctctgagc ccaggcaagt aat 1423
<210> 10
<211> 10686
<212> DNA
<213> Artificial Sequence
<220>
<223> pNC32c-U533
<400> 10
cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc 60
aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt 120
aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta 180
tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg 240
gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga 300
cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt 360
tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg 420
gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc 480
cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg 540
taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat 600
aagcagagct ctctggctaa ctagagaacc cactgttaac tggcttatcg aaattggcgc 660
gccgcgatcg ccctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt 720
gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 780
ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 840
gtatcttatc atgtctggat ctccgcggcc ctccgcgcct acagctcaag ccacatccga 900
agggggaggg agccgggagc tgcgcgcggg gccgccgggg ggaggggtgg caccgcccac 960
gccgggcggc cacgaagggc ggggcagcgg gcgcgcgcgc ggcgggggga ggggccggcg 1020
ccgcgcccgc tgggaattgg ggccctaggg ggagggcgga ggcgccgacg accgcggcac 1080
ttaccgttcg cggcgtggcg cccggtggtc cccaagggga gggaaggggg aggcggggcg 1140
aggacagtga ccggagtctc ctcagcggtg gcttttctgc ttggcagcct cagcggctgg 1200
cgccaaaacc ggactccgcc cacttcctcg cccgccggtg cgagggtgtg gaatcctcca 1260
gacgctgggg gagggggagt tgggagctta aaaactagta cccctttggg accactttca 1320
gcagcgaact ctcctgtaca ccaggggtca gttccacaga cgcgggccag gggtgggtca 1380
ttgcggcgtg aacaataatt tgactagaag ttgattcggg tgtttccgga aggggccgag 1440
tcaatccgcc gagttggggc acggaaaaca aaaagggaag gctactaaga tttttctggc 1500
gggggttatc attggcgtaa ctgcagggac cacctcccgg gttgaggggg ctggatctcc 1560
aggctgcgga ttaagcccct cccgtcggcg ttaatttcaa actgcgcgac gtttctcacc 1620
tgccttcgcc aaggcagggg ccgggaccct attccaagag gtagtaacta gcaggactct 1680
agccttccgc aattcattga gcgcatttac ggaagtaacg tcgggtactg tctctggccg 1740
caagggtggg aggagtacgc atttggcgta aggtggggcg tagagccttc ccgccattgg 1800
cggcggatag ggcgtttacg cgacggcctg acgtagcgga agacgcctta gtggggggga 1860
aggttctaga aaagcggcgg cagcggctct agcggcagta gcagcagcgc cgggtcccgt 1920
gcggaggtgc tcctcgcaga gttgtttctc cagcagcggc agttctcact acagcgccag 1980
gacgagtccg gttcgtgttc gtccgcggag atctctctca tctcgctcgg ctgcgggaaa 2040
tcgggctgaa gcgactgagt ccgcgatgga ggtaacgggt ttgaaatcaa tgagttattg 2100
aaaagggcat ggcgaggccg ttggcgcctc agtggaagtc ggccagccgc ctccgtggga 2160
gagaggcagg aaatcggacc aattcagtag cagtggggct taaggtttat gaacggggtc 2220
ttgagcggag gcctgagcgt acaaacagct tccccaccct cagcctcccg gcgccatttc 2280
ccttcactgg gggtggggga tggggagctt tcacatggcg gacgctgccc cgctggggtg 2340
aaagtggggc gcggaggcgg gacttcttat tccctttcta aagcacgctg cttcgggggc 2400
cacggcgtct cctcggagaa ttccgatgta cgggccagat atacgcgttg acattgatta 2460
ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 2520
ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 2580
ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 2640
cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 2700
atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 2760
cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 2820
attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 2880
cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 2940
caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 3000
cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgtt 3060
aactggctta tcgaaattgc ggccgccctg caggcctccc acacctcccc ctgaacctga 3120
aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca 3180
aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 3240
gtggtttgtc caaactcatc aatgtatctt atcatgtctg gactccgcgg ggccctccgc 3300
gcctacagct caagccacat ccgaaggggg agggagccgg gagctgcgcg cggggccgcc 3360
ggggggaggg gtggcaccgc ccacgccggg cggccacgaa gggcggggca gcgggcgcgc 3420
gcgcggcggg gggaggggcc ggcgccgcgc ccgctgggaa ttggggccct agggggaggg 3480
cggaggcgcc gacgaccgcg gcacttaccg ttcgcggcgt ggcgcccggt ggtccccaag 3540
gggagggaag ggggaggcgg ggcgaggaca gtgaccggag tctcctcagc ggtggctttt 3600
ctgcttggca gcctcagcgg ctggcgccaa aaccggactc cgcccacttc ctcgcccgcc 3660
ggtgcgaggg tgtggaatcc tccagacgct gggggagggg gagttgggag cttaaaaact 3720
agtacccctt tgggaccact ttcagcagcg aactctcctg tacaccaggg gtcagttcca 3780
cagacgcggg ccaggggtgg gtcattgcgg cgtgaacaat aatttgacta gaagttgatt 3840
cgggtgtttc cggaaggggc cgagtcaatc cgccgagttg gggcacggaa aacaaaaagg 3900
gaaggctact aagatttttc tggcgggggt tatcattggc gtaactgcag ggaccacctc 3960
ccgggttgag ggggctggat ctccaggctg cggattaagc ccctcccgtc ggcgttaatt 4020
tcaaactgcg cgacgtttct cacctgcctt cgccaaggca ggggccggga ccctattcca 4080
agaggtagta actagcagga ctctagcctt ccgcaattca ttgagcgcat ttacggaagt 4140
