KR20180035092A

KR20180035092A - A DNA vaccine for preventing and treating HSV-2 infection

Info

Publication number: KR20180035092A
Application number: KR1020160125149A
Authority: KR
Inventors: 서용복; 신주아
Original assignee: 주식회사 에스엘백시젠
Priority date: 2016-09-28
Filing date: 2016-09-28
Publication date: 2018-04-05
Also published as: WO2018009047A2; TWI697558B; TW201814043A; HK1252391A1; WO2018009047A3; KR101962683B1; CN108419437B; CN108419437A

Abstract

The present invention relates to a DNA vaccine for preventing and treating novel HSV-2 infection, which is a polynucleotide encoding shuffled UL39 protein; to a vector comprising the polynucleotide; and to a DNA vaccine composition comprising the vector.

Description

HSV-2 감염 예방 및 치료용 DNA 백신{A DNA vaccine for preventing and treating HSV-2 infection}[0001] DNA vaccines for preventing and treating HSV-2 infection [0002]

본 발명은 신규 DNA 백신에 관한 것으로서, 보다 구체적으로는 HSV-2(herpes simplex virus-2) 감염 예방 및 치료용 DNA 백신에 관한 것이다. The present invention relates to a novel DNA vaccine, and more particularly, to a DNA vaccine for the prevention and treatment of HSV-2 (herpes simplex virus-2) infection.

HSV-2(herpes simplex virus-2)는 피부 병소를 일으키는 DNA 바이러스인 Herpetoviridae의 구성원으로 전 세계적으로 5억명의 사람들이 HSV-2에 감염되어 있으며, 매년 2천 3백만명의 신규 감염자가 발생하는 것으로 예측 되고 있다. 미국의 경우에는 성인 5명중 1명이 감염되어 있는 것으로 보고되고 있고, BRIC 국가(브라질, 러시아, 인도, 중국)를 포함한 주요 국가에서도 감염률이 높은 편이다. HSV-2는 점막이나 피부의 손상된 부위를 통해 감염되며, 바이러스 복제 과정에서 감염 부위의 괴사 등의 병증을 유발한다. 구체적인 병증으로 HSV-2는 생식기 궤양의 주요 원인이 되는데, 이 궤양은 작은 액체가 채워진 수포 집합체로, 파열되면 아픈 상처가 생기게 된다. 또한 열 발생, 전반적인 아픈 느낌, 근육 통증, 배뇨통증, 질 분비물 발생 등의 증상이 유발 된다. HSV-2는 감각 신경 세포의 축색돌기를 따라 배근신경절(dorsal root ganglia)까지 들어가면서 잠복하며 몇 년 또는 평생 동안 잠복하면서 재활성을 반복하게 된다. HSV-2는 무증상 중에서도 사람간의 전이가 가능하기 때문에 바이러스 확산 통제에 많은 어려움이 있다는 것에 그 심각성이 있다. 실제로 혈청학적으로 HSV-2 양성인 개인의 확산 정도가 1976년부터 1994년까지 30%정도 증가 하였으며, 추가적인 감염 확산을 막지 못할 경우 2025년까지 남성은 39%, 여성은 49%로 감염률이 증가할 것으로 예측된다. 더욱이, HSV-2에 감염된 여성이 출산한 신생아의 60%는 사망하거나 20%는 신경 또는 눈에 심각한 후유증이 발생되고, HSV-2 감염자는 HIV에 감염될 확률이 3배 더 높다는 사실은 전세계적 공중 보건에 심각한 문제가 된다. 또한 경제적 측면에서도 HSV-2 감염은 막대한 부담이 되고 있다. 실례로 HSV-2 감염 치료를 위한 미국의 1996년 연간 직접 의료비용은 2억 8천 3백만 달러에서 9억 8천 4백만 달러로 예측 되었다.HSV-2 (herpes simplex virus-2) is a member of Herpetoviridae, a DNA virus that causes skin lesions, with 500 million people infected with HSV-2 worldwide and 23 million new infections every year Is predicted. In the United States, one in five adults is reported to be infected, and infection rates are high in major countries, including BRIC countries (Brazil, Russia, India, and China). HSV-2 is infected through mucous membranes and damaged areas of the skin, causing virus necrosis and other pathologies in the process of viral replication. As a specific disease, HSV-2 is a major cause of genital ulcers. This ulcer is a small, liquid-filled bovine aggregate that causes sore scars when ruptured. In addition, symptoms such as fever, general sickness, muscle pain, urinary pain, vaginal discharge are caused. HSV-2 encapsulates the dorsal root ganglia along the axons of the sensory neurons and loses latency for several years or lifetime. Since HSV-2 is capable of human-to-human transmission among asymptomatic individuals, there are serious difficulties in controlling the spread of viruses. In fact, serologically, the spread of HSV-2-positive individuals increased by 30% between 1976 and 1994, and by 2025, 39% for males and 49% for females Is predicted. Furthermore, the fact that 60% of newborns born to women infected with HSV-2 die, 20% have a serious aftereffect in the nervous system or eye, and HSV-2 infected people are three times more likely to be infected with HIV It becomes a serious problem in public health. In addition, HSV-2 infection is an enormous burden on the economy. For example, the annual direct medical costs for the US in 1996 for the treatment of HSV-2 infections were estimated at $ 283 million to $ 984 million.

불행히도 현재까지 HSV-2 감염을 근본적으로 치료 할 수 있는 치료제는 없을뿐더러 증상들에 대한 치료 선택은 제한적이다. 바이러스 DNA 합성을 방해해서 바이러스 복제를 억제하는 항바이러스 약제[예를 들어, 팜시클로버(famciclovir), 발라시클로버(valaciclovir), 또는 아시클로버(acyclovir)]를 사용하지만, 이 또한 감염 초기에 적용할 경우 임상 증상을 완화하거나 바이러스 전이를 줄일 수 있을 뿐 재발 및 전파 감염을 방지 할 수 없고 완치가 어렵다. 이를 테면 아시클로비어는 HSV-2 전염을 감소시킬 수 있지만, 뉴런 신경절(ganglion)의 잠복 감염을 예방하지는 못한다. 더욱이 항바이러스 요법은 욕지기, 구토발생, 그리고 신장 기능 감소가 포함된 많은 부작용을 일으킨다. 기존 항바이러스 약제에 대한 미충족 의료 수요로 인해 HSV-2에 대한 새롭고 다양한 백신을 이용한 예방 및 치료방법이 시도되었지만 모두 소용없었다. 비활성화된 바이러스, 감소된 생바이러스, 변형된 생바이러스, 그리고 세포배양물-유도된 소단위(subunit)가 포함된 기존 방식의 백신은 대개 성공하지 못하거나 효과가 매우 낮았다. 최근의 대표적인 3가지 후보 백신, 즉 Chiron사가 개발한 백신, GlaxoSmithKline(GSK)사가 개발한 백신과 Vical사가 개발한 백신에 대한 임상시도는 그 치료효과가 미미하였다. 보조제(adjuvant) MF59와 복합된 HSV-2의 gD와 gB2의 절단형(truncated form)으로 구성된 Chiron사의 백신은 HSV-2에 대한 고역가 항체를 생성하였지만, 그 효과는 일시적이었다. 보조제 alum 및 MPL와 gD를 포함한 GSK의 백신 또한 HSV-2 감염 예방 및 치료에 실패하였다. 마지막으로 보조제 Vaxfectin과 gD 또는 UL46 및 UL47을 포함하는 Vical사의 백신은 동물실험에선 고역가의 항체를 생성하였지만, 임상에서는 HSV-2 감염 치료에서 대조군과 차이를 보이지 않았다. 앞의 사례에서 HSV-2에 대한 고역가 항체 반응이 HSV-2 감염 전파 방지 및 치료에는 충분한 요소가 아님을 알 수 있으며, 이로 인해 HSV-2 감염에 대한 새로운 면역 기작에 바탕을 둔 치료제 개발이 요구되고 있는 실정이다. Unfortunately, until now there is no cure to fundamentally treat HSV-2 infection, and treatment options for symptoms are limited. An antiviral agent (e.g., famciclovir, valaciclovir, or acyclovir) that inhibits viral replication by interfering with viral DNA synthesis is used, but also when applied at the initial stage of infection It can relieve clinical symptoms or reduce virus metastasis and can not prevent recurrence and radiofrequency infection and is difficult to cure. For example, acyclovir may reduce HSV-2 transmission, but it does not prevent latent infection of the ganglion. Moreover, antiviral therapy causes many side effects including nausea, vomiting, and loss of kidney function. Due to unmet medical needs for existing antiviral drugs, new and different vaccines for prevention and treatment of HSV-2 have been tried, but not all. Conventional vaccines containing inactivated viruses, reduced live viruses, modified live viruses, and cell culture-derived subunits were largely unsuccessful or very ineffective. The three most recent candidate vaccines, the vaccine developed by Chiron and the vaccine developed by GlaxoSmithKline (GSK), and the vaccine developed by Vical, were ineffective. The Chiron vaccine, composed of a truncated form of gD and gB2 of HSV-2 conjugated with adjuvant MF59, produced a high-banding antibody to HSV-2, but the effect was transient. The vaccine of GSK, including adjuvant alum and MPL and gD, also failed to prevent and treat HSV-2 infection. Finally, vaccines from Vical, including the adjuvants Vaxfectin and gD or UL46 and UL47, produced high antibody in animal studies, but did not differ clinically from the control in HSV-2 infection treatment. In the previous case, high-level antibody response to HSV-2 is not sufficient for prevention and treatment of HSV-2 infection, and it is necessary to develop a therapeutic agent based on new immune mechanism against HSV-2 infection .

본 발명은 상기 문제점을 포함한 다양한 문제점을 해결하기 위한 것으로서, 보다 효과적으로 HSV-2 감염을 예방하고 치료할 수 있는 새로운 DNA 백신을 제조하는 것을 목적으로 한다. The present invention aims at solving various problems including the above problems, and it is an object of the present invention to produce a novel DNA vaccine which can prevent and treat HSV-2 infection more effectively.

그러나, 본 발명이 상기 목적에 의해 한정되는 것은 아니다.However, the present invention is not limited by the above objects.

본 발명의 다른 일 관점에 따르면, HSV-2의 UL39 단백질의 아미노산 서열상 14-154 a.a 위치에 상응하는 펩타이드에서 78-104 a.a 위치에 상응하는 막통과 도메인이 결실된 UL39-N1 펩타이드, 상기 UL39 단백질의 아미노산 서열상 1117-1142 a.a 위치에 상응하는 UL39-C2 펩타이드, 상기 UL39 단백질의 아미노산 서열상 155-227 a.a 위치에 상응하는 UL39-N2 펩타이드, 상기 UL39 단백질의 아미노산 서열상 399-1116 a.a 위치에 상응하는 UL39 N4-C1 펩타이드, 및 상기 UL39 단백질의 아미노산 서열상 208-398 a.a 위치에 상응하는 UL39-N3 펩타이드가 무작위적으로 섞여있고, 원래의 UL39 단백질의 아미노산 서열은 포함하지 않는 셔플드(shuffled) UL39 단백질을 암호화하는 폴리뉴클레오타이드가 제공된다. According to another aspect of the present invention, a UL39-N1 peptide lacking the transmembrane domain corresponding to the 78-104 aa position in the peptide corresponding to the 14-154 aa position on the amino acid sequence of the UL39 protein of HSV-2, A UL39-C2 peptide corresponding to the amino acid sequence position 1117-1142 aa of the protein, a UL39-N2 peptide corresponding to the amino acid sequence position 155-227 aa position of the UL39 protein, an amino acid sequence 399-1116 aa position of the UL39 protein And a UL39-N3 peptide corresponding to the position 208-398 aa on the amino acid sequence of the UL39 protein are randomly mixed and the amino acid sequence of the original UL39 protein is not included in the shuffled ( polynucleotide encoding the shuffled UL39 protein is provided.

본 발명의 다른 일 관점에 따르면, 상기 폴리뉴클레오타이드를 포함하는 벡터가 제공된다.According to another aspect of the present invention, there is provided a vector comprising the polynucleotide.

본 발명의 다른 일 관점에 따르면, 상기 벡터를 포함하는 단리된 숙주세포가 제공된다.According to another aspect of the present invention, there is provided an isolated host cell comprising said vector.

본 발명의 다른 일 관점에 따르면, 상기 폴리뉴클레오타이드 또는 상기 벡터 및 약학적으로 허용가능한 담체를 포함하는 조성물이 제공된다.According to another aspect of the present invention, there is provided a composition comprising the polynucleotide or the vector and a pharmaceutically acceptable carrier.

본 발명의 다른 일 관점에 따르면, 상기 폴리뉴클레오타이드에 의해 암호화되는 재조합 단백질이 제공된다.According to another aspect of the present invention, there is provided a recombinant protein encoded by the polynucleotide.

본 발명의 다른 일 관점에 따르면, gB, gD, UL39, ICP0 및 ICP4 단백질로 구성되는 군으로부터 선택되는 2종 이상의 또는 모든 HSV-2 항원 단백질을 발현하도록 상기 항원 단백질을 암호화하는 폴리뉴클레오타이드를 포함하는 발현벡터가 제공된다.According to another aspect of the present invention there is provided a kit for the treatment of a subject comprising a polynucleotide encoding said antigen protein to express two or more or all of the HSV-2 antigen proteins selected from the group consisting of gB, gD, UL39, ICPO and ICP4 proteins An expression vector is provided.

본 발명의 다른 일 관점에 따르면, 상기 폴리뉴클레오타이드, 상기 벡터 또는 상술한 발현벡터를 포함하는 DNA 백신 조성물이 제공된다.According to another aspect of the present invention, there is provided a DNA vaccine composition comprising the polynucleotide, the vector, or the expression vector described above.

도 1은 본 발명의 일 실시예에 따른 셔플드 UL39 단백질의 구조를 나타내는 개요도이다.
도 2는 본 발명의 일 실시예에 따른 셔플드 UL39 단백질을 발현하도록 고안된 플라스미드 DNA를 이용한 동물실험 순서를 개략적으로 나타낸 개요도이다.
도 3은 본 발명의 일 실시예에 따른 셔플드 UL39 단백질을 발현하도록 고안된 플라스미드 DNA 및 음성대조군인 mock plasmid DNA를 실험동물에 투여하고 HSV-2를 감염시킨 동물의 생존율을 기록한 그래프이다.
도 4는 본 발명의 일 실시예에 따른 tPA-Flt3L-gB-UL39 plasmid DNA 및 tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA를 통해 발현되는 융합단백질의 구조를 각각 나타낸 개요도이다.
도 5는 본 발명의 일 실시예에 따른 상기 tPA-Flt3L-gB-UL39 plasmid DNA 및 tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA를 각각 투여한 그룹 및 tPA-Flt3L-gB-UL39 plasmid DNA 및 tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA를 병용투여한 그룹 그리고 음성대조군으로 mock plasmid DNA를 투여한 그룹의 HSV-2 감염 실험 스케쥴을 나타낸 개요도이다.
도 6은 본 발명의 일 실시예에 따른 상기 tPA-Flt3L-gB-UL39 plasmid DNA 및 tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA를 각각 투여한 그룹 및 tPA-Flt3L-gB-UL39 plasmid DNA 및 tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA를 병용투여한 그룹 그리고 음성대조군으로 mock plasmid DNA를 투여한 그룹의 HSV-2 감염 실험결과를 나타내는 것으로서 생존율(a) 및 병리학 점수(b)를 각각 나타내는 그래프이다.1 is a schematic diagram showing the structure of a shuffled UL39 protein according to an embodiment of the present invention.
FIG. 2 is a schematic diagram showing a procedure of an animal experiment using a plasmid DNA designed to express a shuffled UL39 protein according to an embodiment of the present invention.
FIG. 3 is a graph showing survival rates of animals infected with HSV-2 by administering plasmid DNA designed to express shuffled UL39 protein and mock plasmid DNA as a negative control according to an embodiment of the present invention.
FIG. 4 is a schematic diagram showing the structure of a fusion protein expressed through plasmid DNA tPA-Flt3L-gB-UL39 and tPA-Flt3L-gD-ICP0-ICP4 according to an embodiment of the present invention.
FIG. 5 is a graph showing a comparison between the tPA-Flt3L-gB-UL39 plasmid DNA and tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA and tPA- -Flt3L-gD-ICP0-ICP4 plasmid DNA and mock plasmid DNA as a negative control group.
FIG. 6 is a graph showing a comparison between the tPA-Flt3L-gB-UL39 plasmid DNA and tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA and tPA- (A) and pathology score (b) showing the result of HSV-2 infection test in a group in which plasmid DNA of -Flt3L-gD-ICP0-ICP4 was coadministered and mock plasmid DNA was administered as a negative control group to be.

본 발명의 다른 일 관점에 따르면, HSV-2의 UL39 단백질의 아미노산 서열상 14-154 a.a 위치에 상응하는 펩타이드에서 78-104 a.a 위치에 상응하는 막통과 도메인이 결실된 UL39-N1 펩타이드, 상기 UL39 단백질의 아미노산 서열상 1117-1142 a.a 위치에 상응하는 UL39-C2 펩타이드, 상기 UL39 단백질의 아미노산 서열상 155-227 a.a 위치에 상응하는 UL39-N2 펩타이드, 상기 UL39 단백질의 아미노산 서열상 399-1116 a.a 위치에 상응하는 UL39 N4-C1 펩타이드, 및 상기 UL39 단백질의 아미노산 서열상 208-398 a.a 위치에 상응하는 UL39-N3 펩타이드가 무작위적으로 섞여있고, 원래의 UL39 단백질의 아미노산 서열은 포함하지 않는 셔플드 UL39 단백질을 암호화하는 폴리뉴클레오타이드가 제공된다. According to another aspect of the present invention, a UL39-N1 peptide lacking the transmembrane domain corresponding to the 78-104 aa position in the peptide corresponding to the 14-154 aa position on the amino acid sequence of the UL39 protein of HSV-2, A UL39-C2 peptide corresponding to the amino acid sequence position 1117-1142 aa of the protein, a UL39-N2 peptide corresponding to the amino acid sequence position 155-227 aa position of the UL39 protein, an amino acid sequence 399-1116 aa position of the UL39 protein And the UL39-N3 peptide corresponding to the 208-398 aa position on the amino acid sequence of the UL39 protein are randomly mixed and the amino acid sequence of the original UL39 protein is not included. A polynucleotide encoding the protein is provided.

본 문서에서 사용되는 용어 "셔플드(shuffled)"는 특정 단백질 내에 복수의 도메인(domain)이 존재할 경우 이들 도메인의 순서를 뒤섞는 것을 의미한다. 특히, 본 문서에서 사용되는 용어 "셔플드 단백질"은 다른 의미로는 "에피토프 셔플드 단백질"로서 면역 시스템에 의해 인지되는 복수의 에피토프의 활성은 그대로 유지하면서 에피토프의 순서가 무작위로 뒤섞인 재조합 단백질로서 에피토프의 활성은 그대로 유지하되 원래의 단백질의 기능은 상실한 면역원성 단백질이다.The term "shuffled " as used herein means to shuffle the order of these domains when there are multiple domains within a particular protein. In particular, the term "shuffled protein ", as used herein, refers to an epitope shuffled protein, a recombinant protein in which sequences of epitopes are randomly interspersed while maintaining the activity of multiple epitopes recognized by the immune system It is an immunogenic protein that retains the activity of the epitope, but the function of the original protein is lost.

상기 셔플드 UL39 단백질의 제조에 사용되는 UL39 단백질은 표준서열로서 UniProt 등록번호 P89462로 등록된 RIR1_HHV2H가 사용될 수 있으며, 상기 아미노산 위치는 상기 표준 서열을 기준으로 한 것으로서, UL39 단백질의 기능을 보유하고 있는 다른 다양한 변이체, 예컨대, UniProt 등록번호 G91261, A0A0E3Y5Z7, A0A0E3Y7N5, A0A120I2I0, A0A110B8A6, A0A0E3Y758 등 다양한 변이체가 사용되더라도 무방하며, 이들 변이체 중 상기 표준서열과 길이가 상이한 경우에는 상기 표준서열의 위치에 상응하는 위치가 적용될 것이다.The UL39 protein used for the production of the shuffled UL39 protein may be RIR1_HHV2H registered as UniProt registration number P89462 as a standard sequence, and the amino acid position is based on the above-mentioned standard sequence. A variety of mutants such as UniProt registration number G91261, A0A0E3Y5Z7, A0A0E3Y7N5, A0A120I2I0, A0A110B8A6, A0A0E3Y758 and the like may be used, and when the length of the mutant is different from the standard sequence, a position corresponding to the position of the standard sequence Will be applied.

상기 폴리뉴클레오타이드는 하나 또는 둘 이상의 면역 증진 펩타이드를 암호화하는 폴리뉴클레오타이드를 추가로 포함할 수 있고, 상기 면역 증진 펩타이드는 CD28, ICOS(inducible costimulator), CTLA4(cytotoxic T lymphocyte associated protein 4), PD1(programmed cell death protein 1), BTLA(B and T lymphocyte associated protein), DR3(death receptor 3), 4-1BB, CD2, CD40, CD30, CD27, SLAM(signaling lymphocyte activation molecule), 2B4(CD244), NKG2D(natural-killer group 2, member D)/DAP12(DNAX-activating protein 12), TIM1(T-Cell immunoglobulin and mucin domain containing protein 1), TIM2, TIM3, TIGIT, CD226, CD160, LAG3(lymphocyte activation gene 3), B7-1, B7-H1, GITR(glucocorticoid-induced TNFR family related protein), Flt3 리간드(fms-like tyrosine kinase 3 ligand), 플라젤린(flagellin), HVEM(herpesvirus entry mediator), CD40 L(리간드) 또는 OX40L[ligand for CD134(OX40), CD252]의 세포질 도메인 또는 이들 중 둘 이상의 연결체일 수 있다.The polynucleotide may further comprise a polynucleotide encoding one or more immunity enhancing peptides, wherein the immunity enhancing peptide is selected from the group consisting of CD28, inducible costimulator (ICOS), cytotoxic T lymphocyte associated protein 4 (CTLA4) cell death protein 1), BTLA (B and T lymphocyte associated protein), DR3 (death receptor 3), 4-1BB, CD2, CD40, CD30, CD27, SLAM signaling lymphocyte activation molecule 2B4 CD244, NKG2D (DNAX-activating protein 12), TIM1 (T-cell immunoglobulin and mucin domain containing protein 1), TIM2, TIM3, TIGIT, CD226, CD160, LAG3 (lymphocyte activation gene 3) , B7-1, B7-H1, glucocorticoid-induced TNFR family related protein, fms-like tyrosine kinase 3 ligand, flagellin, herpesvirus entry mediator, CD40 L (ligand) Or the cytoplasmic domain of OX40L [ligand for CD134 (OX40), CD252], or both Of cheil can connect.

본 문서에서 사용되는 용어 "면역 증진 펩타이드"는 면역 반응에 관계하는 세포(예를 들어, 수지상 세포 등)를 활성화시켜 면역 반응을 증가시키는 펩타이드를 의미한다.As used herein, the term " immunopotentiating peptide "refers to a peptide that activates cells involved in an immune response (e. G., Dendritic cells, etc.) and thereby increases the immune response.

상기 폴리뉴클레오타이드는 분비 신호서열을 암호화하는 폴리뉴클레오타이드를 추가로 포함할 수 있으며, 상기 분비 신호서열은 상기 셔플드 UL39 단백질의 세포 밖으로의 분비를 유도하며, tPA(tissue plasminogen activator) 신호서열, HSV gDs(단순포진 바이러스 당단백질 Ds) 신호서열 또는 성장호르몬 신호서열일 수 있다.The polynucleotide may further comprise a polynucleotide encoding a secretion signal sequence, wherein the secretory signal sequence induces secretion of the shuffled UL39 protein out of the cell, and comprises a tPA (tissue plasminogen activator) signal sequence, HSV gDs (Herpes simplex virus glycoprotein Ds) signal sequence or a growth hormone signal sequence.

상기 폴리뉴클레오타이드는 하나 또는 둘 이상의 제2형 단순포진 바이러스(HSV-2)의 항원 단백질을 암호화하는 폴리뉴클레오타이드를 추가로 포함할 수 있다. 상기 항원 단백질은 당단백질 B(gB), 당단백질 D(gD), ICP0 또는 ICP4일 수 있다.The polynucleotide may further comprise a polynucleotide encoding one or more antigenic proteins of type 2 herpes simplex virus (HSV-2). The antigen protein may be glycoprotein B (gB), glycoprotein D (gD), ICPO or ICP4.

상기 폴리뉴클레오타이드에 있어서, 상기 당단백질 B는 1-22 a.a 위치에 상응하는 신호서열 및 772-792 a.a 위치에 상응하는 막통과 도메인이 결실된 것일 수 있고, 상기 당단백질 D는 1-25 a.a 위치에 상응하는 신호서열 및 341-364 a.a 위치에 상응하는 막통과 도메인이 결실된 것일 수 있으며, 상기 ICP0은 510-516 a.a 위치에 상응하는 핵위치화 신호(NLS)가 결실된 것일 수 있고 상기 IPC4는 767-1318 a.a 위치에 상응하는 RS1.3 부위가 결실된 것일 수 있다.In the polynucleotide, the glycoprotein B may have a signal sequence corresponding to the 1-22 aa position and a transmembrane domain corresponding to the 772-792 aa position deleted, and the glycoprotein D may have a 1-25 aa position And the transmembrane domain corresponding to position 341-364 aa may have been deleted, the ICPO may be the deletion of the nuclear localization signal (NLS) corresponding to positions 510-516 aa, and the IPC4 May be the deletion of the RS1.3 site corresponding to position 767-1318 aa.

상기 항원 단백질들 역시 표준서열로 gB의 경우 UniProt 등록번호 P08666인 GB_HHV2H, gD의 경우 UniProt 등록번호 Q69467인 GD_HHV2H, ICP0의 경우 UniProt 등록번호 P28284인 ICP0_HHV2H, ICP4의 경우 UniProt 등록번호 P90493인 ICP4_HHV2H를 사용하였으며, 상기 항원 단백질들의 사용 아미노산 위치 역시 상기 표준서열을 기준으로 표시된 것으로, 상기 항원 단백질들의 활성을 유지하고 있는 다양한 변이체 역시 사용이 가능하며, 이들 변이체의 길이가 상기 표준서열과 상이할 경우에는 상기 표준서열의 아미노산 위치에 상응하는 아미노산 위치가 적용될 것이다. 상기와 같은 다양한 변이체의 예로는 gB의 경우 UniProt 등록번호 P06763, Q69465, D6QV12, 및 D6QV07 등의 변이체가 사용될 수 있고, gD의 경우 UniProt 등록번호 P03172, T1PZZ0, A0A0Y0QWV3, A0A110AVP3, 및 A0A0Y0RM80 등이 사용될 수 있으며, ICP0의 경우 UniProt 등록번호 G9I221, G9I280, W0NW81, D6PUZ6, 및 D6PUZ3 등이 사용될 수 있고, ICP4의 경우 UniProt 등록번호 G9I282, A0A0E3Y5F0, A0A0Y0RBE8, A0A120I2I2, 및 A0A0Y0RAK6 등이 사용될 수 있다.These antigenic proteins were also used as the standard sequence, GB_HHV2H which is the UniProt registration number P08666 for GB, GD_HHV2H for UniProt registration number Q69467 for GD, ICP0_HHV2H for UniProt registration number P28284 for ICP0 and ICP4_HHV2H for Uniprot registration number P90493 for ICP4 , The amino acid positions of the antigenic proteins are also indicated on the basis of the standard sequence, and various mutants which maintain the activity of the antigenic proteins are also usable. When the length of these mutants is different from the standard sequence, The amino acid position corresponding to the amino acid position of the sequence will be applied. Examples of various mutants include giA, giA, giA, giA, giA, giA, giA, giA, giA, giA, giA, giA, giA, giA, giA, giA, In the case of ICP0, UniProt registration numbers G9I282, A0A0E3Y5F0, A0A0Y0RBE8, A0A120I2I2, and A0A0Y0RAK6 can be used for the ICP4, and the UniProt registration numbers G9I221, G9I280, W0NW81, D6PUZ6 and D6PUZ3 can be used.

또한, 상기 폴리뉴클레오타이드는 숙주세포에서 발현 빈도가 높은 코돈으로 치환될 수 있다. 본 발명에서 사용된 "숙주세포에서 발현 빈도가 높은 코돈으로 치환된 것" 또는 "최적화된 코돈"이란 숙주세포 내에서 DNA가 단백질로 전사(transcription) 및 번역(translation)될 때 아미노산을 지령하는 코돈 사이에 숙주에 따라 선호도가 높은 코돈이 존재하는데, 이들 선호도가 높은 코돈으로 치환함으로써 그 핵산이 암호화하는 아미노산 또는 단백질의 발현 효율을 증가시키는 것을 말한다.In addition, the polynucleotide may be replaced with a codon having a high expression frequency in a host cell. Quot; or "optimized codon" as used herein refers to a codon that directs an amino acid when the DNA is transcribed and translated into a protein in a host cell. There is a codon having a high preference according to a host, and the expression efficiency of the amino acid or protein encoded by the nucleic acid is increased by replacing the codon with the higher preference codon.

상기 폴리뉴클레오타이드에 있어서, 상기 UL39-N1 펩타이드는 서열번호 1로 구성되고, 상기 UL39-C2 펩타이드는 서열번호 2로 구성되며, 상기 UL39-N2 펩타이드는 서열번호 3으로 구성되며, 상기 UL39 N4-C1 펩타이드는 서열번호 4로 구성되고, 및 상기 UL39-N3 펩타이드는 서열번호 5로 구성될 수 있다. 아울러 상기 셔플드 UL39는 서열번호 1로 표시되는 UL-39-N1 펩타이드, 서열번호 2로 표시되는 UL39-C2 펩타이드, 서열번호 3으로 표시되는 UL39-N2 펩타이드, 서열번호 4로 표시되는 UL39 N4-C1 펩타이드, 및 서열번호 5로 표시되는 UL39-N3 펩타이드를 순차적으로 포함할 수 있으며, 이 경우 상기 셔플드 UL39 단백질은 서열번호 6으로 표시되는 아미노산 서열로 구성될 수 있다. 그러나, 상기 각각의 펩타이드들의 순서는 원래의 전장 UL39 단백질의 아미노산 서열과 동일하지 않는 한 바뀌어도 무방하다.In the polynucleotide, the UL39-N1 peptide comprises SEQ ID NO: 1, the UL39-C2 peptide comprises SEQ ID NO: 2, the UL39-N2 peptide comprises SEQ ID NO: 3, The peptide is composed of SEQ ID NO: 4, and the UL39-N3 peptide is composed of SEQ ID NO: 5. The shuffled UL39 is a peptide consisting of the UL-39-N1 peptide represented by SEQ ID NO: 1, the UL39-C2 peptide represented by SEQ ID NO: 2, the UL39-N2 peptide represented by SEQ ID NO: 3, the UL39 N4- C1 peptide, and the UL39-N3 peptide represented by SEQ ID NO: 5, wherein the shuffled UL39 protein may be an amino acid sequence represented by SEQ ID NO: 6. However, the sequence of each of the peptides may be changed as long as it is not identical to the amino acid sequence of the original full length UL39 protein.

본 발명의 일 실시예에 따른 상기 벡터는 상기 폴리뉴클레오타이드가 조절서열에 작동가능하게 연결된 유전자 컨스트럭트를 포함하여 숙주세포에서 상기 셔플드 UL39 단백질을 발현하도록 할 수 있는 발현벡터일 수 있으며, 상기 발현벡터는 플라스미드 벡터, 바이러스 벡터, 코스미드 벡터, 파지미드 벡터, 인공 인간 염색체 등 그 어떠한 형태를 나타내더라도 무방하다.The vector according to an embodiment of the present invention may be an expression vector capable of expressing the shuffled UL39 protein in a host cell, wherein the polynucleotide comprises a gene construct operably linked to the regulatory sequence, The expression vector may be any of a plasmid vector, a viral vector, a cosmid vector, a phagemid vector, an artificial human chromosome, and the like.

"작동 가능하게 연결된(operably linked to)"이란 목적으로 하는 핵산서열(예컨대, 시험관내 전사/번역 시스템에서 또는 숙주세포에서)이 그의 발현이 이루어질 수 있도록 하는 방식으로 상기 조절서열에 연결되어 있다는 것을 의미한다.It is contemplated that a nucleic acid sequence that is "operably linked to" (eg, in an in vitro transcription / translation system or in a host cell) is linked to the regulatory sequence in such a way that its expression can be achieved it means.

상기 "조절서열"이란 용어는 프로모터, 인핸서 및 다른 조절 요소(예, 폴리아데닐화 신호)를 포함하는 의미이다. 조절서열에는 많은 숙주세포에서 목적으로 하는 핵산이 항상적으로 발현될 수 있도록 지시하는 것, 특정한 조직세포에서만 목적으로 하는 핵산이 발현될 수 있도록 지시하는 것(예, 조직특이적 조절서열), 그리고 특정 신호에 의해 발현이 유도되도록 지시하는 것(예, 유도성 조절서열)이 포함된다. 발현벡터의 설계는 형질전환될 숙주세포의 선택 및 원하는 단백질 발현의 수준 등과 같은 인자에 따라 달라질 수 있다는 것은 당업자라면 이해할 수 있다. 본 발명의 발현벡터는 숙주 세포에 도입되어 상기 융합 단백질을 발현할 수 있다. 상기 진핵세포 및 원핵세포에서 발현을 가능하게 하는 조절서열들은 당업자에게 잘 알려져 있다. 상술한 바와 같이, 이들은 보통 전사개시를 담당하는 조절서열들 및, 선택적으로 전사물의 전사종결 및 안정화를 담당하는 폴리-A 신호를 포함한다. 추가적인 조절서열들은 전사조절인자 외에도 번역 증진인자 및/또는 천연-조합 또는 이종성 프로모터 영역을 포함할 수 있다. 예를 들어 포유류 숙주 세포에서 발현을 가능하게 하는 가능한 조절서열들은 CMV-HSV 티미딘 키나아제 프로모터, SV40, RSV-프로모터(로우스 육종 바이러스), 인간 신장 요소 1α-프로모터, 글루코코르티코이드-유도성 MMTV-프로모터(몰로니 마우스 종양 바이러스), 메탈로티오네인-유도성 또는 테트라사이클린-유도성 프로모터 또는, CMV 증폭제 또는 SV40-증폭제와 같은 증폭제를 포함한다. 신경 세포 내 발현을 위해, 신경미세섬유-프로모터(neurofilament-promoter), PGDF-프로모터, NSE-프로모터, PrP-프로모터 또는 thy-1-프로모터들이 사용될 수 있다는 것이 고려되고 있다. 상기 프로모터들은 당 분야에 알려져 있으며, 문헌(Charron, J. Biol. Chem. 1995, 270: 25739-25745)에 기술되어 있다. 원핵세포내 발현을 위해, lac-프로모터, tac-프로모터 또는 trp 프로모터를 포함하는 다수의 프로모터들이 개시되어 있다. 전사를 개시할 수 있는 인자들 외에, 상기 조절서열들은 본 발명의 일 실시예에 따른 폴리뉴클레오타이드의 하류(downstream)에 SV40-폴리-A 부위 또는 TK-폴리-A 부위와 같은 전사 종결 신호를 포함할 수도 있다. 본 문서에서, 적당한 발현 벡터들은 당 분야에 알려져 있으며, 그 예로는 오카야마-베르그(Okayama-Berg) cDNA 발현 벡터 pcDV1(Parmacia), pRc/CMV, pcDNA1, pcDNA3(In-vitrogene), pSPORT1(GIBCO BRL), pGX27(특허 제1442254호), pX(Pagano (1992) Science 255, 1144-1147), 효모 2-혼성(two-hybrid) 벡터, 가령 pEG202 및 dpJG4-5(Gyuris (1995) Cell 75, 791-803) 또는 원핵 발현 벡터, 가령 람다 gt11 또는 pGEX(Amersham-Pharmacia)가 있다. 본 발명의 핵산 분자들 외에, 벡터는 분비 신호를 암호화하는 폴리뉴클레오타이드를 추가로 포함할 수 있다. 상기 분비신호들은 당업자에게 잘 알려져 있다. 그리고, 사용된 발현 시스템에 따라, 융합단백질을 세포 구획으로 이끌 수 있는 리더서열(leader sequence)이 본 발명의 일 실시예에 따른 폴리뉴클레오타이드의 코딩 서열에 조합되며, 바람직하게는 해독된 단백질 또는 이의 단백질을 세포질 주변 또는 세포외 매질로 직접 분비할 수 있는 리더 서열이다. The term "regulatory sequence" is intended to include promoters, enhancers and other regulatory elements (e.g., polyadenylation signals). Regulatory sequences include directing the expression of the nucleic acid of interest in many host cells at all times, directing the expression of the nucleic acid of interest in a particular tissue cell (e.g., a tissue-specific regulatory sequence), and And directing expression to be induced by a particular signal (e.g., inducible regulatory sequences). It will be appreciated by those skilled in the art that the design of the expression vector may depend on factors such as the choice of host cell to be transformed and the level of protein expression desired. The expression vector of the present invention may be introduced into a host cell to express the fusion protein. Regulatory sequences enabling expression in the eukaryotic and prokaryotic cells are well known to those skilled in the art. As discussed above, these usually include regulatory sequences responsible for transcription initiation and, optionally, poly-A signals responsible for transcription termination and stabilization of transcripts. Additional regulatory sequences may include translation enhancing factors and / or native-combining or heterologous promoter regions in addition to transcriptional regulatory factors. For example, possible regulatory sequences enabling expression in mammalian host cells include the CMV-HSV thymidine kinase promoter, SV40, the RSV-promoter (low-grade sarcoma virus), the human renal element 1 alpha-promoter, the glucocorticoid- A promoter (Moloney murine tumor virus), a metallothionein-inducible or tetracycline-inducible promoter, or an amplifying agent such as a CMV or SV40-amplification agent. For neuronal expression, it is contemplated that neurofilament-promoter, PGDF-promoter, NSE-promoter, PrP-promoter or thy-1-promoter may be used. Such promoters are known in the art and described in the literature (Charron, J. Biol. Chem. 1995, 270: 25739-25745). For prokaryotic expression, a number of promoters have been disclosed, including lac-promoter, tac-promoter or trp promoter. In addition to factors capable of initiating transcription, the control sequences include a transcription termination signal such as the SV40-poly-A site or the TK-poly-A site downstream of the polynucleotide according to an embodiment of the invention You may. In this document, suitable expression vectors are known in the art, including Okayama-Berg cDNA expression vectors pcDV1 (Parmacia), pRc / CMV, pcDNA1, pcDNA3 (In-vitrogene), pSPORT1 (GIBCO BRL pGX27 (Patent No. 1442254), pX (Pagano (1992) Science 255, 1144-1147), yeast two-hybrid vectors such as pEG202 and dpJG4-5 (Gyuris (1995) Cell 75, 791 -803) or a prokaryotic expression vector, such as lambda gt11 or pGEX (Amersham-Pharmacia). In addition to the nucleic acid molecules of the present invention, the vector may further comprise a polynucleotide encoding a secretion signal. Such secretion signals are well known to those skilled in the art. And, depending on the expression system used, a leader sequence capable of directing the fusion protein to the cell compartment is combined with the coding sequence of the polynucleotide according to one embodiment of the present invention, It is a leader sequence that can secrete proteins directly into the cytoplasmic or extracellular medium.