aacgtcgggt actgtctctg gccgcaaggg tgggaggagt acgcatttgg cgtaaggtgg 4200
ggcgtagagc cttcccgcca ttggcggcgg atagggcgtt tacgcgacgg cctgacgtag 4260
cggaagacgc cttagtgggg gggaaggttc tagaaaagcg gcggcagcgg ctctagcggc 4320
agtagcagca gcgccgggtc ccgtgcggag gtgctcctcg cagagttgtt tctccagcag 4380
cggcagttct cactacagcg ccaggacgag tccggttcgt gttcgtccgc ggagatctct 4440
ctcatctcgc tcggctgcgg gaaatcgggc tgaagcgact gagtccgcga tggaggtaac 4500
gggtttgaaa tcaatgagtt attgaaaagg gcatggcgag gccgttggcg cctcagtgga 4560
agtcggccag ccgcctccgt gggagagagg caggaaatcg gaccaattca gtagcagtgg 4620
ggcttaaggt ttatgaacgg ggtcttgagc ggaggcctga gcgtacaaac agcttcccca 4680
ccctcagcct cccggcgcca tttcccttca ctgggggtgg gggatgggga gctttcacat 4740
ggcggacgct gccccgctgg ggtgaaagtg gggcgcggag gcgggacttc ttattccctt 4800
tctaaagcac gctgcttcgg gggccacggc gtctcctcgg aaccggttgt ggaatgtgtg 4860
tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 4920
tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 4980
gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 5040
gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 5100
ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt 5160
ttttggaggc ctaggctttt gcaaaaaagc tgcagatgat tgaacaagat ggattgcacg 5220
caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa 5280
tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg 5340
tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt 5400
ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa 5460
gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc 5520
ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg 5580
ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg 5640
aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg 5700
aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg 5760
gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact 5820
gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg 5880
ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc 5940
ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gatccgtgac 6000
ataattggac aaactaccta cagagattta aagctctaag gtaaatataa aatttttaag 6060
tgtataatgt gttaaactac tgattctaat tgtttgtgta ttttagattc caacctatgg 6120
aactgatgaa tgggagcagt ggtggaatgc ctttaatgag gaaaacctgt tttgctcaga 6180
agaaatgcca tctagtgatg atgaggctac tgctgactct caacattcta ctcctccaaa 6240
aaagaagaga aaggtagaag accccaagga ctttccttca gaattgctaa gttttttgag 6300
tcatgctgtg tttagtaata gaactcttgc ttgctttgct atttacacca caaaggaaaa 6360
agctgcactg ctatacaaga aaattatgga aaaatattct gtaaccttta taagtaggca 6420
taacagttat aatcataaca tactgttttt tcttactcca cacaggcata gagtgtctgc 6480
tattaataac tatgctcaaa aattgtgtac ctttagcttt ttaatttgta aaggggttaa 6540
taaggaatat ttgatgtata gtgccttgac tagagatcat aatcagccat accacatttg 6600
tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa 6660
tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca 6720
atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 6780
ccaaactcat caatgtatct tatcatgtct gggcccatcg atgccgacgt agcgcctgat 6840
gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag 6900
tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga 6960
cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc 7020
cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg 7080
cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc 7140
aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca 7200
ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa 7260
aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt 7320
ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca 7380
gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag 7440
ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc 7500
ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca 7560
gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt 7620
aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct 7680
gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt 7740
aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga 7800
caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact 7860
tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc 7920
acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga 7980
gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt 8040
agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga 8100
gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact 8160
ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga 8220
taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 8280
agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 8340
aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 8400
ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta 8460
gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 8520
aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 8580
aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 8640
gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 8700
aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 8760
aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 8820
cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 8880
cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 8940
tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt 9000
tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga 9060
ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta 9120
atgcagctgg gccctccgcg cctacagctc aagccacatc cgaaggggga gggagccggg 9180
agctgcgcgc ggggccgccg gggggagggg tggcaccgcc cacgccgggc ggccacgaag 9240
ggcggggcag cgggcgcgcg cgcggcgggg ggaggggccg gcgccgcgcc cgctgggaat 9300
tggggcccta gggggagggc ggaggcgccg acgaccgcgg cacttaccgt tcgcggcgtg 9360
gcgcccggtg gtccccaagg ggagggaagg gggaggcggg gcgaggacag tgaccggagt 9420
ctcctcagcg gtggcttttc tgcttggcag cctcagcggc tggcgccaaa accggactcc 9480
gcccacttcc tcgcccgccg gtgcgagggt gtggaatcct ccagacgctg ggggaggggg 9540
agttgggagc ttaaaaacta gtaccccttt gggaccactt tcagcagcga actctcctgt 9600
acaccagggg tcagttccac agacgcgggc caggggtggg tcattgcggc gtgaacaata 9660
atttgactag aagttgattc gggtgtttcc ggaaggggcc gagtcaatcc gccgagttgg 9720
ggcacggaaa acaaaaaggg aaggctacta agatttttct ggcgggggtt atcattggcg 9780
taactgcagg gaccacctcc cgggttgagg gggctggatc tccaggctgc ggattaagcc 9840
cctcccgtcg gcgttaattt caaactgcgc gacgtttctc acctgccttc gccaaggcag 9900
gggccgggac cctattccaa gaggtagtaa ctagcaggac tctagccttc cgcaattcat 9960
tgagcgcatt tacggaagta acgtcgggta ctgtctctgg ccgcaagggt gggaggagta 10020
cgcatttggc gtaaggtggg gcgtagagcc ttcccgccat tggcggcgga tagggcgttt 10080
acgcgacggc ctgacgtagc ggaagacgcc ttagtggggg ggaaggttct agaaaagcgg 10140
cggcagcggc tctagcggca gtagcagcag cgccgggtcc cgtgcggagg tgctcctcgc 10200
agagttgttt ctccagcagc ggcagttctc actacagcgc caggacgagt ccggttcgtg 10260
ttcgtccgcg gagatctctc tcatctcgct cggctgcggg aaatcgggct gaagcgactg 10320
agtccgcgat ggaggtaacg ggtttgaaat caatgagtta ttgaaaaggg catggcgagg 10380
ccgttggcgc ctcagtggaa gtcggccagc cgcctccgtg ggagagaggc aggaaatcgg 10440
accaattcag tagcagtggg gcttaaggtt tatgaacggg gtcttgagcg gaggcctgag 10500
cgtacaaaca gcttccccac cctcagcctc ccggcgccat ttcccttcac tgggggtggg 10560
ggatggggag ctttcacatg gcggacgctg ccccgctggg gtgaaagtgg ggcgcggagg 10620
cgggacttct tattcccttt ctaaagcacg ctgcttcggg ggccacggcg tctcctcgga 10680
aagctt 10686
<210> 11
<211> 728
<212> DNA
<213> Artificial Sequence
<220>
<223> cDNA encoding human omalizumab antibody (OKLH) light chain to be
substitued for the sequence from No. 658 to No. 668 in
pNC32c-U533
<400> 11
cgcgccacca tgggttggtc ttgtatcatc ttatttttag ttgctactgc tactggtgtt 60
cattctgata tacagctcac ccaaagccca tcatctctgt ctgcaagcgt cggcgacagg 120
gtgaccatta cctgtcgcgc aagccaaagc gttgactacg acggcgacag ctacatgaac 180
tggtaccagc agaagcccgg caaggctcct aagctgctga tctatgccgc ctcctacctt 240
gaatctggag tgccttctcg tttttccggc tcagggtccg gaactgattt taccctgacc 300
atttcctccc tccagcccga ggattttgcc acttactact gtcagcagtc ccacgaggac 360
ccatatacat tcggacaagg tacaaaggta gaaatcaagc gtacggtggc tgccccatcc 420
gtgttcatat ttcctcctag cgacgaacaa ctcaagtccg gtaccgccag cgtggtctgc 480
ctgttgaaca atttttatcc aagagaagct aaggtccagt ggaaggttga caacgccctt 540
cagtccggaa atagccaaga gagcgtcacc gaacaggact ccaaggacag tacatactca 600
ctgagctcta cactgaccct ttctaaggcc gactacgaga agcacaaggt ctacgcatgc 660
gaagtgaccc atcagggact cagtagccct gtaacaaaga gttttaatcg aggcgagtgc 720
taagcgat 728
<210> 12
<211> 1429
<212> DNA
<213> Artificial Sequence
<220>
<223> cDNA encoding human omalizumab antibody (OKLH) heavy chain to be
substitued for the sequence from No. 3081 to No. 3092 in
pNC32c-U533
<400> 12
ggccgccacc atggaatttg gtttatcttg ggttttttta gttgctttat taagaggtgt 60
tcaatgtgag gtgcagttgg tcgaatccgg cggcggactc gtgcaaccag gcggaagttt 120
gcggctgtcc tgcgcagtgt ctggttacag catcacctcc gggtatagct ggaactggat 180
ccgccaggct cctggaaagg ggcttgagtg ggtggcttcc attacctacg acggctccac 240
taactataac ccgagcgtca aaggcagaat caccatctct cgggacgact caaagaatac 300
cttctacctg cagatgaact cactgagggc cgaagatacc gcagtttact actgcgccag 360
ggggtcccac tatttcggcc actggcactt cgccgtgtgg ggacagggca cactcgtgac 420
cgttagtagc gctagcacca aaggcccctc tgtgttccca cttgctccct ccagtaaatc 480
tacctccgga ggaaccgcag ccctcggctg cctggtgaag gattacttcc cagagcccgt 540
caccgtctct tggaactccg gagccttgac tagcggagtg cacactttcc ctgctgtatt 600
gcagtccagc ggcttgtatt cactgagtag cgtcgtcacc gtgccttcaa gcagcctcgg 660
gacacagaca tacatatgta atgtcaacca taagccatca aacactaaag ttgataaaaa 720
ggtggaacct aagagttgcg ataagaccca tacctgtcct ccttgccctg ctcctgagct 780
gctgggaggc cctagcgtgt ttctgttccc ccccaagccc aaagatacac tgatgatttc 840
ccgcacacct gaagtaacat gtgtcgtggt tgatgtgagt cacgaggatc cagaggtcaa 900