또한, 본 발명의 벡터는 예를 들면, 표준 재조합 DNA 기술에 의하여 제조될 수 있으며, 표준 재조합 DNA 기술에는 예를 들면, 평활말단 및 접착말단 라이게이션, 적절한 말단을 제공하기 위한 제한 효소 처리, 부적합한 결합을 방지하기 위하여 알칼리 포스테이즈 처리에 의한 인산기 제거 및 T4 DNA 라이게이즈에 의한 효소적 연결 등이 포함된다. 화학적 합성 또는 유전자 재조합 기술에 의하여 얻어진 신호 펩타이드를 코딩하는 DNA, 본 발명의 HSV-2 항원 단백질을 암호화하는 DNA를 적절한 조절서열이 포함되어 있는 벡터에 재조합함으로써 본 발명의 벡터가 제조될 수 있다. 상기 조절 서열이 포함되어 있는 벡터는 상업적으로 구입 또는 제조할 수 있으며, 본 발명의 일 실시예에서는 DNA 백신 제조용 벡터인 pGX27을 제조하여 사용하였다.In addition, vectors of the present invention can be prepared, for example, by standard recombinant DNA techniques, and standard recombinant DNA techniques include, for example, blunt end and adhesive end ligation, restriction enzyme treatment to provide appropriate ends, Removal of phosphates by alkaline phosphatase treatment and enzymatic linkage by T4 DNA ligation to prevent binding. The vector of the present invention can be prepared by recombining a DNA encoding a signal peptide obtained by chemical synthesis or a recombinant technology, a DNA encoding the HSV-2 antigen protein of the present invention, and a vector containing an appropriate regulatory sequence. The vector containing the regulatory sequence may be purchased or manufactured commercially. In one embodiment of the present invention, pGX27, a vector for preparing a DNA vaccine, was prepared and used.

본 발명에서 용어, "융합단백질(fusion protein)"이란 둘 이상의 단백질 또는 단백질 내 특정 기능을 담당하는 도메인이 각각의 단백질 또는 도메인이 본연의 기능을 담당하도록 연결된 재조합 단백질(recombinant protein)을 의미한다. 상기 둘 이상의 단백질 또는 도메인 사이에는 통상적으로 유연한 구조를 갖는 링커(linker)가 삽입될 수 있다. 이러한 링커로는 GS₄ 등 다양한 종류의 링커가 알려져 있다.In the present invention, the term "fusion protein" refers to a recombinant protein in which two or more proteins or domains having a specific function in the protein are linked so that each protein or domain functions as a primary function. A linker having a generally flexible structure may be inserted between the two or more proteins or domains. Such linkers are known a wide variety of linkers, such as ₄ GS.

본 문서에서 사용되는 용어 "숙주세포"란 원핵 또는 진핵 세포를 포함하며, 진핵세포는 진균, 효모 등을 포함하는 하등 진핵 세포뿐만 아니라 포유동물 등을 포함하는 고등 진핵 세포를 포함한다.As used herein, the term "host cell" includes prokaryotic or eukaryotic cells, and eukaryotic cells include higher eukaryotic cells, including mammals, as well as lower eukaryotic cells including fungi, yeast and the like.

본 발명의 일 실시예에 따른 벡터로 형질감염 또는 형질전환된 숙주세포 또는 비-인간 숙주개체는 상기 벡터에 의해 유전적으로 변형된 숙주세포 또는 숙주개체일 수 있다. 본 문서에서 사용되는 용어 "유전적으로 변형된(genetically modified)"이라는 용어는 숙주세포 또는 숙주개체, 또는 선행종(predecessors)/모종(parents) 중 하나로 도입된 본 발명의 일 실시예에 따른 폴리뉴클레오타이드 또는 벡터를 숙주세포 또는 숙주개체가 자신의 게놈 외에 포함하는 것을 의미한다. 아울러, 본 발명의 일 실시예에 따른 폴리뉴클레오타이드 또는 벡터는 게놈 외부의 독립적 분자, 바람직하게는 복제할 수 있는 분자로서 유전적으로 변형된 숙주세포 또는 숙주개체 내에 존재할 수 있거나, 또는 숙주세포 또는 숙주개체의 게놈으로 안정적으로 삽입될 수 있다.A host cell or non-human host host transfected or transformed with a vector according to an embodiment of the present invention may be a host cell or host host genetically modified by said vector. The term "genetically modified " as used herein refers to a polynucleotide according to one embodiment of the present invention that is introduced into either a host cell or host entity, or predecessors / parents. Or vector is included in the host cell or host entity other than its own genome. In addition, a polynucleotide or vector according to one embodiment of the present invention may be present in a genetically modified host cell or host entity as an independent molecule outside the genome, preferably as a replicable molecule, or may be a host cell or host entity Lt; RTI ID = 0.0 > genomic < / RTI >

본 발명의 일 실시예에 따른 숙주세포는 원핵 또는 진핵세포이다. 적당한 원핵세포들은 대장균(E. coli) 또는 고초균(Bacillus subtilis)와 같이 클로닝에 보통 사용되는 세포들이다. 그리고, 진핵세포들은 진균세포(fungus), 식물세포 또는 동물세포들을 포함한다. 적당한 진균세포들의 예로는 효모, 바람직하게는 사카로마이세스 속(Saccharomyces sp.) 중의 효모 및 가장 바람직하게는 사카로마이세스 세레비지애(S. cerevisiae)일 수 있다. 적당한 동물세포들의 예로는 곤충세포들, 식물세포들, 바람직하게는 포유류 세포들, 예컨대 HEK293, 293T, NSO, CHO, MDCK, U2-OSHela, NIH3T3, MOLT-4, Jurkat, PC-12, PC-3, IMR, NT2N, Sk-n-sh, CaSki, C33A를 들 수 있다. 상기 숙주세포들, 가령 CHO 세포는 리더 펩타이드 제거, H(중)쇄 및 L(경)쇄의 중첩 및 조립, 정확한 위치에서의 분자의 당화(glycosylation) 및 기능 분자의 분비를 포함하는, 본 발명의 일 실시예에 따른 항체분자에 대한 번역후 수식(post-translational modification)을 제공할 수 있다. 또한, 당 분야에 알려져 있는 적당한 세포주들은 American Type Culture Collection(ATCC)와 같은 세포주 기탁물로부터 수득할 수 있다. 본 발명에 따라, 1차배양 세포(primary culture cell)/세포 배양액들은 숙주세포들로서 기능할 수 있다고 고려되고 있다. 상기 세포들은 특히 곤충들(초파리 또는 Blatta 속의 곤충) 또는 포유류들(인간, 돼지, 마우스 또는 쥐)로부터 유도된다. 상술한 바와 같이 상기 1차배양 세포는 대식세포(macrophage), 단핵구(monocyte), 과립구(granulocyte), 조혈모 줄기세포, 림포카인 활성화 킬러세포, gd 세포, NKT 세포(natural killer T cell), T 세포 또는 NK 세포(natural killer cell)와 같은 면역세포일 수 있다.A host cell according to one embodiment of the present invention is prokaryotic or eukaryotic. Suitable prokaryotic cells are those cells commonly used for cloning, such as E. coli or Bacillus subtilis . And eukaryotic cells include fungal cells, plant cells or animal cells. Examples of suitable fungal cells are yeast, preferably yeast in the genus Saccharomyces sp., And most preferably S. cerevisiae . Examples of suitable animal cells include insect cells, plant cells, preferably mammalian cells such as HEK293, 293T, NSO, CHO, MDCK, U2-OSHela, NIH3T3, MOLT-4, Jurkat, PC- 3, IMR, NT2N, Sk-n-sh, CaSki, and C33A. The host cells, such as CHO cells, can be used in the present invention, including leader peptide removal, superposition and assembly of the H (heavy) and L (light) chains, glycosylation of the molecule at the correct location, Translational modification of the antibody molecule according to one embodiment of the invention. In addition, suitable cell lines known in the art can be obtained from cell line deposits such as the American Type Culture Collection (ATCC). According to the present invention, primary culture cells / cell culture fluids are considered to be able to function as host cells. The cells are derived from insects (insects of the genus Drosophila or Blatta ) or mammals (humans, pigs, mice or rats). As described above, the primary cultured cells include macrophages, monocytes, granulocytes, hematopoietic stem cells, lymphokine activated killer cells, gd cells, NKT cells (natural killer T cells) T cells or NK cells (natural killer cells).

상기 조성물은 상기 담체 외에 약학적으로 허용가능한 보조제, 부형제 또는 희석제를 추가적으로 포함할 수 있다.The composition may additionally contain, in addition to the carrier, a pharmaceutically acceptable adjuvant, excipient or diluent.

본 문서에서 사용되는 용어 "약학적으로 허용가능한"이란 생리학적으로 허용되고 인간에게 투여될 때, 통상적으로 위장 장애, 현기증과 같은 알레르기 반응 또는 이와 유사한 반응을 일으키지 않는 조성물을 말한다. 상기 담체, 부형제 및 희석제의 예로는, 락토즈, 덱스트로즈, 수크로즈, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 폴리비닐피롤리돈, 물, 메틸하이드록시벤조에이트, 프로필하이드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 또한, 충진제, 항응집제, 윤활제, 습윤제, 향료, 유화제 및 방부제 등을 추가로 포함할 수 있다. The term " pharmaceutically acceptable "as used herein refers to compositions which are physiologically acceptable and which, when administered to humans, do not normally cause an allergic reaction such as gastrointestinal disorders, dizziness, or similar reactions. Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. Further, it may further include a filler, an anticoagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent and an antiseptic agent.

본 발명의 다른 일관점에 따르면, 상기 폴리뉴클레오타이드에 의해 암호화되는 재조합 단백질이 제공된다.According to another aspect of the present invention, there is provided a recombinant protein encoded by the polynucleotide.

본 발명의 다른 일관점에 따르면, gB, gD, UL39, ICP0 및 ICP4 단백질로 구성되는 군으로부터 선택되는 2종 이상의 또는 모든 HSV-2 항원 단백질을 발현하도록 상기 항원 단백질을 암호화하는 폴리뉴클레오타이드를 포함하는 발현벡터가 제공된다.According to another aspect of the present invention there is provided a kit for the treatment of a subject comprising a polynucleotide encoding said antigen protein to express two or more or all of the HSV-2 antigen proteins selected from the group consisting of gB, gD, UL39, ICPO and ICP4 proteins An expression vector is provided.

상기 발현벡터에 있어서, 상기 gB는 1-22 a.a 위치에 상응하는 신호서열 및 772-792 a.a 위치에 상응하는 막통과 도메인이 결실된 것일 수 있고, 상기 gD는 1-25 a.a 위치에 상응하는 신호서열 및 341-364 a.a 위치에 상응하는 막통과 도메인이 결실된 것일 수 있으며, 상기 ICP0은 510-516 a.a 위치에 상응하는 핵위치화 신호(NLS)가 결실된 것일 수 있고, 상기 IPC4는 767-1318 a.a 위치에 상응하는 RS1.3 부위가 결실된 것일 수 있다.In the above expression vector, gB may be a signal sequence corresponding to the 1-22 aa position and a transmembrane domain corresponding to the 772-792 aa position deleted, and the gD may be a signal corresponding to the 1-25 aa position And the transmembrane domain corresponding to positions 341-364 aa may have been deleted, the ICPO may be a deletion of the nuclear localization signal (NLS) corresponding to positions 510-516 aa, and IPC4 may be a 767- The RS1.3 site corresponding to the 1318 aa position may have been deleted.

상기 발현벡터에 있어서, 상기 UL39는 내부의 도메인들이 뒤섞인 셔플드 UL39 단백질일 수 있고, 상기 셔플드 UL39 단백질은 상술한 폴리뉴클레오타이드 중 어느 하나에 의해 암호화될 수 있다.In the expression vector, the UL39 may be a shuffled UL39 protein mixed with internal domains, and the shuffled UL39 protein may be encoded by any of the above-described polynucleotides.

아울러 상기 발현벡터는 상기 gB, gD, UL39, ICP0 및 ICP4 단백질은 별개의 단백질로 발현되거나 하나의 융합단백질 형태로 발현되도록 제작될 수 있다.In addition, the expression vector may be constructed such that the gB, gD, UL39, ICP0 and ICP4 proteins are expressed as separate proteins or expressed as a single fusion protein.

상기 발현벡터에 있어서, 상기 셔플드 UL39 단백질은 상술한 바와 같이 서열번호 1로 표시되는 UL-39-N1, 서열번호 2로 표시되는 UL39-C2, 서열번호 3으로 표시되는 UL39-N2, 서열번호 4로 표시되는 UL39 N4-C1, 및 서열번호 5로 표시되는 UL39-N3이 무작위적으로 섞인 것이나, 서열번호 1 내지 5의 펩타이드가 순차적으로 연결된 것일 수 있다. 다만 이 경우 상기 셔플드 UL39 단백질은 원래의 전장 UL39 단백질은 제외한다. In the expression vector, the shuffled UL39 protein is preferably selected from the group consisting of UL-39-N1 represented by SEQ ID NO: 1, UL39-C2 represented by SEQ ID NO: 2, UL39-N2 represented by SEQ ID NO: 3, 4 and UL39-N3 represented by SEQ. ID. NO: 5 may be randomly mixed, or the peptides of SEQ ID NOS: 1 to 5 may be sequentially connected. In this case, however, the shuffled UL39 protein excludes the original full length UL39 protein.

상기 발현벡터에 있어서, 상기 gB, gD, UL39, ICP0 및 ICP4 단백질은 별개의 단백질로 발현되거나 하나의 융합단백질 형태로 발현될 수 있다.In the expression vector, the gB, gD, UL39, ICP0 and ICP4 proteins may be expressed as separate proteins or expressed in the form of a single fusion protein.

상기 발현벡터에 있어서, 상기 폴리뉴클레오타이드는 분비 신호서열을 암호화하는 폴리뉴클레오타이드를 추가로 포함할 수 있고, 상기 분비 신호서열은 상술한 바와 같이 tPA(tissue plasminogen activator) 신호서열, HSV gDs(단순포진 바이러스 당단백질 Ds) 신호서열 또는 성장호르몬 신호서열일 수 있다.In the expression vector, the polynucleotide may further comprise a polynucleotide encoding a secretory signal sequence, wherein the secretory signal sequence is selected from the group consisting of a tissue plasminogen activator (tPA) signal sequence, HSV gDs Glycoprotein Ds) signal sequence or a growth hormone signal sequence.

상기 발현벡터에 있어서, 상기 폴리뉴클레오타이드는 면역 증진 펩타이드를 암호화하는 폴리뉴클레오타이드를 추가로 포함할 수 있고, 상기 면역 증진 펩타이드는 CD28, ICOS(inducible costimulator), CTLA4(cytotoxic T lymphocyte associated protein 4), PD1(programmed cell death protein 1), BTLA(B and T lymphocyte associated protein), DR3(death receptor 3), 4-1BB, CD2, CD40, CD30, CD27, SLAM(signaling lymphocyte activation molecule), 2B4(CD244), NKG2D(natural-killer group 2, member D)/DAP12(DNAX-activating protein 12), TIM1(T-Cell immunoglobulin and mucin domain containing protein 1), TIM2, TIM3, TIGIT, CD226, CD160, LAG3(lymphocyte activation gene 3), B7-1, B7-H1, GITR(glucocorticoid-induced TNFR family related protein), Flt3 리간드(fms-like tyrosine kinase 3 ligand), 플라젤린(flagellin), HVEM(herpesvirus entry mediator), CD40 L(리간드) 또는 OX40L[ligand for CD134(OX40), CD252]의 세포질 도메인 또는 이들 중 둘 이상의 연결체일 수 있다. 상기 면역 증진 펩타이드는 바람직하게는 Flt3 리간드일 수 있고 상기 Flt3 리간드는 서열번호 9에 기재된 아미노산 서열을 가질 수 있다.In the expression vector, the polynucleotide may further comprise a polynucleotide encoding an immunity enhancing peptide, wherein the immunity enhancing peptide is selected from the group consisting of CD28, an inducible costimulator (ICOS), cytotoxic T lymphocyte associated protein 4 (CTLA4) (BAT), BAT (B and T lymphocyte associated protein), DR3 (death receptor 3), 4-1BB, CD2, CD40, CD30, CD27, SLAM (signaling lymphocyte activation molecule), 2B4 (DNAX-activating protein 12), TIM1 (T-cell immunoglobulin and mucin domain containing protein 1), TIM2, TIM3, TIGIT, CD226, CD160, LAG3 (lymphocyte activation gene 3), B7-1, B7-H1, glucocorticoid-induced TNFR family related protein, fms-like tyrosine kinase 3 ligand, flagellin, herpesvirus entry mediator, CD40 L Ligand) or the cytoplasmic domain of OX40L [ligand for CD134 (OX40), CD252], or both Or more. The immuno-promoting peptide may preferably be an Flt3 ligand and the Flt3 ligand may have the amino acid sequence set forth in SEQ ID NO: 9.

상기 DNA 백신 조성물은 구체적으로 gB 및 UL39를 포함하는 제1융합단백질을 암호화하는 폴리뉴클레오타이드가 프로모터에 작동가능하게 연결된 제1유전자 컨스트럭트를 포함하는 제1발현벡터 및 gD, ICP0 및 ICP4를 포함하는 제2융합단백질을 암호화하는 폴리뉴클레오타이드가 프로모터에 작동가능하게 연결된 제2유전자 컨스트럭트를 포함하는 제2발현벡터를 포함할 수 있다.The DNA vaccine composition specifically comprises a first expression vector comprising a first gene construct operably linked to a promoter and a polynucleotide encoding a first fusion protein comprising gB and UL39 and gD, ICPO and ICP4 A second expression construct comprising a second gene construct operably linked to a promoter, wherein the polynucleotide encoding the second fusion protein is operably linked to the promoter.

상기 DNA 백신 조성물에 있어서, 상기 제1융합단백질 및/또는 제2융합단백질은 분비 신호서열을 추가로 포함할 수 있으며, 상기 분비 신호서열은 상술한 바와 같다.In the DNA vaccine composition, the first fusion protein and / or the second fusion protein may further comprise a secretory signal sequence, and the secretory signal sequence is as described above.

상기 DNA 백신 조성물에 있어서, 상기 제1융합단백질 및/또는 제2융합단백질은 면역 증진 펩타이드를 추가로 포함할 수 있으며, 상기 면역 증진 펩타이드는 상술한 바와 같다.In the DNA vaccine composition, the first fusion protein and / or the second fusion protein may further comprise an immunity enhancing peptide, and the immunity enhancing peptide is as described above.

상기 DNA 백신 조성물에 있어서, 상기 UL39는 내부의 도메인들이 뒤섞인 셔플드 UL39일 수 있다.In the DNA vaccine composition, the UL39 may be a shuffled UL39 mixed with internal domains.

상기 DNA 백신 조성물은 적어도 하나 이상의 약학적으로 허용가능한 보조제(adjuvant)를 포함할 수 있다.The DNA vaccine composition may comprise at least one pharmaceutically acceptable adjuvant.

본 문서에서 사용되는 "보조제(adjuvant)"는 백신의 면역반응을 향상시킬 목적으로 투여되는 약학적 또는 면역학적 제제를 의미한다. "Adjuvant ", as used herein, refers to a pharmaceutical or immunological preparation that is administered for the purpose of enhancing the immune response of a vaccine.

상기 보조제는 알루미늄 하이드록사이드, 알루미늄 포스페이트, 알룸(포타슘 알루미늄 설페이트), MF59, virosome, AS04[알루미늄 하이드록사이드 및 모노포스포릴 리피드 A(MPL)의 혼합물], AS03(DL-α-tocopherol, squalene 및 유화제인 polysorbate 80의 혼합물), CpG, Flagellin, Poly I:C, AS01,AS02, ISCOMs 또는 ISCOMMATRIX일 수 있다.These adjuvants include aluminum hydroxide, aluminum phosphate, alum (potassium aluminum sulfate), MF59, virosome, AS04 [mixture of aluminum hydroxide and monophosphoryl lipid A (MPL)], AS03 (DL-alpha-tocopherol, squalene And polysorbate 80, an emulsifier), CpG, Flagellin, Poly I: C, AS01, AS02, ISCOMs or ISCOMMATRIX.

또한, 본 발명에 일 실시예에 따른 백신 조성물은 포유동물에 투여시, 활성 성분의 신속한 방출, 또는 지속 또는 지연된 방출이 가능하도록 당업계에 공지된 방법을 사용하여 제형화될 수 있다. 제형은 분말, 과립, 정제, 에멀젼, 시럽, 에어로졸, 연질 또는 경질 젤라틴 캅셀, 멸균 주사용액, 멸균 분말 형태를 포함한다. In addition, the vaccine composition according to one embodiment of the present invention may be formulated using methods known in the art to allow rapid release, or sustained or delayed release, of the active ingredient upon administration to a mammal. Formulations include powders, granules, tablets, emulsions, syrups, aerosols, soft or hard gelatin capsules, sterile injectable solutions, sterile powders.

본 발명의 일 실시예에 따른 백신 조성물은 다양한 경로로 투여될 수 있으며, 예를 들면, 경구, 비경구, 예를 들면 좌제, 경피, 정맥, 복강, 근육내, 병변내, 비강, 피내, 척추관내 투여로 투여될 수 있으며, 또한 서방형 또는 연속적 또는 반복적 방출을 위한 이식장치를 사용하여 투여될 수 있다. 투여횟수는 원하는 범위 내에서 하루에 1회, 또는 수회로 나누어 투여할 수 있으며, 투여 기간도 특별히 한정되지 않는다. The vaccine composition according to an embodiment of the present invention may be administered in various routes such as oral, parenteral, e.g. suppository, transdermal, intravenous, intraperitoneal, intramuscular, lesional, nasal, Intravenous administration, and may also be administered using an implantable device for sustained or continuous or repeated release. The number of administrations can be administered once or several times a day within a desired range, and the administration period is not particularly limited.

본 발명의 일 실시예에 따른 백신 조성물은 일반적인 전신성 투여 또는 국소성 투여, 예컨대, 근육내 주사 또는 정맥 주사 방식으로 투여될 수 있으나, 가장 바람직하게는 전기천공기(electroporator)를 이용하여 주입될 수 있다. 상기 전기천공기는 시판 중인 DNA 약물 체내 주입용 전기천공기, 예컨대, 이탈리아의 IGEA 사의 Glinporator^TM, 한국의 JCBIO사의 CUY21EDIT, 스위스의 Supertech사의 SP-4a 등이 사용될 수 있다.The vaccine composition according to an embodiment of the present invention may be administered by general systemic administration or topical administration such as intramuscular injection or intravenous injection, but most preferably, it may be injected using an electroporator. The electric perforator may be an electric perforator for injecting a commercially available DNA drug into the body, for example, Glinporator ( ^TM) of IGEA of Italy, CUY21EDIT of JCBIO of Korea, or SP-4a of Supertech of Switzerland.

본 발명에 일 실시예에 따른 백신 조성물의 투여 경로는 목적 조직에 도달할 수 있는 한 어떠한 일반적인 경로를 통하여 투여될 수 있다. 이와 같은 투여경로는 비경구 투여, 예를 들어, 복강내 투여, 정맥내 투여, 근육내 투여, 피하 투여, 활막강 내 투여될수 있으나, 이에 제한되지는 않는다. The administration route of the vaccine composition according to an embodiment of the present invention may be administered through any conventional route as long as it can reach the target tissue. Such an administration route may be, but not limited to, parenteral administration, for example, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, and intrathecal administration.

본 발명에 일 실시예에 따른 백신 조성물은 일반적으로 사용되는 약학적으로 허용가능한 담체와 함께 적합한 형태로 제형화될 수 있다. 약학적으로 허용되는 담체로는 예를 들면, 물, 적합한 오일, 식염수, 수성 글루코스 및 글리콜 등과 같은 비경구 투여용 담체 등이 있으며 안정화제 및 보존제를 추가로 포함할 수 있다. 적합한 안정화제로는 아황산수소나트륨, 아황산나트륨 또는 아스코르브산과 같은 항산화제가 있다. 적합한 보존제로는 벤즈알코늄 클로라이드, 메틸- 또는 프로필-파라벤 및 클로로부탄올이 있다. 또한 본 발명에 따른 조성물은 그 투여방법이나 제형에 따라 필요한 경우, 현탁제, 용해보조제, 안정화제, 등장화제, 보존제, 흡착방지제, 계면활성화제, 희석제, 부형제, pH 조정제, 무통화제, 완충제, 산화방지제 등을 적절히 포함할 수 있다. 상기에 예시된 것들을 비롯하여 본 발명에 적합한 약학적으로 허용되는 담체 및 제제는 문헌[Remington's Pharmaceutical Sciences, 최신판]에 상세히 기재되어 있다. The vaccine composition according to an embodiment of the present invention may be formulated into a suitable form together with a commonly used pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers include, for example, water, suitable oils, saline, aqueous carriers for parenteral administration such as aqueous glucose and glycols, etc., and may additionally contain stabilizers and preservatives. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol. In addition, the composition according to the present invention may contain various additives such as a suspending agent, a solubilizer, a stabilizer, an isotonic agent, a preservative, an adsorption inhibitor, an interface activator, a diluent, an excipient, a pH adjuster, An antioxidant, and the like. Pharmaceutically acceptable carriers and formulations suitable for the present invention, including those exemplified above, are described in detail in Remington ' s Pharmaceutical Sciences, Current Edition.

상기 백신 조성물의 환자에 대한 투여량은 환자의 신장, 체표면적, 연령, 투여되는 특정 화합물, 성별, 투여 시간 및 경로, 일반적인 건강, 및 동시에 투여되는 다른 약물들을 포함하는 많은 요소들에 따라 다르다. 약학적으로 활성인 DNA는 100 ng/체중(kg) - 10 ㎎/체중(㎏)의 양으로 투여될 수 있고, 더 바람직하게는 1 내지 500 ㎍/kg(체중)으로 투여될 수 있으며, 가장 바람직하게는 5 내지 50 ㎍/kg(체중)으로 투여될 수 있고, 10 ㎍, 100 ㎍, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg 또는 100 mg과 같은 단위 용량으로 투여될 수도 있으며, 상기 요소들을 고려하여 투여량이 조절될 수 있다.The dosage for the patient of the vaccine composition will depend on a number of factors including the patient's height, body surface area, age, the particular compound being administered, sex, time and route of administration, general health, and other drugs being administered concurrently. The pharmaceutically active DNA may be administered in an amount of 100 ng / body weight (kg) - 10 mg / body weight (kg), more preferably 1 to 500 μg / kg (body weight) Preferably 10 mg, 100 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 20 mg, 30 mg, 40 mg , 50 mg, 60 mg, 70 mg, 80 mg, 90 mg or 100 mg, and the dosage can be adjusted in consideration of the above factors.

이하, 실시예 및 실험예를 통하여 본 발명을 더 상세히 설명한다. 그러나 본 발명은 이하에서 개시되는 실시예 및 실험예에 한정되는 것이 아니라 서로 다른 다양한 형태로 구현될 수 있는 것으로, 이하의 실시예 및 실험예는 본 발명의 개시가 완전하도록 하며, 본 발명이 속한 기술분야의 통상의 지식을 가진 자에게 발명의 범주를 완전하게 알려주기 위해 제공되는 것이다. Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples. It should be understood, however, that the invention is not limited to the disclosed embodiments and examples, but may be embodied in many different forms and should not be construed as being limited to the embodiments set forth herein. And is provided to fully disclose the scope of the invention to a person having ordinary skill in the art.

실시예 1: Shuffled-UL39 plasmid DNA 제조Example 1: Preparation of Shuffled-UL39 plasmid DNA

본 발명자들은 HSV-2 항원 중의 하나인 UL39(ICP10)가 CD4⁺ T 세포 및 CD8⁺ T 세포 반응을 유도한다는 선행연구 결과를 바탕으로(Posavad et al., Mucosal Immunol. 8(1): 115-126, 2015), UL39의 내부 도메인을 뒤섞어 놓은 shuffled construct가 면역반응을 유발하는 백신으로서 기능할 수 있는지 여부를 조사하였다.(Posavad et al ., Mucosal Immunol. 8 (1): 115-7), which is one of the HSV-2 antigens, induces CD4 ⁺ T cell and CD8 ⁺ 126, 2015), and investigated whether a shuffled construct that interspersed with the internal domain of UL39 could function as a vaccine to induce an immune response.

이를 위해 발현능이 향상 되도록 개선된 pGX27 plasmid vector(특허 제1442254호)을 기반으로 하여 5개의 조각(N1: UL39_{14-154(Δ78-104)}, C2: UL39_1117-1142, N2: UL39_155-227, N4-C1: UL39_399-1116, N3: UL39_208-398)으로 나누어 뒤섞여 배치된 HSV-2의 셔플드 UL-39 항원 및 상기 셔플드 UL-39를 암호화하는 유전자 컨스트럭트를 포함하는 플라스미드를 제조하였다.To this end, five fragments (N1: UL39 _{14-154 (Δ78-104)} , C2: UL39 _1117-1142 , N2: UL39 _155-227 (based on patent No. 1442254) improved the pGX27 plasmid vector , A shuffled UL-39 antigen of HSV-2, and a gene construct encoding the shuffled UL-39, which are shuffled and divided into N4-Cl: UL39 _399-1116 and N3: UL39 _208-398 .

구체적으로 shuffled-UL39 plasmid DNA는 UL39-N1(서열번호 1), UL39-C2(서열번호 2), UL39-N2(서열번호 3), UL39-N4-C1(서열번호 4), 및 UL39-N3(서열번호 5) 부분으로 나눠진 형태를 기반으로, UL39-N1, UL39-C2, UL39-N2, UL39-N4-C1, 및 UL39-N3 순서로 순차적으로 연결된 형태(서열번호 6)을 암호화하는 폴리뉴클레오타이드(서열번호 7)를 포함하는 유전자 컨스트럭트를 상기 pGX27 플라스미드 벡터에 삽입함으로써 제작되었다(도 1).Specifically, the shuffled-UL39 plasmid DNA comprises at least one of UL39-N1 (SEQ ID NO: 1), UL39-C2 (SEQ ID NO: 2), UL39-N2 (SEQ ID NO: 3), UL39-N4- (SEQ ID NO: 6) coding sequence (SEQ ID NO: 6) in the order of UL39-N1, UL39-C2, UL39-N2, UL39-N4-C1 and UL39- Nucleotide (SEQ ID NO: 7) into the pGX27 plasmid vector (Fig. 1).

실험예 1: Shuffled-UL39 plasmid DNA에 의한 HSV-2 감염 방어 효능 확인Experimental Example 1: Shuffled-UL39 plasmid DNA was used to confirm HSV-2 defense efficacy

본 발명자들은 본 발명의 일 실시예에 따른 shuffled-UL39 plasmid DNA가 HSV-2 감염 방어에 효능이 있는 지를 in vivo 감염동물 모델상에서 확인하기 위하여, 상기 plasmid DNA 백신 투여 후에 HSV-2 감염 방어능을 평가 하였다.The inventors of the present invention found that the shuffled-UL39 plasmid DNA according to an embodiment of the present invention is effective against the HSV-2 infection in an in vivo animal model of infection, Respectively.

구체적으로, C57BL/6 mouse에 대하여 mock plasmid DNA 및 shuffled-UL39 plasmid DNA를 각각 투여한 그룹으로 나누었다. 각 투여 군에 해당 plasmid DNA 4㎍을 2 주 간격으로 2회 in vivo 전기천공(electroporation) 전달 방법으로 근육내 투여 하였으며, 최종 투여 2주 후에 1 x 10⁴pfu의 HSV-2 바이러스를 질내(intravaginal route)로 감염을 시켰다(도 2 및 표 1). 감염 후, 10일간 매일 각 투여군 마우스의 생존여부를 추적 관찰하여 HSV-2 감염 방어능을 평가 하였다(도 3).Specifically, C57BL / 6 mice were divided into mock plasmid DNA and shuffled-UL39 plasmid DNA. 4 μg of the corresponding plasmid DNA was administered intramuscularly by in vivo electroporation twice at intervals of 2 weeks and 1 × 10 ⁴ pfu of HSV-2 virus was injected intravaginally (Fig. 2 and Table 1). After infection, the viability of HSV-2 infection was evaluated by monitoring the survival of mice in each treatment group daily for 10 days (FIG. 3).

그룹group 백신vaccine 실험동물 수Number of experimental animals 투여량 (μg)Dose (μg) 경로(투여방법)Route (method of administration) 대조군Control group mock plasmidmock plasmid 88 4 4 근육내(전기천공법)Intramuscular (electroporation) 실험군Experimental group 셔플드 UL-39Shuffled UL-39 88 44 근육내(전기천공법)Intramuscular (electroporation)

그 결과, 도 3에서 확인되는 바와 같이 mock plasmid DNA 투여군에서는 HSV-2 바이러스 감염 8일에 대부분의 마우스가 죽은 반면, shuffled-UL39 plasmid DNA 투여군에서는 10일까지 40%의 생존율을 보여 유의미하게 개선된 생존율을 관찰할 수 있었다.As a result, as shown in FIG. 3, in the mock plasmid DNA-treated group, most of the mice died on the 8th day of the HSV-2 virus infection whereas in the shuffled-UL39 plasmid DNA treated group, the survival rate was 40% Survival rate was observed.

실시예 2: HSV-2 복합 항원 발현 plamid DNA 제조Example 2: Preparation of plasmid DNA expressing HSV-2 complex antigen

본 발명자들은 상기 실험예 1의 결과로부터 UL39의 DNA 백신으로서의 가능성을 확인하여, 다른 항원 단백질과 조합된 플라스미드 DNA를 제조하였다.The present inventors confirmed the possibility of UL39 as a DNA vaccine from the results of Experimental Example 1, and prepared plasmid DNA in combination with other antigen proteins.

구체적으로, 상기 실시예 1에서 사용한 pGX27 plasmid vector을 기반으로 하여 여러 종류의 HSV-2 항원 즉, 신호서열(gB_1-22) 및 막통과 도메인(gB_772-792)이 제거된 Glycoprotein B(gB_{23-904(Δ772-792)}, 서열번호 10), 및 상기 실시예 1에서 사용한 shuffled UL-39(서열번호 6)가 연결된 tPA-Flt3L-gB-UL39 융합단백질(서열번호 11)를 암호화하는 폴리뉴클레오타이드(서열번호 12)가 포함된 tPA-Flt3L-gB-UL39 plasmid DNA와 신호서열(gD_1-25) 및 막통과 도메인(gD_341-364)이 제거된 Glycoprotein D(gD_{6-393(Δ341-364)}, 서열번호 13), 핵위치화 서열(NLS, ICP0_510-516)이 제거된 infected cell polypeptide 0(ICP0_Δ510-516, 서열번호 14) 및 RS1.3 부분(ICP_767-1318)이 제거된 infected cell polypeptide 4(ICP4_Δ767-1318, 서열번호 15)가 연결된 tPA-Flt3L-gD-IPC0-ICP4 융합단백질(서열번호 16)을 암호화하는 폴리뉴클레오타이드(서열번호 17)가 포함된 tPA-Flt3L-gD-IPC0-ICP4 plasmid DNA를 각각 제작하였다. 상기 두 플라스미드 모두 효율적인 발현을 위하여 코돈 최적화된 tPA 분비 신호서열(서열번호 8)과 면역계 활성 단백질인 FMS-like tyrosine kinase 3 ligand(Flt3L, 서열번호 9)이 N-말단에 부가된 형태의 plasmid DNA이다(도 4).Specifically, based on the pGX27 plasmid vector used in Example 1, various kinds of HSV-2 antigens, ie, the glycoprotein B (gB _1-22 ) and the transmembrane domain (gB _772-792 ) _Encoding the tPA-Flt3L-gB-UL39 fusion protein (SEQ ID NO: 11) to which the shuffled UL-39 (SEQ ID NO: 6) the nucleotide (SEQ ID NO: 12) contains the tPA-Flt3L-gB-UL39 plasmid DNA with signal sequence (gD _1-25) and the transmembrane domain (gD _341-364) is removed Glycoprotein D _{(gD 6-393 (Δ341- 364),} SEQ ID NO: 13), a nuclear localization sequence (NLS, ICP0 _510-516) that the removed infected cell polypeptide 0 (ICP0 _Δ510-516, SEQ ID NO: 14) and RS1.3 part (ICP _767-1318) is removed, (SEQ ID NO: 17) coding for the tPA-Flt3L-gD-IPC0-ICP4 fusion protein (SEQ ID NO: 16) to which the infected cell polypeptide 4 (ICP4 _Δ767-1318 , SEQ ID NO: 15) tPA-Flt3L-gD-IPC0-ICP4 plasmid DNA were prepared. In order to efficiently express both plasmids, a plasmid DNA in which a codon-optimized tPA secretion signal sequence (SEQ ID NO: 8) and an immune system activating protein FMS-like tyrosine kinase 3 ligand (Flt3L, SEQ ID NO: 9) (Fig. 4).

실험예 2: tPA-Flt3L-gB-UL39 및 tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA에 의한 HSV-2 감염 방어 효능 확인Experimental Example 2: Effect of tPA-Flt3L-gB-UL39 and tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA against HSV-2 infection

본 발명자들은 상기 실시예 2에서 제조된 tPA-Flt3L-gB-UL39 및 tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA가 HSV-2 감염 방어에 효능이 있는 지를 in vivo 감염동물 모델상에서 확인하기 위하여, 백신 투여 후에 HSV-2 감염 방어능을 평가 하였다.To the inventors to verify in the second embodiment the tPA-Flt3L-gB-UL39 and tPA-Flt3L-gD-ICP0- ICP4 plasmid whether in vivo infection animal model DNA is operative in HSV-2 infection defense manufactured, After vaccination, the ability to protect against HSV-2 infection was evaluated.

구체적으로, C57BL/6 mouse에 대하여 mock plasmid DNA, tPA-Flt3L-gB-UL39 plasmid DNA, 및 tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA를 각각 투여한 그룹과 tPA-Flt3L-gB-UL39 plasmid DNA 및 tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA를 병용투여한 군으로 나누었다. 각 투여 군에 해당 plasmid DNA 4 ㎍(병용투여의 경우 각각 4 ㎍씩 투여함)을 2 주 간격으로 2회 생체내 전기천공법(in vivo electroporation)으로 근육내 투여 하였으며, 최종 투여 2 주 후에 1 x 10⁴pfu의 HSV-2 바이러스를 질내투여(intravaginal route)로 감염시켰다(도 5 및 표 2). 감염 후 20일간 매일 각 투여군 마우스의 생존률 및 병리학 점수(pathology score)를 추적 관찰하여 HSV-2 감염 방어능을 평가 하였다(도 6). 병리학 점수는 Oh 등이 발표한 논문에 기재된 바에 따라 매겼다(Oh et al., Proc. Natl. Acad. Sci. USA. 113(6): E762-E771, 2016; 병리학적 점수 '0', 무증상; '1', 약한 정도의 생식기 홍반 및 부종; '2', 중간 정도의 생식기 염증; '3', 화농성 생식기 손상; '4', 뒷다리 마비; '5', 폐사).Specifically, the tPA-Flt3L-gB-UL39 plasmid DNA and the tPA-Flt3L-gB-UL39 plasmid DNA and the tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA were respectively administered to C57BL / And tPA-Flt3L-gD-ICP0-ICP4 plasmid DNAs. In each group, 4 μg of the corresponding plasmid DNA (4 μg each for co-administration) was administered intramuscularly by in vivo electroporation twice at intervals of 2 weeks. After 2 weeks of the final administration, 1 x 10 ⁴ pfu of the HSV-2 virus were infected with an intravaginal route (Figure 5 and Table 2). The survival rate and pathology score of each group of mice were monitored daily for 20 days after the infection to assess the ability to protect against HSV-2 infection (FIG. 6). Pathologic scores were scored according to the criteria described by Oh et al . (Oh et al ., Proc Natl Acad Sci USA 113 (6): E762-E771, 2016, pathological score '0', asymptomatic; '1', weak genital erythema and edema; '2', moderate genital inflammation; '3', purulent genital damage; '4', hind palsy;

그룹group 백신vaccine 마우스 수Number of mice 투여량 (μg)Dose (μg) 투여경로(투여방법)Administration route (administration method) 대조군Control group mockmock 1010 8 8 근육내(전기천공법)Intramuscular (electroporation) 실험군 1Experiment 1 gB-UL39 (BD-02B)gB-UL39 (BD-02B) 1010 4 + 4 (mock)4 + 4 (mock) 근육내(전기천공법)Intramuscular (electroporation) 실험군 2Experiment 2 gD-ICP0/ICP4gD-ICP0 / ICP4 1010 4 + 4 (mock)4 + 4 (mock) 근육내(전기천공법)Intramuscular (electroporation) 실험군 3Experiment group 3 gD-ICP0/ICP4 +
gB-UL39gD-ICP0 / ICP4 +
gB-UL39 1010 4 + 44 + 4 근육내(전기천공법)Intramuscular (electroporation)

그 결과, 도 6a에서 확인되는 바와 같이 mock plasmid DNA 투여군에서는 HSV-2 바이러스 감염 10일째 모든 동물이 폐사하였다. 이와는 반대로 tPA-Flt3L-gB-UL39 plasmid DNA 또는 tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA 투여군에서는 상당히 개선된 HSV-2 바이러스에 대한 방어효능을 확인할 수 있었다. tPA-Flt3L-gB-UL39 plasmid DNA에서는 40%의 생존율을 보였으며, tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA 투여군에서는 100%의 생존율을 보였다. tPA-Flt3L-gB-UL39 plasmid DNA 및 tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA 병용 투여군 또한 100% 생존률이 관찰되었다. As a result, as shown in Fig. 6A, all animals died from the mock plasmid DNA-treated group on the 10th day of HSV-2 virus infection. In contrast, the tPA-Flt3L-gB-UL39 plasmid DNA or the tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA-treated group showed a significantly improved defense against the HSV-2 virus. The survival rate of tPA-Flt3L-gB-UL39 plasmid DNA was 40% and the survival rate of tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA was 100%. 100% survival rate was also observed in the tPA-Flt3L-gB-UL39 plasmid DNA and tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA combination group.

아울러, 도 6b에서 확인되는 바와 같이 mock plasmid DNA 투여군에서는 병리학 점수가 매우 높게 나타났고, tPA-Flt3L-gB-UL39 plasmid DNA의 경우 mock plasmid DNA 보다는 현저하게 낮은 병리학 점수를 나타냈으나, tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA 투여군보다는 병리학 점수가 높게 나타났다. 반면, tPA-Flt3L-gB-UL39 plasmid DNA 및 tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA 병용 투여 군에서는 각각의 투여군과 비교하여 유의미하게 개선된 병리학 점수가 기록되었다. In addition, as shown in FIG. 6B, the mock plasmid DNA-treated group had a high pathological score, and the tPA-Flt3L-gB-UL39 plasmid DNA showed a significantly lower pathological score than the mock plasmid DNA, but tPA-Flt3L -GD-ICP0-ICP4 plasmid DNA. On the other hand, significantly improved pathology scores were recorded in the tPA-Flt3L-gB-UL39 plasmid DNA and tPA-Flt3L-gD-ICP0-ICP4 plasmid DNA combination groups compared with the respective administration groups.

상술한 바와 같이, 본 발명의 일 실시예에 따른 셔플드 UL39를 항원으로 발현시킬 수 있는 DNA 및 상기 셔플드 UL39 항원과 종래의 HSV-2 항원을 조합시킨 융합단백질을 발현시킬 수 있는 DNA는 HSV-2에 대한 효과적인 감염 방어활성을 나타냈으며, 따라서, HSV-2에 대한 예방 및 치료를 위한 백신으로 매우 효율적으로 사용될 수 있다.As described above, the DNA capable of expressing the shuffled UL39 according to an embodiment of the present invention as an antigen, and the DNA capable of expressing the fusion protein in which the shuffled UL39 antigen and the conventional HSV-2 antigen are combined, -2, and thus can be used effectively as a vaccine for the prevention and treatment of HSV-2.

본 발명은 상술한 실시예 및 실험예를 참고로 설명되었으나 이는 예시적인 것에 불과하며, 당해 기술분야에서 통상의 지식을 가진 자라면 이로부터 다양한 변형 및 균등한 다른 실시예가 가능하다는 점을 이해할 것이다. 따라서 본 발명의 진정한 기술적 보호 범위는 첨부된 특허청구범위의 기술적 사상에 의하여 정해져야 할 것이다.While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. Accordingly, the true scope of the present invention should be determined by the technical idea of the appended claims.