gtttaattgg tacgtggacg gagtggaggt gcacaacgct aagactaagc ctcgggagga 960
acagtacaac agcacatacc gcgtggtcag cgttttgact gtgctgcatc aagactggct 1020
caatggaaag gaatacaagt gcaaggtctc taataaagcc ctccccgctc ctattgagaa 1080
gactatttct aaagccaagg gccagcctcg cgaacctcag gtatatactt tgccaccctc 1140
tcgcgaagaa atgacaaaga atcaggtctc actcacttgc ctcgtcaaag ggttttaccc 1200
ttctgacatc gctgtcgaat gggaaagtaa tggtcagcca gaaaacaatt acaagactac 1260
tccaccagtg ctcgattctg atggaagttt ctttctctac agtaagctca ctgtggacaa 1320
atctcgctgg cagcagggta acgtattctc atgctccgtg atgcatgaag ccctccacaa 1380
ccattacacc cagaagagcc tgtctctgag cccaggcaag taacctgca 1429
<210> 13
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> BstBI recognition site-containing sequence to be substituted for
the sequence from No. 5309 to No. 5311 in UCOE-Hu-P
<400> 13
cctagtagta gtagtagttc gaag 24
<210> 14
<211> 3400
<212> DNA
<213> Artificial Sequence
<220>
<223> cDNA encoding human omalizumab antibody light chain (OML) linking
to cDNA encoding Simian Virus 40 polyadenylation signal (SV40pA),
guinea pig cytomegalovirus promoter (PgpCMV) and human omalizumab
antibody heavy chain (OMH)
<400> 14
cccaccatgg gttggtcttg tatcatctta tttttagttg ctactgctac tggtgttcat 60
tctgatatac agctcaccca aagcccatca tctctgtctg caagcgtcgg cgacagggtg 120
accattacct gtcgcgcaag ccaaagcgtt gactacgacg gcgacagcta catgaactgg 180
taccagcaga agcccggcaa ggctcctaag ctgctgatct atgccgcctc ctaccttgaa 240
tctggagtgc cttctcgttt ttccggctca gggtccggaa ctgattttac cctgaccatt 300
tcctccctcc agcccgagga ttttgccact tactactgtc agcagtccca cgaggaccca 360
tatacattcg gacaaggtac aaaggtagaa atcaagcgta cggtggctgc cccatccgtg 420
ttcatatttc ctcctagcga cgaacaactc aagtccggta ccgccagcgt ggtctgcctg 480
ttgaacaatt tttatccaag agaagctaag gtccagtgga aggttgacaa cgcccttcag 540
tccggaaata gccaagagag cgtcaccgaa caggactcca aggacagtac atactcactg 600
agctctacac tgaccctttc taaggccgac tacgagaagc acaaggtcta cgcatgcgaa 660
gtgacccatc agggactcag tagccctgta acaaagagtt ttaatcgagg cgagtgctaa 720
gcgatcgcgc tagcgcggcc acgtagtcga ctacgtagag ctcggtaccc ggggatcctc 780
tagagtcgac ctgcaggcat gcaagctggc cgcgactcta gatcataatc agccatacca 840
catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac 900
ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat 960
aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 1020
gtttgtccaa actcatcaat gtatcttaac cggttatgtt acttggcaga ggccgcatgg 1080
aaagtccctg gacgtgggac atctgattaa tacgtgagga ggtcagccat gttctttttg 1140
gcaaaggact acggtcattg gacgtttgat tggcatggga tagggtcagc cagagttaac 1200
agtgttcttt tggcaaaggg atacgtggaa agtcccgggc catttacagt aaactgatac 1260
ggggacaaag cacagccata tttagtcatg tattgcttgg cagagggtct atggaaagtc 1320
cctggacgtg ggacgtctga ttaatatgaa agaaggtcag ccagaggtag ctgtgtcctt 1380
tttggcaaag ggatacggtt atgggacgtt tgattggact gggatagggt cagccagagt 1440
taacagtgtt cttttggcaa aggaaacgtg gaaagtcccg ggccatttac agtaaactga 1500
tactgggaca aagtacaccc atatttagtc atgttctttt tggcaaagag catctggaaa 1560
gtcccgggca gcattatagt cacttggcag agggaaaggg tcactcagag ttaagtacat 1620
ctttccaggg ccaatattcc agtaaattac acttagtttt atgcaaatca gccacaaagg 1680
ggattttccc ggtcaattat gactttttcc ttagtcatgc ggtatccaat tactgccaaa 1740
ttggcagtac atactaggtg attcactgac atttggccgt cctctggaaa gtccctggaa 1800
accgctcaag tactgtatca tggtgacttt gcatttttgg agagcacgcc ccactccacc 1860
attggtccac gtaccctatg ggggagtggt ttatgagtat ataaggggct ccggtttaga 1920
agccgggcag agcggaattc gagctccctg caggttagtt aagttaacgg cgcgccacca 1980
tggaatttgg tttatcttgg gtttttttag ttgctttatt aagaggtgtt caatgtgagg 2040
tgcagttggt cgaatccggc ggcggactcg tgcaaccagg cggaagtttg cggctgtcct 2100
gcgcagtgtc tggttacagc atcacctccg ggtatagctg gaactggatc cgccaggctc 2160
ctggaaaggg gcttgagtgg gtggcttcca ttacctacga cggctccact aactataacc 2220
cgagcgtcaa aggcagaatc accatctctc gggacgactc aaagaatacc ttctacctgc 2280
agatgaactc actgagggcc gaagataccg cagtttacta ctgcgccagg gggtcccact 2340
atttcggcca ctggcacttc gccgtgtggg gacagggcac actcgtgacc gttagtagcg 2400
ctagcaccaa aggcccctct gtgttcccac ttgctccctc cagtaaatct acctccggag 2460
gaaccgcagc cctcggctgc ctggtgaagg attacttccc agagcccgtc accgtctctt 2520
ggaactccgg agccttgact agcggagtgc acactttccc tgctgtattg cagtccagcg 2580
gcttgtattc actgagtagc gtcgtcaccg tgccttcaag cagcctcggg acacagacat 2640
acatatgtaa tgtcaaccat aagccatcaa acactaaagt tgataaaaag gtggaaccta 2700
agagttgcga taagacccat acctgtcctc cttgccctgc tcctgagctg ctgggaggcc 2760
ctagcgtgtt tctgttcccc cccaagccca aagatacact gatgatttcc cgcacacctg 2820
aagtaacatg tgtcgtggtt gatgtgagtc acgaggatcc agaggtcaag tttaattggt 2880
acgtggacgg agtggaggtg cacaacgcta agactaagcc tcgggaggaa cagtacaaca 2940
gcacataccg cgtggtcagc gttttgactg tgctgcatca agactggctc aatggaaagg 3000
aatacaagtg caaggtctct aataaagccc tccccgctcc tattgagaag actatttcta 3060
aagccaaggg ccagcctcgc gaacctcagg tatatacttt gccaccctct cgcgaagaaa 3120