<110> BioDion Co., LTD. <120> A DNA vaccine for preventing and treating HSV-2 infection <130> PD16-5410 <160> 17 <170> KoPatentIn 3.0 <210> 1 <211> 114 <212> PRT <213> Artificial Sequence <220> <223> UL39-N1 <400> 1 Arg Ser Pro Ser Glu Arg Gln Glu Pro Arg Glu Pro Glu Val Ala Pro 1 5 10 15 Pro Gly Gly Asp His Val Phe Cys Arg Lys Val Ser Gly Val Met Val 20 25 30 Leu Ser Ser Asp Pro Pro Gly Pro Ala Ala Tyr Arg Ile Ser Asp Ser 35 40 45 Ser Phe Val Gln Cys Gly Ser Asn Cys Ser Met Ile Ile Asp Gly Asp 50 55 60 Gly Asp Gly Arg Thr Ala Val Val Ala Leu Gly Gly Thr Ser Gly Pro 65 70 75 80 Ser Ala Thr Thr Ser Val Gly Thr Gln Thr Ser Gly Glu Phe Leu His 85 90 95 Gly Asn Pro Arg Thr Pro Glu Pro Gln Gly Pro Gln Ala Val Pro Pro 100 105 110 Pro Pro <210> 2 <211> 26 <212> PRT <213> Artificial Sequence <220> <223> UL39-C2 <400> 2 Tyr Cys Lys Val Arg Lys Ala Thr Asn Ser Gly Val Phe Ala Gly Asp 1 5 10 15 Asp Asn Ile Val Cys Thr Ser Cys Ala Leu 20 25 <210> 3 <211> 73 <212> PRT <213> Artificial Sequence <220> <223> UL39-N2 <400> 3 Pro Pro Pro Phe Pro Trp Gly His Glu Cys Cys Ala Arg Arg Asp Ala 1 5 10 15 Arg Gly Gly Ala Glu Lys Asp Val Gly Ala Ala Glu Ser Trp Ser Asp 20 25 30 Gly Pro Ser Ser Asp Ser Glu Thr Glu Asp Ser Asp Ser Ser Asp Glu 35 40 45 Asp Thr Gly Ser Glu Thr Leu Ser Arg Ser Ser Ser Ile Trp Ala Ala 50 55 60 Gly Ala Thr Asp Asp Asp Asp Ser Asp 65 70 <210> 4 <211> 718 <212> PRT <213> Artificial Sequence <220> <223> UL39-N4-C1 <400> 4 Pro Asn Ala Tyr Thr Pro Tyr His Leu Arg Glu Tyr Ala Thr Arg Leu 1 5 10 15 Val Asn Gly Phe Lys Pro Leu Val Arg Arg Ser Ala Arg Leu Tyr Arg 20 25 30 Ile Leu Gly Val Leu Val His Leu Arg Ile Arg Thr Arg Glu Ala Ser 35 40 45 Phe Glu Glu Trp Met Arg Ser Lys Glu Val Asp Leu Asp Phe Gly Leu 50 55 60 Thr Glu Arg Leu Arg Glu His Glu Ala Gln Leu Met Ile Leu Ala Gln 65 70 75 80 Ala Leu Asn Pro Tyr Asp Cys Leu Ile His Ser Thr Pro Asn Thr Leu 85 90 95 Val Glu Arg Gly Leu Gln Ser Ala Leu Lys Tyr Glu Glu Phe Tyr Leu 100 105 110 Lys Arg Phe Gly Gly His Tyr Met Glu Ser Val Phe Gln Met Tyr Thr 115 120 125 Arg Ile Ala Gly Phe Leu Ala Cys Arg Ala Thr Arg Gly Met Arg His 130 135 140 Ile Ala Leu Gly Arg Gln Gly Ser Trp Trp Glu Met Phe Lys Phe Phe 145 150 155 160 Phe His Arg Leu Tyr Asp His Gln Ile Val Pro Ser Thr Pro Ala Met 165 170 175 Leu Asn Leu Gly Thr Arg Asn Tyr Tyr Thr Ser Ser Cys Tyr Leu Val 180 185 190 Asn Pro Gln Ala Thr Thr Asn Gln Ala Thr Leu Arg Ala Ile Thr Gly 195 200 205 Asn Val Ser Ala Ile Leu Ala Arg Asn Gly Gly Ile Gly Leu Cys Met 210 215 220 Gln Ala Phe Asn Asp Ala Ser Pro Gly Thr Ala Ser Ile Met Pro Ala 225 230 235 240 Leu Lys Val Leu Asp Ser Leu Val Ala Ala His Asn Lys Gln Ser Thr 245 250 255 Arg Pro Thr Gly Ala Cys Val Tyr Leu Glu Pro Trp His Ser Asp Val 260 265 270 Arg Ala Val Leu Arg Met Lys Gly Val Leu Ala Gly Glu Glu Ala Gln 275 280 285 Arg Cys Asp Asn Ile Phe Ser Ala Leu Trp Met Pro Asp Leu Phe Phe 290 295 300 Lys Arg Leu Ile Arg His Leu Asp Gly Glu Lys Asn Val Thr Trp Ser 305 310 315 320 Leu Phe Asp Arg Asp Thr Ser Met Ser Leu Ala Asp Phe His Gly Glu 325 330 335 Glu Phe Glu Lys Leu Tyr Glu His Leu Glu Ala Met Gly Phe Gly Glu 340 345 350 Thr Ile Pro Ile Gln Asp Leu Ala Tyr Ala Ile Val Arg Ser Ala Ala 355 360 365 Thr Thr Gly Ser Pro Phe Ile Met Phe Lys Asp Ala Val Asn Arg His 370 375 380 Tyr Ile Tyr Asp Thr Gln Gly Ala Ala Ile Ala Gly Ser Asn Leu Cys 385 390 395 400 Thr Glu Ile Val His Pro Ala Ser Lys Arg Ser Ser Gly Val Cys Asn 405 410 415 Leu Gly Ser Val Asn Leu Ala Arg Cys Val Ser Arg Gln Thr Phe Asp 420 425 430 Phe Gly Arg Leu Arg Asp Ala Val Gln Ala Cys Val Leu Met Val Asn 435 440 445 Ile Met Ile Asp Ser Thr Leu Gln Pro Thr Pro Gln Cys Thr Arg Gly 450 455 460 Asn Asp Asn Leu Arg Ser Met Gly Ile Gly Met Gln Gly Leu His Thr 465 470 475 480 Ala Cys Leu Lys Met Gly Leu Asp Leu Glu Ser Ala Glu Phe Arg Asp 485 490 495 Leu Asn Thr His Ile Ala Glu Val Met Leu Leu Ala Ala Met Lys Thr 500 505 510 Ser Asn Ala Leu Cys Val Arg Gly Ala Arg Pro Phe Ser His Phe Lys 515 520 525 Arg Ser Met Tyr Arg Ala Gly Arg Phe His Trp Glu Arg Phe Ser Asn 530 535 540 Ala Ser Pro Arg Tyr Glu Gly Glu Trp Glu Met Leu Arg Gln Ser Met 545 550 555 560 Met Lys His Gly Leu Arg Asn Ser Gln Phe Ile Ala Leu Met Pro Thr 565 570 575 Ala Ala Ser Ala Gln Ile Ser Asp Val Ser Glu Gly Phe Ala Pro Leu 580 585 590 Phe Thr Asn Leu Phe Ser Lys Val Thr Arg Asp Gly Glu Thr Leu Arg 595 600 605 Pro Asn Thr Leu Leu Leu Lys Glu Leu Glu Arg Thr Phe Gly Gly Lys 610 615 620 Arg Leu Leu Asp Ala Met Asp Gly Leu Glu Ala Lys Gln Trp Ser Val 625 630 635 640 Ala Gln Ala Leu Pro Cys Leu Asp Pro Ala His Pro Leu Arg Arg Phe 645 650 655 Lys Thr Ala Phe Asp Tyr Asp Gln Glu Leu Leu Ile Asp Leu Cys Ala 660 665 670 Asp Arg Ala Pro Tyr Val Asp His Ser Gln Ser Met Thr Leu Tyr Val 675 680 685 Thr Glu Lys Ala Asp Gly Thr Leu Pro Ala Ser Thr Leu Val Arg Leu 690 695 700 Leu Val His Ala Tyr Lys Arg Gly Leu Lys Thr Gly Met Tyr 705 710 715 <210> 5 <211> 191 <212> PRT <213> Artificial Sequence <220> <223> UL39-N3 <400> 5 Thr Leu Ser Arg Ser Ser Ser Ile Trp Ala Ala Gly Ala Thr Asp Asp 1 5 10 15 Asp Asp Ser Asp Ser Asp Ser Arg Ser Asp Asp Ser Val Gln Pro Asp 20 25 30 Val Val Val Arg Arg Arg Trp Ser Asp Gly Pro Ala Pro Val Ala Phe 35 40 45 Pro Lys Pro Arg Arg Pro Gly Asp Ser Pro Gly Asn Pro Gly Leu Gly 50 55 60 Ala Gly Thr Gly Pro Gly Ser Ala Thr Asp Pro Arg Ala Ser Ala Asp 65 70 75 80 Ser Asp Ser Ala Ala His Ala Ala Ala Pro Gln Ala Asp Val Ala Pro 85 90 95 Val Leu Asp Ser Gln Pro Thr Val Gly Thr Asp Pro Gly Tyr Pro Val 100 105 110 Pro Leu Glu Leu Thr Pro Glu Asn Ala Glu Ala Val Ala Arg Phe Leu 115 120 125 Gly Asp Ala Val Asp Arg Glu Pro Ala Leu Met Leu Glu Tyr Phe Cys 130 135 140 Arg Cys Ala Arg Glu Glu Ser Lys Arg Val Pro Pro Arg Thr Phe Gly 145 150 155 160 Ser Ala Pro Arg Leu Thr Glu Asp Asp Phe Gly Leu Leu Asn Tyr Ala 165 170 175 Leu Ala Glu Met Arg Arg Leu Cys Leu Asp Leu Pro Pro Val Pro 180 185 190 <210> 6 <211> 1122 <212> PRT <213> Artificial Sequence <220> <223> shuffled UL39 protein <400> 6 Arg Ser Pro Ser Glu Arg Gln Glu Pro Arg Glu Pro Glu Val Ala Pro 1 5 10 15 Pro Gly Gly Asp His Val Phe Cys Arg Lys Val Ser Gly Val Met Val 20 25 30 Leu Ser Ser Asp Pro Pro Gly Pro Ala Ala Tyr Arg Ile Ser Asp Ser 35 40 45 Ser Phe Val Gln Cys Gly Ser Asn Cys Ser Met Ile Ile Asp Gly Asp 50 55 60 Gly Asp Gly Arg Thr Ala Val Val Ala Leu Gly Gly Thr Ser Gly Pro 65 70 75 80 Ser Ala Thr Thr Ser Val Gly Thr Gln Thr Ser Gly Glu Phe Leu His 85 90 95 Gly Asn Pro Arg Thr Pro Glu Pro Gln Gly Pro Gln Ala Val Pro Pro 100 105 110 Pro Pro Tyr Cys Lys Val Arg Lys Ala Thr Asn Ser Gly Val Phe Ala 115 120 125 Gly Asp Asp Asn Ile Val Cys Thr Ser Cys Ala Leu Pro Pro Pro Phe 130 135 140 Pro Trp Gly His Glu Cys Cys Ala Arg Arg Asp Ala Arg Gly Gly Ala 145 150 155 160 Glu Lys Asp Val Gly Ala Ala Glu Ser Trp Ser Asp Gly Pro Ser Ser 165 170 175 Asp Ser Glu Thr Glu Asp Ser Asp Ser Ser Asp Glu Asp Thr Gly Ser 180 185 190 Glu Thr Leu Ser Arg Ser Ser Ser Ile Trp Ala Ala Gly Ala Thr Asp 195 200 205 Asp Asp Asp Ser Asp Pro Asn Ala Tyr Thr Pro Tyr His Leu Arg Glu 210 215 220 Tyr Ala Thr Arg Leu Val Asn Gly Phe Lys Pro Leu Val Arg Arg Ser 225 230 235 240 Ala Arg Leu Tyr Arg Ile Leu Gly Val Leu Val His Leu Arg Ile Arg 245 250 255 Thr Arg Glu Ala Ser Phe Glu Glu Trp Met Arg Ser Lys Glu Val Asp 260 265 270 Leu Asp Phe Gly Leu Thr Glu Arg Leu Arg Glu His Glu Ala Gln Leu 275 280 285 Met Ile Leu Ala Gln Ala Leu Asn Pro Tyr Asp Cys Leu Ile His Ser 290 295 300 Thr Pro Asn Thr Leu Val Glu Arg Gly Leu Gln Ser Ala Leu Lys Tyr 305 310 315 320 Glu Glu Phe Tyr Leu Lys Arg Phe Gly Gly His Tyr Met Glu Ser Val 325 330 335 Phe Gln Met Tyr Thr Arg Ile Ala Gly Phe Leu Ala Cys Arg Ala Thr 340 345 350 Arg Gly Met Arg His Ile Ala Leu Gly Arg Gln Gly Ser Trp Trp Glu 355 360 365 Met Phe Lys Phe Phe Phe His Arg Leu Tyr Asp His Gln Ile Val Pro 370 375 380 Ser Thr Pro Ala Met Leu Asn Leu Gly Thr Arg Asn Tyr Tyr Thr Ser 385 390 395 400 Ser Cys Tyr Leu Val Asn Pro Gln Ala Thr Thr Asn Gln Ala Thr Leu 405 410 415 Arg Ala Ile Thr Gly Asn Val Ser Ala Ile Leu Ala Arg Asn Gly Gly 420 425 430 Ile Gly Leu Cys Met Gln Ala Phe Asn Asp Ala Ser Pro Gly Thr Ala 435 440 445 Ser Ile Met Pro Ala Leu Lys Val Leu Asp Ser Leu Val Ala Ala His 450 455 460 Asn Lys Gln Ser Thr Arg Pro Thr Gly Ala Cys Val Tyr Leu Glu Pro 465 470 475 480 Trp His Ser Asp Val Arg Ala Val Leu Arg Met Lys Gly Val Leu Ala 485 490 495 Gly Glu Glu Ala Gln Arg Cys Asp Asn Ile Phe Ser Ala Leu Trp Met 500 505 510 Pro Asp Leu Phe Phe Lys Arg Leu Ile Arg His Leu Asp Gly Glu Lys 515 520 525 Asn Val Thr Trp Ser Leu Phe Asp Arg Asp Thr Ser Met Ser Leu Ala 530 535 540 Asp Phe His Gly Glu Glu Phe Glu Lys Leu Tyr Glu His Leu Glu Ala 545 550 555 560 Met Gly Phe Gly Glu Thr Ile Pro Ile Gln Asp Leu Ala Tyr Ala Ile 565 570 575 Val Arg Ser Ala Ala Thr Thr Gly Ser Pro Phe Ile Met Phe Lys Asp 580 585 590 Ala Val Asn Arg His Tyr Ile Tyr Asp Thr Gln Gly Ala Ala Ile Ala 595 600 605 Gly Ser Asn Leu Cys Thr Glu Ile Val His Pro Ala Ser Lys Arg Ser 610 615 620 Ser Gly Val Cys Asn Leu Gly Ser Val Asn Leu Ala Arg Cys Val Ser 625 630 635 640 Arg Gln Thr Phe Asp Phe Gly Arg Leu Arg Asp Ala Val Gln Ala Cys 645 650 655 Val Leu Met Val Asn Ile Met Ile Asp Ser Thr Leu Gln Pro Thr Pro 660 665 670 Gln Cys Thr Arg Gly Asn Asp Asn Leu Arg Ser Met Gly Ile Gly Met 675 680 685 Gln Gly Leu His Thr Ala Cys Leu Lys Met Gly Leu Asp Leu Glu Ser 690 695 700 Ala Glu Phe Arg Asp Leu Asn Thr His Ile Ala Glu Val Met Leu Leu 705 710 715 720 Ala Ala Met Lys Thr Ser Asn Ala Leu Cys Val Arg Gly Ala Arg Pro 725 730 735 Phe Ser His Phe Lys Arg Ser Met Tyr Arg Ala Gly Arg Phe His Trp 740 745 750 Glu Arg Phe Ser Asn Ala Ser Pro Arg Tyr Glu Gly Glu Trp Glu Met 755 760 765 Leu Arg Gln Ser Met Met Lys His Gly Leu Arg Asn Ser Gln Phe Ile 770 775 780 Ala Leu Met Pro Thr Ala Ala Ser Ala Gln Ile Ser Asp Val Ser Glu 785 790 795 800 Gly Phe Ala Pro Leu Phe Thr Asn Leu Phe Ser Lys Val Thr Arg Asp 805 810 815 Gly Glu Thr Leu Arg Pro Asn Thr Leu Leu Leu Lys Glu Leu Glu Arg 820 825 830 Thr Phe Gly Gly Lys Arg Leu Leu Asp Ala Met Asp Gly Leu Glu Ala 835 840 845 Lys Gln Trp Ser Val Ala Gln Ala Leu Pro Cys Leu Asp Pro Ala His 850 855 860 Pro Leu Arg Arg Phe Lys Thr Ala Phe Asp Tyr Asp Gln Glu Leu Leu 865 870 875 880 Ile Asp Leu Cys Ala Asp Arg Ala Pro Tyr Val Asp His Ser Gln Ser 885 890 895 Met Thr Leu Tyr Val Thr Glu Lys Ala Asp Gly Thr Leu Pro Ala Ser 900 905 910 Thr Leu Val Arg Leu Leu Val His Ala Tyr Lys Arg Gly Leu Lys Thr 915 920 925 Gly Met Tyr Thr Leu Ser Arg Ser Ser Ser Ile Trp Ala Ala Gly Ala 930 935 940 Thr Asp Asp Asp Asp Ser Asp Ser Asp Ser Arg Ser Asp Asp Ser Val 945 950 955 960 Gln Pro Asp Val Val Val Arg Arg Arg Trp Ser Asp Gly Pro Ala Pro 965 970 975 Val Ala Phe Pro Lys Pro Arg Arg Pro Gly Asp Ser Pro Gly Asn Pro 980 985 990 Gly Leu Gly Ala Gly Thr Gly Pro Gly Ser Ala Thr Asp Pro Arg Ala 995 1000 1005 Ser Ala Asp Ser Asp Ser Ala Ala His Ala Ala Ala Pro Gln Ala Asp 1010 1015 1020 Val Ala Pro Val Leu Asp Ser Gln Pro Thr Val Gly Thr Asp Pro Gly 1025 1030 1035 1040 Tyr Pro Val Pro Leu Glu Leu Thr Pro Glu Asn Ala Glu Ala Val Ala 1045 1050 1055 Arg Phe Leu Gly Asp Ala Val Asp Arg Glu Pro Ala Leu Met Leu Glu 1060 1065 1070 Tyr Phe Cys Arg Cys Ala Arg Glu Glu Ser Lys Arg Val Pro Pro Arg 1075 1080 1085 Thr Phe Gly Ser Ala Pro Arg Leu Thr Glu Asp Asp Phe Gly Leu Leu 1090 1095 1100 Asn Tyr Ala Leu Ala Glu Met Arg Arg Leu Cys Leu Asp Leu Pro Pro 1105 1110 1115 1120 Val Pro <210> 7 <211> 3369 <212> DNA <213> Artificial Sequence <220> <223> polynucleotide encoding shuffled UL39 protein <400> 7 agaagtccaa gcgaaaggca ggaacctaga gagccagaag tggcaccacc tggaggggac 60 cacgtcttct gcaggaaggt gtctggggtc atggtgctga gctccgatcc accaggacct 120 gcagcttacc ggatcagtga ctctagtttt gtgcagtgcg ggagcaactg ttccatgatc 180 attgacggcg atggggacgg aagaacagct gtggtcgcac tgggaggcac ttctggaccc 240 agtgccacca catccgtggg cacccagaca tctggggaat ttctgcacgg aaatccccga 300 acccctgagc cacagggacc tcaggccgtg cctccacccc cttattgtaa ggtcagaaaa 360 gctaccaaca gtggagtgtt cgcaggcgac gataatattg tctgcacatc atgtgccctg 420 ccaccccctt ttccatgggg ccatgagtgc tgtgcccgga gagatgctag ggggggagca 480 gaaaaagacg tgggggcagc cgagagttgg tcagatggac cctcaagcga cagcgagacc 540 gaagattccg actcctctga tgaagacact ggcagtgaga ccctgtcacg cagttcaagc 600 atctgggctg caggggccac agacgatgac gattccgacc caaacgctta cacaccctat 660 cacctgaggg aatacgcaac tcgcctggtg aatgggttca agcctctggt caggcgcagc 720 gcccgcctgt atagaatcct gggagtcctg gtgcatctga gaatcaggac acgcgaggct 780 agcttcgagg aatggatgag gtccaaagaa gtggatctgg actttggcct gactgagcga 840 ctgcgggagc acgaagccca gctgatgatt ctggcacagg ccctgaaccc atacgattgc 900 ctgatccatt caactcccaa taccctggtg gaacgaggac tgcagagcgc cctgaagtac 960 gaggagttct acctgaaacg gtttggcggg cactacatgg agagcgtgtt ccagatgtat 1020 accagaattg ccggctttct ggcctgtcgg gctacaagag ggatgaggca tatcgctctg 1080 ggccgccagg ggagctggtg ggagatgttt aagttctttt tccaccggct gtacgaccat 1140 cagatcgtgc catccacccc cgccatgctg aacctgggca ctcggaatta ctatacctcc 1200 tcttgctatc tggtgaaccc ccaggccact accaatcagg ccacactgcg cgctatcact 1260 gggaacgtga gcgcaattct ggcccgaaat ggaggcatcg gactgtgcat gcaggcattc 1320 aacgatgcct ctcctggcac cgccagtatc atgccagctc tgaaggtcct ggactccctg 1380 gtggccgctc acaacaagca gtctacaagg cctactggcg cctgcgtgta cctggagcca 1440 tggcattctg atgtcagagc tgtgctgagg atgaaggggg tgctggctgg agaggaagca 1500 cagcggtgcg ataacatttt cagcgccctg tggatgcctg acctgttttt caagcgcctg 1560 atccgacacc tggatggaga gaaaaatgtg acctggagcc tgtttgatcg ggacacaagc 1620 atgtccctgg ccgacttcca cggcgaggaa tttgaaaaac tgtacgaaca tctggaggct 1680 atgggcttcg gggagaccat cccaattcag gacctggctt atgcaattgt gaggagtgca 1740 gccacaactg gctcaccctt tatcatgttc aaggatgccg tcaaccgcca ctacatctac 1800 gacactcagg gcgctgcaat cgctgggtct aatctgtgca ccgagatcgt gcatccagca 1860 agtaaaagaa gttcaggagt ctgcaacctg ggctcagtga atctggcacg atgcgtgagc 1920 cggcagacct tcgacttcgg aagactgagg gacgctgtgc aggcatgcgt cctgatggtg 1980 aacatcatga ttgatagcac actgcagccc actcctcagt gtacccgagg caacgacaat 2040 ctgcggtcca tgggaattgg catgcaggga ctgcacacag cctgcctgaa gatgggcctg 2100 gatctggaat ctgccgagtt ccgggacctg aacacacata tcgctgaagt gatgctgctg 2160 gccgctatga agactagcaa tgcactgtgc gtgcggggag ccagaccttt ttcacacttc 2220 aaaagaagca tgtacagggc aggccgcttc cattgggagc ggttttcaaa cgccagccca 2280 agatatgagg gggaatggga gatgctgaga cagagtatga tgaagcacgg actgcggaac 2340 agccagttta ttgcactgat gcccactgca gcctctgccc agatcagcga cgtgagcgaa 2400 ggctttgccc ctctgttcac caacctgttt agcaaagtca ccagagacgg ggagacactg 2460 aggcctaata ctctgctgct gaaggaactg gagcgcactt tcgggggaaa acgactgctg 2520 gatgctatgg acggcctgga ggcaaagcag tggtccgtgg cacaggctct gccatgcctg 2580 gaccctgctc atccactgcg acggttcaaa acagcatttg attacgacca ggaactgctg 2640 atcgacctgt gcgccgacag agctccctac gtggatcact ctcagagtat gacactgtat 2700 gtcactgaga aggccgacgg caccctgcct gctagcacac tggtcaggct gctggtgcat 2760 gcctacaagc gcggcctgaa aaccgggatg tatacactgt ccaggagctc ctctatctgg 2820 gccgccgggg ccaccgacga tgacgattca gatagcgact cccgctctga cgattccgtg 2880 cagccagatg tggtcgtgag aaggcgctgg tctgacggac cagcaccagt ggcattccct 2940 aaaccacgac ggcccggaga tagcccaggc aaccctggac tgggagcagg cactgggcct 3000 ggatctgcta ccgacccaag ggcaagtgcc gatagtgact cagccgctca cgcagccgct 3060 ccacaggctg atgtcgcacc tgtgctggac tcccagccaa ccgtggggac agaccccgga 3120 taccccgtcc ctctggagct gacccctgaa aatgctgagg cagtggcccg attcctgggc 3180 gatgcagtgg accgggaacc agccctgatg ctggagtatt tttgccgatg tgcccgggag 3240 gaaagcaagc gggtgccacc aagaactttc ggctccgctc cacgactgac cgaggacgat 3300 tttgggctgc tgaactatgc cctggccgaa atgcggagac tgtgcctgga cctgcccccc 3360 gtgccctaa 3369 <210> 8 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> tPA signal sequence <400> 8 Met Asp Ala Met Lys Arg Gly Leu Cys Cys Val Leu Leu Leu Cys Gly 1 5 10 15 Ala Val Phe Val Ser Pro Ser His Ala 20 25 <210> 9 <211> 156 <212> PRT <213> Artificial Sequence <220> <223> Flt3L <400> 9 Thr Gln Asp Cys Ser Phe Gln His Ser Pro Ile Ser Ser Asp Phe Ala 1 5 10 15 Val Lys Ile Arg Glu Leu Ser Asp Tyr Leu Leu Gln Asp Tyr Pro Val 20 25 30 Thr Val Ala Ser Asn Leu Gln Asp Glu Glu Leu Cys Gly Gly Leu Trp 35 40 45 Arg Leu Val Leu Ala Gln Arg Trp Met Glu Arg Leu Lys Thr Val Ala 50 55 60 Gly Ser Lys Met Gln Gly Leu Leu Glu Arg Val Asn Thr Glu Ile His 65 70 75 80 Phe Val Thr Lys Cys Ala Phe Gln Pro Pro Pro Ser Cys Leu Arg Phe 85 90 95 Val Gln Thr Asn Ile Ser Arg Leu Leu Gln Glu Thr Ser Glu Gln Leu 100 105 110 Val Ala Leu Lys Pro Trp Ile Thr Arg Gln Asn Phe Ser Arg Cys Leu 115 120 125 Glu Leu Gln Cys Gln Pro Asp Ser Ser Thr Leu Pro Pro Pro Trp Ser 130 135 140 Pro Arg Pro Leu Glu Ala Thr Ala Pro Thr Ala Pro 145 150 155 <210> 10 <211> 861 <212> PRT <213> Artificial Sequence <220> <223> HSV-2 gB <400> 10 Ala Pro Ala Ala Pro Ala Ala Pro Arg Ala Ser Gly Gly Val Ala Ala 1 5 10 15 Thr Val Ala Ala Asn Gly Gly Pro Ala Ser Arg Pro Pro Pro Val Pro 20 25 30 Ser Pro Ala Thr Thr Lys Ala Arg Lys Arg Lys Thr Lys Lys Pro Pro 35 40 45 Lys Arg Pro Glu Ala Thr Pro Pro Pro Asp Ala Asn Ala Thr Val Ala 50 55 60 Ala Gly His Ala Thr Leu Arg Ala His Leu Arg Glu Ile Lys Val Glu 65 70 75 80 Asn Ala Asp Ala Gln Phe Tyr Val Cys Pro Pro Pro Thr Gly Ala Thr 85 90 95 Val Val Gln Phe Glu Gln Pro Arg Arg Cys Pro Thr Arg Pro Glu Gly 100 105 110 Gln Asn Tyr Thr Glu Gly Ile Ala Val Val Phe Lys Glu Asn Ile Ala 115 120 125 Pro Tyr Lys Phe Lys Ala Thr Met Tyr Tyr Lys Asp Val Thr Val Ser 130 135 140 Gln Val Trp Phe Gly His Arg Tyr Ser Gln Phe Met Gly Ile Phe Glu 145 150 155 160 Asp Arg Ala Pro Val Pro Phe Glu Glu Val Ile Asp Lys Ile Asn Thr 165 170 175 Lys Gly Val Cys Arg Ser Thr Ala Lys Tyr Val Arg Asn Asn Met Glu 180 185 190 Thr Thr Ala Phe His Arg Asp Asp His Glu Thr Asp Met Glu Leu Lys 195 200 205 Pro Ala Lys Val Ala Thr Arg Thr Ser Arg Gly Trp His Thr Thr Asp 210 215 220 Leu Lys Tyr Asn Pro Ser Arg Val Glu Ala Phe His Arg Tyr Gly Thr 225 230 235 240 Thr Val Asn Cys Ile Val Glu Glu Val Asp Ala Arg Ser Val Tyr Pro 245 250 255 Tyr Asp Glu Phe Val Leu Ala Thr Gly Asp Phe Val Tyr Met Ser Pro 260 265 270 Phe Tyr Gly Tyr Arg Glu Gly Ser His Thr Glu His Thr Ser Tyr Ala 275 280 285 Ala Asp Arg Phe Lys Gln Val Asp Gly Phe Tyr Ala Arg Asp Leu Thr 290 295 300 Thr Lys Ala Arg Ala Thr Ser Pro Thr Thr Arg Asn Leu Leu Thr Thr 305 310 315 320 Pro Lys Phe Thr Val Ala Trp Asp Trp Val Pro Lys Arg Pro Ala Val 325 330 335 Cys Thr Met Thr Lys Trp Gln Glu Val Asp Glu Met Leu Arg Ala Glu 340 345 350 Tyr Gly Gly Ser Phe Arg Phe Ser Ser Asp Ala Ile Ser Thr Thr Phe 355 360 365 Thr Thr Asn Leu Thr Glu Tyr Ser Leu Ser Arg Val Asp Leu Gly Asp 370 375 380 Cys Ile Gly Arg Asp Ala Arg Glu Ala Ile Asp Arg Met Phe Ala Arg 385 390 395 400 Lys Tyr Asn Ala Thr His Ile Lys Val Gly Gln Pro Gln Tyr Tyr Leu 405 410 415 Ala Thr Gly Gly Phe Leu Ile Ala Tyr Gln Pro Leu Leu Ser Asn Thr 420 425 430 Leu Ala Glu Leu Tyr Val Arg Glu Tyr Met Arg Glu Gln Asp Arg Lys 435 440 445 Pro Arg Asn Ala Thr Pro Ala Pro Leu Arg Glu Ala Pro Ser Ala Asn 450 455 460 Ala Ser Val Glu Arg Ile Lys Thr Thr Ser Ser Ile Glu Phe Ala Arg 465 470 475 480 Leu Gln Phe Thr Tyr Asn His Ile Gln Arg His Val Asn Asp Met Leu 485 490 495 Gly Arg Ile Ala Val Ala Trp Cys Glu Leu Gln Asn His Glu Leu Thr 500 505 510 Leu Trp Asn Glu Ala Arg Lys Leu Asn Pro Asn Ala Ile Ala Ser Ala 515 520 525 Thr Val Gly Arg Arg Val Ser Ala Arg Met Leu Gly Asp Val Met Ala 530 535 540 Val Ser Thr Cys Val Pro Val Ala Pro Asp Asn Val Ile Val Gln Asn 545 550 555 560 Ser Met Arg Val Ser Ser Arg Pro Gly Thr Cys Tyr Ser Arg Pro Leu 565 570 575 Val Ser Phe Arg Tyr Glu Asp Gln Gly Pro Leu Ile Glu Gly Gln Leu 580 585 590 Gly Glu Asn Asn Glu Leu Arg Leu Thr Arg Asp Ala Leu Glu Pro Cys 595 600 605 Thr Val Gly His Arg Arg Tyr Phe Ile Phe Gly Gly Gly Tyr Val Tyr 610 615 620 Phe Glu Glu Tyr Ala Tyr Ser His Gln Leu Ser Arg Ala Asp Val Thr 625 630 635 640 Thr Val Ser Thr Phe Ile Asp Leu Asn Ile Thr Met Leu Glu Asp His 645 650 655 Glu Phe Val Pro Leu Glu Val Tyr Thr Arg His Glu Ile Lys Asp Ser 660 665 670 Gly Leu Leu Asp Tyr Thr Glu Val Gln Arg Arg Asn Gln Leu His Asp 675 680 685 Leu Arg Phe Ala Asp Ile Asp Thr Val Ile Arg Ala Asp Ala Asn Ala 690 695 700 Ala Met Phe Ala Gly Leu Cys Ala Phe Phe Glu Gly Met Gly Asp Leu 705 710 715 720 Gly Arg Ala Val Gly Lys Val Val Met Gly Val Val Gly Gly Val Val 725 730 735 Ser Ala Val Ser Gly Val Ser Ser Phe Met Ser Asn Pro Arg Tyr Val 740 745 750 Leu Gln Leu Gln Arg Asn Pro Met Lys Ala Leu Tyr Pro Leu Thr Thr 755 760 765 Lys Glu Leu Lys Thr Ser Asp Pro Gly Gly Val Gly Gly Glu Gly Glu 770 775 780 Glu Gly Ala Glu Gly Gly Gly Phe Asp Glu Ala Lys Leu Ala Glu Ala 785 790 795 800 Arg Glu Met Ile Arg Tyr Met Ala Leu Val Ser Ala Met Glu Arg Thr 805 810 815 Glu His Lys Ala Arg Lys Lys Gly Thr Ser Ala Leu Leu Ser Ser Lys 820 825 830 Val Thr Asn Met Val Leu Arg Lys Arg Asn Lys Ala Arg Tyr Ser Pro 835 840 845 Leu His Asn Glu Asp Glu Ala Gly Asp Glu Asp Glu Leu 850 855 860 <210> 11 <211> 2199 <212> PRT <213> Artificial Sequence <220> <223> tPA-Flt3L-gB-shuffled UL39 fusion protein <400> 11 Met Asp Ala Met Lys Arg Gly Leu Cys Cys Val Leu Leu Leu Cys Gly 1 5 10 15 Ala Val Phe Val Ser Pro Ser His Ala Thr Gln Asp Cys Ser Phe Gln 20 25 30 His Ser Pro Ile Ser Ser Asp Phe Ala Val Lys Ile Arg Glu Leu Ser 35 40 45 Asp Tyr Leu Leu Gln Asp Tyr Pro Val Thr Val Ala Ser Asn Leu Gln 50 55 60 Asp Glu Glu Leu Cys Gly Gly Leu Trp Arg Leu Val Leu Ala Gln Arg 65 70 75 80 Trp Met Glu Arg Leu Lys Thr Val Ala Gly Ser Lys Met Gln Gly Leu 85 90 95 Leu Glu Arg Val Asn Thr Glu Ile His Phe Val Thr Lys Cys Ala Phe 100 105 110 Gln Pro Pro Pro Ser Cys Leu Arg Phe Val Gln Thr Asn Ile Ser Arg 115 120 125 Leu Leu Gln Glu Thr Ser Glu Gln Leu Val Ala Leu Lys Pro Trp Ile 130 135 140 Thr Arg Gln Asn Phe Ser Arg Cys Leu Glu Leu Gln Cys Gln Pro Asp 145 150 155 160 Ser Ser Thr Leu Pro Pro Pro Trp Ser Pro Arg Pro Leu Glu Ala Thr 165 170 175 Ala Pro Thr Ala Pro Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Ala 180 185 190 Pro Ala Ala Pro Ala Ala Pro Arg Ala Ser Gly Gly Val Ala Ala Thr 195 200 205 Val Ala Ala Asn Gly Gly Pro Ala Ser Arg Pro Pro Pro Val Pro Ser 210 215 220 Pro Ala Thr Thr Lys Ala Arg Lys Arg Lys Thr Lys Lys Pro Pro Lys 225 230 235 240 Arg Pro Glu Ala Thr Pro Pro Pro Asp Ala Asn Ala Thr Val Ala Ala 245 250 255 Gly His Ala Thr Leu Arg Ala His Leu Arg Glu Ile Lys Val Glu Asn 260 265 270 Ala Asp Ala Gln Phe Tyr Val Cys Pro Pro Pro Thr Gly Ala Thr Val 275 280 285 Val Gln Phe Glu Gln Pro Arg Arg Cys Pro Thr Arg Pro Glu Gly Gln 290 295 300 Asn Tyr Thr Glu Gly Ile Ala Val Val Phe Lys Glu Asn Ile Ala Pro 305 310 315 320 Tyr Lys Phe Lys Ala Thr Met Tyr Tyr Lys Asp Val Thr Val Ser Gln 325 330 335 Val Trp Phe Gly His Arg Tyr Ser Gln Phe Met Gly Ile Phe Glu Asp 340 345 350 Arg Ala Pro Val Pro Phe Glu Glu Val Ile Asp Lys Ile Asn Thr Lys 355 360 365 Gly Val Cys Arg Ser Thr Ala Lys Tyr Val Arg Asn Asn Met Glu Thr 370 375 380 Thr Ala Phe His Arg Asp Asp His Glu Thr Asp Met Glu Leu Lys Pro 385 390 395 400 Ala Lys Val Ala Thr Arg Thr Ser Arg Gly Trp His Thr Thr Asp Leu 405 410 415 Lys Tyr Asn Pro Ser Arg Val Glu Ala Phe His Arg Tyr Gly Thr Thr 420 425 430 Val Asn Cys Ile Val Glu Glu Val Asp Ala Arg Ser Val Tyr Pro Tyr 435 440 445 Asp Glu Phe Val Leu Ala Thr Gly Asp Phe Val Tyr Met Ser Pro Phe 450 455 460 Tyr Gly Tyr Arg Glu Gly Ser His Thr Glu His Thr Ser Tyr Ala Ala 465 470 475 480 Asp Arg Phe Lys Gln Val Asp Gly Phe Tyr Ala Arg Asp Leu Thr Thr 485 490 495 Lys Ala Arg Ala Thr Ser Pro Thr Thr Arg Asn Leu Leu Thr Thr Pro 500 505 510 Lys Phe Thr Val Ala Trp Asp Trp Val Pro Lys Arg Pro Ala Val Cys 515 520 525 Thr Met Thr Lys Trp Gln Glu Val Asp Glu Met Leu Arg Ala Glu Tyr 530 535 540 Gly Gly Ser Phe Arg Phe Ser Ser Asp Ala Ile Ser Thr Thr Phe Thr 545 550 555 560 Thr Asn Leu Thr Glu Tyr Ser Leu Ser Arg Val Asp Leu Gly Asp Cys 565 570 575 Ile Gly Arg Asp Ala Arg Glu Ala Ile Asp Arg Met Phe Ala Arg Lys 580 585 590 Tyr Asn Ala Thr His Ile Lys Val Gly Gln Pro Gln Tyr Tyr Leu Ala 595 600 605 Thr Gly Gly Phe Leu Ile Ala Tyr Gln Pro Leu Leu Ser Asn Thr Leu 610 615 620 Ala Glu Leu Tyr Val Arg Glu Tyr Met Arg Glu Gln Asp Arg Lys Pro 625 630 635 640 Arg Asn Ala Thr Pro Ala Pro Leu Arg Glu Ala Pro Ser Ala Asn Ala 645 650 655 Ser Val Glu Arg Ile Lys Thr Thr Ser Ser Ile Glu Phe Ala Arg Leu 660 665 670 Gln Phe Thr Tyr Asn His Ile Gln Arg His Val Asn Asp Met Leu Gly 675 680 685 Arg Ile Ala Val Ala Trp Cys Glu Leu Gln Asn His Glu Leu Thr Leu 690 695 700 Trp Asn Glu Ala Arg Lys Leu Asn Pro Asn Ala Ile Ala Ser Ala Thr 705 710 715 720 Val Gly Arg Arg Val Ser Ala Arg Met Leu Gly Asp Val Met Ala Val 725 730 735 Ser Thr Cys Val Pro Val Ala Pro Asp Asn Val Ile Val Gln Asn Ser 740 745 750 Met Arg Val Ser Ser Arg Pro Gly Thr Cys Tyr Ser Arg Pro Leu Val 755 760 765 Ser Phe Arg Tyr Glu Asp Gln Gly Pro Leu Ile Glu Gly Gln Leu Gly 770 775 780 Glu Asn Asn Glu Leu Arg Leu Thr Arg Asp Ala Leu Glu Pro Cys Thr 785 790 795 800 Val Gly His Arg Arg Tyr Phe Ile Phe Gly Gly Gly Tyr Val Tyr Phe 805 810 815 Glu Glu Tyr Ala Tyr Ser His Gln Leu Ser Arg Ala Asp Val Thr Thr 820 825 830 Val Ser Thr Phe Ile Asp Leu Asn Ile Thr Met Leu Glu Asp His Glu 835 840 845 Phe Val Pro Leu Glu Val Tyr Thr Arg His Glu Ile Lys Asp Ser Gly 850 855 860 Leu Leu Asp Tyr Thr Glu Val Gln Arg Arg Asn Gln Leu His Asp Leu 865 870 875 880 Arg Phe Ala Asp Ile Asp Thr Val Ile Arg Ala Asp Ala Asn Ala Ala 885 890 895 Met Phe Ala Gly Leu Cys Ala Phe Phe Glu Gly Met Gly Asp Leu Gly 900 905 910 Arg Ala Val Gly Lys Val Val Met Gly Val Val Gly Gly Val Val Ser 915 920 925 Ala Val Ser Gly Val Ser Ser Phe Met Ser Asn Pro Arg Tyr Val Leu 930 935 940 Gln Leu Gln Arg Asn Pro Met Lys Ala Leu Tyr Pro Leu Thr Thr Lys 945 950 955 960 Glu Leu Lys Thr Ser Asp Pro Gly Gly Val Gly Gly Glu Gly Glu Glu 965 970 975 Gly Ala Glu Gly Gly Gly Phe Asp Glu Ala Lys Leu Ala Glu Ala Arg 980 985 990 Glu Met Ile Arg Tyr Met Ala Leu Val Ser Ala Met Glu Arg Thr Glu 995 1000 1005 His Lys Ala Arg Lys Lys Gly Thr Ser Ala Leu Leu Ser Ser Lys Val 1010 1015 1020 Thr Asn Met Val Leu Arg Lys Arg Asn Lys Ala Arg Tyr Ser Pro Leu 1025 1030 1035 1040 His Asn Glu Asp Glu Ala Gly Asp Glu Asp Glu Leu Gly Gly Gly Gly 1045 1050 1055 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 1060 1065 1070 Gly Gly Gly Gly Ser Arg Ser Pro Ser Glu Arg Gln Glu Pro Arg Glu 1075 1080 1085 Pro Glu Val Ala Pro Pro Gly Gly Asp His Val Phe Cys Arg Lys Val 1090 1095 1100 Ser Gly Val Met Val Leu Ser Ser Asp Pro Pro Gly Pro Ala Ala Tyr 1105 1110 1115 1120 Arg Ile Ser Asp Ser Ser Phe Val Gln Cys Gly Ser Asn Cys Ser Met 1125 1130 1135 Ile Ile Asp Gly Asp Gly Asp Gly Arg Thr Ala Val Val Ala Leu Gly 1140 1145 1150 Gly Thr Ser Gly Pro Ser Ala Thr Thr Ser Val Gly Thr Gln Thr Ser 1155 1160 1165 Gly Glu Phe Leu His Gly Asn Pro Arg Thr Pro Glu Pro Gln Gly Pro 1170 1175 1180 Gln Ala Val Pro Pro Pro Pro Tyr Cys Lys Val Arg Lys Ala Thr Asn 1185 1190 1195 1200 Ser Gly Val Phe Ala Gly Asp Asp Asn Ile Val Cys Thr Ser Cys Ala 1205 1210 1215 Leu Pro Pro Pro Phe Pro Trp Gly His Glu Cys Cys Ala Arg Arg Asp 1220 1225 1230 Ala Arg Gly Gly Ala Glu Lys Asp Val Gly Ala Ala Glu Ser Trp Ser 1235 1240 1245 Asp Gly Pro Ser Ser Asp Ser Glu Thr Glu Asp Ser Asp Ser Ser Asp 1250 1255 1260 Glu Asp Thr Gly Ser Glu Thr Leu Ser Arg Ser Ser Ser Ile Trp Ala 1265 1270 1275 1280 Ala Gly Ala Thr Asp Asp Asp Asp Ser Asp Pro Asn Ala Tyr Thr Pro 1285 1290 1295 Tyr His Leu Arg Glu Tyr Ala Thr Arg Leu Val Asn Gly Phe Lys Pro 1300 1305 1310 Leu Val Arg Arg Ser Ala Arg Leu Tyr Arg Ile Leu Gly Val Leu Val 1315 1320 1325 His Leu Arg Ile Arg Thr Arg Glu Ala Ser Phe Glu Glu Trp Met Arg 1330 1335 1340 Ser Lys Glu Val Asp Leu Asp Phe Gly Leu Thr Glu Arg Leu Arg Glu 1345 1350 1355 1360 His Glu Ala Gln Leu Met Ile Leu Ala Gln Ala Leu Asn Pro Tyr Asp 1365 1370 1375 Cys Leu Ile His Ser Thr Pro Asn Thr Leu Val Glu Arg Gly Leu Gln 1380 1385 1390 Ser Ala Leu Lys Tyr Glu Glu Phe Tyr Leu Lys Arg Phe Gly Gly His 1395 1400 1405 Tyr Met Glu Ser Val Phe Gln Met Tyr Thr Arg Ile Ala Gly Phe Leu 1410 1415 1420 Ala Cys Arg Ala Thr Arg Gly Met Arg His Ile Ala Leu Gly Arg Gln 1425 1430 1435 1440 Gly Ser Trp Trp Glu Met Phe Lys Phe Phe Phe His Arg Leu Tyr Asp 1445 1450 1455 His Gln Ile Val Pro Ser Thr Pro Ala Met Leu Asn Leu Gly Thr Arg 1460 1465 1470 Asn Tyr Tyr Thr Ser Ser Cys Tyr Leu Val Asn Pro Gln Ala Thr Thr 1475 1480 1485 Asn Gln Ala Thr Leu Arg Ala Ile Thr Gly Asn Val Ser Ala Ile Leu 1490 1495 1500 Ala Arg Asn Gly Gly Ile Gly Leu Cys Met Gln Ala Phe Asn Asp Ala 1505 1510 1515 1520 Ser Pro Gly Thr Ala Ser Ile Met Pro Ala Leu Lys Val Leu Asp Ser 1525 1530 1535 Leu Val Ala Ala His Asn Lys Gln Ser Thr Arg Pro Thr Gly Ala Cys 1540 1545 1550 Val Tyr Leu Glu Pro Trp His Ser Asp Val Arg Ala Val Leu Arg Met 1555 1560 1565 Lys Gly Val Leu Ala Gly Glu Glu Ala Gln Arg Cys Asp Asn Ile Phe 1570 1575 1580 Ser Ala Leu Trp Met Pro Asp Leu Phe Phe Lys Arg Leu Ile Arg His 1585 1590 1595 1600 Leu Asp Gly Glu Lys Asn Val Thr Trp Ser Leu Phe Asp Arg Asp Thr 1605 1610 1615 Ser Met Ser Leu Ala Asp Phe His Gly Glu Glu Phe Glu Lys Leu Tyr 1620 1625 1630 Glu His Leu Glu Ala Met Gly Phe Gly Glu Thr Ile Pro Ile Gln Asp 1635 1640 1645 Leu Ala Tyr Ala Ile Val Arg Ser Ala Ala Thr Thr Gly Ser Pro Phe 1650 1655 1660 Ile Met Phe Lys Asp Ala Val Asn Arg His Tyr Ile Tyr Asp Thr Gln 1665 1670 1675 1680 Gly Ala Ala Ile Ala Gly Ser Asn Leu Cys Thr Glu Ile Val His Pro 1685 1690 1695 Ala Ser Lys Arg Ser Ser Gly Val Cys Asn Leu Gly Ser Val Asn Leu 1700 1705 1710 Ala Arg Cys Val Ser Arg Gln Thr Phe Asp Phe Gly Arg Leu Arg Asp 1715 1720 1725 Ala Val Gln Ala Cys Val Leu Met Val Asn Ile Met Ile Asp Ser Thr 1730 1735 1740 Leu Gln Pro Thr Pro Gln Cys Thr Arg Gly Asn Asp Asn Leu Arg Ser 1745 1750 1755 1760 Met Gly Ile Gly Met Gln Gly Leu His Thr Ala Cys Leu Lys Met Gly 1765 1770 1775 Leu Asp Leu Glu Ser Ala Glu Phe Arg Asp Leu Asn Thr His Ile Ala 1780 1785 1790 Glu Val Met Leu Leu Ala Ala Met Lys Thr Ser Asn Ala Leu Cys Val 1795 1800 1805 Arg Gly Ala Arg Pro Phe Ser His Phe Lys Arg Ser Met Tyr Arg Ala 1810 1815 1820 Gly Arg Phe His Trp Glu Arg Phe Ser Asn Ala Ser Pro Arg Tyr Glu 1825 1830 1835 1840 Gly Glu Trp Glu Met Leu Arg Gln Ser Met Met Lys His Gly Leu Arg 1845 1850 1855 Asn Ser Gln Phe Ile Ala Leu Met Pro Thr Ala Ala Ser Ala Gln Ile 1860 1865 1870 Ser Asp Val Ser Glu Gly Phe Ala Pro Leu Phe Thr Asn Leu Phe Ser 1875 1880 1885 Lys Val Thr Arg Asp Gly Glu Thr Leu Arg Pro Asn Thr Leu Leu Leu 1890 1895 1900 Lys Glu Leu Glu Arg Thr Phe Gly Gly Lys Arg Leu Leu Asp Ala Met 1905 1910 1915 1920 Asp Gly Leu Glu Ala Lys Gln Trp Ser Val Ala Gln Ala Leu Pro Cys 1925 1930 1935 Leu Asp Pro Ala His Pro Leu Arg Arg Phe Lys Thr Ala Phe Asp Tyr 1940 1945 1950 Asp Gln Glu Leu Leu Ile Asp Leu Cys Ala Asp Arg Ala Pro Tyr Val 1955 1960 1965 Asp His Ser Gln Ser Met Thr Leu Tyr Val Thr Glu Lys Ala Asp Gly 1970 1975 1980 Thr Leu Pro Ala Ser Thr Leu Val Arg Leu Leu Val His Ala Tyr Lys 1985 1990 1995 2000 Arg Gly Leu Lys Thr Gly Met Tyr Thr Leu Ser Arg Ser Ser Ser Ile 2005 2010 2015 Trp Ala Ala Gly Ala Thr Asp Asp Asp Asp Ser Asp Ser Asp Ser Arg 2020 2025 2030 Ser Asp Asp Ser Val Gln Pro Asp Val Val Val Arg Arg Arg Trp Ser 2035 2040 2045 Asp Gly Pro Ala Pro Val Ala Phe Pro Lys Pro Arg Arg Pro Gly Asp 2050 2055 2060 Ser Pro Gly Asn Pro Gly Leu Gly Ala Gly Thr Gly Pro Gly Ser Ala 2065 2070 2075 2080 Thr Asp Pro Arg Ala Ser Ala Asp Ser Asp Ser Ala Ala His Ala Ala 2085 2090 2095 Ala Pro Gln Ala Asp Val Ala Pro Val Leu Asp Ser Gln Pro Thr Val 2100 2105 2110 Gly Thr Asp Pro Gly Tyr Pro Val Pro Leu Glu Leu Thr Pro Glu Asn 2115 2120 2125 Ala Glu Ala Val Ala Arg Phe Leu Gly Asp Ala Val Asp Arg Glu Pro 2130 2135 2140 Ala Leu Met Leu Glu Tyr Phe Cys Arg Cys Ala Arg Glu Glu Ser Lys 2145 2150 2155 2160 Arg Val Pro Pro Arg Thr Phe Gly Ser Ala Pro Arg Leu Thr Glu Asp 2165 2170 2175 Asp Phe Gly Leu Leu Asn Tyr Ala Leu Ala Glu Met Arg Arg Leu Cys 2180 2185 2190 Leu Asp Leu Pro Pro Val Pro 2195 <210> 12 <211> 6600 <212> DNA <213> Artificial Sequence <220> <223> polynucleotide encoding tPA-Flt3L-gB-shuffled UL39 fusion protein <400> 12 atggacgcca tgaagagagg cctgtgctgc gtgctgctgc tgtgcggcgc cgtgttcgtg 60 agccccagcc acgccaccca ggactgcagc ttccagcaca gccccatcag cagcgacttc 120 gccgtgaaga tcagagagct gagcgactac ctgctgcagg actaccccgt gaccgtggcc 180 agcaacctgc aggacgagga gctgtgcggc ggcctgtgga gactggtgct ggcccagaga 240 tggatggaga gactgaagac cgtggccggc agcaagatgc agggcctgct ggagagagtg 300 aacaccgaga tccacttcgt gaccaagtgc gccttccagc ctccccccag ctgcctgagg 360 ttcgtgcaga ccaacatcag cagactgctg caggagacca gcgagcagct ggtggccctg 420 aagccctgga tcaccagaca gaacttcagc agatgcctgg agctgcagtg ccagcccgac 480 agcagcaccc tgccccctcc ctggagcccc agacccctgg aggccaccgc tcccacagcc 540 cctggcagcg ggtccggaag tgggtctgga tctgcaccag cagcaccagc cgctccaagg 600 gcatcagggg gcgtcgcagc aactgtggca gcaaatgggg gaccagcctc cagacctcca 660 ccagtgccca gccctgcaac cacaaaggcc cgcaagcgaa aaactaagaa accacctaaa 720 cgacctgagg caaccccacc acctgacgca aacgctactg tggcagctgg acacgctacc 780 ctgcgagcac atctgagaga gatcaaggtc gaaaatgcag atgcccagtt ctacgtgtgc 840 ccacccccta caggagccac tgtggtccag tttgagcagc ctcggagatg tcccacaaga 900 cctgaggggc agaactacac tgaaggaatc gctgtggtct tcaaggaaaa catcgcccca 960 tacaagttta aagctacaat gtactacaaa gacgtgactg tctcacaagt gtggttcggc 1020 cacagataca gccagttcat ggggattttt gaggacaggg ccccagtgcc ctttgaggaa 1080 gtgatcgaca agattaacac caaaggcgtc tgcagatcca cagccaaata tgtgaggaac 1140 aatatggaaa ctaccgcttt ccacagggac gatcatgaga cagacatgga actgaagccc 1200 gcaaaagtgg ccaccaggac aagtcgcgga tggcacacaa ctgatctgaa gtacaaccct 1260 tcacgggtcg aggccttcca tagatatggc accacagtga attgtatcgt ggaggaagtc 1320 gatgcacgga gcgtgtaccc ttatgacgaa tttgtcctgg ccaccggcga tttcgtgtac 1380 atgagcccct tctacgggta tagggaggga agccacaccg aacatacatc ctacgcagcc 1440 gaccgcttca agcaggtgga tgggttttat gcccgcgacc tgactaccaa agcccgggcc 1500 accagcccta caactcggaa cctgctgacc acaccaaagt tcaccgtggc ttgggactgg 1560 gtcccaaagc gaccagcagt ctgcactatg accaaatggc aggaggtgga cgaaatgctg 1620 cgagctgagt acggaggcag cttccggttc agcagcgatg caatttctac tacctttaca 1680 actaatctga ccgagtatag cctgtccaga gtggacctgg gcgattgtat cgggcgagat 1740 gctcgggaag caattgacag gatgttcgct cgcaagtaca acgcaacaca catcaaagtg 1800 ggccagccac agtactatct ggcaactgga ggctttctga ttgcctatca gcccctgctg 1860 agcaatacac tggccgagct gtacgtgcga gagtatatgc gggaacagga cagaaagcca 1920 aggaacgcaa cccctgctcc actgcgagaa gcaccttcag ccaatgctag cgtggagcgg 1980 atcaaaacca catctagtat tgagttcgct agactgcagt ttacctacaa ccacatccag 2040 agacatgtca atgatatgct gggcaggatt gcagtggcct ggtgcgagct gcagaaccat 2100 gaactgacac tgtggaatga ggcccggaag ctgaacccaa atgctatcgc atctgccact 2160 gtgggaaggc gcgtcagtgc cagaatgctg ggcgacgtga tggctgtcag cacatgcgtg 2220 cctgtcgcac cagataacgt gattgtccag aacagcatga gagtgtcaag caggccaggc 2280 acctgttaca gtaggcccct ggtgtcattc cgctatgaag accagggacc actgatcgag 2340 ggacagctgg gggagaacaa tgaactgcga ctgacccgag atgccctgga gccatgcaca 2400 gtgggccacc gacggtattt catttttggg ggaggctacg tgtattttga ggaatacgcc 2460 tattcccatc agctgtctag ggctgacgtg actaccgtct ctactttcat cgacctgaac 2520 attaccatgc tggaggatca cgaatttgtg cccctggagg tctacacccg ccatgaaatc 2580 aaggacagcg gactgctgga ttatacagag gtgcagagaa ggaaccagct gcacgacctg 2640 cgcttcgccg acatcgacac cgtgattcgg gctgatgcaa atgctgcaat gtttgccggg 2700 ctgtgcgctt tctttgaggg aatgggcgac ctgggacggg ccgtgggcaa ggtggtcatg 2760 ggagtggtcg gaggagtggt ctctgctgtg agtggagtct cctctttcat gagcaacccc 2820 agatacgtgc tgcagctgca gaggaatccc atgaaagccc tgtatcctct gacaactaag 2880 gagctgaaaa cctccgaccc tggaggagtg ggaggagagg gagaggaagg cgcagaaggg 2940 ggaggcttcg atgaggccaa gctggccgag gctcgcgaaa tgatccgata catggcactg 3000 gtgtctgcca tggagcgaac tgaacacaag gctcggaaga aaggcaccag tgcactgctg 3060 agttcaaaag tgacaaacat ggtcctgcgg aagagaaaca aggcccggta ttctccactg 3120 cataatgaag acgaggctgg cgacgaagac gaactggggg gcggaggatc tgggggcggg 3180 gggagcgggg gcgggggaag cggagggggg ggaagcggag ggggagggag cagaagtcca 3240 agcgaaaggc aggaacctag agagccagaa gtggcaccac ctggagggga ccacgtcttc 3300 tgcaggaagg tgtctggggt catggtgctg agctccgatc caccaggacc tgcagcttac 3360 cggatcagtg actctagttt tgtgcagtgc gggagcaact gttccatgat cattgacggc 3420 gatggggacg gaagaacagc tgtggtcgca ctgggaggca cttctggacc cagtgccacc 3480 acatccgtgg gcacccagac atctggggaa tttctgcacg gaaatccccg aacccctgag 3540 ccacagggac ctcaggccgt gcctccaccc ccttattgta aggtcagaaa agctaccaac 3600 agtggagtgt tcgcaggcga cgataatatt gtctgcacat catgtgccct gccaccccct 3660 tttccatggg gccatgagtg ctgtgcccgg agagatgcta gggggggagc agaaaaagac 3720 gtgggggcag ccgagagttg gtcagatgga ccctcaagcg acagcgagac cgaagattcc 3780 gactcctctg atgaagacac tggcagtgag accctgtcac gcagttcaag catctgggct 3840 gcaggggcca cagacgatga cgattccgac ccaaacgctt acacacccta tcacctgagg 3900 gaatacgcaa ctcgcctggt gaatgggttc aagcctctgg tcaggcgcag cgcccgcctg 3960 tatagaatcc tgggagtcct ggtgcatctg agaatcagga cacgcgaggc tagcttcgag 4020 gaatggatga ggtccaaaga agtggatctg gactttggcc tgactgagcg actgcgggag 4080 cacgaagccc agctgatgat tctggcacag gccctgaacc catacgattg cctgatccat 4140 tcaactccca ataccctggt ggaacgagga ctgcagagcg ccctgaagta cgaggagttc 4200 tacctgaaac ggtttggcgg gcactacatg gagagcgtgt tccagatgta taccagaatt 4260 gccggctttc tggcctgtcg ggctacaaga gggatgaggc atatcgctct gggccgccag 4320 gggagctggt gggagatgtt taagttcttt ttccaccggc tgtacgacca tcagatcgtg 4380 ccatccaccc ccgccatgct gaacctgggc actcggaatt actatacctc ctcttgctat 4440 ctggtgaacc cccaggccac taccaatcag gccacactgc gcgctatcac tgggaacgtg 4500 agcgcaattc tggcccgaaa tggaggcatc ggactgtgca tgcaggcatt caacgatgcc 4560 tctcctggca ccgccagtat catgccagct ctgaaggtcc tggactccct ggtggccgct 4620 cacaacaagc agtctacaag gcctactggc gcctgcgtgt acctggagcc atggcattct 4680 gatgtcagag ctgtgctgag gatgaagggg gtgctggctg gagaggaagc acagcggtgc 4740 gataacattt tcagcgccct gtggatgcct gacctgtttt tcaagcgcct gatccgacac 4800 ctggatggag agaaaaatgt gacctggagc ctgtttgatc gggacacaag catgtccctg 4860 gccgacttcc acggcgagga atttgaaaaa ctgtacgaac atctggaggc tatgggcttc 4920 ggggagacca tcccaattca ggacctggct tatgcaattg tgaggagtgc agccacaact 4980 ggctcaccct ttatcatgtt caaggatgcc gtcaaccgcc actacatcta cgacactcag 5040 ggcgctgcaa tcgctgggtc taatctgtgc accgagatcg tgcatccagc aagtaaaaga 5100 agttcaggag tctgcaacct gggctcagtg aatctggcac gatgcgtgag ccggcagacc 5160 ttcgacttcg gaagactgag ggacgctgtg caggcatgcg tcctgatggt gaacatcatg 5220 attgatagca cactgcagcc cactcctcag tgtacccgag gcaacgacaa tctgcggtcc 5280 atgggaattg gcatgcaggg actgcacaca gcctgcctga agatgggcct ggatctggaa 5340 tctgccgagt tccgggacct gaacacacat atcgctgaag tgatgctgct ggccgctatg 5400 aagactagca atgcactgtg cgtgcgggga gccagacctt tttcacactt caaaagaagc 5460 atgtacaggg caggccgctt ccattgggag cggttttcaa acgccagccc aagatatgag 5520 ggggaatggg agatgctgag acagagtatg atgaagcacg gactgcggaa cagccagttt 5580 attgcactga tgcccactgc agcctctgcc cagatcagcg acgtgagcga aggctttgcc 5640 cctctgttca ccaacctgtt tagcaaagtc accagagacg gggagacact gaggcctaat 5700 actctgctgc tgaaggaact ggagcgcact ttcgggggaa aacgactgct ggatgctatg 5760 gacggcctgg aggcaaagca gtggtccgtg gcacaggctc tgccatgcct ggaccctgct 5820 catccactgc gacggttcaa aacagcattt gattacgacc aggaactgct gatcgacctg 5880 tgcgccgaca gagctcccta cgtggatcac tctcagagta tgacactgta tgtcactgag 5940 aaggccgacg gcaccctgcc tgctagcaca ctggtcaggc tgctggtgca tgcctacaag 6000 cgcggcctga aaaccgggat gtatacactg tccaggagct cctctatctg ggccgccggg 6060 gccaccgacg atgacgattc agatagcgac tcccgctctg acgattccgt gcagccagat 6120 gtggtcgtga gaaggcgctg gtctgacgga ccagcaccag tggcattccc taaaccacga 6180 cggcccggag atagcccagg caaccctgga ctgggagcag gcactgggcc tggatctgct 6240 accgacccaa gggcaagtgc cgatagtgac tcagccgctc acgcagccgc tccacaggct 6300 gatgtcgcac ctgtgctgga ctcccagcca accgtgggga cagaccccgg ataccccgtc 6360 cctctggagc tgacccctga aaatgctgag gcagtggccc gattcctggg cgatgcagtg 6420 gaccgggaac cagccctgat gctggagtat ttttgccgat gtgcccggga ggaaagcaag 6480 cgggtgccac caagaacttt cggctccgct ccacgactga ccgaggacga ttttgggctg 6540 ctgaactatg ccctggccga aatgcggaga ctgtgcctgg acctgccccc cgtgccctaa 6600 6600 <210> 13 <211> 344 <212> PRT <213> Artificial Sequence <220> <223> HSV-2 gD <400> 13 Lys Tyr Ala Leu Ala Asp Pro Ser Leu Lys Met Ala Asp Pro Asn Arg 1 5 10 15 Phe Arg Gly Lys Asn Leu Pro Val Leu Asp Gln Leu Thr Asp Pro Pro 20 25 30 Gly Val Lys Arg Val Tyr His Ile Gln Pro Ser Leu Glu Asp Pro Phe 35 40 45 Gln Pro Pro Ser Ile Pro Ile Thr Val Tyr Tyr Ala Val Leu Glu Arg 50 55 60 Ala Cys Arg Ser Val Leu Leu His Ala Pro Ser Glu Ala Pro Gln Ile 65 70 75 80 Val Arg Gly Ala Ser Asp Glu Ala Arg Lys His Thr Tyr Asn Leu Thr 85 90 95 Ile Ala Trp Tyr Arg Met Gly Asp Asn Cys Ala Ile Pro Ile Thr Val 100 105 110 Met Glu Tyr Thr Glu Cys Pro Tyr Asn Lys Ser Leu Gly Val Cys Pro 115 120 125 Ile Arg Thr Gln Pro Arg Trp Ser Tyr Tyr Asp Ser Phe Ser Ala Val 130 135 140 Ser Glu Asp Asn Leu Gly Phe Leu Met His Ala Pro Ala Phe Glu Thr 145 150 155 160 Ala Gly Thr Tyr Leu Arg Leu Val Lys Ile Asn Asp Trp Thr Glu Ile 165 170 175 Thr Gln Phe Ile Leu Glu His Arg Ala Arg Ala Ser Cys Lys Tyr Ala 180 185 190 Leu Pro Leu Arg Ile Pro Pro Ala Ala Cys Leu Thr Ser Lys Ala Tyr 195 200 205 Gln Gln Gly Val Thr Val Asp Ser Ile Gly Met Leu Pro Arg Phe Ile 210 215 220 Pro Glu Asn Gln Arg Thr Val Ala Leu Tyr Ser Leu Lys Ile Ala Gly 225 230 235 240 Trp His Gly Pro Lys Pro Pro Tyr Thr Ser Thr Leu Leu Pro Pro Glu 245 250 255 Leu Ser Asp Thr Thr Asn Ala Thr Gln Pro Glu Leu Val Pro Glu Asp 260 265 270 Pro Glu Asp Ser Ala Leu Leu Glu Asp Pro Ala Gly Thr Val Ser Ser 275 280 285 Gln Ile Pro Pro Asn Trp His Ile Pro Ser Ile Gln Asp Val Ala Pro 290 295 300 His His Ala Pro Ala Ala Pro Ser Asn Pro Arg Arg Arg Ala Gln Met 305 310 315 320 Ala Pro Lys Arg Leu Arg Leu Pro His Ile Arg Asp Asp Asp Ala Pro 325 330 335 Pro Ser His Gln Pro Leu Phe Tyr 340 <210> 14 <211> 818 <212> PRT <213> Artificial Sequence <220> <223> HSV-2 ICP0 variant <400> 14 Met Glu Pro Arg Pro Gly Thr Ser Ser Arg Ala Asp Pro Gly Pro Glu 1 5 10 15 Arg Pro Pro Arg Gln Thr Pro Gly Thr Gln Pro Ala Ala Pro His Ala 20 25 30 Trp Gly Met Leu Asn Asp Met Gln Trp Leu Ala Ser Ser Asp Ser Glu 35 40 45 Glu Glu Thr Glu Val Gly Ile Ser Asp Asp Asp Leu His Arg Asp Ser 50 55 60 Thr Ser Glu Ala Gly Ser Thr Asp Thr Glu Met Phe Glu Ala Gly Leu 65 70 75 80 Met Asp Ala Ala Thr Pro Pro Ala Arg Pro Pro Ala Glu Arg Gln Gly 85 90 95 Ser Pro Thr Pro Ala Asp Ala Gln Gly Ser Cys Gly Gly Gly Pro Val 100 105 110 Gly Glu Glu Glu Ala Glu Ala Gly Gly Gly Gly Asp Val Cys Ala Val 115 120 125 Cys Thr Asp Glu Ile Ala Pro Pro Leu Arg Cys Gln Ser Phe Pro Cys 130 135 140 Leu His Pro Phe Cys Ile Pro Cys Met Lys Thr Trp Ile Pro Leu Arg 145 150 155 160 Asn Thr Cys Pro Leu Cys Asn Thr Pro Val Ala Tyr Leu Ile Val Gly 165 170 175 Val Thr Ala Ser Gly Ser Phe Ser Thr Ile Pro Ile Val Asn Asp Pro 180 185 190 Arg Thr Arg Val Glu Ala Glu Ala Ala Val Arg Ala Gly Thr Ala Val 195 200 205 Asp Phe Ile Trp Thr Gly Asn Pro Arg Thr Ala Pro Arg Ser Leu Ser 210 215 220 Leu Gly Gly His Thr Val Arg Ala Leu Ser Pro Thr Pro Pro Trp Pro 225 230 235 240 Gly Thr Asp Asp Glu Asp Asp Asp Leu Ala Asp Val Asp Tyr Val Pro 245 250 255 Pro Ala Pro Arg Arg Ala Pro Arg Arg Gly Gly Gly Gly Ala Gly Ala 260 265 270 Thr Arg Gly Thr Ser Gln Pro Ala Ala Thr Arg Pro Ala Pro Pro Gly 275 280 285 Ala Pro Arg Ser Ser Ser Ser Gly Gly Ala Pro Leu Arg Ala Gly Val 290 295 300 Gly Ser Gly Ser Gly Gly Gly Pro Ala Val Ala Ala Val Val Pro Arg 305 310 315 320 Val Ala Ser Leu Pro Pro Ala Ala Gly Gly Gly Arg Ala Gln Ala Arg 325 330 335 Arg Val Gly Glu Asp Ala Ala Ala Ala Glu Gly Arg Thr Pro Pro Ala 340 345 350 Arg Gln Pro Arg Ala Ala Gln Glu Pro Pro Ile Val Ile Ser Asp Ser 355 360 365 Pro Pro Pro Ser Pro Arg Arg Pro Ala Gly Pro Gly Pro Leu Ser Phe 370 375 380 Val Ser Ser Ser Ser Ala Gln Val Ser Ser Gly Pro Gly Gly Gly Gly 385 390 395 400 Leu Pro Gln Ser Ser Gly Arg Ala Ala Arg Pro Arg Ala Ala Val Ala 405 410 415 Pro Arg Val Arg Ser Pro Pro Arg Ala Ala Ala Ala Pro Val Val Ser 420 425 430 Ala Ser Ala Asp Ala Ala Gly Pro Ala Pro Pro Ala Val Pro Val Asp 435 440 445 Ala His Arg Ala Pro Arg Ser Arg Met Thr Gln Ala Gln Thr Asp Thr 450 455 460 Gln Ala Gln Ser Leu Gly Arg Ala Gly Ala Thr Asp Ala Arg Gly Ser 465 470 475 480 Gly Gly Pro Gly Ala Glu Gly Gly Pro Gly Val Pro Arg Gly Thr Asn 485 490 495 Thr Pro Gly Ala Ala Pro His Ala Ala Glu Gly Ala Ala Gly Ser Asp 500 505 510 Ser Gly Pro Ala Ala Ser Ser Ser Ala Ser Ser Ser Ala Ala Pro Arg 515 520 525 Ser Pro Leu Ala Pro Gln Gly Val Gly Ala Lys Arg Ala Ala Pro Arg 530 535 540 Arg Ala Pro Asp Ser Asp Ser Gly Asp Arg Gly His Gly Pro Leu Ala 545 550 555 560 Pro Ala Ser Ala Gly Ala Ala Pro Pro Ser Ala Ser Pro Ser Ser Gln 565 570 575 Ala Ala Val Ala Ala Ala Ser Ser Ser Ser Ala Ser Ser Ser Ser Ala 580 585 590 Ser Ser Ser Ser Ala Ser Ser Ser Ser Ala Ser Ser Ser Ser Ala Ser 595 600 605 Ser Ser Ser Ala Ser Ser Ser Ser Ala Ser Ser Ser Ala Gly Gly Ala 610 615 620 Gly Gly Ser Val Ala Ser Ala Ser Gly Ala Gly Glu Arg Arg Glu Thr 625 630 635 640 Ser Leu Gly Pro Arg Ala Ala Ala Pro Arg Gly Pro Arg Lys Cys Ala 645 650 655 Arg Lys Thr Arg His Ala Glu Gly Gly Pro Glu Pro Gly Ala Arg Asp 660 665 670 Pro Ala Pro Gly Leu Thr Arg Tyr Leu Pro Ile Ala Gly Val Ser Ser 675 680 685 Val Val Ala Leu Ala Pro Tyr Val Asn Lys Thr Val Thr Gly Asp Cys 690 695 700 Leu Pro Val Leu Asp Met Glu Thr Gly His Ile Gly Ala Tyr Val Val 705 710 715 720 Leu Val Asp Gln Thr Gly Asn Val Ala Asp Leu Leu Arg Ala Ala Ala 725 730 735 Pro Ala Trp Ser Arg Arg Thr Leu Leu Pro Glu His Ala Arg Asn Cys 740 745 750 Val Arg Pro Pro Asp Tyr Pro Thr Pro Pro Ala Ser Glu Trp Asn Ser 755 760 765 Leu Trp Met Thr Pro Val Gly Asn Met Leu Phe Asp Gln Gly Thr Leu 770 775 780 Val Gly Ala Leu Asp Phe His Gly Leu Arg Ser Arg His Pro Trp Ser 785 790 795 800 Arg Glu Gln Gly Ala Pro Ala Pro Ala Gly Asp Ala Pro Ala Gly His 805 810 815 Gly Glu <210> 15 <211> 766 <212> PRT <213> Artificial Sequence <220> <223> HSV-2 ICP4 variant <400> 15 Met Ser Ala Glu Gln Arg Lys Lys Lys Lys Thr Thr Thr Thr Thr Gln 1 5 10 15 Gly Arg Gly Ala Glu Val Ala Met Ala Asp Glu Asp Gly Gly Arg Leu 20 25 30 Arg Ala Ala Ala Glu Thr Thr Gly Gly Pro Gly Ser Pro Asp Pro Ala 35 40 45 Asp Gly Pro Pro Pro Thr Pro Asn Pro Asp Arg Arg Pro Ala Ala Arg 50 55 60 Pro Gly Phe Gly Trp His Gly Gly Pro Glu Glu Asn Glu Asp Glu Ala 65 70 75 80 Asp Asp Ala Ala Ala Asp Ala Asp Ala Asp Glu Ala Ala Pro Ala Ser 85 90 95 Gly Glu Ala Val Asp Glu Pro Ala Ala Asp Gly Val Val Ser Pro Arg 100 105 110 Gln Leu Ala Leu Leu Ala Ser Met Val Asp Glu Ala Val Arg Thr Ile 115 120 125 Pro Ser Pro Pro Pro Glu Arg Asp Gly Ala Gln Glu Glu Ala Ala Arg 130 135 140 Ser Pro Ser Pro Pro Arg Thr Pro Ser Met Arg Ala Asp Tyr Gly Glu 145 150 155 160 Glu Asn Asp Asp Asp Asp Asp Asp Asp Asp Asp Asp Asp Arg Asp Ala 165 170 175 Gly Arg Trp Val Arg Gly Pro Glu Thr Thr Ser Ala Val Arg Gly Ala 180 185 190 Tyr Pro Asp Pro Met Ala Ser Leu Ser Pro Arg Pro Pro Ala Pro Arg 195 200 205 Arg His His His His His His His Arg Arg Arg Arg Ala Pro Arg Arg 210 215 220 Arg Ser Ala Ala Ser Asp Ser Ser Lys Ser Gly Ser Ser Ser Ser Ala 225 230 235 240 Ser Ser Ala Ser Ser Ser Ala Ser Ser Ser Ser Ser Ala Ser Ala Ser 245 250 255 Ser Ser Asp Asp Asp Asp Asp Asp Asp Ala Ala Arg Ala Pro Ala Ser 260 265 270 Ala Ala Asp His Ala Ala Gly Gly Thr Leu Gly Ala Asp Asp Glu Glu 275 280 285 Ala Gly Val Pro Ala Arg Ala Pro Gly Ala Ala Pro Arg Pro Ser Pro 290 295 300 Pro Arg Ala Glu Pro Ala Pro Ala Arg Thr Pro Ala Ala Thr Ala Gly 305 310 315 320 Arg Leu Glu Arg Arg Arg Ala Arg Ala Ala Val Ala Gly Arg Asp Ala 325 330 335 Thr Gly Arg Phe Thr Ala Gly Arg Pro Arg Arg Val Glu Leu Asp Ala 340 345 350 Asp Ala Ala Ser Gly Ala Phe Tyr Ala Arg Tyr Arg Asp Gly Tyr Val 355 360 365 Ser Gly Glu Pro Trp Pro Gly Ala Gly Pro Pro Pro Pro Gly Arg Val 370 375 380 Leu Tyr Gly Gly Leu Gly Asp Ser Arg Pro Gly Leu Trp Gly Ala Pro 385 390 395 400 Glu Ala Glu Glu Ala Arg Ala Arg Phe Glu Ala Ser Gly Ala Pro Ala 405 410 415 Pro Val Trp Ala Pro Glu Leu Gly Asp Ala Ala Gln Gln Tyr Ala Leu 420 425 430 Ile Thr Arg Leu Leu Tyr Thr Pro Asp Ala Glu Ala Met Gly Trp Leu 435 440 445 Gln Asn Pro Arg Val Ala Pro Gly Asp Val Ala Leu Asp Gln Ala Cys 450 455 460 Phe Arg Ile Ser Gly Ala Ala Arg Asn Ser Ser Ser Phe Ile Ser Gly 465 470 475 480 Ser Val Ala Arg Ala Val Pro His Leu Gly Tyr Ala Met Ala Ala Gly 485 490 495 Arg Phe Gly Trp Gly Leu Ala His Val Ala Ala Ala Val Ala Met Ser 500 505 510 Arg Arg Tyr Asp Arg Ala Gln Lys Gly Phe Leu Leu Thr Ser Leu Arg 515 520 525 Arg Ala Tyr Ala Pro Leu Leu Ala Arg Glu Asn Ala Ala Leu Thr Gly 530 535 540 Ala Arg Thr Pro Asp Asp Gly Gly Asp Ala Asn Arg His Asp Gly Asp 545 550 555 560 Asp Ala Arg Gly Lys Pro Ala Ala Ala Ala Ala Pro Leu Pro Ser Ala 565 570 575 Ala Ala Ser Pro Ala Asp Glu Arg Ala Val Pro Ala Gly Tyr Gly Ala 580 585 590 Ala Gly Val Leu Ala Ala Leu Gly Arg Leu Ser Ala Ala Pro Ala Ser 595 600 605 Ala Pro Ala Gly Ala Asp Asp Asp Asp Asp Asp Asp Gly Ala Gly Gly 610 615 620 Gly Gly Gly Gly Arg Arg Ala Glu Ala Gly Arg Val Ala Val Glu Cys 625 630 635 640 Leu Ala Ala Cys Arg Gly Ile Leu Glu Ala Leu Ala Glu Gly Phe Asp 645 650 655 Gly Asp Leu Ala Ala Val Pro Gly Leu Ala Gly Ala Arg Pro Ala Ala 660 665 670 Pro Pro Arg Pro Gly Pro Ala Gly Ala Ala Ala Pro Pro His Ala Asp 675 680 685 Ala Pro Arg Leu Arg Ala Trp Leu Arg Glu Leu Arg Phe Val Arg Asp 690 695 700 Ala Leu Val Leu Met Arg Leu Arg Gly Asp Leu Arg Val Ala Gly Gly 705 710 715 720 Ser Glu Ala Ala Val Ala Ala Val Arg Ala Val Ser Leu Val Ala Gly 725 730 735 Ala Leu Gly Pro Ala Leu Pro Arg Ser Pro Arg Leu Leu Ser Ser Ala 740 745 750 Ala Ala Ala Ala Ala Asp Leu Leu Phe Gln Asn Gln Ser Leu 755 760 765 <210> 16 <211> 2172 <212> PRT <213> Artificial Sequence <220> <223> tPA-Flt3L-gD-IPC0-IPC4 fusion protein <400> 16 Met Asp Ala Met Lys Arg Gly Leu Cys Cys Val Leu Leu Leu Cys Gly 1 5 10 15 Ala Val Phe Val Ser Pro Ser His Ala Thr Gln Asp Cys Ser Phe Gln 20 25 30 His Ser Pro Ile Ser Ser Asp Phe Ala Val Lys Ile Arg Glu Leu Ser 35 40 45 Asp Tyr Leu Leu Gln Asp Tyr Pro Val Thr Val Ala Ser Asn Leu Gln 50 55 60 Asp Glu Glu Leu Cys Gly Gly Leu Trp Arg Leu Val Leu Ala Gln Arg 65 70 75 80 Trp Met Glu Arg Leu Lys Thr Val Ala Gly Ser Lys Met Gln Gly Leu 85 90 95 Leu Glu Arg Val Asn Thr Glu Ile His Phe Val Thr Lys Cys Ala Phe 100 105 110 Gln Pro Pro Pro Ser Cys Leu Arg Phe Val Gln Thr Asn Ile Ser Arg 115 120 125 Leu Leu Gln Glu Thr Ser Glu Gln Leu Val Ala Leu Lys Pro Trp Ile 130 135 140 Thr Arg Gln Asn Phe Ser Arg Cys Leu Glu Leu Gln Cys Gln Pro Asp 145 150 155 160 Ser Ser Thr Leu Pro Pro Pro Trp Ser Pro Arg Pro Leu Glu Ala Thr 165 170 175 Ala Pro Thr Ala Pro Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly 180 185 190 Arg Ala Lys Tyr Ala Leu Ala Asp Pro Ser Leu Lys Met Ala Asp Pro 195 200 205 Asn Arg Phe Arg Gly Lys Asn Leu Pro Val Leu Asp Gln Leu Thr Asp 210 215 220 Pro Pro Gly Val Lys Arg Val Tyr His Ile Gln Pro Ser Leu Glu Asp 225 230 235 240 Pro Phe Gln Pro Pro Ser Ile Pro Ile Thr Val Tyr Tyr Ala Val Leu 245 250 255 Glu Arg Ala Cys Arg Ser Val Leu Leu His Ala Pro Ser Glu Ala Pro 260 265 270 Gln Ile Val Arg Gly Ala Ser Asp Glu Ala Arg Lys His Thr Tyr Asn 275 280 285 Leu Thr Ile Ala Trp Tyr Arg Met Gly Asp Asn Cys Ala Ile Pro Ile 290 295 300 Thr Val Met Glu Tyr Thr Glu Cys Pro Tyr Asn Lys Ser Leu Gly Val 305 310 315 320 Cys Pro Ile Arg Thr Gln Pro Arg Trp Ser Tyr Tyr Asp Ser Phe Ser 325 330 335 Ala Val Ser Glu Asp Asn Leu Gly Phe Leu Met His Ala Pro Ala Phe 340 345 350 Glu Thr Ala Gly Thr Tyr Leu Arg Leu Val Lys Ile Asn Asp Trp Thr 355 360 365 Glu Ile Thr Gln Phe Ile Leu Glu His Arg Ala Arg Ala Ser Cys Lys 370 375 380 Tyr Ala Leu Pro Leu Arg Ile Pro Pro Ala Ala Cys Leu Thr Ser Lys 385 390 395 400 Ala Tyr Gln Gln Gly Val Thr Val Asp Ser Ile Gly Met Leu Pro Arg 405 410 415 Phe Ile Pro Glu Asn Gln Arg Thr Val Ala Leu Tyr Ser Leu Lys Ile 420 425 430 Ala Gly Trp His Gly Pro Lys Pro Pro Tyr Thr Ser Thr Leu Leu Pro 435 440 445 Pro Glu Leu Ser Asp Thr Thr Asn Ala Thr Gln Pro Glu Leu Val Pro 450 455 460 Glu Asp Pro Glu Asp Ser Ala Leu Leu Glu Asp Pro Ala Gly Thr Val 465 470 475 480 Ser Ser Gln Ile Pro Pro Asn Trp His Ile Pro Ser Ile Gln Asp Val 485 490 495 Ala Pro His His Ala Pro Ala Ala Pro Ser Asn Pro Arg Arg Arg Ala 500 505 510 Gln Met Ala Pro Lys Arg Leu Arg Leu Pro His Ile Arg Asp Asp Asp 515 520 525 Ala Pro Pro Ser His Gln Pro Leu Phe Tyr Gly Gly Gly Gly Ser Gly 530 535 540 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 545 550 555 560 Gly Gly Ser Met Glu Pro Arg Pro Gly Thr Ser Ser Arg Ala Asp Pro 565 570 575 Gly Pro Glu Arg Pro Pro Arg Gln Thr Pro Gly Thr Gln Pro Ala Ala 580 585 590 Pro His Ala Trp Gly Met Leu Asn Asp Met Gln Trp Leu Ala Ser Ser 595 600 605 Asp Ser Glu Glu Glu Thr Glu Val Gly Ile Ser Asp Asp Asp Leu His 610 615 620 Arg Asp Ser Thr Ser Glu Ala Gly Ser Thr Asp Thr Glu Met Phe Glu 625 630 635 640 Ala Gly Leu Met Asp Ala Ala Thr Pro Pro Ala Arg Pro Pro Ala Glu 645 650 655 Arg Gln Gly Ser Pro Thr Pro Ala Asp Ala Gln Gly Ser Cys Gly Gly 660 665 670 Gly Pro Val Gly Glu Glu Glu Ala Glu Ala Gly Gly Gly Gly Asp Val 675 680 685 Cys Ala Val Cys Thr Asp Glu Ile Ala Pro Pro Leu Arg Cys Gln Ser 690 695 700 Phe Pro Cys Leu His Pro Phe Cys Ile Pro Cys Met Lys Thr Trp Ile 705 710 715 720 Pro Leu Arg Asn Thr Cys Pro Leu Cys Asn Thr Pro Val Ala Tyr Leu 725 730 735 Ile Val Gly Val Thr Ala Ser Gly Ser Phe Ser Thr Ile Pro Ile Val 740 745 750 Asn Asp Pro Arg Thr Arg Val Glu Ala Glu Ala Ala Val Arg Ala Gly 755 760 765 Thr Ala Val Asp Phe Ile Trp Thr Gly Asn Pro Arg Thr Ala Pro Arg 770 775 780 Ser Leu Ser Leu Gly Gly His Thr Val Arg Ala Leu Ser Pro Thr Pro 785 790 795 800 Pro Trp Pro Gly Thr Asp Asp Glu Asp Asp Asp Leu Ala Asp Val Asp 805 810 815 Tyr Val Pro Pro Ala Pro Arg Arg Ala Pro Arg Arg Gly Gly Gly Gly 820 825 830 Ala Gly Ala Thr Arg Gly Thr Ser Gln Pro Ala Ala Thr Arg Pro Ala 835 840 845 Pro Pro Gly Ala Pro Arg Ser Ser Ser Ser Gly Gly Ala Pro Leu Arg 850 855 860 Ala Gly Val Gly Ser Gly Ser Gly Gly Gly Pro Ala Val Ala Ala Val 865 870 875 880 Val Pro Arg Val Ala Ser Leu Pro Pro Ala Ala Gly Gly Gly Arg Ala 885 890 895 Gln Ala Arg Arg Val Gly Glu Asp Ala Ala Ala Ala Glu Gly Arg Thr 900 905 910 Pro Pro Ala Arg Gln Pro Arg Ala Ala Gln Glu Pro Pro Ile Val Ile 915 920 925 Ser Asp Ser Pro Pro Pro Ser Pro Arg Arg Pro Ala Gly Pro Gly Pro 930 935 940 Leu Ser Phe Val Ser Ser Ser Ser Ala Gln Val Ser Ser Gly Pro Gly 945 950 955 960 Gly Gly Gly Leu Pro Gln Ser Ser Gly Arg Ala Ala Arg Pro Arg Ala 965 970 975 Ala Val Ala Pro Arg Val Arg Ser Pro Pro Arg Ala Ala Ala Ala Pro 980 985 990 Val Val Ser Ala Ser Ala Asp Ala Ala Gly Pro Ala Pro Pro Ala Val 995 1000 1005 Pro Val Asp Ala His Arg Ala Pro Arg Ser Arg Met Thr Gln Ala Gln 1010 1015 1020 Thr Asp Thr Gln Ala Gln Ser Leu Gly Arg Ala Gly Ala Thr Asp Ala 1025 1030 1035 1040 Arg Gly Ser Gly Gly Pro Gly Ala Glu Gly Gly Pro Gly Val Pro Arg 1045 1050 1055 Gly Thr Asn Thr Pro Gly Ala Ala Pro His Ala Ala Glu Gly Ala Ala 1060 1065 1070 Gly Ser Asp Ser Gly Pro Ala Ala Ser Ser Ser Ala Ser Ser Ser Ala 1075 1080 1085 Ala Pro Arg Ser Pro Leu Ala Pro Gln Gly Val Gly Ala Lys Arg Ala 1090 1095 1100 Ala Pro Arg Arg Ala Pro Asp Ser Asp Ser Gly Asp Arg Gly His Gly 1105 1110 1115 1120 Pro Leu Ala Pro Ala Ser Ala Gly Ala Ala Pro Pro Ser Ala Ser Pro 1125 1130 1135 Ser Ser Gln Ala Ala Val Ala Ala Ala Ser Ser Ser Ser Ala Ser Ser 1140 1145 1150 Ser Ser Ala Ser Ser Ser Ser Ala Ser Ser Ser Ser Ala Ser Ser Ser 1155 1160 1165 Ser Ala Ser Ser Ser Ser Ala Ser Ser Ser Ser Ala Ser Ser Ser Ala 1170 1175 1180 Gly Gly Ala Gly Gly Ser Val Ala Ser Ala Ser Gly Ala Gly Glu Arg 1185 1190 1195 1200 Arg Glu Thr Ser Leu Gly Pro Arg Ala Ala Ala Pro Arg Gly Pro Arg 1205 1210 1215 Lys Cys Ala Arg Lys Thr Arg His Ala Glu Gly Gly Pro Glu Pro Gly 1220 1225 1230 Ala Arg Asp Pro Ala Pro Gly Leu Thr Arg Tyr Leu Pro Ile Ala Gly 1235 1240 1245 Val Ser Ser Val Val Ala Leu Ala Pro Tyr Val Asn Lys Thr Val Thr 1250 1255 1260 Gly Asp Cys Leu Pro Val Leu Asp Met Glu Thr Gly His Ile Gly Ala 1265 1270 1275 1280 Tyr Val Val Leu Val Asp Gln Thr Gly Asn Val Ala Asp Leu Leu Arg 1285 1290 1295 Ala Ala Ala Pro Ala Trp Ser Arg Arg Thr Leu Leu Pro Glu His Ala 1300 1305 1310 Arg Asn Cys Val Arg Pro Pro Asp Tyr Pro Thr Pro Pro Ala Ser Glu 1315 1320 1325 Trp Asn Ser Leu Trp Met Thr Pro Val Gly Asn Met Leu Phe Asp Gln 1330 1335 1340 Gly Thr Leu Val Gly Ala Leu Asp Phe His Gly Leu Arg Ser Arg His 1345 1350 1355 1360 Pro Trp Ser Arg Glu Gln Gly Ala Pro Ala Pro Ala Gly Asp Ala Pro 1365 1370 1375 Ala Gly His Gly Glu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 1380 1385 1390 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Met Ser 1395 1400 1405 Ala Glu Gln Arg Lys Lys Lys Lys Thr Thr Thr Thr Thr Gln Gly Arg 1410 1415 1420 Gly Ala Glu Val Ala Met Ala Asp Glu Asp Gly Gly Arg Leu Arg Ala 1425 1430 1435 1440 Ala Ala Glu Thr Thr Gly Gly Pro Gly Ser Pro Asp Pro Ala Asp Gly 1445 1450 1455 Pro Pro Pro Thr Pro Asn Pro Asp Arg Arg Pro Ala Ala Arg Pro Gly 1460 1465 1470 Phe Gly Trp His Gly Gly Pro Glu Glu Asn Glu Asp Glu Ala Asp Asp 1475 1480 1485 Ala Ala Ala Asp Ala Asp Ala Asp Glu Ala Ala Pro Ala Ser Gly Glu 1490 1495 1500 Ala Val Asp Glu Pro Ala Ala Asp Gly Val Val Ser Pro Arg Gln Leu 1505 1510 1515 1520 Ala Leu Leu Ala Ser Met Val Asp Glu Ala Val Arg Thr Ile Pro Ser 1525 1530 1535 Pro Pro Pro Glu Arg Asp Gly Ala Gln Glu Glu Ala Ala Arg Ser Pro 1540 1545 1550 Ser Pro Pro Arg Thr Pro Ser Met Arg Ala Asp Tyr Gly Glu Glu Asn 1555 1560 1565 Asp Asp Asp Asp Asp Asp Asp Asp Asp Asp Asp Arg Asp Ala Gly Arg 1570 1575 1580 Trp Val Arg Gly Pro Glu Thr Thr Ser Ala Val Arg Gly Ala Tyr Pro 1585 1590 1595 1600 Asp Pro Met Ala Ser Leu Ser Pro Arg Pro Pro Ala Pro Arg Arg His 1605 1610 1615 His His His His His His Arg Arg Arg Arg Ala Pro Arg Arg Arg Ser 1620 1625 1630 Ala Ala Ser Asp Ser Ser Lys Ser Gly Ser Ser Ser Ser Ala Ser Ser 1635 1640 1645 Ala Ser Ser Ser Ala Ser Ser Ser Ser Ser Ala Ser Ala Ser Ser Ser 1650 1655 1660 Asp Asp Asp Asp Asp Asp Asp Ala Ala Arg Ala Pro Ala Ser Ala Ala 1665 1670 1675 1680 Asp His Ala Ala Gly Gly Thr Leu Gly Ala Asp Asp Glu Glu Ala Gly 1685 1690 1695 Val Pro Ala Arg Ala Pro Gly Ala Ala Pro Arg Pro Ser Pro Pro Arg 1700 1705 1710 Ala Glu Pro Ala Pro Ala Arg Thr Pro Ala Ala Thr Ala Gly Arg Leu 1715 1720 1725 Glu Arg Arg Arg Ala Arg Ala Ala Val Ala Gly Arg Asp Ala Thr Gly 1730 1735 1740 Arg Phe Thr Ala Gly Arg Pro Arg Arg Val Glu Leu Asp Ala Asp Ala 1745 1750 1755 1760 Ala Ser Gly Ala Phe Tyr Ala Arg Tyr Arg Asp Gly Tyr Val Ser Gly 1765 1770 1775 Glu Pro Trp Pro Gly Ala Gly Pro Pro Pro Pro Gly Arg Val Leu Tyr 1780 1785 1790 Gly Gly Leu Gly Asp Ser Arg Pro Gly Leu Trp Gly Ala Pro Glu Ala 1795 1800 1805 Glu Glu Ala Arg Ala Arg Phe Glu Ala Ser Gly Ala Pro Ala Pro Val 1810 1815 1820 Trp Ala Pro Glu Leu Gly Asp Ala Ala Gln Gln Tyr Ala Leu Ile Thr 1825 1830 1835 1840 Arg Leu Leu Tyr Thr Pro Asp Ala Glu Ala Met Gly Trp Leu Gln Asn 1845 1850 1855 Pro Arg Val Ala Pro Gly Asp Val Ala Leu Asp Gln Ala Cys Phe Arg 1860 1865 1870 Ile Ser Gly Ala Ala Arg Asn Ser Ser Ser Phe Ile Ser Gly Ser Val 1875 1880 1885 Ala Arg Ala Val Pro His Leu Gly Tyr Ala Met Ala Ala Gly Arg Phe 1890 1895 1900 Gly Trp Gly Leu Ala His Val Ala Ala Ala Val Ala Met Ser Arg Arg 1905 1910 1915 1920 Tyr Asp Arg Ala Gln Lys Gly Phe Leu Leu Thr Ser Leu Arg Arg Ala 1925 1930 1935 Tyr Ala Pro Leu Leu Ala Arg Glu Asn Ala Ala Leu Thr Gly Ala Arg 1940 1945 1950 Thr Pro Asp Asp Gly Gly Asp Ala Asn Arg His Asp Gly Asp Asp Ala 1955 1960 1965 Arg Gly Lys Pro Ala Ala Ala Ala Ala Pro Leu Pro Ser Ala Ala Ala 1970 1975 1980 Ser Pro Ala Asp Glu Arg Ala Val Pro Ala Gly Tyr Gly Ala Ala Gly 1985 1990 1995 2000 Val Leu Ala Ala Leu Gly Arg Leu Ser Ala Ala Pro Ala Ser Ala Pro 2005 2010 2015 Ala Gly Ala Asp Asp Asp Asp Asp Asp Asp Gly Ala Gly Gly Gly Gly 2020 2025 2030 Gly Gly Arg Arg Ala Glu Ala Gly Arg Val Ala Val Glu Cys Leu Ala 2035 2040 2045 Ala Cys Arg Gly Ile Leu Glu Ala Leu Ala Glu Gly Phe Asp Gly Asp 2050 2055 2060 Leu Ala Ala Val Pro Gly Leu Ala Gly Ala Arg Pro Ala Ala Pro Pro 2065 2070 2075 2080 Arg Pro Gly Pro Ala Gly Ala Ala Ala Pro Pro His Ala Asp Ala Pro 2085 2090 2095 Arg Leu Arg Ala Trp Leu Arg Glu Leu Arg Phe Val Arg Asp Ala Leu 2100 2105 2110 Val Leu Met Arg Leu Arg Gly Asp Leu Arg Val Ala Gly Gly Ser Glu 2115 2120 2125 Ala Ala Val Ala Ala Val Arg Ala Val Ser Leu Val Ala Gly Ala Leu 2130 2135 2140 Gly Pro Ala Leu Pro Arg Ser Pro Arg Leu Leu Ser Ser Ala Ala Ala 2145 2150 2155 2160 Ala Ala Ala Asp Leu Leu Phe Gln Asn Gln Ser Leu 2165 2170 <210> 17 <211> 6519 <212> DNA <213> Artificial Sequence <220> <223> polynucleotide encoding tPA-Flt3L-gD-IPC0-IPC4 fusion protein <400> 17 atggacgcca tgaagagagg cctgtgctgc gtgctgctgc tgtgcggcgc cgtgttcgtg 60 agccccagcc acgccaccca ggactgcagc ttccagcaca gccccatcag cagcgacttc 120 gccgtgaaga tcagagagct gagcgactac ctgctgcagg actaccccgt gaccgtggcc 180 agcaacctgc aggacgagga gctgtgcggc ggcctgtgga gactggtgct ggcccagaga 240 tggatggaga gactgaagac cgtggccggc agcaagatgc agggcctgct ggagagagtg 300 aacaccgaga tccacttcgt gaccaagtgc gccttccagc ctccccccag ctgcctgagg 360 ttcgtgcaga ccaacatcag cagactgctg caggagacca gcgagcagct ggtggccctg 420 aagccctgga tcaccagaca gaacttcagc agatgcctgg agctgcagtg ccagcccgac 480 agcagcaccc tgccccctcc ctggagcccc agacccctgg aggccaccgc tcccacagcc 540 cctggcagcg ggtccggaag tgggtctgga tccgggcgcg ccaagtatgc tctggccgac 600 ccaagcctga agatggccga ccctaataga ttccggggca agaacctgcc tgtcctggac 660 cagctgaccg atccccctgg cgtgaagcgc gtctaccaca tccagccttc actggaggac 720 ccattccagc cacccagcat ccctattacc gtgtactatg ctgtcctgga acgcgcatgc 780 cgaagtgtgc tgctgcacgc tccctcagag gcacctcaga tcgtcagagg agcctccgat 840 gaagctagga agcatactta caacctgacc attgcctggt atcggatggg cgacaattgt 900 gctatcccca ttacagtgat ggagtacact gaatgccctt ataacaaatc tctgggcgtc 960 tgtcccattc gaacacagcc tcggtggtcc tactatgatt cattcagcgc cgtgtctgag 1020 gacaacctgg gcttcctgat gcacgctcca gcatttgaaa cagccgggac ttatctgaga 1080 ctggtgaaaa tcaatgattg gaccgagatc acacagttta ttctggaaca tagggcccgc 1140 gctagctgca agtacgcact gccactgagg attcctccag cagcttgtct gaccagtaaa 1200 gcttatcagc agggagtgac agtggactca atcggcatgc tgccacgctt cattcccgag 1260 aaccagcgaa ccgtggcact gtacagcctg aagatcgctg ggtggcacgg acccaaaccc 1320 ccttatacta gcaccctgct gccacccgag ctgtccgata ccacaaatgc cacacagccc 1380 gaactggtgc ctgaggaccc agaagacagc gcactgctgg aggaccctgc cggcactgtg 1440 agctcccaga tccctccaaa ctggcatatc cctagcattc aggacgtggc accacaccat 1500 gcaccagcag caccttccaa tccacggaga agggctcaga tggcaccaaa gcgactgcgg 1560 ctgccccaca ttagagacga tgacgccccc ccttcccatc agcctctgtt ttacggagga 1620 ggaggctccg gaggaggagg atctggaggc gggggaagtg gcgggggagg ctcaggggga 1680 ggcggatcta tggaacccag accaggaact tcaagcagag ccgacccagg accagaaaga 1740 cctccccgac agacaccagg cactcagcca gccgcaccac acgcctgggg catgctgaat 1800 gatatgcagt ggctggccag ctccgactct gaggaggaga cagaggtggg catctccgat 1860 gacgatctgc acagggactc cacaagcgag gccggctcca ccgataccga gatgtttgag 1920 gccggcctga tggacgccgc cacccctcca gcccggcccc cagccgagag gcagggctct 1980 ccaacacccg ccgatgccca gggctcctgc ggcggcggcc ctgtgggcga ggaggaggcc 2040 gaggccggcg gcggcggcga cgtgtgcgcc gtgtgcacag acgagatcgc cccacctctg 2100 agatgccaga gctttccatg tctgcaccca ttctgcatcc catgcatgaa gacctggata 2160 cctctgagga atacctgtcc tctgtgcaat accccagtgg cctacctgat cgtgggcgtg 2220 accgcctctg gctcctttag cacaatcccc atcgtgaacg atcctagaac ccgggtggag 2280 gccgaggccg ccgtgcgggc cggcaccgcc gtggatttca tctggaccgg caacccacgg 2340 acagccccta ggtctctgag cctgggcggc cacaccgtga gagccctgag ccccacaccc 2400 ccttggccag gcacagacga tgaggatgac gacctggccg acgtggatta tgtgccccct 2460 gcccccagaa gagccccccg gagaggcggc ggcggcgccg gcgccacaag aggcacatcc 2520 cagcctgccg ccacaaggcc tgccccacct ggcgccccac ggtcctcctc cagcggcggc 2580 gccccactgc gggccggcgt gggctccggc tccggcggcg gcccagccgt ggccgccgtg 2640 gtgcccagag tggccagcct gcctccagcc gccggcggcg gcagagccca ggccagacgg 2700 gtgggcgagg atgccgccgc cgccgagggc cggacacccc ctgccagaca gccaagagcc 2760 gcccaggagc cccctatcgt gatcagcgac tctccacctc cctccccaag aaggccagcc 2820 ggcccaggcc ctctgtcctt tgtgagctct tcttccgccc aggtgtcttc cggcccaggc 2880 ggcggcggcc tgcctcagtc ctccggcaga gccgccagac cacgggccgc cgtggcccct 2940 agagtgagga gccccccaag ggccgccgcc gccccagtgg tgtccgcctc cgccgatgcc 3000 gccggcccag ccccacccgc cgtgccagtg gacgcccaca gggccccacg gagcagaatg 3060 acccaggccc agaccgatac ccaggcccag agcctgggca gggccggcgc caccgatgcc 3120 agaggctccg gcggcccagg cgccgagggc ggccccggcg tgcccagggg cacaaataca 3180 ccaggcgccg ccccccacgc cgccgagggc gccgccggca gcgattccgg ccctgccgcc 3240 tcctcttctg cctccagcag cgccgcccca cgctctcccc tggccccaca gggcgtgggc 3300 gccaagcggg ccgccccaag acgggcccct gactccgatt ccggcgatcg gggccacggc 3360 ccactggccc ctgcctctgc cggcgccgcc cctcctagcg cctccccaag ctctcaggcc 3420 gccgtggccg ccgccagctc cagctccgcc agctcttcct ctgccagctc ctcctctgcc 3480 tcctcctcct ctgcctcttc cagctctgcc agctctagca gcgcctcctc tagctccgcc 3540 tcttctagcg ccggcggcgc cggcggctcc gtggcctctg cctccggcgc cggcgagaga 3600 agagagacct ctctgggccc aagagccgcc gcccccagag gccctagaaa gtgtgccagg 3660 aagaccaggc acgccgaggg cggcccagag ccaggcgccc gcgaccctgc cccaggcctg 3720 acacggtatc tgccaatcgc cggcgtgagc tctgtggtgg ccctggcccc atatgtgaac 3780 aagacagtga caggcgactg tctgccagtg ctggacatgg agacaggcca catcggcgcc 3840 tatgtggtgc tggtggacca gacaggcaat gtggccgatc tgctgagagc cgccgcccct 3900 gcctggtcca gaaggacact gctgccagag cacgccagaa actgcgtgag accacctgac 3960 tatcccacac cacccgcctc tgagtggaat agcctgtgga tgacaccagt gggcaacatg 4020 ctgtttgatc agggcacact ggtgggcgcc ctggatttcc acggcctgag atcccgccac 4080 ccttggtcca gagagcaggg cgcccccgcc ccagccggcg atgccccagc cggccacggc 4140 gagggcggcg gcggctcagg gggggggggc agcggcggag gtggctcagg aggtggaggt 4200 agcggaggtg gcggatctat gtcagcagaa cagaggaaga agaagaaaac aacaacaact 4260 actcagggaa ggggagcaga agtggcaatg gcagacgagg atggaggcag actgagggcc 4320 gccgccgaga caaccggcgg cccaggctct cctgacccag ccgatggccc accaccaaca 4380 cccaacccag accgcagacc agccgccaga cctggctttg gctggcacgg cggcccagag 4440 gagaacgagg atgaggccga cgatgccgcc gccgatgccg acgccgatga ggccgccccc 4500 gcctccggcg aggccgtgga cgagccagcc gccgacggcg tggtgtctcc tagacagctg 4560 gccctgctgg cctccatggt ggacgaggcc gtgaggacca tcccatctcc tcctccagag 4620 agagatggcg cccaggagga ggccgccagg agcccatccc ccccaagaac ccctagcatg 4680 agagccgact atggcgagga gaacgatgat gacgacgacg atgacgatga tgatgacagg 4740 gacgccggcc gctgggtgcg gggcccagag accacatctg ccgtgagagg cgcctatcct 4800 gacccaatgg cctctctgag cccaagacct cccgccccaa gacggcacca ccaccaccac 4860 caccacagga gacggagggc cccaagaagg cggagcgccg ccagcgactc ctctaagtct 4920 ggctccagca gctctgcctc cagcgccagc tcctctgcca gctcctcctc tagcgccagc 4980 gcctcttcct ccgacgatga tgacgatgat gacgccgcca gggcccctgc ctccgccgcc 5040 gatcacgccg ccggcggcac cctgggcgcc gatgacgagg aggccggcgt gccagccaga 5100 gccccaggcg ccgcccccag gccttctcca ccaagagccg agcccgcccc agccaggacc 5160 ccagccgcca cagccggcag actggagcgg agacgcgcca gagccgccgt ggccggcaga 5220 gatgccacag gcaggtttac cgccggcagg cccagaaggg tggagctgga tgccgatgcc 5280 gcctccggcg ccttttacgc cagatacagg gacggctacg tgagcggcga gccctggcca 5340 ggcgccggcc ctcccccacc tggcagggtg ctgtatggcg gcctgggcga tagcagacct 5400 ggcctgtggg gcgcccctga ggccgaggag gccagagcca gattcgaggc ctctggcgcc 5460 ccagcccccg tgtgggcccc agagctgggc gacgccgccc agcagtacgc cctgatcacc 5520 agactgctgt ataccccaga cgccgaggcc atgggctggc tgcagaaccc tagagtggcc 5580 ccaggcgacg tggccctgga tcaggcctgc ttccggatct ctggcgccgc ccggaacagc 5640 tctagcttta tctccggctc tgtggccaga gccgtgccac acctgggcta cgccatggcc 5700 gccggcaggt tcggctgggg cctggcccac gtagccgccg ccgtggccat gtccagacgg 5760 tatgacagag cccagaaggg ctttctgctg acatctctga ggcgggccta tgcccctctg 5820 ctggccaggg agaatgccgc cctgacaggc gccagaaccc ctgacgacgg cggcgatgcc 5880 aacagacacg atggcgacga tgccagaggc aagccagccg ccgccgccgc ccccctgcca 5940 tccgccgccg cctcccccgc cgatgagaga gccgtgcctg ccggctatgg cgccgccggc 6000 gtgctggccg ccctgggccg gctgtctgcc gccccagcct ccgcccccgc cggcgccgac 6060 gatgatgacg acgatgatgg cgccggcggc ggcggcggcg gcagacgggc cgaggccggc 6120 agggtggccg tggagtgtct ggccgcctgc cgcggcatcc tggaggccct ggccgagggc 6180 tttgatggcg acctggccgc cgtgcccggc ctggccggcg ccagacctgc cgccccacct 6240 agacctggcc ccgccggcgc cgccgccccc cctcacgccg acgccccacg gctgagagcc 6300 tggctgagag agctgagatt cgtgcgggac gccctggtgc tgatgcggct gagaggcgat 