tgacaaagaa tcaggtctca ctcacttgcc tcgtcaaagg gttttaccct tctgacatcg 3180
ctgtcgaatg ggaaagtaat ggtcagccag aaaacaatta caagactact ccaccagtgc 3240
tcgattctga tggaagtttc tttctctaca gtaagctcac tgtggacaaa tctcgctggc 3300
agcagggtaa cgtattctca tgctccgtga tgcatgaagc cctccacaac cattacaccc 3360
agaagagcct gtctctgagc ccaggcaagt aatctagatt 3400
<210> 15
<211> 394
<212> PRT
<213> Canis lupus
<400> 15
Met Ala Gly Phe Gly Phe Arg Arg His Gly Ala Gln Pro Asp Leu Ala
1 5 10 15
Ser Arg Thr Trp Pro Cys Thr Ala Leu Phe Ser Leu Leu Phe Ile Pro
20 25 30
Val Phe Ser Lys Gly Met His Val Ala Gln Pro Ala Val Val Leu Ala
35 40 45
Ser Ser Arg Gly Val Ala Ser Phe Val Cys Glu Tyr Gly Ser Ser Gly
50 55 60
Asn Ala Ala Glu Val Arg Val Thr Val Leu Arg Gln Ala Gly Ser Gln
65 70 75 80
Met Thr Glu Val Cys Ala Ala Thr Tyr Thr Val Glu Asp Glu Leu Ala
85 90 95
Phe Leu Asp Asp Ser Thr Cys Thr Gly Thr Ser Ser Gly Asn Lys Val
100 105 110
Asn Leu Thr Ile Gln Gly Leu Arg Ala Met Asp Thr Gly Leu Tyr Ile
115 120 125
Cys Lys Val Glu Leu Met Tyr Pro Pro Pro Tyr Tyr Val Gly Met Gly
130 135 140
Asn Gly Thr Gln Ile Tyr Val Ile Asp Pro Glu Pro Cys Pro Asp Ser
145 150 155 160
Asp Pro Lys Glu Ser Thr Cys Lys Cys Ile Ser Pro Cys Pro Val Pro
165 170 175
Glu Ser Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys
180 185 190
Asp Ile Leu Arg Ile Thr Arg Thr Pro Glu Ile Thr Cys Val Val Leu
195 200 205
Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp
210 215 220
Gly Lys Glu Val His Thr Ala Lys Thr Gln Pro Arg Glu Gln Gln Phe
225 230 235 240
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile Glu His Gln Asp
245 250 255
Trp Leu Thr Gly Lys Glu Phe Lys Cys Arg Val Asn His Ile Gly Leu
260 265 270
Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His
275 280 285
Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Pro Lys Glu Leu Ser Ser
290 295 300
Ser Asp Thr Val Thr Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro
305 310 315 320
Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Pro Glu Pro Glu Ser
325 330 335
Lys Tyr His Thr Thr Ala Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe
340 345 350
Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Gln Gly Asp
355 360 365
Thr Phe Thr Cys Ala Val Met His Glu Ala Leu Gln Asn His Tyr Thr
370 375 380
Asp Leu Ser Leu Ser His Ser Pro Gly Lys
385 390
<210> 16
<211> 1185
<212> DNA
<213> Artificial Sequence
<220>
<223> Codon-optimized nucleotide sequence encoding canis lupus
CTLA-4-Ig
<400> 16
atggctggat ttggattcag aaggcacgga gcccagcccg acctggcatc tcgcacttgg 60
ccctgtaccg cactgttttc actgctgttc atcccagtgt tcagcaaggg aatgcacgtg 120
gctcagccag ctgtggtgct ggcttccagc agaggcgtgg cttccttcgt gtgcgagtac 180
ggctcttccg gcaacgccgc tgaggtgaga gtgaccgtgc tgaggcaggc tggctcccag 240
atgacagagg tgtgcgccgc tacctataca gtggaggacg agctggcttt cctggacgat 300
agcacctgta caggcaccag ctctggcaac aaggtcaatc tgaccatcca gggcctgcgc 360
gccatggata caggcctgta catctgtaag gtggagctga tgtatccccc tccatactat 420
gtgggcatgg gcaatggcac ccagatctac gtgatcgacc ccgagccttg cccagactct 480
gatccaaagg agtcgacatg caagtgtatc tctccatgtc ctgtgccaga gagcctggga 540
ggaccttccg tgttcatctt tccccctaag ccaaaggata tcctgaggat cacacggacc 600
cctgagatca cctgcgtggt gctggacctg ggaagggagg atccagaggt gcagatctcc 660
tggttcgtgg acggcaagga ggtgcatacc gctaagacac agcccagaga gcagcagttt 720
aactccacct atcgcgtggt gagcgtgctg cctatcgagc accaggattg gctgacaggc 780
aaggagttta agtgccgggt gaatcatatc ggcctgccct ctcctatcga gaggaccatc 840
tccaaggcta ggggacaggc tcaccagcca agcgtgtacg tgctgccacc ctctcctaag 900
gagctgtcca gctctgacac agtgaccctg acatgtctga tcaaggactt ctttcctcca 960
gagatcgacg tggagtggca gtccaacggc cagccagagc ccgagagcaa gtatcatacc 1020
acagcccccc agctggacga ggatggctct tacttcctgt attccaagct gagcgtggac 1080
aagtccaggt ggcagcaggg cgataccttt acatgtgctg tgatgcacga agccctgcag 1140
aatcattaca ccgacctgtc actgtcccac tcccctggca aataa 1185
<210> 17
<211> 8444
<212> DNA
<213> Artificial Sequence
<220>
<223> pDC62c5-U533 containing one cloning site
<400> 17
cgatgtacgg gccagatata cgcgttgaca ttgattattg actagtcgcg ttacataact 60
tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat 120
gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta 180
tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc 240
tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg 300
ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg 360
gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct 420
ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa 480
atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt 540
ctatataagc agagctcgtt tagtgaaccg tcagatcctc actctcttcc gcatcgctgt 600
ctgcgagggc cagctgttgg gctcgcggtt gaggacaaac tcttcgcggt ctttccagta 660
ctcttggatc ggaaacccgt cggcctccga acggtactcc gccaccgagg gacctgagcg 720