6360 ctgcgggtgg ccggcggctc tgaggccgcc gtggccgccg tgagagccgt gtccctggtg 6420 gccggcgccc tgggcccagc cctgcctaga tccccaagac tgctgtcctc cgccgctgcc 6480 gctgccgccg acctgctgtt tcagaaccag agcctgtaa 6519 <110> BioDion Co., LTD. <120> A DNA vaccine for preventing and treating HSV-2 infection <130> PD16-5410 <160> 17 <170> KoPatentin 3.0 <210> 1 <211> 114 <212> PRT <213> Artificial Sequence <220> <223> UL39-N1 <400> 1 Arg Ser Ser Glu Arg Glu Glu Pro Arg Glu Pro Glu Val Ala Pro 1 5 10 15 Pro Gly Gly Asp His Val Phe Cys Arg Lys Val Ser Gly Val Met Val 20 25 30 Leu Ser Ser Asp Pro Pro Gly Pro Ala Ala Tyr Arg Ile Ser Asp Ser 35 40 45 Ser Phe Val Gln Cys Gly Ser Asn Cys Ser Met Ile Asp Gly Asp 50 55 60 Gly Asp Gly Arg Thr Ala Val Val Ala Leu Gly Gly Thr Ser Gly Pro 65 70 75 80 Ser Ala Thr Ser Val Gly Thr Gln Thr Ser Gly Glu Phe Leu His 85 90 95 Gly Asn Pro Arg Thr Pro Glu Pro Gln Gly Pro Gln Ala Val Pro Pro 100 105 110 Pro Pro <210> 2 <211> 26 <212> PRT <213> Artificial Sequence <220> <223> UL39-C2 <400> 2 Tyr Cys Lys Val Arg Lys Ala Thr Asn Ser Gly Val Phe Ala Gly Asp 1 5 10 15 Asp Asn Ile Val Cys Thr Ser Cys Ala Leu 20 25 <210> 3 <211> 73 <212> PRT <213> Artificial Sequence <220> <223> UL39-N2 <400> 3 Pro Pro Pro Phe Pro Trp Gly His Glu Cys Cys Ala Arg Arg Asp Ala 1 5 10 15 Arg Gly Gly Ala Glu Lys Asp Val Gly Ala Ala Glu Ser Trp Ser Asp 20 25 30 Gly Pro Ser Ser Asp Ser Glu Thr Glu Asp Ser Asp Ser Ser Asp Glu 35 40 45 Asp Thr Gly Ser Glu Thr Leu Ser Arg Ser Ser Ser Ile Trp Ala Ala 50 55 60 Gly Ala Thr Asp Asp Asp Asp Ser Asp 65 70 <210> 4 <211> 718 <212> PRT <213> Artificial Sequence <220> <223> UL39-N4-C1 <400> 4 Pro Asn Ala Tyr Thr Pro Tyr His Leu Arg Glu Tyr Ala Thr Arg Leu 1 5 10 15 Val Asn Gly Phe Lys Pro Leu Val Arg Arg Ser Ala Arg Leu Tyr Arg 20 25 30 Ile Leu Gly Val Leu Val His Leu Arg Ile Arg Thr Arg Glu Ala Ser 35 40 45 Phe Glu Glu Trp Met Arg Ser Lys Glu Val Asp Leu Asp Phe Gly Leu 50 55 60 Thr Glu Arg Leu Arg Glu His Glu Ala Gln Leu Met Ile Leu Ala Gln 65 70 75 80 Ala Leu Asn Pro Tyr Asp Cys Leu Ile His Ser Thr Pro Asn Thr Leu 85 90 95 Val Glu Arg Gly Leu Gln Ser Ala Leu Lys Tyr Glu Glu Phe Tyr Leu 100 105 110 Lys Arg Phe Gly Gly His Tyr Met Glu Ser Val Phe Gln Met Tyr Thr 115 120 125 Arg Ile Ala Gly Phe Leu Ala Cys Arg Ala Thr Arg Gly Met Arg His 130 135 140 Ile Ala Leu Gly Arg Gln Gly Ser Trp Trp Glu Met Phe Lys Phe Phe 145 150 155 160 Phe His Arg Leu Tyr Asp His Gln Ile Val Pro Ser Thr Pro Ala Met 165 170 175 Leu Asn Leu Gly Thr Arg Asn Tyr Tyr Thr Ser Ser Cys Tyr Leu Val 180 185 190 Asn Pro Gln Ala Thr Thr Asn Gln Ala Thr Leu Arg Ala Ile Thr Gly 195 200 205 Asn Val Ser Ala Ile Leu Ala Arg Asn Gly Gly Ile Gly Leu Cys Met 210 215 220 Gln Ala Phe Asn Asp Ala Ser Pro Gly Thr Ala Ser Ile Met Pro Ala 225 230 235 240 Leu Lys Val Leu Asp Ser Leu Val Ala Ala His Asn Lys Gln Ser Thr 245 250 255 Arg Pro Thr Gly Ala Cys Val Tyr Leu Glu Pro Trp His Ser Asp Val 260 265 270 Arg Ala Val Leu Arg Met Lys Gly Val Leu Ala Gly Glu Glu Ala Gln 275 280 285 Arg Cys Asp Asn Ile Phe Ser Ala Leu Trp Met Pro Asp Leu Phe Phe 290 295 300 Lys Arg Leu Ile Arg His Leu Asp Gly Glu Lys Asn Val Thr Trp Ser 305 310 315 320 Leu Phe Asp Arg Asp Thr Ser Met Ser Leu Ala Asp Phe His Gly Glu 325 330 335 Glu Phe Glu Lys Leu Tyr Glu His Leu Glu Ala Met Gly Phe Gly Glu 340 345 350 Thr Ile Pro Ile Gln Asp Leu Ala Tyr Ala Ile Val Arg Ser Ala Ala 355 360 365 Thr Thr Gly Ser Pro Phe Ile Met Phe Lys Asp Ala Val Asn Arg His 370 375 380 Tyr Ile Tyr Asp Thr Gln Gly Ala Ala Ile Ala Gly Ser Asn Leu Cys 385 390 395 400 Thr Glu Ile Val His Pro Ala Ser Lys Arg Ser Ser Gly Val Cys Asn 405 410 415 Leu Gly Ser Val Asn Leu Ala Arg Cys Val Ser Ser Gln Thr Phe Asp 420 425 430 Phe Gly Arg Leu Arg Asp Ala Val Gln Ala Cys Val Leu Met Val Asn 435 440 445 Ile Met Ile Asp Ser Thr Leu Gln Pro Thr Pro Gln Cys Thr Arg Gly 450 455 460 Asn Asp Leu Arg Ser Met Gly Ile Gly Met Gln Gly Leu His Thr 465 470 475 480 Ala Cys Leu Lys Met Gly Leu Asp Leu Glu Ser Ala Glu Phe Arg Asp 485 490 495 Leu Asn Thr His Ile Ala Glu Val Met Leu Leu Ala Ala Met Lys Thr 500 505 510 Ser Asn Ala Leu Cys Val Arg Gly Ala Arg Pro Phe Ser His Phe Lys 515 520 525 Arg Ser Met Tyr Arg Ala Gly Arg Phe His Trp Glu Arg Phe Ser Asn 530 535 540 Ala Ser Pro Arg Tyr Glu Gly Glu Trp Glu Met Leu Arg Gln Ser Met 545 550 555 560 Met Lys His Gly Leu Arg Asn Ser Gln Phe Ile Ala Leu Met Pro Thr 565 570 575 Ala Ala Ser Ala Gln Ile Ser Asp Val Ser Glu Gly Phe Ala Pro Leu 580 585 590 Phe Thr Asn Leu Phe Ser Lys Val Thr Arg Asp Gly Glu Thr Leu Arg 595 600 605 Pro Asn Thr Leu Leu Leu Lys Glu Leu Glu Arg Thr Phe Gly Gly Lys 610 615 620 Arg Leu Leu Asp Ala Met Asp Gly Leu Glu Ala Lys Gln Trp Ser Val 625 630 635 640 Ala Gln Ala Leu Pro Cys Leu Asp Pro Ala His Pro Leu Arg Arg Phe 645 650 655 Lys Thr Ala Phe Asp Tyr Asp Gln Glu Leu Leu Ile Asp Leu Cys Ala 660 665 670 Asp Arg Ala Pro Tyr Val Asp His Ser Gln Ser Met Thr Leu Tyr Val 675 680 685 Thr Glu Lys Ala Asp Gly Thr Leu Pro Ala Ser Thr Leu Val Arg Leu 690 695 700 Leu Val His Ala Tyr Lys Arg Gly Leu Lys Thr Gly Met Tyr 705 710 715 <210> 5 <211> 191 <212> PRT <213> Artificial Sequence <220> <223> UL39-N3 <400> 5 Thr Leu Ser Arg Ser Ser Ser Ile Trp Ala Ala Gly Ala Thr Asp Asp 1 5 10 15 Asp Asp Ser Asp Ser Asp Ser Arg Ser Asp Asp Ser Val Gln Pro Asp 20 25 30 Val Val Val Arg Arg Arg Trp Ser Asp Gly Pro Ala Pro Val Ala Phe 35 40 45 Pro Lys Pro Arg Arg Pro Gly Asp Ser Pro Gly Asn Pro Gly Leu Gly 50 55 60 Ala Gly Thr Gly Pro Gly Ser Ala Thr Asp Pro Arg Ala Ser Ala Asp 65 70 75 80 Ser Asp Ser Ala Ala His Ala Ala Ala Pro Gln Ala Asp Val Ala Pro 85 90 95 Val Leu Asp Ser Gln Pro Thr Val Gly Thr Asp Pro Gly Tyr Pro Val 100 105 110 Pro Leu Glu Leu Thr Pro Glu Asn Ala Glu Ala Val Ala Arg Phe Leu 115 120 125 Gly Asp Ala Val Asp Arg Glu Pro Ala Leu Met Leu Glu Tyr Phe Cys 130 135 140 Arg Cys Ala Arg Glu Glu Ser Lys Arg Val Pro Pro Arg Thr Phe Gly 145 150 155 160 Ser Ala Pro Arg Leu Thr Glu Asp Asp Phe Gly Leu Leu Asn Tyr Ala 165 170 175 Leu Ala Glu Met Arg Arg Leu Cys Leu Asp Leu Pro Pro Val Pro 180 185 190 <210> 6 <211> 1122 <212> PRT <213> Artificial Sequence <220> <223> shuffled UL39 protein <400> 6 Arg Ser Ser Glu Arg Glu Glu Pro Arg Glu Pro Glu Val Ala Pro 1 5 10 15 Pro Gly Gly Asp His Val Phe Cys Arg Lys Val Ser Gly Val Met Val 20 25 30 Leu Ser Ser Asp Pro Pro Gly Pro Ala Ala Tyr Arg Ile Ser Asp Ser 35 40 45 Ser Phe Val Gln Cys Gly Ser Asn Cys Ser Met Ile Asp Gly Asp 50 55 60 Gly Asp Gly Arg Thr Ala Val Val Ala Leu Gly Gly Thr Ser Gly Pro 65 70 75 80 Ser Ala Thr Ser Val Gly Thr Gln Thr Ser Gly Glu Phe Leu His 85 90 95 Gly Asn Pro Arg Thr Pro Glu Pro Gln Gly Pro Gln Ala Val Pro Pro 100 105 110 Pro Pro Tyr Cys Lys Val Arg Lys Ala Thr Asn Ser Gly Val Phe Ala 115 120 125 Gly Asp Asp Asn Ile Val Cys Thr Ser Cys Ala Leu Pro Pro Pro Phe 130 135 140 Pro Trp Gly His Glu Cys Cys Ala Arg Arg Asp Ala Arg Gly Gly Ala 145 150 155 160 Glu Lys Asp Val Gly Ala Ala Glu Ser Trp Ser Asp Gly Pro Ser Ser 165 170 175 Asp Ser Glu Thr Glu Asp Ser Asp Ser Ser Asp Glu Asp Thr Gly Ser 180 185 190 Glu Thr Leu Ser Arg Ser Ser Ser Ile Trp Ala Ala Gly Ala Thr Asp 195 200 205 Asp Asp Ser Asp Pro Asn Ala Tyr Thr Pro Tyr His Leu Arg Glu 210 215 220 Tyr Ala Thr Arg Leu Val Asn Gly Phe Lys Pro Leu Val Arg Arg Ser 225 230 235 240 Ala Arg Leu Tyr Arg Ile Leu Gly Val Leu Val His Leu Arg Ile Arg 245 250 255 Thr Arg Glu Ala Ser Phe Glu Glu Trp Met Arg Ser Ser Lys Glu Val Asp 260 265 270 Leu Asp Phe Gly Leu Thr Glu Arg Leu Arg Glu His Glu Ala Gln Leu 275 280 285 Met Ile Leu Ala Gln Ala Leu Asn Pro Tyr Asp Cys Leu Ile His Ser 290 295 300 Thr Pro Asn Thr Leu Val Glu Arg Gly Leu Gln Ser Ala Leu Lys Tyr 305 310 315 320 Glu Glu Phe Tyr Leu Lys Arg Phe Gly Gly His Tyr Met Glu Ser Val 325 330 335 Phe Gln Met Tyr Thr Arg Ile Ala Gly Phe Leu Ala Cys Arg Ala Thr 340 345 350 Arg Gly Met Arg His Ile Ala Leu Gly Arg Gln Gly Ser Trp Trp Glu 355 360 365 Met Phe Lys Phe Phe Phe His Arg Leu Tyr Asp His Gln Ile Val Pro 370 375 380 Ser Thr Pro Ala Met Leu Asn Leu Gly Thr Arg Asn Tyr Tyr Thr Ser 385 390 395 400 Ser Cys Tyr Leu Val Asn Pro Gln Ala Thr Thr Asn Gln Ala Thr Leu 405 410 415 Arg Ala Ile Thr Gly Asn Val Ser Ala Ile Leu Ala Arg Asn Gly Gly 420 425 430 Ile Gly Leu Cys Met Gln Ala Phe Asn Asp Ala Ser Pro Gly Thr Ala 435 440 445 Ser Ile Met Pro Ala Leu Lys Val Leu Asp Ser Leu Val Ala Ala His 450 455 460 Asn Lys Gln Ser Thr Arg Pro Thr Gly Ala Cys Val Tyr Leu Glu Pro 465 470 475 480 Trp His Ser Asp Val Arg Ala Val Leu Arg Met Lys Gly Val Leu Ala 485 490 495 Gly Glu Glu Ala Gln Arg Cys Asp Asn Ile Phe Ser Ala Leu Trp Met 500 505 510 Pro Asp Leu Phe Phe Lys Arg Leu Ile Arg His Leu Asp Gly Glu Lys 515 520 525 Asn Val Thr Trp Ser Leu Phe Asp Arg Asp Thr Ser Met Ser Leu Ala 530 535 540 Asp Phe His Gly Glu Glu Phe Glu Lys Leu Tyr Glu His Leu Glu Ala 545 550 555 560 Met Gly Phe Gly Glu Thr Ile Pro Ile Gln Asp Leu Ala Tyr Ala Ile 565 570 575 Val Arg Ser Ala Ala Thr Thr Gly Ser Pro Phe Ile Met Phe Lys Asp 580 585 590 Ala Val Asn Arg His Tyr Ile Tyr Asp Thr Gln Gly Ala Ala Ile Ala 595 600 605 Gly Ser Asn Leu Cys Thr Glu Ile Val His Pro Ala Ser Lys Arg Ser 610 615 620 Ser Gly Val Cys Asn Leu Gly Ser Val Asn Leu Ala Arg Cys Val Ser 625 630 635 640 Arg Gln Thr Phe Asp Phe Gly Arg Leu Arg Asp Ala Val Gln Ala Cys 645 650 655 Val Leu Met Val Asn Ile Met Ile Asp Ser Thr Leu Gln Pro Thr Pro 660 665 670 Gln Cys Thr Arg Gly Asn Asp Asn Leu Arg Ser Met Gly Ile Gly Met 675 680 685 Gln Gly Leu His Thr Ala Cys Leu Lys Met Gly Leu Asp Leu Glu Ser 690 695 700 Ala Glu Phe Arg Asp Leu Asn Thr His Ile Ala Glu Val Met Leu Leu 705 710 715 720 Ala Ala Met Lys Thr Ser Asn Ala Leu Cys Val Arg Gly Ala Arg Pro 725 730 735 Phe Ser His Phe Lys Arg Ser Met Tyr Arg Ala Gly Arg Phe His Trp 740 745 750 Glu Arg Phe Ser Asn Ala Ser Pro Arg Tyr Glu Gly Glu Trp Glu Met 755 760 765 Leu Arg Gln Ser Met Met Lys His Gly Leu Arg Asn Ser Gln Phe Ile 770 775 780 Ala Leu Met Pro Thr Ala Ala Ser Ala Gln Ile Ser Asp Val Ser Glu 785 790 795 800 Gly Phe Ala Pro Leu Phe Thr Asn Leu Phe Ser Lys Val Thr Arg Asp 805 810 815 Gly Glu Thr Leu Arg Pro Asn Thr Leu Leu Leu Lys Glu Leu Glu Arg 820 825 830 Thr Phe Gly Gly Lys Arg Leu Leu Asp Ala Met Asp Gly Leu Glu Ala 835 840 845 Lys Gln Trp Ser Val Ala Gln Ala Leu Pro Cys Leu Asp Pro Ala His 850 855 860 Pro Leu Arg Arg Phe Lys Thr Ala Phe Asp Tyr Asp Gln Glu Leu Leu 865 870 875 880 Ile Asp Leu Cys Ala Asp Arg Ala Pro Tyr Val Asp His Ser Gln Ser 885 890 895 Met Thr Leu Tyr Val Thr Glu Lys Ala Asp Gly Thr Leu Pro Ala Ser 900 905 910 Thr Leu Val Arg Leu Leu Val His Ala Tyr Lys Arg Gly Leu Lys Thr 915 920 925 Gly Met Tyr Thr Leu Ser Arg Ser Ser Ser Ile Trp Ala Ala Gly Ala 930 935 940 Thr Asp Asp Asp Asp Ser Asp Ser Asp Ser Arg Ser Asp Asp Ser Val 945 950 955 960 Gln Pro Asp Val Val Val Arg Arg Arg Trp Ser Asp Gly Pro Ala Pro 965 970 975 Val Ala Phe Pro Lys Pro Arg Arg Pro Gly Asp Ser Pro Gly Asn Pro 980 985 990 Gly Leu Gly Aly Gly Thr Gly Pro Gly Ser Ala Thr Asp Pro Arg Ala 995 1000 1005 Ser Ala Asp Ser Asp Ser Ala Ala His Ala Ala Ala Pro Gln Ala Asp 1010 1015 1020 Val Ala Pro Val Leu Asp Ser Gln Pro Thr Val Gly Thr Asp Pro Gly 1025 1030 1035 1040 Tyr Pro Val Pro Leu Glu Leu Thr Pro Glu Asn Ala Glu Ala Val Ala 1045 1050 1055 Arg Phe Leu Gly Asp Ala Val Asp Arg Glu Pro Ala Leu Met Leu Glu 1060 1065 1070 Tyr Phe Cys Arg Cys Ala Arg Glu Glu Ser Lys Arg Val Pro Pro Arg 1075 1080 1085 Thr Phe Gly Ser Ala Pro Arg Leu Thr Glu Asp Asp Phe Gly Leu Leu 1090 1095 1100 Asn Tyr Ala Leu Ala Glu Met Arg Arg Leu Cys Leu Asp Leu Pro Pro 1105 1110 1115 1120 Val Pro <210> 7 <211> 3369 <212> DNA <213> Artificial Sequence <220> <223> polynucleotide encoding shuffled UL39 protein <400> 7 agaagtccaa gcgaaaggca ggaacctaga gagccagaag tggcaccacc tggaggggac 60 cacgtcttct gcaggaaggt gtctggggtc atggtgctga gctccgatcc accaggacct 120 gcagcttacc ggatcagtga ctctagtttt gtgcagtgcg ggagcaactg ttccatgatc 180 attgacggcg atggggacgg aagaacyct gtggtcgcac tgggaggcac ttctggaccc 240 agtgccacca catccgtggg cacccagaca tctggggaat ttctgcacgg aaatccccga 300 acccctgagc cacagggacc tcaggccgtg cctccacccc cttattgtaa ggtcagaaaa 360 gctaccaaca gtggagtgtt cgcaggcgac gataatattg tctgcacatc atgtgccctg 420 ccaccccctt ttccatgggg ccatgagtgc tgtgcccgga gagatgctag ggggggagca 480 gaaaaagacg tgggggcagc cgagagttgg tcagatggac cctcaagcga cagcgagacc 540 gaagattccg actcctctga tgaagacact ggcagtgaga ccctgtcacg cagttcaagc 600 atctgggctg caggggccac agacgatgac gattccgacc caaacgctta cacaccctat 660 cacctgaggg aatacgcaac tcgcctggtg aatgggttca agcctctggt caggcgcagc 720 gcccgcctgt atagaatcct gggagtcctg gtgcatctga gaatcaggac acgcgaggct 780 agcttcgagg aatggatgag gtccaaagaa gtggatctgg actttggcct gactgagcga 840 ctgcgggagc acgaagccca gctgatgatt ctggcacagg ccctgaaccc atacgattgc 900 ctgatccatt caactcccaa taccctggtg gaacgaggac tgcagagcgc cctgaagtac 960 gaggagttct acctgaaacg gtttggcggg cactacatgg agagcgtgtt ccagatgtat 1020 accagaattg ccggctttct ggcctgtcgg gctacaagag ggatgaggca tatcgctctg 1080 ggccgccagg ggagctggtg ggagatgttt aagttctttt tccaccggct gtacgaccat 1140 cagatcgtgc catccacccc cgccatgctg aacctgggca ctcggaatta ctatacctcc 1200 tcttgctatc tggtgaaccc ccaggccact accaatcagg ccacactgcg cgctatcact 1260 gggaacgtga gcgcaattct ggcccgaaat ggaggcatcg gactgtgcat gcaggcattc 1320 aacgatgcct ctcctggcac cgccagtatc atgccagctc tgaaggtcct ggactccctg 1380 gtggccgctc acaacaagca gtctacaagg cctactggcg cctgcgtgta cctggagcca 1440 tggcattctg atgtcagagc tgtgctgagg atgaaggggg tgctggctgg agaggaagca 1500 cagcggtgcg ataacatttt cagcgccctg tggatgcctg acctgttttt caagcgcctg 1560 atccgacacc tggatggaga gaaaaatgtg acctggagcc tgtttgatcg ggacacaagc 1620 atgtccctgg ccgacttcca cggcgaggaa tttgaaaaac tgtacgaaca tctggaggct 1680 atgggcttcg gggagaccat cccaattcag gacctggctt atgcaattgt gaggagtgca 1740 gccacaactg gctcaccctt tatcatgttc aaggatgccg tcaaccgcca ctacatctac 1800 gacactcagg gcgctgcaat cgctgggtct aatctgtgca ccgagatcgt gcatccagca 1860 agtaaaagaa gttcaggagt ctgcaacctg ggctcagtga atctggcacg atgcgtgagc 1920 cggcagacct tcgacttcgg aagactgagg gacgctgtgc aggcatgcgt cctgatggtg 1980 aacatcatga ttgatagcac actgcagccc actcctcagt gtacccgagg caacgacaat 2040 ctgcggtcca tgggaattgg catgcaggga ctgcacacag cctgcctgaa gatgggcctg 2100 gatctggaat ctgccgagtt ccgggacctg aacacacata tcgctgaagt gatgctgctg 2160 gccgctatga agactagcaa tgcactgtgc gtgcggggag ccagaccttt ttcacacttc 2220 aaaagaagca tgtacagggc aggccgcttc cattgggagc ggttttcaaa cgccagccca 2280 agatatgagg gggaatggga gatgctgaga cagagtatga tgaagcacgg actgcggaac 2340 agccagttta ttgcactgat gcccactgca gcctctgccc agatcagcga cgtgagcgaa 2400 ggctttgccc ctctgttcac caacctgttt agcaaagtca ccagagacgg ggagacactg 2460 aggcctaata ctctgctgct gaaggaactg gagcgcactt tcgggggaaa acgactgctg 2520 gatgctatgg acggcctgga ggcaaagcag tggtccgtgg cacaggctct gccatgcctg 2580 gaccctgctc atccactgcg acggttcaaa acagcatttg attacgacca ggaactgctg 2640 atcgacctgt gcgccgacag agctccctac gtggatcact ctcagagtat gacactgtat 2700 gtcactgaga aggccgacgg caccctgcct gctagcacac tggtcaggct gctggtgcat 2760 gcctacaagc gcggcctgaa aaccgggatg tatacactgt ccaggagctc ctctatctgg 2820 gccgccgggg ccaccgacga tgacgattca gatagcgact cccgctctga cgattccgtg 2880 cagccagatg tggtcgtgag aaggcgctgg tctgacggac cagcaccagt ggcattccct 2940 aaaccacgac ggcccggaga tagcccaggc aaccctggac tgggagcagg cactgggcct 3000 ggatctgcta ccgacccaag ggcaagtgcc gatagtgact cagccgctca cgcagccgct 3060 ccacaggctg atgtcgcacc tgtgctggac tcccagccaa ccgtggggac agaccccgga 3120 taccccgtcc ctctggagct gacccctgaa aatgctgagg cagtggcccg attcctgggc 3180 gggcagtgg accgggaacc agccctgatg ctggagtatt tttgccgatg tgcccgggag 3240 gaaagcaagc gggtgccacc aagaactttc ggctccgctc cacgactgac cgaggacgat 3300 tttgggctgc tgaactatgc cctggccgaa atgcggagac tgtgcctgga cctgcccccc 3360 gtgccctaa 3369 <210> 8 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> tPA signal sequence <400> 8 Met Asp Ala Met Lys Arg Gly Leu Cys Cys Val Leu Leu Leu Cys Gly 1 5 10 15 Ala Val Phe Val Ser Ser Ser Ala 20 25 <210> 9 <211> 156 <212> PRT <213> Artificial Sequence <220> <223> Flt3L <400> 9 Thr Gln Asp Cys Ser Phe Gln His Ser Pro Ile Ser Ser Asp Phe Ala 1 5 10 15 Val Lys Ile Arg Glu Leu Ser Asp Tyr Leu Leu Gln Asp Tyr Pro Val 20 25 30 Thr Val Ala Ser Asn Leu Gln Asp Glu Glu Leu Cys Gly Gly Leu Trp 35 40 45 Arg Leu Val Leu Ala Gln Arg Trp Met Glu Arg Leu Lys Thr Val Ala 50 55 60 Gly Ser Lys Met Gln Gly Leu Leu Glu Arg Val Asn Thr Glu Ile His 65 70 75 80 Phe Val Thr Lys Cys Ala Phe Gln Pro Pro Pro Ser Cys Leu Arg Phe 85 90 95 Val Gln Thr Asn Ile Ser Arg Leu Gln Glu Thr Ser Glu Gln Leu 100 105 110 Val Ala Leu Lys Pro Trp Ile Thr Arg Gln Asn Phe Ser Arg Cys Leu 115 120 125 Glu Leu Gln Cys Gln Pro Asp Ser Ser Thr Leu Pro Pro Pro Trp Ser 130 135 140 Pro Arg Pro Leu Glu Ala Thr Ala Pro Thr Ala Pro 145 150 155 <210> 10 <211> 861 <212> PRT <213> Artificial Sequence <220> <223> HSV-2 gB <400> 10 Ala Pro Ala Ala Pro Ala Ala Pro Arg Ala Ser Gly Gly Val Ala Ala 1 5 10 15 Thr Val Ala Asn Gly Gly Pro Ala Ser Arg Pro Pro Pro Val Pro 20 25 30 Ser Pro Ala Thr Thr Lys Ala Arg Lys Arg Lys Thr Lys Lys Pro Pro 35 40 45 Lys Arg Pro Glu Ala Thr Pro Pro Pro Asp Ala Asn Ala Thr Val Ala 50 55 60 Ala Gly His Ala Thr Leu Arg Ala His Leu Arg Glu Ile Lys Val Glu 65 70 75 80 Asn Ala Asp Ala Gln Phe Tyr Val Cys Pro Pro Thr Gly Ala Thr 85 90 95 Val Val Gln Phe Glu Gln Pro Arg Arg Cys Pro Thr Arg Pro Glu Gly 100 105 110 Gln Asn Tyr Thr Glu Gly Ile Ala Val Val Phe Lys Glu Asn Ile Ala 115 120 125 Pro Tyr Lys Phe Lys Ala Thr Met Tyr Tyr Lys Asp Val Thr Val Ser 130 135 140 Gln Val Trp Phe Gly His Arg Tyr Ser Gln Phe Met Gly Ile Phe Glu 145 150 155 160 Asp Arg Ala Pro Val Phe Glu Glu Val Ile Asp Lys Ile Asn Thr 165 170 175 Lys Gly Val Cys Arg Ser Thr Ala Lys Tyr Val Arg Asn Asn Met Glu 180 185 190 Thr Thr Ala Phe His Arg Asp Asp His Glu Thr Asp Met Glu Leu Lys 195 200 205 Pro Ala Lys Val Ala Thr Arg Thr Ser Arg Gly Trp His Thr Thr Asp 210 215 220 Leu Lys Tyr Asn Pro Ser Arg Val Glu Ala Phe His Arg Tyr Gly Thr 225 230 235 240 Thr Val Asn Cys Ile Val Glu Glu Val Asp Ala Arg Ser Val Tyr Pro 245 250 255 Tyr Asp Glu Phe Val Leu Ala Thr Gly Asp Phe Val Tyr Met Ser Pro 260 265 270 Phe Tyr Gly Tyr Arg Glu Gly Ser His Thr Glu His Thr Ser Tyr Ala 275 280 285 Ala Asp Arg Phe Lys Gln Val Asp Gly Phe Tyr Ala Arg Asp Leu Thr 290 295 300 Thr Lys Ala Arg Ala Thr Ser Pro Thr Thr Arg Asn Leu Leu Thr Thr 305 310 315 320 Pro Lys Phe Thr Val Ala Trp Asp Trp Val Pro Lys Arg Pro Ala Val 325 330 335 Cys Thr Met Thr Lys Trp Gln Glu Val Asp Glu Met Leu Arg Ala Glu 340 345 350 Tyr Gly Gly Ser Phe Arg Phe Ser Ser Asp Ala Ile Ser Thr Thr Phe 355 360 365 Thr Asn Leu Thr Glu Tyr Ser Leu Ser Arg Val Asp Leu Gly Asp 370 375 380 Cys Ile Gly Arg Asp Ala Arg Glu Ala Ile Asp Arg Met Phe Ala Arg 385 390 395 400 Lys Tyr Asn Ala Thr His Ile Lys Val Gly Gln Pro Gln Tyr Tyr Leu 405 410 415 Ala Thr Gly Gly Phe Leu Ile Ala Tyr Gln Pro Leu Leu Ser Asn Thr 420 425 430 Leu Ala Glu Leu Tyr Val Arg Glu Tyr Met Arg Glu Gln Asp Arg Lys 435 440 445 Pro Arg Asn Ala Thr Pro Ala Pro Leu Arg Glu Ala Pro Ser Ala Asn 450 455 460 Ala Ser Val Glu Arg Ile Lys Thr Thr Ser Ser Ile Glu Phe Ala Arg 465 470 475 480 Leu Gln Phe Thr Tyr Asn His Ile Gln Arg His Val Asn Asp Met Leu 485 490 495 Gly Arg Ile Ala Val Ala Trp Cys Glu Leu Gln Asn His Glu Leu Thr 500 505 510 Leu Trp Asn Glu Ala Arg Lys Leu Asn Pro Asn Ale Ile Ala Ser Ala 515 520 525 Thr Val Gly Arg Val Val Ser Ala Arg Met Leu Gly Asp Val Met Ala 530 535 540 Val Ser Thr Cys Val Pro Val Ala Pro Asp Asn Val Ile Val Gln Asn 545 550 555 560 Ser Met Arg Val Ser Ser Arg Pro Gly Thr Cys Tyr Ser Arg Pro Leu 565 570 575 Val Ser Phe Arg Tyr Glu Asp Gln Gly Pro Leu Ile Glu Gly Gln Leu 580 585 590 Gly Glu Asn Asn Glu Leu Arg Leu Thr Arg Asp Ala Leu Glu Pro Cys 595 600 605 Thr Val Gly His Arg Arg Tyr Phe Ile Phe Gly Gly Gly Tyr Val Tyr 610 615 620 Phe Glu Glu Tyr Ala Tyr Ser His Gln Leu Ser Arg Ala Asp Val Thr 625 630 635 640 Thr Val Ser Thr Phe Ile Asp Leu Asn Ile Thr Met Leu Glu Asp His 645 650 655 Glu Phe Val Pro Leu Glu Val Tyr Thr Arg His Glu Ile Lys Asp Ser 660 665 670 Gly Leu Leu Asp Tyr Thr Glu Val Gln Arg Arg Asn Gln Leu His Asp 675 680 685 Leu Arg Phe Ala Asp Ale Asp Thr Val Ile Arg Asa Asp Ala Asn Ala 690 695 700 Ala Met Phe Ala Gly Leu Cys Ala Phe Phe Glu Gly Met Gly Asp Leu 705 710 715 720 Gly Arg Ala Val Gly Lys Val Val Met Gly Val Val Gly Gly Val Val 725 730 735 Ser Ala Val Ser Gly Val Ser Ser Phe Met Ser Asn Pro Arg Tyr Val 740 745 750 Leu Gln Leu Gln Arg Asn Pro Met Lys Ala Leu Tyr Pro Leu Thr Thr 755 760 765 Lys Glu Leu Lys Thr Ser Asp Pro Gly Gly Val Gly Gly Glu Gly Glu 770 775 780 Glu Gly Ala Glu Gly Gly Gly Phe Asp Glu Ala Lys Leu Ala Glu Ala 785 790 795 800 Arg Glu Met Ile Arg Tyr Met Ala Leu Val Ser Ala Met Glu Arg Thr 805 810 815 Glu His Lys Ala Arg Lys Lys Gly Thr Ser Ala Leu Leu Ser Ser Lys 820 825 830 Val Thr Asn Met Val Leu Arg Lys Arg Asn Lys Ala Arg Tyr Ser Pro 835 840 845 Leu His Asn Glu Asp Glu Ala Gly Asp Glu Asp Glu Leu 850 855 860 <210> 11 <211> 2199 <212> PRT <213> Artificial Sequence <220> <223> tPA-Flt3L-gB-shuffled UL39 fusion protein <400> 11 Met Asp Ala Met Lys Arg Gly Leu Cys Cys Val Leu Leu Leu Cys Gly 1 5 10 15 Ala Val Phe Val Ser Ser Ser His Ala Thr Gln Asp Cys Ser Phe Gln 20 25 30 His Ser Pro Ile Ser Ser Asp Phe Ala Val Lys Ile Arg Glu Leu Ser 35 40 45 Asp Tyr Leu Leu Gln Asp Tyr Pro Val Thr Val Ala Ser Asn Leu Gln 50 55 60 Asp Glu Glu Leu Cys Gly Gly Leu Trp Arg Leu Val Leu Ala Gln Arg 65 70 75 80 Trp Met Glu Arg Leu Lys Thr Val Ala Gly Ser Lys Met Gln Gly Leu 85 90 95 Leu Glu Arg Val Asn Thr Glu Ile His Phe Val Thr Lys Cys Ala Phe 100 105 110 Gln Pro Pro Pro Ser Cys Leu Arg Phe Val Gln Thr Asn Ile Ser Arg 115 120 125 Leu Leu Gln Glu Thr Ser Glu Gln Leu Val Ala Leu Lys Pro Trp Ile 130 135 140 Thr Arg Gln Asn Phe Ser Arg Cys Leu Glu Leu Gln Cys Gln Pro Asp 145 150 155 160 Ser Ser Thr Leu Pro Pro Pro Trp Ser Pro Arg Pro Leu Glu Ala Thr 165 170 175 Ala Pro Thr Ala Pro Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Ala 180 185 190 Pro Ala Ala Pro Ala Ala Pro Arg Ala Ser Gly Gly Val Ala Ala Thr 195 200 205 Val Ala Ala Asn Gly Gly Pro Ala Ser Arg Pro Pro Pro Val Ser Ser 210 215 220 Pro Ala Thr Thr Lys Ala Arg Lys Arg Lys Thr Lys Lys Pro Pro Lys 225 230 235 240 Arg Pro Glu Ala Thr Pro Pro Pro Asp Ala Asn Ala Thr Val Ala Ala 245 250 255 Gly His Ala Thr Leu Arg Ala His Leu Arg Glu Ile Lys Val Glu Asn 260 265 270 Ala Asp Ala Gln Phe Tyr Val Cys Pro Pro Thr Gly Ala Thr Val 275 280 285 Val Gln Phe Glu Gln Pro Arg Arg Cys Pro Thr Arg Pro Glu Gly Gln 290 295 300 Asn Tyr Thr Glu Gly Ile Ala Val Val Phe Lys Glu Asn Ile Ala Pro 305 310 315 320 Tyr Lys Phe Lys Ala Thr Met Tyr Tyr Lys Asp Val Thr Val Ser Gln 325 330 335 Val Trp Phe Gly His Arg Tyr Ser Gln Phe Met Gly Ile Phe Glu Asp 340 345 350 Arg Ala Pro Val Phe Glu Glu Val Ile Asp Lys Ile Asn Thr Lys 355 360 365 Gly Val Cys Arg Ser Thr Ala Lys Tyr Val Arg Asn As Met Glu Thr 370 375 380 Thr Ala Phe His Arg Asp Asp His Glu Thr Asp Met Glu Leu Lys Pro 385 390 395 400 Ala Lys Val Ala Thr Arg Thr Ser Arg Gly Trp His Thr Thr Asp Leu 405 410 415 Lys Tyr Asn Pro Ser Arg Val Glu Ala Phe His Arg Tyr Gly Thr Thr 420 425 430 Val Asn Cys Ile Val Glu Glu Val Asp Ala Arg Ser Val Tyr Pro Tyr 435 440 445 Asp Glu Phe Val Leu Ala Thr Gly Asp Phe Val Tyr Met Ser Pro Phe 450 455 460 Tyr Gly Tyr Arg Glu Gly Ser His Thr Glu His Thr Ser Tyr Ala Ala 465 470 475 480 Asp Arg Phe Lys Gln Val Asp Gly Phe Tyr Ala Arg Asp Leu Thr Thr 485 490 495 Lys Ala Arg Ala Thr Ser Pro Thr Thr Arg Asn Leu Leu Thr Thr Pro 500 505 510 Lys Phe Thr Val Ala Trp Asp Trp Val Pro Lys Arg Pro Ala Val Cys 515 520 525 Thr Met Thr Lys Trp Gln Glu Val Asp Glu Met Leu Arg Ala Glu Tyr 530 535 540 Gly Gly Ser Phe Arg Phe Ser Ser Asp Ala Ile Ser Thr Thr Phe Thr 545 550 555 560 Thr Asn Leu Thr Glu Tyr Ser Leu Ser Arg Val Asp Leu Gly Asp Cys 565 570 575 Ile Gly Arg Asp Ala Arg Glu Ala Ile Asp Arg Met Phe Ala Arg Lys 580 585 590 Tyr Asn Ala Thr His Ile Lys Val Gly Gln Pro Gln Tyr Tyr Leu Ala 595 600 605 Thr Gly Gly Phe Leu Ile Ala Tyr Gln Pro Leu Leu Ser Asn Thr Leu 610 615 620 Ala Glu Leu Tyr Val Arg Glu Tyr Met Arg Glu Gln Asp Arg Lys Pro 625 630 635 640 Arg Asn Ala Thr Pro Ala Pro Leu Arg Glu Ala Pro Ser Ala Asn Ala 645 650 655 Ser Val Glu Arg Ile Lys Thr Thr Ser Ser Ile Glu Phe Ala Arg Leu 660 665 670 Gln Phe Thr Tyr Asn His Ile Gln Arg His Val Asn Asp Met Leu Gly 675 680 685 Arg Ile Ala Val Ala Trp Cys Glu Leu Gln Asn His Glu Leu Thr Leu 690 695 700 Trp Asn Glu Ala Arg Lys Leu Asn Pro Asn Ala Ile Ala Ser Ala Thr 705 710 715 720 Val Gly Arg Val Val Ser Ala Arg Met Leu Gly Asp Val Met Ala Val 725 730 735 Ser Thr Cys Val Pro Ala Pro Asp Asn Val Ile Val Gln Asn Ser 740 745 750 Met Arg Val Ser Ser Arg Pro Gly Thr Cys Tyr Ser Arg Pro Leu Val 755 760 765 Ser Phe Arg Tyr Glu Asp Gln Gly Pro Leu Ile Glu Gly Gln Leu Gly 770 775 780 Glu Asn Asn Glu Leu Arg Leu Thr Arg Asp Ala Leu Glu Pro Cys Thr 785 790 795 800 Val Gly His Arg Arg Tyr Phe Ile Phe Gly Gly Gly Tyr Val Tyr Phe 805 810 815 Glu Glu Tyr Ala Tyr Ser His Gln Leu Ser Arg Ala Asp Val Thr Thr 820 825 830 Val Ser Thr Phe Ile Asp Leu Asn Ile Thr Met Leu Glu Asp His Glu 835 840 845 Phe Val Pro Leu Glu Val Tyr Thr Arg His Glu Ile Lys Asp Ser Gly 850 855 860 Leu Leu Asp Tyr Thr Glu Val Gln Arg Arg Asn Gln Leu His Asp Leu 865 870 875 880 Arg Phe Ala Asp Ile Asp Thr Val Ile Arg Ala Asp Ala Asn Ala Ala 885 890 895 Met Phe Ala Gly Leu Cys Ala Phe Phe Glu Gly Met Gly Asp Leu Gly 900 905 910 Arg Ala Val Gly Lys Val Val Met Gly Val Val Gly Gly Val Val Ser 915 920 925 Ala Val Ser Gly Val Ser Ser Phe Met Ser Asn Pro Arg Tyr Val Leu 930 935 940 Gln Leu Gln Arg Asn Pro Met Lys Ala Leu Tyr Pro Leu Thr Thr Lys 945 950 955 960 Glu Leu Lys Thr Ser Asp Pro Gly Gly Val Gly Gly Glu Gly Glu Glu 965 970 975 Gly Ala Glu Gly Gly Gly Phe Asp Glu Ala Lys Leu Ala Glu Ala Arg 980 985 990 Glu Met Ile Arg Tyr Met Ala Leu Val Ser Ala Met Glu Arg Thr Glu 995 1000 1005 His Lys Ala Arg Lys Lys Gly Thr Ser Ala Leu Leu Ser Ser Lys Val 1010 1015 1020 Thr Asn Met Val Leu Arg Lys Arg Asn Lys Ala Arg Tyr Ser Pro Leu 1025 1030 1035 1040 His Asn Glu Asp Glu Ala Gly Asp Glu Asp Glu Leu Gly Gly Gly Gly 1045 1050 1055 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser 1060 1065 1070 Gly Gly Gly Ser Ser Ser Ser Ser Glu Arg Gln Glu Pro Arg Glu 1075 1080 1085 Pro Glu Val Ala Pro Pro Gly Gly Asp His Val Phe Cys Arg Lys Val 1090 1095 1100 Ser Gly Val Met Val Leu Ser Ser Asp Pro Pro Gly Pro Ala Ala Tyr 1105 1110 1115 1120 Arg Ile Ser Asp Ser Ser Phe Val Gln Cys Gly Ser Asn Cys Ser Met 1125 1130 1135 Ile Ile Asp Gly Asp Gly Asp Gly Arg Thr Ala Val Val Ala Leu Gly 1140 1145 1150 Gly Thr Ser Gly Pro Ser Ala Thr Thr Ser Val Gly Thr Gln Thr Ser 1155 1160 1165 Gly Glu Phe Leu His Gly Asn Pro Arg Thr Pro Glu Pro Gln Gly Pro 1170 1175 1180 Gln Ala Val Pro Pro Pro Pro Tyr Cys Lys Val Arg Lys Ala Thr Asn 1185 1190 1195 1200 Ser Gly Val Phe Ala Gly Asp Asp Asn Ile Val Cys Thr Ser Cys Ala 1205 1210 1215 Leu Pro Pro Pro Phe Pro Trp Gly His Glu Cys Cys Ala Arg Arg Asp 1220 1225 1230 Ala Arg Gly Gly Ala Glu Lys Asp Val Gly Ala Ala Glu Ser Trp Ser 1235 1240 1245 Asp Gly Pro Ser Ser Asp Ser Glu Thr Glu Asp Ser Asp Ser Ser Asp 1250 1255 1260 Glu Asp Thr Gly Ser Glu Thr Leu Ser Arg Ser Ser Ser Ile Trp Ala 1265 1270 1275 1280 Ala Gly Ala Thr Asp Asp Asp Asp Ser Asp Pro Asn Ala Tyr Thr Pro 1285 1290 1295 Tyr His Leu Arg Glu Tyr Ala Thr Arg Leu Val Asn Gly Phe Lys Pro 1300 1305 1310 Leu Val Arg Ser Ser Ala Arg Leu Tyr Arg Ile Leu Gly Val Leu Val 1315 1320 1325 His Leu Arg Ile Arg Thr Arg Glu Ala Ser Phe Glu Glu Trp Met Arg 1330 1335 1340 Ser Lys Glu Val Asp Leu Asp Phe Gly Leu Thr Glu Arg Leu Arg Glu 1345 1350 1355 1360 His Glu Ala Gln Leu Met Ile Leu Ala Gln Ala Leu Asn Pro Tyr Asp 1365 1370 1375 Cys Leu Ile His Ser Thr Pro Asn Thr Leu Val Glu Arg Gly Leu Gln 1380 1385 1390 Ser Ala Leu Lys Tyr Glu Glu Phe Tyr Leu Lys Arg Phe Gly Gly His 1395 1400 1405 Tyr Met Glu Ser Val Phe Gln Met Tyr Thr Arg Ile Ala Gly Phe Leu 1410 1415 1420 Ala Cys Arg Ala Thr Arg Gly Met Arg His Ile Ala Leu Gly Arg Gln 1425 1430 1435 1440 Gly Ser Trp Trp Glu Met Phe Lys Phe Phe Phe His Arg Leu Tyr Asp 1445 1450 1455 His Gln Ile Val Ser Thr Pro Ala Met Leu Asn Leu Gly Thr Arg 1460 1465 1470 Asn Tyr Tyr Thr Ser Ser Cys Tyr Leu Val Asn Pro Gln Ala Thr Thr 1475 1480 1485 Asn Gln Ala Thr Leu Arg Ala Ile Thr Gly Asn Val Ser Ala Ile Leu 1490 1495 1500 Ala Arg Asn Gly Gly Ile Gly Leu Cys Met Gln Ala Phe Asn Asp Ala 1505 1510 1515 1520 Ser Pro Gly Thr Ala Ser Ile Met Pro Ala Leu Lys Val Leu Asp Ser 1525 1530 1535 Leu Val Ala Ala His Asn Lys Gln Ser Thr Arg Pro Thr Gly Ala Cys 1540 1545 1550 Val Tyr Leu Glu Pro Trp His Ser Asp Val Arg Ala Val Leu Arg Met 1555 1560 1565 Lys Gly Val Leu Ala Gly Glu Glu Ala Gln Arg Cys Asp Asn Ile Phe 1570 1575 1580 Ser Ala Leu Trp Met Pro Asp Leu Phe Phe Lys Arg Leu Ile Arg His 1585 1590 1595 1600 Leu Asp Gly Glu Lys Asn Val Thr Trp Ser Leu Phe Asp Arg Asp Thr 1605 1610 1615 Ser Met Ser Leu Ala Asp Phe His Gly Glu Glu Phe Glu Lys Leu Tyr 1620 1625 1630 Glu His Leu Glu Ala Met Gly Phe Gly Glu Thr Ile Pro Ile Gln Asp 1635 1640 1645 Leu Ala Tyr Ala Ile Val Arg Ser Ala Ala Thr Thr Gly Ser Pro Phe 1650 1655 1660 Ile Met Phe Lys Asp Ala Val Asn Arg His Tyr Ile Tyr Asp Thr Gln 1665 1670 1675 1680 Gly Ala Ala Ile Ala Gly Ser Asn Leu Cys Thr Glu Ile Val His Pro 1685 1690 1695 Ala Ser Lys Arg Ser Ser Gly Val Cys Asn Leu Gly Ser Val Asn Leu 1700 1705 1710 Ala Arg Cys Val Ser Ser Gln Thr Phe Asp Phe Gly Arg Leu Arg Asp 1715 1720 1725 Ala Val Gln Ala Cys Val Leu Met Val Asn Ile Met Ile Asp Ser Thr 1730 1735 1740 Leu Gln Pro Thr Pro Gln Cys Thr Arg Gly Asn Asp Asn Leu Arg Ser 1745 1750 1755 1760 Met Gly Ile Gly Met Gln Gly Leu His Thr Ala Cys Leu Lys Met Gly 1765 1770 1775 Leu Asp Leu Glu Ser Ala Glu Phe Arg Asp Leu Asn Thr His Ile Ala 1780 1785 1790 Glu Val Met Leu Leu Ala Ala Met Lys Thr Ser Asn Ala Leu Cys Val 1795 1800 1805 Arg Gly Ala Arg Pro Phe Ser His Phe Lys Arg Ser Met Tyr Arg Ala 1810 1815 1820 Gly Arg Phe His Trp Glu Arg Phe Ser Asn Ala Ser Pro Arg Tyr Glu 1825 1830 1835 1840 Gly Glu Trp Glu Met Leu Arg Gln Ser Met Met Lys His Gly Leu Arg 1845 1850 1855 Asn Ser Gln Phe Ile Ala Leu Met Pro Thr Ala Ala Ser Ala Gln Ile 1860 1865 1870 Ser Asp Val Ser Glu Gly Phe Ala Pro Leu Phe Thr Asn Leu Phe Ser 1875 1880 1885 Lys Val Thr Arg Asp Gly Glu Thr Leu Arg Pro Asn Thr Leu Leu Leu 1890 1895 1900 Lys Glu Leu Glu Arg Thr Phe Gly Gly Lys Arg Leu Leu Asp Ala Met 1905 1910 1915 1920 Asp Gly Leu Glu Ala Lys Gln Trp Ser Val Ala Gln Ala Leu Pro Cys 1925 1930 1935 Leu Asp Pro Ala His Pro Leu Arg Arg Phe Lys Thr Ala Phe Asp Tyr 1940 1945 1950 Asp Gln Glu Leu Leu Ile Asp Leu Cys Ala Asp Arg Ala Pro Tyr Val 1955 1960 1965 Asp His Ser Gln Ser Met Thr Leu Tyr Val Thr Glu Lys Ala Asp Gly 1970 1975 1980 Thr Leu Pro Ala Ser Thr Leu Val Arg Leu Leu Val His Ala Tyr Lys 1985 1990 1995 2000 Arg Gly Leu Lys Thr Gly Met Tyr Thr Leu Ser Arg Ser Ser Ser Ile 2005 2010 2015 Trp Ala Ala Gly Ala Thr Asp Asp Asp Asp Ser Asp Ser Asp Ser Arg 2020 2025 2030 Ser Asp Ser Val Gln Pro Asp Val Val Val Arg Arg Arg Trp Ser 2035 2040 2045 Asp Gly Pro Ala Pro Val Ala Phe Pro Lys Pro Arg Arg Pro Gly Asp 2050 2055 2060 Ser Pro Gly Asn Pro Gly Leu Gly Ala Gly Thr Gly Pro Gly Ser Ala 2065 2070 2075 2080 Thr Asp Pro Arg Ala Ser Ala Asp Ser Asp Ser Ala Ala His Ala Ala 2085 2090 2095 Ala Pro Gln Ala Asp Val Ala Pro Val Leu Asp Ser Gln Pro Thr Val 2100 2105 2110 Gly Thr Asp Pro Gly Tyr Pro Val Leu Glu Leu Thr Pro Glu Asn 2115 2120 2125 Ala Glu Ala Val Ala Arg Phe Leu Gly Asp Ala Val Asp Arg Glu Pro 2130 2135 2140 Ala Leu Met Leu Glu Tyr Phe Cys Arg Cys Ala Arg Glu Glu Ser Lys 2145 2150 2155 2160 Arg Val Pro Pro Arg Thr Phe Gly Ser Ala Pro Arg Leu Thr Glu Asp 2165 2170 2175 Asp Phe Gly Leu Leu Asn Tyr Ala Leu Ala Glu Met Arg Arg Leu Cys 2180 2185 2190 Leu Asp Leu Pro Pro Val Pro 2195 <210> 12 <211> 6600 <212> DNA <213> Artificial Sequence <220> <223> Polynucleotide encoding tPA-Flt3L-gB-shuffled UL39 fusion protein <400> 12 atggacgcca tgaagagagg cctgtgctgc gtgctgctgc tgtgcggcgc cgtgttcgtg 60 agccccagcc acgccaccca ggactgcagc ttccagcaca gccccatcag cagcgacttc 120 gccgtgaaga tcagagagct gagcgactac ctgctgcagg actaccccgt gaccgtggcc 180 agcaacctgc aggacgagga gctgtgcggc ggcctgtgga gactggtgct ggcccagaga 240 tggatggaga gactgaagac cgtggccggc agcaagatgc agggcctgct ggagagagtg 300 aacaccgaga tccacttcgt gaccaagtgc gccttccagc ctccccccag ctgcctgagg 360 ttcgtgcaga ccaacatcag cagactgctg caggagacca gcgagcagct ggtggccctg 420 aagccctgga tcaccagaca gaacttcagc agatgcctgg agctgcagtg ccagcccgac 480 agcagcaccc tgccccctcc ctggagcccc agacccctgg aggccaccgc tcccacagcc 540 cctggcagcg ggtccggaag tgggtctgga tctgcaccag cagcaccagc cgctccaagg 600 gcatcagggg gcgtcgcagc aactgtggca gcaaatgggg gaccagcctc cagacctcca 660 ccagtgccca gccctgcaac cacaaaggcc cgcaagcgaa aaactaagaa accacctaaa 720 cgacctgagg caaccccacc acctgacgca aacgctactg tggcagctgg acacgctacc 780 ctgcgagcac atctgagaga gatcaaggtc gaaaatgcag atgcccagtt ctacgtgtgc 840 ccacccccta caggagccac tgtggtccag tttgagcagc ctcggagatg tcccacaaga 900 cctgaggggc agaactacac tgaaggaatc gctgtggtct tcaaggaaaa catcgcccca 960 tacaagttta aagctacaat gtactacaaa gacgtgactg tctcacaagt gtggttcggc 1020 cacagataca gccagttcat ggggattttt gaggacaggg ccccagtgcc ctttgaggaa 1080 gtgatcgaca agattaacac caaaggcgtc tgcagatcca cagccaaata tgtgaggaac 1140 aatatggaaa ctaccgcttt ccacagggac gatcatgaga cagacatgga actgaagccc 1200 gcaaaagtgg ccaccaggac aagtcgcgga tggcacacaa ctgatctgaa gtacaaccct 1260 tcacgggtcg aggccttcca tagatatggc accacagtga attgtatcgt ggaggaagtc 1320 gatgcacgga gcgtgtaccc ttatgacgaa tttgtcctgg ccaccggcga tttcgtgtac 1380 atgagcccct tctacgggta tagggaggga agccacaccg aacatacatc ctacgcagcc 1440 gccgcttca agcaggtgga tgggttttat gcccgcgacc tgactaccaa agcccgggcc 1500 accagcccta caactcggaa cctgctgacc acaccaaagt tcaccgtggc ttgggactgg 1560 gtcccaaagc gaccagcagt ctgcactatg accaaatggc aggaggtgga cgaaatgctg 1620 cgagctgagt acggaggcag cttccggttc agcagcgatg caatttctac tacctttaca 1680 actaatctga ccgagtatag cctgtccaga gtggacctgg gcgattgtat cgggcgagat 1740 gctcgggaag caattgacag gatgttcgct cgcaagtaca acgcaacaca catcaaagtg 1800 ggccagccac agtactatct ggcaactgga ggctttctga ttgcctatca gcccctgctg 1860 agcaatacac tggccgagct gtacgtgcga gagtatatgc gggaacagga cagaaagcca 1920 aggaacgcaa cccctgctcc actgcgagaa gcaccttcag ccaatgctag cgtggagcgg 1980 atcaaaacca catctagtat tgagttcgct agactgcagt ttacctacaa ccacatccag 2040 agacatgtca atgatatgct gggcaggatt gcagtggcct ggtgcgagct gcagaaccat 2100 gaactgacac tgtggaatga ggcccggaag ctgaacccaa atgctatcgc atctgccact 2160 gtgggaaggc gcgtcagtgc cagaatgctg ggcgacgtga tggctgtcag cacatgcgtg 2220 cctgtcgcac cagataacgt gattgtccag aacagcatga gagtgtcaag caggccaggc 2280 acctgttaca gtaggcccct ggtgtcattc cgctatgaag accagggacc actgatcgag 2340 ggacagctgg gggagaacaa tgaactgcga ctgacccgag atgccctgga gccatgcaca 2400 gtgggccacc gacggtattt catttttggg ggaggctacg tgtattttga ggaatacgcc 2460 tattcccatc agctgtctag ggctgacgtg actaccgtct ctactttcat cgacctgaac 2520 attaccatgc tggaggatca cgaatttgtg cccctggagg tctacacccg ccatgaaatc 2580 aaggacagcg gactgctgga ttatacagag gtgcagagaa ggaaccagct gcacgacctg 2640 cgcttcgccg acatcgacac cgtgattcgg gctgatgcaa atgctgcaat gtttgccggg 2700 ctgtgcgctt tctttgaggg aatgggcgac ctgggacggg ccgtgggcaa ggtggtcatg 2760 ggagtggtcg gaggagtggt ctctgctgtg agtggagtct cctctttcat gagcaacccc 2820 agatacgtgc tgcagctgca gaggaatccc atgaaagccc tgtatcctct gacaactaag 2880 gagctgaaaa cctccgaccc tggaggagtg ggaggagagg gagaggaagg cgcagaaggg 2940 ggaggcttcg atgaggccaa gctggccgag gctcgcgaaa tgatccgata catggcactg 3000 gtgtctgcca tggagcgaac tgaacacaag gctcggaaga aaggcaccag tgcactgctg 3060 agttcaaaag tgacaaacat ggtcctgcgg aagagaaaca aggcccggta ttctccactg 3120 cataatgaag acgaggctgg cgacgaagac gaactggggg gcggaggatc tgggggcggg 3180 gggagcgggg gcgggggaag cggagggggg ggaagcggag ggggagggag cagaagtcca 3240 agcgaaaggc aggaacctag agagccagaa gtggcaccac ctggagggga ccacgtcttc 3300 tgcaggaagg tgtctggggt catggtgctg agctccgatc caccaggacc tgcagcttac 3360 cggatcagtg actctagttt tgtgcagtgc gggagcaact gttccatgat cattgacggc 3420 gatggggacg gaagaacagc tgtggtcgca ctgggaggca cttctggacc cagtgccacc 3480 acatccgtgg gcacccagac atctggggaa tttctgcacg gaaatccccg aacccctgag 3540 ccacagggac ctcaggccgt gcctccaccc ccttattgta aggtcagaaa agctaccaac 3600 agtggagtgt tcgcaggcga cgataatatt gtctgcacat catgtgccct gccaccccct 3660 tttccatggg gccatgagtg ctgtgcccgg agagatgcta gggggggagc agaaaaagac 3720 gtgggggcag ccgagagttg gtcagatgga ccctcaagcg acagcgagac cgaagattcc 3780 gactcctctg atgaagacac tggcagtgag accctgtcac gcagttcaag catctgggct 3840 gcaggggcca cagacgatga cgattccgac ccaaacgctt acacacccta tcacctgagg 3900 gaatacgcaa ctcgcctggt gaatgggttc aagcctctgg tcaggcgcag cgcccgcctg 3960 tatagaatcc tgggagtcct ggtgcatctg agaatcagga cacgcgaggc tagcttcgag 4020 gaatggatga ggtccaaaga agtggatctg gactttggcc tgactgagcg actgcgggag 4080 cacgaagccc agctgatgat tctggcacag gccctgaacc catacgattg cctgatccat 4140 tcaactccca ataccctggt ggaacgagga ctgcagagcg ccctgaagta cgaggagttc 4200 tacctgaaac ggtttggcgg gcactacatg gagagcgtgt tccagatgta taccagaatt 4260 gccggctttc tggcctgtcg ggctacaaga gggatgaggc atatcgctct gggccgccag 4320 gggagctggt gggagatgtt taagttcttt ttccaccggc tgtacgacca tcagatcgtg 4380 ccatccaccc ccgccatgct gaacctgggc actcggaatt actatacctc ctcttgctat 4440 ctggtgaacc cccaggccac taccaatcag gccacactgc gcgctatcac tgggaacgtg 4500 agcgcaattc tggcccgaaa tggaggcatc ggactgtgca tgcaggcatt caacgatgcc 4560 tctcctggca ccgccagtat catgccagct ctgaaggtcc tggactccct ggtggccgct 4620 cacaacaagc agtctacaag gcctactggc gcctgcgtgt acctggagcc atggcattct 4680 gatgtcagag ctgtgctgag gatgaagggg gtgctggctg gagaggaagc acagcggtgc 4740 gataacattt tcagcgccct gtggatgcct gacctgtttt tcaagcgcct gatccgacac 4800 ctggatggag agaaaaatgt gacctggagc ctgtttgatc gggacacaag catgtccctg 4860 gccgacttcc acggcgagga atttgaaaaa ctgtacgaac atctggaggc tatgggcttc 4920 ggggagacca tcccaattca ggacctggct tatgcaattg tgaggagtgc agccacaact 4980 ggctcaccct ttatcatgtt caaggatgcc gtcaaccgcc actacatcta cgacactcag 5040 ggcgctgcaa tcgctgggtc taatctgtgc accgagatcg tgcatccagc aagtaaaaga 5100 agttcaggag tctgcaacct gggctcagtg aatctggcac gatgcgtgag ccggcagacc 5160 ttcgacttcg gaagactgag ggacgctgtg caggcatgcg tcctgatggt gaacatcatg 5220 attgatagca cactgcagcc cactcctcag tgtacccgag gcaacgacaa tctgcggtcc 5280 atgggaattg gcatgcaggg actgcacaca gcctgcctga agatgggcct ggatctggaa 5340 tctgccgagt tccgggacct gaacacacat atcgctgaag tgatgctgct ggccgctatg 5400 aagactagca atgcactgtg cgtgcgggga gccagacctt tttcacactt caaaagaagc 5460 atgtacaggg caggccgctt ccattgggag cggttttcaa acgccagccc aagatatgag 5520 ggggaatggg agatgctgag acagagtatg atgaagcacg gactgcggaa cagccagttt 5580 attgcactga tgcccactgc agcctctgcc cagatcagcg acgtgagcga aggctttgcc 5640 cctctgttca ccaacctgtt tagcaaagtc accagagacg gggagacact gaggcctaat 5700 actctgctgc tgaaggaact ggagcgcact ttcgggggaa aacgactgct ggatgctatg 5760 gcggcctgg aggcaaagca gtggtccgtg gcacaggctc tgccatgcct ggaccctgct 5820 catccactgc gacggttcaa aacagcattt gattacgacc aggaactgct gatcgacctg 5880 tgcgccgaca gagctcccta cgtggatcac tctcagagta tgacactgta tgtcactgag 5940 aaggccgacg gcaccctgcc tgctagcaca ctggtcaggc tgctggtgca tgcctacaag 6000 cgcggcctga aaaccgggat gtatacactg tccaggagct cctctatctg ggccgccggg 6060 gccaccgacg atgacgattc agatagcgac tcccgctctg acgattccgt gcagccagat 6120 gtggtcgtga gaaggcgctg gtctgacgga ccagcaccag tggcattccc taaaccacga 6180 cggcccggag atagcccagg caaccctgga ctgggagcag gcactgggcc tggatctgct 6240 accgacccaa gggcaagtgc cgatagtgac tcagccgctc acgcagccgc tccacaggct 6300 gatgtcgcac ctgtgctgga ctcccagcca accgtgggga cagaccccgg ataccccgtc 6360 cctctggagc tgacccctga aaatgctgag gcagtggccc gattcctggg cgatgcagtg 6420 gaccgggaac cagccctgat gctggagtat ttttgccgat gtgcccggga ggaaagcaag 6480 cgggtgccac caagaacttt cggctccgct ccacgactga ccgaggacga ttttgggctg 6540 ctgaactatg ccctggccga aatgcggaga ctgtgcctgg acctgccccc cgtgccctaa 6600 6600 <210> 13 <211> 344 <212> PRT <213> Artificial Sequence <220> <223> HSV-2 gD <400> 13 Lys Tyr Ala Leu Ala Asp Pro Ser Leu Lys Met Ala Asp Pro Asn Arg 1 5 10 15 Phe Arg Gly Lys Asn Leu Pro Val Leu Asp Gln Leu Thr Asp Pro Pro 20 25 30 Gly Val Lys Arg Val Tyr His Ile Gln Pro Ser Leu Glu Asp Pro Phe 35 40 45 Gln Pro Pro Ser Ile Pro Ile Thr Val Tyr Tyr Ala Val Leu Glu Arg 50 55 60 Ala Cys Arg Ser Val Leu Leu His Ala Pro Ser Glu Ala Pro Gln Ile 65 70 75 80 Val Arg Gly Ala Ser Asp Glu Ala Arg Lys His Thr Tyr Asn Leu Thr 85 90 95 Ile Ala Trp Tyr Arg Met Gly Asp Asn Cys Ala Ile Pro Ile Thr Val 100 105 110 Met Glu Tyr Thr Glu Cys Pro Tyr Asn Lys Ser Leu Gly Val Cys Pro 115 120 125 Ile Arg Thr Gln Pro Arg Trp Ser Tyr Tyr Asp Ser Phe Ser Ala Val 130 135 140 Ser Glu Asp Asn Leu Gly Phe Leu Met His Ala Pro Ala Phe Glu Thr 145 150 155 160 Ala Gly Thr Tyr Leu Arg Leu Val Lys Ile Asn Asp Trp Thr Glu Ile 165 170 175 Thr Gln Phe Ile Leu Glu His Arg Ala Arg Ala Ser Cys Lys Tyr Ala 180 185 190 Leu Pro Leu Arg Ile Pro Pro Ala Ala Cys Leu Thr Ser Lys Ala Tyr 195 200 205 Gln Gln Gly Val Thr Val Asp Ser Ile Gly Met Leu Pro Arg Phe Ile 210 215 220 Pro Glu Asn Gln Arg Thr Val Ala Leu Tyr Ser Leu Lys Ile Ala Gly 225 230 235 240 Trp His Gly Pro Lys Pro Pro Tyr Thr Ser Thr Leu Leu Pro Pro Glu 245 250 255 Leu Ser Asp Thr Thr Asn Ala Thr Gln Pro Glu Leu Val Pro Glu Asp 260 265 270 Pro Glu Asp Ser Ala Leu Leu Glu Asp Pro Ala Gly Thr Val Ser Ser 275 280 285 Gln Ile Pro Pro Asn Trp His Ile Pro Ser Ile Gln Asp Val Ala Pro 290 295 300 His His Ala Pro Ala Ala Pro Ser Asn Pro Arg Arg Ala Gln Met 305 310 315 320 Ala Pro Lys Arg Leu Arg Leu Pro His Ile Arg Asp Asp Asp Ala Pro 325 330 335 Pro Ser His Gln Pro Leu Phe Tyr 340 <210> 14 <211> 818 <212> PRT <213> Artificial Sequence <220> <223> HSV-2 ICP0 variant <400> 14 Met Glu Pro Arg Pro Gly Thr Ser Ser Ala Asp Pro Gly Pro Glu 1 5 10 15 Arg Pro Pro Arg Gln Thr Pro Gly Thr Gln Pro Ala Ala Pro His Ala 20 25 30 Trp Gly Met Leu Asn Asp Met Gln Trp Leu Ala Ser Ser Asp Ser Glu 35 40 45 Glu Glu Thr Glu Val Gly Ile Ser Asp Asp Asp Leu His Arg Asp Ser 50 55 60 Thr Ser Glu Ala Gly Ser Thr Asp Thr Glu Met Phe Glu Ala Gly Leu 65 70 75 80 Met Asp Ala Ala Thr Pro Pro Ala Arg Pro Pro Ala Glu Arg Gln Gly 85 90 95 Ser Pro Thr Pro Ala Asp Ala Gln Gly Ser Cys Gly Gly Gly Pro Val 100 105 110 Gly Glu Glu Gly Aly Gly Aly Gly Gly Gly Gly Asp Val Cys Ala Val 115 120 125 Cys Thr Asp Glu Ile Ala Pro Pro Leu Arg Cys Gln Ser Phe Pro Cys 130 135 140 Leu His Pro Phe Cys Ile Pro Cys Met Lys Thr Trp Ile Pro Leu Arg 145 150 155 160 Asn Thr Cys Pro Leu Cys Asn Thr Pro Val Ala Tyr Leu Ile Val Gly 165 170 175 Val Thr Ala Ser Gly Ser Phe Ser Thr Ile Pro Ile Val Asn Asp Pro 180 185 190 Arg Thr Arg Val Glu Ala Gla Ala Ala Val Arg Ala Gly Thr Ala Val 195 200 205 Asp Phe Ile Trp Thr Gly Asn Pro Arg Thr Ala Pro Arg Ser Leu Ser 210 215 220 Leu Gly Gly His Thr Val Arg Ala Leu Ser Pro Thr Pro Pro Trp Pro 225 230 235 240 Gly Thr Asp Asp Glu Asp Asp Asp Leu Ala Asp Val Asp Tyr Val Pro 245 250 255 Pro Ala Pro Arg Arg Ala Pro Arg Arg Gly Gly Gly Gly Ala Gly Ala 260 265 270 Thr Arg Gly Thr Ser Gln Pro Ala Ala Thr Arg Pro Ala Pro Pro Gly 275 280 285 Ala Pro Arg Ser Ser Ser Ser Gly Gly Ala Pro Leu Arg Ala Gly Val 290 295 300 Gly Ser Gly Ser Gly Gly Gly Pro Ala Val Ala Ala Val Val Pro Arg 305 310 315 320 Val Ala Ser Leu Pro Pro Ala Ala Gly Gly Gly Arg Ala Gln Ala Arg 325 330 335 Arg Val Gly Glu Asp Ala Ala Ala Glu Gly Arg Thr Pro Pro Ala 340 345 350 Arg Gln Pro Arg Ala Gln Glu Pro Pro Ile Val Ile Ser Asp Ser 355 360 365 Pro Pro Ser Pro Arg Arg Pro Ala Gly Pro Gly Pro Leu Ser Phe 370 375 380 Val Ser Ser Ser Ala Gln Val Ser Ser Gly Pro Gly Gly Gly Gly 385 390 395 400 Leu Pro Gln Ser Ser Gly Arg Ala Ala Arg Pro Ala Ala Val Ala 405 410 415 Pro Arg Val Ser Ser Pro Pro Arg Ala Ala Ala Pro Val Val Ser 420 425 430 Ala Ser Ala Asp Ala Ala Gly Pro Ala Pro Pro Ala Val Pro Val Asp 435 440 445 Ala His Arg Ala Pro Arg Ser Arg Met Thr Gln Ala Gln Thr Asp Thr 450 455 460 Gln Ala Gln Ser Leu Gly Arg Ala Gly Ala Thr Asp Ala Arg Gly Ser 465 470 475 480 Gly Gly Pro Gly Ala Gly Gly Gly Pro Gly Val Pro Arg Gly Thr Asn 485 490 495 Thr Pro Gly Ala Ala Pro His Ala Ala Glu Gly Ala Ala Gly Ser Asp 500 505 510 Ser Gly Pro Ala Ala Ser Ser Ala Ser Ser Ser Ala Ala Pro Arg 515 520 525 Ser Pro Leu Ala Pro Gln Gly Val Gly Ala Lys Arg Ala Ala Pro Arg 530 535 540 Arg Ala Pro Asp Ser Asp Ser Gly Asp Arg Gly His Gly Pro Leu Ala 545 550 555 560 Pro Ala Ser Ala Gly Ala Pro Pro Ser Ala Ser Pro Ser Ser Gln 565 570 575 Ala Ala Val Ala Ala Ala Ser Ser Ser Ala Ser Ser Ser Ala 580 585 590 Ser Ser Ser Ala Ser Ser Ser Ala Ser Ser Ser Ser Ala Ser 595 600 605 Ser Ser Ala Ser Ser Ser Ser Ala Ser Ser Ser Ala Gly Gly Ala 610 615 620 Gly Gly Ser Val Ala Ser A Gly Ala Gly Glu Arg Arg Glu Thr 625 630 635 640 Ser Leu Gly Pro Arg Ala Ala Ala Pro Arg Gly Pro Arg Lys Cys Ala 645 650 655 Arg Lys Thr Arg His Ala Glu Gly Gly Pro Glu Pro Gly Ala Arg Asp 660 665 670 Pro Ala Pro Gly Leu Thr Arg Tyr Leu Pro Ile Ala Gly Val Ser Ser 675 680 685 Val Val Ala Leu Ala Pro Tyr Val Asn Lys Thr Val Thr Gly Asp Cys 690 695 700 Leu Pro Val Leu Asp Met Glu Thr Gly His Ile Gly Ala Tyr Val Val 705 710 715 720 Leu Val Asp Gln Thr Gly Asn Val Ala Asp Leu Leu Arg Ala Ala Ala 725 730 735 Pro Ala Trp Ser Arg Arg Thr Leu Leu Pro Glu His Ala Arg Asn Cys 740 745 750 Val Arg Pro Pro Asp Tyr Pro Thr Pro Pro Ala Ser Glu Trp Asn Ser 755 760 765 Leu Trp Met Thr Pro Val Gly Asn Met Leu Phe Asp Gln Gly Thr Leu 770 775 780 Val Gly Ala Leu Asp Phe His Gly Leu Arg Ser Ser His His Trp Ser 785 790 795 800 Arg Glu Gln Gly Ala Pro Ala Pro Ala Gly Asp Ala Pro Ala Gly His 805 810 815 Gly Glu <210> 15 <211> 766 <212> PRT <213> Artificial Sequence <220> <223> HSV-2 ICP4 variant <400> 15 Met Ser Ala Glu Gln Arg Lys Lys Lys Lys Thr Thr Thr Thr Thr Gln 1 5 10 15 Gly Arg Gly Ala Glu Val Ala Met Ala Asp Glu Asp Gly Gly Arg Leu 20 25 30 Arg Ala Ala Ala Glu Thr Thr Gly Gly Pro Gly Ser Pro Asp Pro Ala 35 40 45 Asp Gly Pro Pro Thr Pro Asn Pro Asp Arg Arg Pro Ala Ala Arg 50 55 60 Pro Gly Phe Gly Trp His Gly Gly Pro Glu Glu Asn Glu Asp Glu Ala 65 70 75 80 Asp Asp Ala Asa Asp Asp Asp Asp Asp Asp Glu 85 90 95 Gly Glu Ala Val Asp Glu Pro Ala Ala Asp Gly Val Val Ser Pro Arg 100 105 110 Gln Leu Ala Leu Leu Ala Ser Met Val Asp Glu Ala Val Arg Thr Ile 115 120 125 Pro Ser Pro Pro Glu Arg Asp Gly Ala Glu Glu Glu Ala Ala Arg 130 135 140 Ser Pro Ser Pro Pro Arg Thr Pro Ser Met Arg Ala Asp Tyr Gly Glu 145 150 155 160 Glu Asn Asp Asp Asp Asp Asp Asp Asp Asp Asp Asp Asp Arg Asp Ala 165 170 175 Gly Arg Trp Val Arg Gly Pro Glu Thr Thr Ser Ala Val Arg Gly Ala 180 185 190 Tyr Pro Asp Pro Met Ala Ser Leu Ser Pro Arg Pro Pro Ala Pro Arg 195 200 205 Arg His His His His His His His Arg Arg Arg Arg Ala Pro Arg Arg 210 215 220 Arg Ser Ala Ala Ser Asp Ser Ser Lys Ser Ser Ser Ser Ser Ser Ala 225 230 235 240 Ser Ser Ala Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ala Ser 245 250 255 Ser Ser Asp Asp Asp Asp Asp Asp Asp Ala Ala Arg Ala Pro Ala Ser 260 265 270 Ala Ala Asp His Ala Ala Gly Gly Thr Leu Gly Ala Asp Asp Glu Glu 275 280 285 Ala Gly Val Pro Ala Arg Ala Pro Gly Ala Ala Pro Arg Pro Ser Pro 290 295 300 Pro Arg Ala Glu Pro Ala Pro Ala Arg Thr Pro Ala Ala Thr Ala Gly 305 310 315 320 Arg Leu Glu Arg Arg Ala Arg Ala Ala Val Ala Gly Arg Asp Ala 325 330 335 Thr Gly Arg Phe Thr Ala Gly Arg Pro Arg Arg Val Glu Leu Asp Ala 340 345 350 Asp Ala Ala Ser Gly Ala Phe Tyr Ala Arg Tyr Arg Asp Gly Tyr Val 355 360 365 Ser Gly Glu Pro Trp Pro Gly Ala Gly Pro Pro Pro Pro Gly Arg Val 370 375 380 Leu Tyr Gly Gly Leu Gly Asp Ser Arg Pro Gly Leu Trp Gly Ala Pro 385 390 395 400 Glu Ala Glu Glu Ala Arg Ala Arg Phe Glu Ala Ser Gly Ala Pro Ala 405 410 415 Pro Val Trp Ala Pro Glu Leu Gly Asp Ala Ala Gln Gln Tyr Ala Leu 420 425 430 Ile Thr Arg Leu Leu Tyr Thr Pro Asp Ala Glu Ala Met Gly Trp Leu 435 440 445 Gln Asn Pro Arg Val Ala Pro Gly Asp Val Ala Leu Asp Gln Ala Cys 450 455 460 Phe Arg Ile Ser Gly Ala Ala Arg Asn Ser Ser Ser Phe Ile Ser Gly 465 470 475 480 Ser Val Ala Arg Ala Val Pro His Leu Gly Tyr Ala Met Ala Ala Gly 485 490 495 Arg Phe Gly Trp Gly Leu Ala His Val Ala Ala Ala Val Ala Met Ser 500 505 510 Arg Arg Tyr Asp Arg Ala Gln Lys Gly Phe Leu Leu Thr Ser Leu Arg 515 520 525 Arg Ala Tyr Ala Pro Leu Leu Ala Arg Glu Asn Ala Ala Leu Thr Gly 530 535 540 Ala Arg Thr Pro Asp Asp Gly Gly Asp Ala Asn Arg His Asp Gly Asp 545 550 555 560 Asp Ala Arg Gly Lys Pro Ala Ala Ala Ala Pro Leu Pro Ser Ala 565 570 575 Ala Ala Ser Pro Ala Asp Glu Ala Ala Val Ala Gly Tyr Gly Ala 580 585 590 Ala Gly Val Leu Ala Ala Leu 595 600 605 Ala Pro Ala Gly Ala Asp Asp Asp Asp Asp Asp Asp Gly Ala Gly Gly 610 615 620 Gly Gly Gly Arg Arg Ala Glu Ala Gly Arg Val Ala Val Glu Cys 625 630 635 640 Leu Ala Ala Cys Arg Gly Ile Leu Glu Ala Leu Ala Glu Gly Phe Asp 645 650 655 Gly Asp Leu Ala Val Pro Gly Leu Ala Gly Ala Arg Pro Ala Ala 660 665 670 Pro Pro Arg Pro Gly Pro Ala Gly Ala Ala Ala Pro Pro His Ala Asp 675 680 685 Ala Pro Arg Leu Arg Ala Trp Leu Arg Glu Leu Arg Phe Val Arg Asp 690 695 700 Ala Leu Val Leu Met Arg Leu Arg Gly Asp Leu Arg Val Ala Gly Gly 705 710 715 720 Ser Glu Ala Ala Val Ala Ala Val Ala Val Ser Seru Val Ala Gly 725 730 735 Ala Leu Gly Pro Ala Leu Pro Arg Ser Pro Arg Leu Leu Ser Ser Ala 740 745 750 Ala Ala Ala Ala Asp Leu Leu Phe Asn Gln Ser Leu 755 760 765 <210> 16 <211> 2172 <212> PRT <213> Artificial Sequence <220> TPA-Flt3L-gD-IPC0-IPC4 fusion protein <400> 16 Met Asp Ala Met Lys Arg Gly Leu Cys Cys Val Leu Leu Leu Cys Gly 1 5 10 15 Ala Val Phe Val Ser Ser Ser His Ala Thr Gln Asp Cys Ser Phe Gln 20 25 30 His Ser Pro Ile Ser Ser Asp Phe Ala Val Lys Ile Arg Glu Leu Ser 35 40 45 Asp Tyr Leu Leu Gln Asp Tyr Pro Val Thr Val Ala Ser Asn Leu Gln 50 55 60 Asp Glu Glu Leu Cys Gly Gly Leu Trp Arg Leu Val Leu Ala Gln Arg 65 70 75 80 Trp Met Glu Arg Leu Lys Thr Val Ala Gly Ser Lys Met Gln Gly Leu 85 90 95 Leu Glu Arg Val Asn Thr Glu Ile His Phe Val Thr Lys Cys Ala Phe 100 105 110 Gln Pro Pro Pro Ser Cys Leu Arg Phe Val Gln Thr Asn Ile Ser Arg 115 120 125 Leu Leu Gln Glu Thr Ser Glu Gln Leu Val Ala Leu Lys Pro Trp Ile 130 135 140 Thr Arg Gln Asn Phe Ser Arg Cys Leu Glu Leu Gln Cys Gln Pro Asp 145 150 155 160 Ser Ser Thr Leu Pro Pro Pro Trp Ser Pro Arg Pro Leu Glu Ala Thr 165 170 175 Ala Pro Thr Ala Pro Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly 180 185 190 Arg Ala Lys Tyr Ala Leu Ala Asp Pro Ser Leu Lys Met Ala Asp Pro 195 200 205 Asn Arg Phe Arg Gly Lys Asn Leu Pro Val Leu Asp Gln Leu Thr Asp 210 215 220 Pro Pro Gly Val Lys Arg Val Tyr His Ile Gln Pro Ser Leu Glu Asp 225 230 235 240 Pro Phe Gln Pro Pro Ser Ile Pro Ile Thr Val Tyr Tyr Ala Val Leu 245 250 255 Glu Arg Ala Cys Arg Ser Val Leu Leu His Ala Pro Ser Glu Ala Pro 260 265 270 Gln Ile Val Arg Gly Ala Ser Asp Glu Ala Arg Lys His Thr Tyr Asn 275 280 285 Leu Thr Ile Ala Trp Tyr Arg Met Gly Asp Asn Cys Ala Ile Pro Ile 290 295 300 Thr Val Met Glu Tyr Thr Glu Cys Pro Tyr Asn Lys Ser Leu Gly Val 305 310 315 320 Cys Pro Ile Arg Thr Gln Pro Arg Trp Ser Tyr Tyr Asp Ser Phe Ser 325 330 335 Ala Val Ser Glu Asp Asn Leu Gly Phe Leu Met His Ala Pro Ala Phe 340 345 350 Glu Thr Ala Gly Thr Tyr Leu Arg Leu Val Lys Ile Asn Asp Trp Thr 355 360 365 Glu Ile Thr Gln Phe Ile Leu Glu His Arg Ala Arg Ala Ser Cys Lys 370 375 380 Tyr Ala Leu Pro Leu Arg Ile Pro Pro Ala Ala Cys Leu Thr Ser Lys 385 390 395 400 Ala Tyr Gln Gln Gly Val Thr Val Asp Ser Ile Gly Met Leu Pro Arg 405 410 415 Phe Ile Pro Glu Asn Gln Arg Thr Val Ala Leu Tyr Ser Leu Lys Ile 420 425 430 Ala Gly Trp His Gly Pro Lys Pro Pro Tyr Thr Ser Thr Leu Leu Pro 435 440 445 Pro Glu Leu Ser Asp Thr Thr Asn Ala Thr Gln Pro Glu Leu Val Pro 450 455 460 Glu Asp Pro Glu Asp Ser Ala Leu Leu Glu Asp Pro Ala Gly Thr Val 465 470 475 480 Ser Ser Gln Ile Pro Pro Asn Trp His Ile Pro Ser Ile Gln Asp Val 485 490 495 Ala Pro His His Ala Pro Ala Ala Pro Ser Asn Pro Arg Arg Arg Ala 500 505 510 Gln Met Ala Pro Lys Arg Leu Arg Leu Pro His Ile Arg Asp Asp Asp 515 520 525 Ala Pro Pro Ser His Gln Pro Leu Phe Tyr Gly Gly Gly Gly Ser Gly 530 535 540 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 545 550 555 560 Gly Gly Ser Met Glu Pro Arg Pro Gly Thr Ser Ser Ala Ala Asp Pro 565 570 575 Gly Pro Glu Arg Pro Pro Arg Gln Thr Pro Gly Thr Gln Pro Ala Ala 580 585 590 Pro His Ala Trp Gly Met Leu Asn Asp Met Gln Trp Leu Ala Ser Ser 595 600 605 Asp Ser Glu Glu Glu Thr Glu Val Gly Ile Ser Asp Asp Asp Leu His 610 615 620 Arg Asp Ser Thr Ser Glu Ala Gly Ser Thr Asp Thr Glu Met Phe Glu 625 630 635 640 Ala Gly Leu Met Asp Ala Ala Thr Pro Pro Ala Arg Pro Pro Ala Glu 645 650 655 Arg Gln Gly Ser Pro Thr Pro Ala Asp Ala Gln Gly Ser Cys Gly Gly 660 665 670 Gly Pro Val Gly Glu Glu Gly Ala Glu Ala Gly Gly Gly Gly Asp Val 675 680 685 Cys Ala Val Cys Thr Asp Glu Ile Ala Pro Pro Leu Arg Cys Gln Ser 690 695 700 Phe Pro Cys Leu His Pro Phe Cys Ile Pro Cys Met Lys Thr Trp Ile 705 710 715 720 Pro Leu Arg Asn Thr Cys Pro Leu Cys Asn Thr Pro Val Ala Tyr Leu 725 730 735 Ile Val Gly Val Thr Ala Ser Gly Ser Phe Ser Thr Ile Pro Ile Val 740 745 750 Asn Asp Pro Arg Thr Arg Val Glu Ala Glu Ala Ala Val Arg Ala Gly 755 760 765 Thr Ala Val Asp Phe Ile Trp Thr Gly Asn Pro Arg Thr Ala Pro Arg 770 775 780 Ser Leu Ser Leu Gly Gly His Thr Val Arg Ala Leu Ser Pro Thr Pro 785 790 795 800 Pro Trp Pro Gly Thr Asp Asp Glu Asp Asp Asp Leu Ala Asp Val Asp 805 810 815 Tyr Val Pro Pro Ala Pro Arg Arg Ala Pro Arg Arg Gly Gly Gly Gly 820 825 830 Ala Gly Ala Thr Arg Gly Thr Ser Gln Pro Ala Ala Thr Arg Pro Ala 835 840 845 Pro Pro Gly Ala Pro Arg Ser Ser Ser Ser Gly Gly Ala Pro Leu Arg 850 855 860 Ala Gly Val Gly Ser Gly Ser Gly Gly Gly Pro Ala Val Ala Ala Val 865 870 875 880 Val Pro Arg Val Ala Ser Leu Pro Pro Ala Ala Gly Gly Gly Arg Ala 885 890 895 Gln Ala Arg Arg Val Gly Glu Asp Ala Ala Ala Glu Gly Arg Thr 900 905 910 Pro Pro Ala Arg Gln Pro Arg Ala Gln Glu Pro Pro Ile Val Ile 915 920 925 Ser Asp Ser Pro Pro Ser Pro Arg Arg Pro Ala Gly Pro Gly Pro 930 935 940 Leu Ser Phe Val Ser Ser Ser Ala Gln Val Ser Ser Gly Pro Gly 945 950 955 960 Gly Gly Gly Leu Pro Gln Ser Ser Gly Arg Ala Ala Arg Pro Ala 965 970 975 Ala Val Ala Pro Arg Val Ala Pro Ala Ala Pro 980 985 990 Val Val Ser Ala Ser Ala Asp Ala Ala Gly Ala Pro Pro Ala Val 995 1000 1005 Pro Val Asp His Arg Ala Pro Arg Ser Ser Met Thr Gln Ala Gln 1010 1015 1020 Thr Asp Thr Gln Ala Gln Ser Leu Gly Arg Ala Gly Ala Thr Asp Ala 1025 1030 1035 1040 Arg Gly Ser Gly Gly Pro Gly Ala Glu Gly Gly Pro Gly Val Pro Arg 1045 1050 1055 Gly Thr Asn Thr Pro Gly Ala Ala Pro His Ala Ala Glu Gly Ala Ala 1060 1065 1070 Gly Ser Asp Ser Gly Pro Ala Ala Ser Ser Ser Ala Ser Ser Ala 1075 1080 1085 Ala Pro Arg Ser Pro Leu Ala Pro Gln Gly Val Gly Ala Lys Arg Ala 1090 1095 1100 Ala Pro Arg Arg Ala Pro Asp Ser Asp Ser Gly Asp Arg Gly His Gly 1105 1110 1115 1120 Pro Leu Ala Pro Ala Ser Ala Gly Ala Ala Pro Pro Ser Ala Ser Pro 1125 1130 1135 Ser Ser Gln Ser Ser Ala Ser Ser Ser Ser Ala Ser Ser 1140 1145 1150 Ser Ser Ala Ser Ser Ser Ser Ser Ser Ser Ser Ala Ser Ser Ser 1155 1160 1165 Ser Ala Ser Ser Ser Ala Ser Ser Ser Ala Ser Ser Ala 1170 1175 1180 Gly Gly Ala Gly Gly Ser Val Ala Ser Ala Ser Gly Ala Gly Glu Arg 1185 1190 1195 1200 Arg Glu Thr Ser Leu Gly Pro Arg Ala Ala Pro Arg Gly Pro Arg 1205 1210 1215 Lys Cys Ala Arg Lys Thr Arg His Ala Glu Gly Gly Pro Glu Pro Gly 1220 1225 1230 Ala Arg Asp Pro Ala Pro Gly Leu Thr Arg Tyr Leu Pro Ile Ala Gly 1235 1240 1245 Val Ser Ser Val Val Ala Leu Ala Pro Tyr Val Asn Lys Thr Val Thr 1250 1255 1260 Gly Asp Cys Leu Pro Val Leu Asp Met Glu Thr Gly His Ile Gly Ala 1265 1270 1275 1280 Tyr Val Val Leu Val Asp Gln Thr Gly Asn Val Ala Asp Leu Leu Arg 1285 1290 1295 Ala Ala Ala Pro Ala Trp Ser Arg Arg Thr Leu Leu Pro Glu His Ala 1300 1305 1310 Arg Asn Cys Val Arg Pro Pro Asp Tyr Pro Thr Pro Pro Ala Ser Glu 1315 1320 1325 Trp Asn Ser Leu Trp Met Thr Pro Val Gly Asn Met Leu Phe Asp Gln 1330 1335 1340 Gly Thr Leu Val Gly Ala Leu Asp Phe His Gly Leu Arg Ser Ser His 1345 1350 1355 1360 Pro Trp Ser Arg Glu Gln Gly Ala Pro Ala Pro Ala Gly Asp Ala Pro 1365 1370 1375 Ala Gly His Gly Gly Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ser Gly 1380 1385 1390 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Met Ser 1395 1400 1405 Ala Glu Gln Arg Lys Lys Lys Lys Thr Thr Thr Thr Gln Gly Arg 1410 1415 1420 Gly Ala Glu Val Ala Met Ala Asp Glu Asp Gly Gly Arg Leu Arg Ala 1425 1430 1435 1440 Ala Ala Glu Thr Thr Gly Gly Pro Gly Ser Pro Asp Pro Ala Asp Gly 1445 1450 1455 Pro Pro Thr Pro Asn Pro Asp Arg Arg Pro Ala Ala Arg Pro Gly 1460 1465 1470 Phe Gly Trp His Gly Gly Pro Glu Glu Asn Glu Asp Glu Ala Asp Asp 1475 1480 1485 Ala Ala Ala Asp Ala Asp Ala Asp Glu Ala Ala Pro Ala Ser Gly Glu 1490 1495 1500 Ala Val Asp Glu Pro Ala Ala Asp Gly Val Val Ser Pro Arg Gln Leu 1505 1510 1515 1520 Ala Leu Leu Ala Ser Met Val Asp Glu Ala Val Arg Thr Ile Pro Ser 1525 1530 1535 Pro Pro Glu Arg Asp Gly Ala Gln Glu Glu Ala Ala Arg Ser Pro 1540 1545 1550 Ser Pro Pro Arg Thr Pro Ser Met Arg Ala Asp Tyr Gly Glu Glu Asn 1555 1560 1565 Asp Asp Asp Asp Asp Asp Asp Asp Asp Asp Asp Arg Asp Ala Gly Arg 1570 1575 1580 Trp Val Arg Gly Pro Glu Thr Thr Ser Ala Val Arg Gly Ala Tyr Pro 1585 1590 1595 1600 Asp Pro Met Ala Ser Leu Ser Pro Arg Pro Pro Ala Pro Arg Arg His 1605 1610 1615 His His His His His His Arg Arg Arg Ala Pro Arg Arg Arg Ser 1620 1625 1630 Ala Ala Ser Asp Ser Ser Lys Ser Gly Ser Ser Ser Ser Ala Ser Ser 1635 1640 1645 Ala Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser A Ser Ser Ser 1650 1655 1660 Asp Asp Asp Asp Asp Asp Asp Ala Ala Arg Ala Pro Ala Ser Ala Ala 1665 1670 1675 1680 Asp His Ala Ala Gly Gly Thr Leu Gly Ala Asp Asp Glu Glu Ala Gly 1685 1690 1695 Val Pro Ala Arg Ala Pro Gly Ala Ala Pro Arg Pro Ser Pro Pro Arg 1700 1705 1710 Ala Glu Pro Ala Pro Ala Arg Thr Pro Ala Ala Thr Ala Gly Arg Leu 1715 1720 1725 Glu Arg Arg Ala Arg Ala Ala Val Ala Gly Arg Asp Ala Thr Gly 1730 1735 1740 Arg Phe Thr Ala Gly Arg Pro Arg Arg Val Glu Leu Asp Ala Asp Ala 1745 1750 1755 1760 Ala Ser Gly Ala Phe Tyr Ala Arg Tyr Arg Asp Gly Tyr Val Ser Gly 1765 1770 1775 Glu Pro Trp Pro Gly Ala Gly Pro Pro Pro Pro Gly Arg Val Leu Tyr 1780 1785 1790 Gly Gly Leu Gly Asp Ser Arg Pro Gly Leu Trp Gly Ala Pro Glu Ala 1795 1800 1805 Glu Glu Ala Arg Ala Arg Phe Glu Ala Ser Gly Ala Pro Ala Pro Val 1810 1815 1820 Trp Ala Pro Glu Leu Gly Asp Ala Ala Gln Gln Tyr Ala Leu Ile Thr 1825 1830 1835 1840 Arg Leu Leu Tyr Thr Pro Asp Ala Glu Ala Met Gly Trp Leu Gln Asn 1845 1850 1855 Pro Arg Val Ala Pro Gly Asp Val Ala Leu Asp Gln Ala Cys Phe Arg 1860 1865 1870 Ile Ser Gly Ala Ala Arg Asn Ser Ser Ser Phe Ile Ser Gly Ser Ser 1875 1880 1885 Ala Arg Ala Val Pro His Leu Gly Tyr Ala Met Ala Ala Gly Arg Phe 1890 1895 1900 Gly Trp Gly Leu Ala His Val Ala Ala Ala Val Ala Met Ser Arg Arg 1905 1910 1915 1920 Tyr Asp Arg Ala Gln Lys Gly Phe Leu Leu Thr Ser Leu Arg Arg Ala 1925 1930 1935 Tyr Ala Pro Leu Leu Ala Arg Glu Asn Ala Ala Leu Thr Gly Ala Arg 1940 1945 1950 Thr Pro Asp Asp Gly Gly Asp Ala Asn Arg His Asp Gly Asp Asp Ala 1955 1960 1965 Arg Gly Lys Pro Ala Ala Ala Ala Ala Pro Ala Ala 1970 1975 1980 Ser Pro Ala Asp Glu Arg Ala Val Pro Ala Gly Tyr Gly Ala Ala Gly 1985 1990 1995 2000 Val Leu Ala Ala Leu Gly Arg Leu Ser Ala Ala Pro Ala Ser Ala Pro 2005 2010 2015 Ala Gly Ala Asp Asp Asp Asp Asp Asp Asp Gly Ala Gly Gly Gly Gly 2020 2025 2030 Gly Gly Arg Arg Ala Glu Ala Gly Arg Val Ala Val Glu Cys Leu Ala 2035 2040 2045 Ala Cys Arg Gly Ile Leu Glu Ala Leu Ala Glu Gly Phe Asp Gly Asp 2050 2055 2060 Leu Ala Ala Val Pro Gly Leu Ala Gly Ala Arg Pro Ala Ala Pro Pro 2065 2070 2075 2080 Arg Pro Gly Pro Ala Gly Ala Ala Ala Pro Pro His Ala Asp Ala Pro 2085 2090 2095 Arg Leu Arg Ala Trp Leu Arg Glu Leu Arg Phe Val Arg Asp Ala Leu 2100 2105 2110 Val Leu Met Arg Leu Arg Gly Asp Leu Arg Val Ala Gly Gly Ser Glu 2115 2120 2125 Ala Ala Val Ala Ala Val Ala Val Ala Val Ala Gly Ala Leu 2130 2135 2140 Gly Pro Ala Leu Pro Ser Ser Ala Ala 2145 2150 2155 2160 Ala Ala Asp Leu Leu Phe Asn Gln Ser Leu 2165 2170 <210> 17 <211> 6519 <212> DNA <213> Artificial Sequence <220> <223> Polynucleotide encoding tPA-Flt3L-gD-IPC0-IPC4 fusion protein <400> 17 atggacgcca tgaagagagg cctgtgctgc gtgctgctgc tgtgcggcgc cgtgttcgtg 60 agccccagcc acgccaccca ggactgcagc ttccagcaca gccccatcag cagcgacttc 120 gccgtgaaga tcagagagct gagcgactac ctgctgcagg actaccccgt gaccgtggcc 180 agcaacctgc aggacgagga gctgtgcggc ggcctgtgga gactggtgct ggcccagaga 240 tggatggaga gactgaagac cgtggccggc agcaagatgc agggcctgct ggagagagtg 300 aacaccgaga tccacttcgt gaccaagtgc gccttccagc ctccccccag ctgcctgagg 360 ttcgtgcaga ccaacatcag cagactgctg caggagacca gcgagcagct ggtggccctg 420 aagccctgga tcaccagaca gaacttcagc agatgcctgg agctgcagtg ccagcccgac 480 agcagcaccc tgccccctcc ctggagcccc agacccctgg aggccaccgc tcccacagcc 540 cctggcagcg ggtccggaag tgggtctgga tccgggcgcg ccaagtatgc tctggccgac 600 ccaagcctga agatggccga ccctaataga ttccggggca agaacctgcc tgtcctggac 660 cagctgaccg atccccctgg cgtgaagcgc gtctaccaca tccagccttc actggaggac 720 ccattccagc cacccagcat ccctattacc gtgtactatg ctgtcctgga acgcgcatgc 780 cgaagtgtgc tgctgcacgc tccctcagag gcacctcaga tcgtcagagg agcctccgat 840 gaagctagga agcatactta caacctgacc attgcctggt atcggatggg cgacaattgt 900 gctatcccca ttacagtgat ggagtacact gaatgccctt ataacaaatc tctgggcgtc 960 tgtcccattc gaacacagcc tcggtggtcc tactatgatt cattcagcgc cgtgtctgag 1020 gacaacctgg gcttcctgat gcacgctcca gcatttgaaa cagccgggac ttatctgaga 1080 ctggtgaaaa tcaatgattg gaccgagatc acacagttta ttctggaaca tagggcccgc 1140 gctagctgca agtacgcact gccactgagg attcctccag cagcttgtct gaccagtaaa 1200 gcttatcagc agggagtgac agtggactca atcggcatgc tgccacgctt cattcccgag 1260 aaccagcgaa ccgtggcact gtacagcctg aagatcgctg ggtggcacgg acccaaaccc 1320 ccttatacta gcaccctgct gccacccgag ctgtccgata ccacaaatgc cacacagccc 1380 gaactggtgc ctgaggaccc agaagacagc gcactgctgg aggaccctgc cggcactgtg 1440 agctcccaga tccctccaaa ctggcatatc cctagcattc aggacgtggc accacaccat 1500 gcaccagcag caccttccaa tccacggaga agggctcaga tggcaccaaa gcgactgcgg 1560 ctgccccaca ttagagacga tgacgccccc ccttcccatc agcctctgtt ttacggagga 1620 ggaggctgg gaggaggagg atctggaggc gggggaagtg gcgggggagg ctcaggggga 1680 ggcggatcta tggaacccag accaggaact tcaagcagag ccgacccagg accagaaaga 1740 cctccccgac agacaccagg cactcagcca gccgcaccac acgcctgggg catgctgaat 1800 gatatgcagt ggctggccag ctccgactct gaggaggaga cagaggtggg catctccgat 1860 gacgatctgc acagggactc cacaagcgag gccggctcca ccgataccga gatgtttgag 1920 gccggcctga tggacgccgc cacccctcca gcccggcccc cagccgagag gcagggctct 1980 ccaacacccg ccgatgccca gggctcctgc ggcggcggcc ctgtgggcga ggaggaggcc 2040 gaggccggcg gcggcggcga cgtgtgcgcc gtgtgcacag acgagatcgc cccacctctg 2100 agatgccaga gctttccatg tctgcaccca ttctgcatcc catgcatgaa gacctggata 2160 cctctgagga atacctgtcc tctgtgcaat accccagtgg cctacctgat cgtgggcgtg 2220 accgcctctg gctcctttag cacaatcccc atcgtgaacg atcctagaac ccgggtggag 2280 gccgaggccg ccgtgcgggc cggcaccgcc gtggatttca tctggaccgg caacccacgg 2340 acagccccta ggtctctgag cctgggcggc cacaccgtga gagccctgag ccccacaccc 2400 ccttggccag gcacagacga tgaggatgac gacctggccg acgtggatta tgtgccccct 2460 gcccccagaa gagccccccg gagaggcggc ggcggcgccg gcgccacaag aggcacatcc 2520 cagcctgccg ccacaaggcc tgccccacct ggcgccccac ggtcctcctc cagcggcggc 2580 gccccactgc gggccggcgt gggctccggc tccggcggcg gcccagccgt ggccgccgtg 2640 gtgcccagag tggccagcct gcctccagcc gccggcggcg gcagagccca ggccagacgg 2700 gtgggcgagg atgccgccgc cgccgagggc cggacacccc ctgccagaca gccaagagcc 2760 gcccaggagc cccctatcgt gatcagcgac tctccacctc cctccccaag aaggccagcc 2820 ggcccaggcc ctctgtcctt tgtgagctct tcttccgccc aggtgtcttc cggcccaggc 2880 ggcggcggcc tgcctcagtc ctccggcaga gccgccagac cacgggccgc cgtggcccct 2940 agagtgagga gccccccaag ggccgccgcc gccccagtgg tgtccgcctc cgccgatgcc 3000 gccggcccag ccccacccgc cgtgccagtg gacgcccaca gggccccacg gagcagaatg 3060 acccaggccc agaccgatac ccaggcccag agcctgggca gggccggcgc caccgatgcc 3120 agaggctccg gcggcccagg cgccgagggc ggccccggcg tgcccagggg cacaaataca 3180 ccaggcgccg ccccccacgc cgccgagggc gccgccggca gcgattccgg ccctgccgcc 3240 tcctcttctg cctccagcag cgccgcccca cgctctcccc tggccccaca gggcgtgggc 3300 gccaagcggg ccgccccaag acgggcccct gactccgatt ccggcgatcg gggccacggc 3360 ccactggccc ctgcctctgc cggcgccgcc cctcctagcg cctccccaag ctctcaggcc 3420 gccgtggccg ccgccagctc cagctccgcc agctcttcct ctgccagctc ctcctctgcc 3480 tcctcctcct ctgcctcttc cagctctgcc agctctagca gcgcctcctc tagctccgcc 3540 tcttctagcg ccggcggcgc cggcggctcc gtggcctctg cctccggcgc cggcgagaga 3600 agagagacct ctctgggccc aagagccgcc gcccccagag gccctagaaa gtgtgccagg 3660 aagaccaggc acgccgaggg cggcccagag ccaggcgccc gcgaccctgc cccaggcctg 3720 acacggtatc tgccaatcgc cggcgtgagc tctgtggtgg ccctggcccc atatgtgaac 3780 aagacagtga caggcgactg tctgccagtg ctggacatgg agacaggcca catcggcgcc 3840 tatgtggtgc tggtggacca gacaggcaat gtggccgatc tgctgagagc cgccgcccct 3900 gcctggtcca gaaggacact gctgccagag cacgccagaa actgcgtgag accacctgac 3960 tatcccacac cacccgcctc tgagtggaat agcctgtgga tgacaccagt gggcaacatg 4020 ctgtttgatc agggcacact ggtgggcgcc ctggatttcc acggcctgag atcccgccac 4080 ccttggtcca gagagcaggg cgcccccgcc ccagccggcg atgccccagc cggccacggc 4140 gagggcggcg gcggctcagg gggggggggc agcggcggag gtggctcagg aggtggaggt 4200 agcggaggtg gcggatctat gtcagcagaa cagaggaaga agaagaaaac aacaacaact 4260 actcagggaa ggggagcaga agtggcaatg gcagacgagg atggaggcag actgagggcc 4320 gccgccgaga caaccggcgg cccaggctct cctgacccag ccgatggccc accaccaaca 4380 cccaacccag accgcagacc agccgccaga cctggctttg gctggcacgg cggcccagag 4440 gagaacgagg atgaggccga cgatgccgcc gccgatgccg acgccgatga ggccgccccc 4500 gcctccggcg aggccgtgga cgagccagcc gccgacggcg tggtgtctcc tagacagctg 4560 gccctgctgg cctccatggt ggacgaggcc gtgaggacca tcccatctcc tcctccagag 4620 agagatggcg cccaggagga ggccgccagg agcccatccc ccccaagaac ccctagcatg 4680 agagccgact atggcgagga gaacgatgat gacgacgacg atgacgatga tgatgacagg 4740 gacgccggcc gctgggtgcg gggcccagag accacatctg ccgtgagagg cgcctatcct 4800 gcccaatgg cctctctgag cccaagacct cccgccccaa gacggcacca ccaccaccac 4860 caccagga gacggagggc cccaagaagg cggagcgccg ccagcgactc ctctaagtct 4920 ggctccagca gctctgcctc cagcgccagc tcctctgcca gctcctcctc tagcgccagc 4980 gcctcttcct ccgacgatga tgacgatgat gacgccgcca gggcccctgc ctccgccgcc 5040 gatcacgccg ccggcggcac cctgggcgcc gatgacgagg aggccggcgt gccagccaga 5100 gccccaggcg ccgcccccag gccttctcca ccaagagccg agcccgcccc agccaggacc 5160 ccagccgcca cagccggcag actggagcgg agacgcgcca gagccgccgt ggccggcaga 5220 gatgccacag gcaggtttac cgccggcagg cccagaaggg tggagctgga tgccgatgcc 5280 gcctccggcg ccttttacgc cagatacagg gacggctacg tgagcggcga gccctggcca 5340 ggcgccggcc ctcccccacc tggcagggtg ctgtatggcg gcctgggcga tagcagacct 5400 ggcctgtggg gcgcccctga ggccgaggag gccagagcca gattcgaggc ctctggcgcc 5460 ccagcccccg tgtgggcccc agagctgggc gacgccgccc agcagtacgc cctgatcacc 5520 agactgctgt ataccccaga cgccgaggcc atgggctggc tgcagaaccc tagagtggcc 5580 ccaggcgacg tggccctgga tcaggcctgc ttccggatct ctggcgccgc ccggaacagc 5640 tctagcttta tctccggctc tgtggccaga gccgtgccac acctgggcta cgccatggcc 5700 gccggcaggt tcggctgggg cctggcccac gtagccgccg ccgtggccat gtccagacgg 5760 tatgacagag cccagaaggg ctttctgctg acatctctga ggcgggccta tgcccctctg 5820 ctggccaggg agaatgccgc cctgacaggc gccagaaccc ctgacgacgg cggcgatgcc 5880 aacagacacg atggcgacga tgccagaggc aagccagccg ccgccgccgc ccccctgcca 5940 tccgccgccg cctcccccgc cgatgagaga gccgtgcctg ccggctatgg cgccgccggc 6000 gtgctggccg ccctgggccg gctgtctgcc gccccagcct ccgcccccgc cggcgccgac 6060 gggatgacg acgatgatgg cgccggcggc ggcggcggcg gcagacgggc cgaggccggc 6120 agggtggccg tggagtgtct ggccgcctgc cgcggcatcc tggaggccct ggccgagggc 6180 tttgatggcg acctggccgc cgtgcccggc ctggccggcg ccagacctgc cgccccacct 6240 agacctggcc ccgccggcgc cgccgccccc cctcacgccg acgccccacg gctgagagcc 6300 tggctgagag agctgagatt cgtgcgggac gccctggtgc tgatgcggct gagaggcgat 6360 ctgcgggtgg ccggcggctc tgaggccgcc gtggccgccg tgagagccgt gtccctggtg 6420 gccggcgccc tgggcccagc cctgcctaga tccccaagac tgctgtcctc cgccgctgcc 6480 gctgccgccg acctgctgtt tcagaaccag agcctgtaa 6519