agtccgcatc gaccggatcg gaaaacctct cgagaaaggc gtctaaccag tcacagtcgc 780
aaggtaggct gagcaccgtg gcgggcggca gcgggtggcg gtcggggttg tttctggcgg 840
aggtgctgct gatgatgtaa ttaaagtagg cggtcttgag acggcggatg gtcgaggtga 900
ggtgtggcag gcttgagatc cagctgttgg ggtgagtact ccctctcaaa agcgggcatg 960
acttctgcgc taagattgtc agtttccaaa aacgaggagg atttgatatt cacctggccc 1020
gatctggcca tacacttgag tgacaatgac atccactttg cctttctctc cacaggtgtc 1080
cactcccagg tccaaggcgc gccgcgatcg cgcctcgact gtgccttcta gttgccagcc 1140
atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt 1200
cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct 1260
ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc 1320
tggggaggat ctccgcgggg ccctccgcgc ctacagctca agccacatcc gaagggggag 1380
ggagccggga gctgcgcgcg gggccgccgg ggggaggggt ggcaccgccc acgccgggcg 1440
gccacgaagg gcggggcagc gggcgcgcgc gcggcggggg gaggggccgg cgccgcgccc 1500
gctgggaatt ggggccctag ggggagggcg gaggcgccga cgaccgcggc acttaccgtt 1560
cgcggcgtgg cgcccggtgg tccccaaggg gagggaaggg ggaggcgggg cgaggacagt 1620
gaccggagtc tcctcagcgg tggcttttct gcttggcagc ctcagcggct ggcgccaaaa 1680
ccggactccg cccacttcct cgcccgccgg tgcgagggtg tggaatcctc cagacgctgg 1740
gggaggggga gttgggagct taaaaactag tacccctttg ggaccacttt cagcagcgaa 1800
ctctcctgta caccaggggt cagttccaca gacgcgggcc aggggtgggt cattgcggcg 1860
tgaacaataa tttgactaga agttgattcg ggtgtttccg gaaggggccg agtcaatccg 1920
ccgagttggg gcacggaaaa caaaaaggga aggctactaa gatttttctg gcgggggtta 1980
tcattggcgt aactgcaggg accacctccc gggttgaggg ggctggatct ccaggctgcg 2040
gattaagccc ctcccgtcgg cgttaatttc aaactgcgcg acgtttctca cctgccttcg 2100
ccaaggcagg ggccgggacc ctattccaag aggtagtaac tagcaggact ctagccttcc 2160
gcaattcatt gagcgcattt acggaagtaa cgtcgggtac tgtctctggc cgcaagggtg 2220
ggaggagtac gcatttggcg taaggtgggg cgtagagcct tcccgccatt ggcggcggat 2280
agggcgttta cgcgacggcc tgacgtagcg gaagacgcct tagtgggggg gaaggttcta 2340
gaaaagcggc ggcagcggct ctagcggcag tagcagcagc gccgggtccc gtgcggaggt 2400
gctcctcgca gagttgtttc tccagcagcg gcagttctca ctacagcgcc aggacgagtc 2460
cggttcgtgt tcgtccgcgg agatctctct catctcgctc ggctgcggga aatcgggctg 2520
aagcgactga gtccgcgatg gaggtaacgg gtttgaaatc aatgagttat tgaaaagggc 2580
atggcgaggc cgttggcgcc tcagtggaag tcggccagcc gcctccgtgg gagagaggca 2640
ggaaatcgga ccaattcagt agcagtgggg cttaaggttt atgaacgggg tcttgagcgg 2700
aggcctgagc gtacaaacag cttccccacc ctcagcctcc cggcgccatt tcccttcact 2760
gggggtgggg gatggggagc tttcacatgg cggacgctgc cccgctgggg tgaaagtggg 2820
gcgcggaggc gggacttctt attccctttc taaagcacgc tgcttcgggg gccacggcgt 2880
ctcctcggag aattcaaatg ggaccggttg tggaatgtgt gtcagttagg gtgtggaaag 2940
tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 3000
atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 3060
ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 3120
gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaaagct 3180
gcagatggta cgaccattaa attgtattgt agcagtatca caaaatatgg gtattggtaa 3240
aaatggtgat ttaccatggc caccattacg aaatgaattt aaatattttc aacgaatgac 3300
tactacttca tcagtagaag gtaaacaaaa tttagtaatt atgggtcgaa aaacttggtt 3360
ttcaattcct gagaagaatc gacctttaaa ggacagaatt aatatagttc tcagtagaga 3420
actcaaagaa ccaccacgag gagctcattt tcttgccaaa agtttggatg atgccttaag 3480
acttattgaa caaccggaat tggcaagtaa agtagacatg gtttggatag tcggaggcag 3540
ttctgtttac caggaagcca tgaatcaacc aggccacctc agactctttg tgacaaggat 3600
catgcaggaa tttgaaagtg acacgttttt cccagaaatt gatttgggga aatataaact 3660
tctcccagaa tacccaggcg tcctctctga ggtccaggag gaaaaaggca tcaagtataa 3720
gtttgaagtc tacgagaaga aagactaaag atccgtgaca taattggaca aactacctac 3780
agagatttaa agctctaagg taaatataaa atttttaagt gtataatgtg ttaaactact 3840
gattctaatt gtttgtgtat tttagattcc aacctatgga actgatgaat gggagcagtg 3900
gtggaatgcc tttaatgagg aaaacctgtt ttgctcagaa gaaatgccat ctagtgatga 3960
tgaggctact gctgactctc aacattctac tcctccaaaa aagaagagaa aggtagaaga 4020
ccccaaggac tttccttcag aattgctaag ttttttgagt catgctgtgt ttagtaatag 4080
aactcttgct tgctttgcta tttacaccac aaaggaaaaa gctgcactgc tatacaagaa 4140
aattatggaa aaatattctg taacctttat aagtaggcat aacagttata atcataacat 4200
actgtttttt cttactccac acaggcatag agtgtctgct attaataact atgctcaaaa 4260
attgtgtacc tttagctttt taatttgtaa aggggttaat aaggaatatt tgatgtatag 4320
tgccttgact agagatcata atcagccata ccacatttgt agaggtttta cttgctttaa 4380
aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta 4440
acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 4500
ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 4560
atcatgtctg ggcccatcga tgccgacgta gcgctgatgc ggtattttct ccttacgcat 4620
ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca 4680
tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg 4740
ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg 4800
ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta 4860
taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat 4920
gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg 4980
agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa 5040
catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac 5100
ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac 5160
atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt 5220
ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc 5280
gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca 5340
ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc 5400
ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag 5460
gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa 5520
ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg 5580
gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa 5640
ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg 5700
gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt 5760
gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt 5820
caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag 5880
cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat 5940
ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct 6000
taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct 6060
tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 6120
gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc 6180
agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc 6240
aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct 6300
gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag 6360
gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc 6420
tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg 6480
agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag 6540
cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt 6600
gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac 6660
gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg 6720
ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc 6780
cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata 6840
cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctgggc cctccgcgcc 6900
tacagctcaa gccacatccg aagggggagg gagccgggag ctgcgcgcgg ggccgccggg 6960
gggaggggtg gcaccgccca cgccgggcgg ccacgaaggg cggggcagcg ggcgcgcgcg 7020
cggcgggggg aggggccggc gccgcgcccg ctgggaattg gggccctagg gggagggcgg 7080
aggcgccgac gaccgcggca cttaccgttc gcggcgtggc gcccggtggt ccccaagggg 7140
agggaagggg gaggcggggc gaggacagtg accggagtct cctcagcggt ggcttttctg 7200
cttggcagcc tcagcggctg gcgccaaaac cggactccgc ccacttcctc gcccgccggt 7260
gcgagggtgt ggaatcctcc agacgctggg ggagggggag ttgggagctt aaaaactagt 7320
acccctttgg gaccactttc agcagcgaac tctcctgtac accaggggtc agttccacag 7380
acgcgggcca ggggtgggtc attgcggcgt gaacaataat ttgactagaa gttgattcgg 7440
gtgtttccgg aaggggccga gtcaatccgc cgagttgggg cacggaaaac aaaaagggaa 7500
ggctactaag atttttctgg cgggggttat cattggcgta actgcaggga ccacctcccg 7560
ggttgagggg gctggatctc caggctgcgg attaagcccc tcccgtcggc gttaatttca 7620
aactgcgcga cgtttctcac ctgccttcgc caaggcaggg gccgggaccc tattccaaga 7680
ggtagtaact agcaggactc tagccttccg caattcattg agcgcattta cggaagtaac 7740
gtcgggtact gtctctggcc gcaagggtgg gaggagtacg catttggcgt aaggtggggc 7800
gtagagcctt cccgccattg gcggcggata gggcgtttac gcgacggcct gacgtagcgg 7860
aagacgcctt agtggggggg aaggttctag aaaagcggcg gcagcggctc tagcggcagt 7920
agcagcagcg ccgggtcccg tgcggaggtg ctcctcgcag agttgtttct ccagcagcgg 7980
cagttctcac tacagcgcca ggacgagtcc ggttcgtgtt cgtccgcgga gatctctctc 8040
atctcgctcg gctgcgggaa atcgggctga agcgactgag tccgcgatgg aggtaacggg 8100
tttgaaatca atgagttatt gaaaagggca tggcgaggcc gttggcgcct cagtggaagt 8160
cggccagccg cctccgtggg agagaggcag gaaatcggac caattcagta gcagtggggc 8220
ttaaggttta tgaacggggt cttgagcgga ggcctgagcg tacaaacagc ttccccaccc 8280
tcagcctccc ggcgccattt cccttcactg ggggtggggg atggggagct ttcacatggc 8340
ggacgctgcc ccgctggggt gaaagtgggg cgcggaggcg ggacttctta ttccctttct 8400
aaagcacgct gcttcggggg ccacggcgtc tcctcggaaa gctt 8444
<210> 18
<211> 724
<212> DNA
<213> Artificial Sequence
<220>
<223> Antibody (TRLH) light chain gene cDNA with optimized Kozak
sequence added upstream of the initiation codon
<400> 18
cgcgccccgc cgccaccatg ggttggtctt gtatcatctt atttttagtt gctactgcta 60
ctggtgttca ttctgatata cagatgaccc aaagcccatc atctctgtct gcaagcgtcg 120
gcgacagggt gaccattacc tgtcgcgcaa gccaagacgt taatacagca gtggcttggt 180
accagcagaa gcccggcaag gctcctaagc tgctgatcta tagcgcctcc tttctttatt 240
ctggagtgcc ttctcgtttt tccggctcaa ggtccggaac tgattttacc ctgaccattt 300
cctccctcca gcccgaggat tttgccactt actactgtca gcagcactat accacaccac 360
ctacattcgg acaaggtaca aaggtagaaa tcaagcgtac ggtggctgcc ccatccgtgt 420
tcatatttcc tcctagcgac gaacaactca agtccggtac cgccagcgtg gtctgcctgt 480
tgaacaattt ttatccaaga gaagctaagg tccagtggaa ggttgacaac gcccttcagt 540
ccggaaatag ccaagagagc gtcaccgaac aggactccaa ggacagtaca tactcactga 600
gctctacact gaccctttct aaggccgact acgagaagca caaggtctac gcatgcgaag 660
tgacccatca gggactcagt agccctgtaa caaagagttt taatcgaggc gagtgctaag 720
cgat 724
<210> 19
<211> 1430
<212> DNA
<213> Artificial Sequence
<220>
<223> Antibody (TRLH) heavy chain gene cDNA with optimized Kozak
sequence added upstream of the initiation codon
<400> 19
ggccgcccgc cgccaccatg gaatttggtt tatcttgggt ttttttagtt gctttattaa 60
gaggtgttca atgtgaggtg cagttggtcg aatccggcgg cggactcgtg caaccaggcg 120
gaagtttgcg gctgtcctgc gcagcctctg gttttaacat caaagatacc tatattcatt 180
gggtacgcca ggctcctgga aaggggcttg agtgggtggc tcgaatttac ccaaccaatg 240
gctacactcg gtatgccgac agcgtcaaag gcagattcac catctctgca gacacatcaa 300
agaataccgc ttacctgcag atgaactcac tgagggccga agataccgca gtttactact 360
gctccaggtg ggggggggac ggcttctacg ccatggatta ctggggacag ggcacactcg 420
tgaccgttag tagcgctagc accaaaggcc cctctgtgtt cccacttgct ccctccagta 480
aatctacctc cggaggaacc gcagccctcg gctgcctggt gaaggattac ttcccagagc 540
ccgtcaccgt ctcttggaac tccggagcct tgactagcgg agtgcacact ttccctgctg 600
tattgcagtc cagcggcttg tattcactga gtagcgtcgt caccgtgcct tcaagcagcc 660
tcgggacaca gacatacata tgtaatgtca accataagcc atcaaacact aaagttgata 720
aaaaggtgga acctaagagt tgcgataaga cccatacctg tcctccttgc cctgctcctg 780
agctgctggg aggccctagc gtgtttctgt tcccccccaa gcccaaagat acactgatga 840
tttcccgcac acctgaagta acatgtgtcg tggttgatgt gagtcacgag gatccagagg 900
tcaagtttaa ttggtacgtg gacggagtgg aggtgcacaa cgctaagact aagcctcggg 960
aggaacagta caacagcaca taccgcgtgg tcagcgtttt gactgtgctg catcaagact 1020
ggctcaatgg aaaggaatac aagtgcaagg tctctaataa agccctcccc gctcctattg 1080
agaagactat ttctaaagcc aagggccagc ctcgcgaacc tcaggtatat actttgccac 1140
cctctcgcga agaaatgaca aagaatcagg tctcactcac ttgcctcgtc aaagggtttt 1200
acccttctga catcgctgtc gaatgggaaa gtaatggtca gccagaaaac aattacaaga 1260
ctactccacc agtgctcgat tctgatggaa gtttctttct ctacagtaag ctcactgtgg 1320
acaaatctcg ctggcagcag ggtaacgtat tctcatgctc cgtgatgcat gaagccctcc 1380
acaaccatta cacccagaag agcctgtctc tgagcccagg ctaacctgca 1430
Claims (9)
- 하기 (a), (b) 및 (c)를 포함하는 발현 벡터:
(a) 코돈을 개변한 번역 장해성 디히드로 엽산 환원효소 유전자 카세트(번역 장해성 DHFR 유전자 카세트)로서, 번역 장해성 DHFR 유전자 카세트의 코돈이 알라닌은 GCA, 아르기닌은 CGA, 아스파라긴은 AAU, 아스파라긴산은 GAU, 시스테인은 UGU, 글루타민은 CAA, 글루탐산은 GAA, 글리신은 GGU, 히스티딘은 CAU, 루이신은 UUA, 리신은 AAA, 프롤린은 CCA, 페닐알라닌은 UUU, 세린은 UCA, 트레오닌은 ACU, 티로신은 UAU, 및/또는 발린은 GUA로 개변되어 있고, 코돈의 개변 영역이 DHFR 유전자의 5' 말단으로부터 DHFR 유전자의 전장의 30% 이상인 번역 장해성 DHFR 유전자 카세트;
(b) 전사 활성 프로모터와 안정성 폴리아데닐화 신호의 사이에 외래 유전자 조립용 클로닝 사이트를 포함하는 유전자 카세트; 및
(c) 편재성 크로마틴 오프닝 엘리먼트(UCOE). - 청구항 1에 있어서,
청구항 1의 (a)에 기재된 번역 장해성 DHFR 유전자 카세트가 프로모터로서 비포유동물 세포의 유전자를 유래로 하는 프로모터, 또는 인핸서 부분을 제거한 프로모터를 사용하고 있는 것을 특징으로 하는 발현 벡터. - 청구항 1 또는 청구항 2에 있어서,
UCOE가 서열번호 1의 뉴클레오티드 서열을 포함하는 발현 벡터. - 청구항 1 내지 청구항 3 중 어느 하나의 항에 기재된 발현 벡터에 외래 유전자를 조립하는 공정, 및 상기 발현 벡터에 의해 디히드로 엽산 환원효소 유전자 결손 숙주 세포를 형질전환시키는 공정을 포함하는, 외래 유전자 유래 단백질을 생산하는 형질전환체의 제조 방법.
- 이하 (a)∼(d)의 공정을 포함하는 외래 유전자 유래 단백질의 생산 방법:
(a) 청구항 1 내지 청구항 3 중 어느 하나의 항에 기재된 발현 벡터에 외래 유전자를 조립하는 공정,
(b) 상기 발현 벡터에 의해 디히드로 엽산 환원효소 유전자 결손 숙주 세포를 형질전환시키는 공정,
(c) 상기 형질전환체를 히포크산틴 티미딘 불포함 배지에서 배양하는 공정,
(d) 배양된 형질전환체로부터 외래 유전자 유래 단백질을 회수하는 공정. - 청구항 5에 있어서,
청구항 5의 (c)의 공정에 있어서, 화학적 구명 배지(CD 배지) 혹은 CD 배지에 비동물성의 첨가물을 가한 배지에서 배양하는 것을 특징으로 하는 방법. - 이하 (a)∼(c)의 공정을 포함하는 외래 유전자 유래 단백질을 생산하는 형질전환체의 스크리닝 방법:
(a) 청구항 1 내지 청구항 3 중 어느 하나의 항에 기재된 발현 벡터에 외래 유전자를 조립하는 공정,
(b) 상기 발현 벡터에 의해 디히드로 엽산 환원효소 유전자 결손 숙주 세포를 형질전환시키는 공정,
(c) 상기 형질전환체를 히포크산틴 티미딘 불포함 배지에서 배양하는 공정. - 청구항 1 내지 청구항 3 중 어느 하나의 항에 기재된 발현 벡터에 외래 유전자가 조립된 외래 유전자 발현 벡터.
- 청구항 8에 기재된 외래 유전자 발현 벡터에 의해 형질전환된 숙주 세포.
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| JPJP-P-2018-099704 | 2018-05-24 | ||
| JP2018099704 | 2018-05-24 | ||
| JPJP-P-2018-168591 | 2018-09-10 | ||
| JP2018168591 | 2018-09-10 | ||
| PCT/JP2019/018899 WO2019225372A1 (ja) | 2018-05-24 | 2019-05-13 | 新規ベクターおよびその利用 |
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| Country | Link |
|---|---|
| US (1) | US11781146B2 (ko) |
| EP (1) | EP3805396A4 (ko) |
| JP (1) | JP7264353B2 (ko) |
| KR (1) | KR20210013036A (ko) |
| CN (1) | CN112135909A (ko) |
| AU (1) | AU2019273200A1 (ko) |
| CA (1) | CA3099955A1 (ko) |
| WO (1) | WO2019225372A1 (ko) |
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Also Published As
| Publication number | Publication date |
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| WO2019225372A1 (ja) | 2019-11-28 |
| CN112135909A (zh) | 2020-12-25 |
| JP7264353B2 (ja) | 2023-04-25 |
| JPWO2019225372A1 (ja) | 2021-06-10 |
| AU2019273200A1 (en) | 2020-12-03 |
| US11781146B2 (en) | 2023-10-10 |
| US20210254079A1 (en) | 2021-08-19 |
| EP3805396A4 (en) | 2022-03-23 |
| EP3805396A1 (en) | 2021-04-14 |
| CA3099955A1 (en) | 2019-11-28 |
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