Claims

HSV-2의 UL39 단백질의 아미노산 서열상 14-154 a.a 위치에 상응하는 펩타이드에서 78-104 a.a 위치에 상응하는 막통과 도메인이 결실된 UL39-N1 펩타이드, 상기 UL39 단백질의 아미노산 서열상 1117-1142 a.a 위치에 상응하는 UL39-C2 펩타이드, 상기 UL39 단백질의 아미노산 서열상 155-227 a.a 위치에 상응하는 UL39-N2 펩타이드, 상기 UL39 단백질의 아미노산 서열상 399-1116 a.a 위치에 상응하는 UL39 N4-C1 펩타이드, 및 상기 UL39 단백질의 아미노산 서열상 208-398 a.a 위치에 상응하는 UL39-N3 펩타이드가 무작위적으로 섞여있고, 원래의 UL39 단백질의 아미노산 서열은 포함하지 않는 셔플드 UL39 단백질을 암호화하는 폴리뉴클레오타이드.A UL39-N1 peptide lacking the transmembrane domain corresponding to the 78-104 aa position in the peptide corresponding to the 14-154 aa position on the amino acid sequence of the UL39 protein of HSV-2, the amino acid sequence of the UL39 protein of 1117-1142 aa , A UL39-N2 peptide corresponding to positions 155-227 aa on the amino acid sequence of said UL39 protein, a UL39 N4-C1 peptide corresponding to positions 399-1116 aa on the amino acid sequence of said UL39 protein, And a polynucleotide encoding a shuffled UL39 protein randomly mixed with a UL39-N3 peptide corresponding to the position 208-398 aa in the amino acid sequence of the UL39 protein and not containing the amino acid sequence of the original UL39 protein.

제1항에 있어서,
하나 또는 둘 이상의 면역 증진 펩타이드를 암호화하는 폴리뉴클레오타이드를 추가로 포함하는, 폴리뉴클레오타이드.The method according to claim 1,
A polynucleotide further comprising a polynucleotide encoding one or more immunity enhancing peptides.

제2항에 있어서,
상기 면역 증진 펩타이드는 CD28, ICOS(inducible costimulator), CTLA4(cytotoxic T lymphocyte associated protein 4), PD1(programmed cell death protein 1), BTLA(B and T lymphocyte associated protein), DR3(death receptor 3), 4-1BB, CD2, CD40, CD30, CD27, SLAM(signaling lymphocyte activation molecule), 2B4(CD244), NKG2D(natural-killer group 2, member D)/DAP12(DNAX-activating protein 12), TIM1(T-Cell immunoglobulin and mucin domain containing protein 1), TIM2, TIM3, TIGIT, CD226, CD160, LAG3(lymphocyte activation gene 3), B7-1, B7-H1, GITR(glucocorticoid-induced TNFR family related protein), Flt3 리간드(fms-like tyrosine kinase 3 ligand), 플라젤린(flagellin), HVEM(herpesvirus entry mediator), CD40 L(리간드) 또는 OX40L[ligand for CD134(OX40), CD252]의 세포질 도메인 또는 이들 중 둘 이상의 연결체인, 폴리뉴클레오타이드.3. The method of claim 2,
The immunopotentiating peptides include CD28, inducible costimulator (ICOS), cytotoxic T lymphocyte associated protein 4 (CTLA4), programmed cell death protein 1 (PD1), B and T lymphocyte associated protein (BTLA), DR3 1BB, CD2, CD40, CD30, CD27, SLAM (signaling lymphocyte activation molecule), 2B4 (CD244), NKG2D / DNAX- activating protein 12, TIM1 immunoglobulin and mucin domain containing protein 1), TIM2, TIM3, TIGIT, CD226, CD160, lymphocyte activation gene 3, B7-1, B7-H1, glucocorticoid-induced TNFR family related protein, Flt3 ligand a tyrosine kinase 3 ligand, a flagellin, a herpesvirus entry mediator (HVEM), a CD40 L (ligand), or a cytoplasmic domain of OX40L [ligand for CD134 (OX40), CD252] Nucleotides.

제1항에 있어서,
분비 신호서열을 암호화하는 폴리뉴클레오타이드를 추가로 포함하는, 폴리뉴클레오타이드.The method according to claim 1,
Wherein the polynucleotide further comprises a polynucleotide encoding a secretory signal sequence.

제4항에 있어서,
상기 분비 신호서열은 tPA(tissue plasminogen activator) 신호서열, HSV gD(단순포진 바이러스 당단백질 D) 신호서열 또는 성장호르몬 신호서열인, 폴리뉴클레오타이드.5. The method of claim 4,
Wherein the secretory signal sequence is a tissue plasminogen activator (tPA) signal sequence, a HSV gD (herpes simplex virus glycoprotein D) signal sequence or a growth hormone signal sequence.

제1항에 있어서,
하나 또는 둘 이상의 제2형 단순포진 바이러스(HSV-2)의 항원 단백질을 암호화하는 폴리뉴클레오타이드를 추가로 포함하는, 폴리뉴클레오타이드.The method according to claim 1,
Wherein the polynucleotide further comprises a polynucleotide encoding an antigenic protein of one or more of the Type 2 herpes simplex virus (HSV-2).

제6항에 있어서,
상기 항원 단백질은 당단백질 B(gB), 당단백질 D(gD), ICP0 또는 ICP4인, 폴리뉴클레오타이드.The method according to claim 6,
Wherein the antigen protein is glycoprotein B (gB), glycoprotein D (gD), ICPO or ICP4.

제7항에 있어서,
상기 당단백질 B는 1-22 a.a 위치에 상응하는 신호서열 및 772-792 a.a 위치에 상응하는 막통과 도메인이 결실된 것인, 폴리뉴클레오타이드.8. The method of claim 7,
Wherein the glycoprotein B has a signal sequence corresponding to the 1-22 aa position and a transmembrane domain corresponding to the 772-792 aa position deleted.

제7항에 있어서,
상기 당단백질 D는 1-25 a.a 위치에 상응하는 신호서열 및 341-364 a.a 위치에 상응하는 막통과 도메인이 결실된 것인, 폴리뉴클레오타이드.8. The method of claim 7,
Wherein the glycoprotein D has a signal sequence corresponding to the 1-25 aa position and a transmembrane domain corresponding to the 341-364 aa position deleted.

제7항에 있어서,
상기 ICP0은 510-516 a.a 위치에 상응하는 핵위치화 신호(NLS)가 결실된 것인, 폴리뉴클레오타이드.8. The method of claim 7,
Wherein the ICPO is a nucleotide localization signal (NLS) corresponding to position 510-516 aa deleted.

제7항에 있어서,
상기 ICP4는 767-1318 a.a 위치에 상응하는 RS1.3 부위가 결실된 것인, 폴리뉴클레오타이드.8. The method of claim 7,
Wherein said ICP4 is deleted from the RS1.3 site corresponding to position 767-1318 aa.

제1항에 있어서,
상기 셔플드 UL39 단백질은 상기 UL-39-N1 펩타이드, 상기 UL39-C2 펩타이드, 상기 UL39-N2 펩타이드, 상기 UL39 N4-C1 펩타이드, 및 상기 UL39-N3 펩타이드가 순차적으로 연결된, 폴리뉴클레오타이드. The method according to claim 1,
Wherein the shuffled UL39 protein is a polynucleotide in which the UL-39-N1 peptide, the UL39-C2 peptide, the UL39-N2 peptide, the UL39 N4-C1 peptide, and the UL39-N3 peptide are sequentially connected.

제1항 내지 제12항 중 어느 한 항에 있어서,
코돈 최적화된, 폴리뉴클레오타이드.13. The method according to any one of claims 1 to 12,
Codon-optimized, polynucleotide.

제1항 내지 제12항 중 어느 한 항에 있어서,
상기 UL-39-N1 펩타이드는 서열번호 1,
상기 UL39-C2 펩타이드는 서열번호 2,
상기 UL39-N2 펩타이드는 서열번호 3,
상기 UL39 N4-C1 펩타이드는 서열번호 4, 및
상기 UL39-N3 펩타이드는 서열번호 5로 구성되는, 폴리뉴클레오타이드.13. The method according to any one of claims 1 to 12,
The UL-39-N1 peptide comprises SEQ ID NO: 1,
The UL39-C2 peptide comprises SEQ ID NO: 2,
The UL39-N2 peptide comprises SEQ ID NO: 3,
The UL39 N4-C1 peptide comprises SEQ ID NO: 4, and
Wherein said UL39-N3 peptide consists of SEQ ID NO: 5.

제1항 내지 제12항 중 어느 한 항에 있어서,
상기 셔플드 UL39 단백질은 서열번호 6으로 기재되는 아미노산 서열로 구성되는, 폴리뉴클레오타이드.13. The method according to any one of claims 1 to 12,
Wherein the shuffled UL39 protein is composed of the amino acid sequence shown in SEQ ID NO: 6.

제1항 내지 제12항 중 어느 한 항의 폴리뉴클레오타이드를 포함하는 벡터.12. A vector comprising the polynucleotide of any one of claims 1 to 12.

제16항에 있어서, 상기 폴리뉴클레오타이드가 프로모터에 작동가능하게 연결된 발현벡터인, 벡터.17. The vector of claim 16, wherein the polynucleotide is an expression vector operably linked to a promoter.

제15항 또는 제16항의 벡터를 포함하는 단리된 숙주 세포.An isolated host cell comprising the vector of claim 15 or 16.

제1항 내지 제12항 중 어느 한 항의 폴리뉴클레오타이드 또는 제16항 또는 제17항의 벡터 및 약학적으로 허용가능한 담체를 포함하는 조성물.13. A composition comprising a polynucleotide of any one of claims 1 to 12 or a vector of claims 16 or 17 and a pharmaceutically acceptable carrier.

제1항 내지 제12항 중 어느 한 항의 폴리뉴클레오타이드에 의해 암호화되는 재조합 단백질.12. A recombinant protein encoded by the polynucleotide of any one of claims 1 to 12.

gB, gD, UL39, ICP0 및 ICP4 단백질로 구성되는 군으로부터 선택되는 2종 이상의 또는 모든 HSV-2 항원 단백질을 발현하도록 상기 항원 단백질을 암호화하는 폴리뉴클레오타이드를 포함하는 발현벡터. an antigenic protein encoding the antigen protein to express two or more or all HSV-2 antigen proteins selected from the group consisting of gB, gD, UL39, ICPO and ICP4 proteins.

제21항에 있어서,
상기 gB는 1-22 a.a 위치에 상응하는 신호서열 및 772-792 a.a 위치에 상응하는 막통과 도메인이 결실된 것인, 발현벡터.22. The method of claim 21,
Wherein the gB has a signal sequence corresponding to the 1-22 aa position and a transmembrane domain corresponding to the 772-792 aa position deleted.

제21항에 있어서,
상기 gD는 1-25 a.a 위치에 상응하는 신호서열 및 341-364 a.a 위치에 상응하는 막통과 도메인이 결실된 것인, 발현벡터.22. The method of claim 21,
Wherein the gD is a signal sequence corresponding to the 1-25 aa position and the transmembrane domain corresponding to the 341-364 aa position is deleted.

제21항에 있어서,
상기 ICP0은 510-516 a.a 위치에 상응하는 핵위치화 신호(NLS)가 결실된 것인, 발현벡터.22. The method of claim 21,
Wherein said ICPO is a deletion of a nuclear localization signal (NLS) corresponding to position 510-516 aa.

제21항에 있어서,
상기 ICP4는 767-1318 a.a 위치에 상응하는 RS1.3 부위가 결실된 것인, 발현벡터.22. The method of claim 21,
Wherein said ICP4 is deleted from the RS1.3 site corresponding to position 767-1318 aa.

제21항에 있어서,
상기 UL39는 내부의 도메인들이 뒤섞인 셔플드 UL39인, 발현벡터.22. The method of claim 21,
The UL39 is an expression vector having a shuffled UL39 mixed with internal domains.

제26항에 있어서,
상기 셔플드 UL39는 HSV-2의 UL39 단백질의 아미노산 서열상 14-154 a.a 위치에 상응하는 펩타이드에서 78-104 a.a 위치에 상응하는 막통과 도메인이 결실된 UL39-N1 펩타이드, 상기 UL39 단백질의 아미노산 서열상 1117-1142 a.a 위치에 상응하는 UL39-C2 펩타이드, 상기 UL39 단백질의 아미노산 서열상 155-227 a.a 위치에 상응하는 UL39-N2 펩타이드, 상기 UL39 단백질의 아미노산 서열상 399-1116 a.a 위치에 상응하는 UL39 N4-C1 펩타이드, 및 상기 UL39 단백질의 아미노산 서열상 208-398 a.a 위치에 상응하는 UL39-N3 펩타이드가 무작위적으로 섞여있고, 원래의 UL39 단백질의 아미노산 서열은 포함하지 않는 것인, 발현벡터27. The method of claim 26,
The shuffled UL39 is a UL39-N1 peptide in which the transmembrane domain corresponding to the 78-104 aa position in the peptide corresponding to the 14-154 aa position on the amino acid sequence of the UL39 protein of HSV-2 is deleted, the amino acid sequence of the UL39 protein UL39-C2 peptide corresponding to position aa of positions 1117-1142, UL39-N2 peptide corresponding to positions 155-227 aa on the amino acid sequence of said UL39 protein, UL39-N2 peptide corresponding to position 399-1116 aa on amino acid sequence of said UL39 protein, N4-Cl peptide and UL39-N3 peptide corresponding to position 208-398 aa on the amino acid sequence of said UL39 protein are randomly mixed and do not contain the amino acid sequence of the original UL39 protein.

제26항에 있어서,
상기 셔플드 UL39는 제1항의 폴리뉴클레오타이드에 의해 암호화되는, 발현벡터.27. The method of claim 26,
Wherein the shuffled UL39 is encoded by the polynucleotide of claim 1.

제21항 내지 제28항 중 어느 한 항에 있어서,
상기 gB, gD, UL39, ICP0 및 ICP4 단백질은 별개의 단백질로 발현되거나 하나의 융합단백질 형태로 발현되는, 발현벡터.29. The method according to any one of claims 21 to 28,
Wherein the gB, gD, UL39, ICPO and ICP4 proteins are expressed as separate proteins or expressed in the form of a fusion protein.

제21항 내지 제28항 중 어느 한 항에 있어서,
상기 폴리뉴클레오타이드는 분비 신호서열 암호화하는 폴리뉴클레오타이드를 추가로 포함하는, 발현벡터.29. The method according to any one of claims 21 to 28,
Wherein the polynucleotide further comprises a polynucleotide encoding a secretory signal sequence.

제30항에 있어서,
상기 분비 신호서열은 tPA(tissue plasminogen activator) 신호서열, HSV gDs(단순포진 바이러스 당단백질 Ds) 신호서열 또는 성장호르몬 신호서열인, 발현벡터.31. The method of claim 30,
Wherein the secretory signal sequence is a tissue plasminogen activator (tPA) signal sequence, a HSV gDs (herpes simplex virus glycoprotein Ds) signal sequence or a growth hormone signal sequence.

제21항 내지 제28항 중 어느 한 항에 있어서,
상기 폴리뉴클레오타이드는 면역 증진 펩타이드를 암호화하는 폴리뉴클레오타이드를 추가로 포함하는, 발현벡터.29. The method according to any one of claims 21 to 28,
Wherein the polynucleotide further comprises a polynucleotide encoding an immunity enhancing peptide.

제32항에 있어서,
상기 면역 증진 펩타이드는 CD28, ICOS(inducible costimulator), CTLA4(cytotoxic T lymphocyte associated protein 4), PD1(programmed cell death protein 1), BTLA(B and T lymphocyte associated protein), DR3(death receptor 3), 4-1BB, CD2, CD40, CD30, CD27, SLAM(signaling lymphocyte activation molecule), 2B4(CD244), NKG2D(natural-killer group 2, member D)/DAP12(DNAX-activating protein 12), TIM1(T-Cell immunoglobulin and mucin domain containing protein 1), TIM2, TIM3, TIGIT, CD226, CD160, LAG3(lymphocyte activation gene 3), B7-1, B7-H1, GITR(glucocorticoid-induced TNFR family related protein), Flt3 리간드(fms-like tyrosine kinase 3 ligand), 플라젤린(flagellin), HVEM(herpesvirus entry mediator), CD40 L(리간드) 또는 OX40L[ligand for CD134(OX40), CD252]의 세포질 도메인 또는 이들 중 둘 이상의 연결체인, 발현벡터.33. The method of claim 32,
The immunopotentiating peptides include CD28, inducible costimulator (ICOS), cytotoxic T lymphocyte associated protein 4 (CTLA4), programmed cell death protein 1 (PD1), B and T lymphocyte associated protein (BTLA), DR3 1BB, CD2, CD40, CD30, CD27, SLAM (signaling lymphocyte activation molecule), 2B4 (CD244), NKG2D / DNAX- activating protein 12, TIM1 immunoglobulin and mucin domain containing protein 1), TIM2, TIM3, TIGIT, CD226, CD160, lymphocyte activation gene 3, B7-1, B7-H1, glucocorticoid-induced TNFR family related protein, Flt3 ligand a tyrosine kinase 3 ligand, a flagellin, a herpesvirus entry mediator (HVEM), CD40 L (ligand) or OX40L [ligand for CD134 (OX40), CD252] vector.

제21항 내지 제28항 중 어느 한 항의 발현벡터를 포함하는 DNA 백신 조성물.28. A DNA vaccine composition comprising an expression vector according to any one of claims 21 to 28.

제34항에 있어서,
gB 및 UL39를 포함하는 제1융합단백질을 암호화하는 폴리뉴클레오타이드가 프로모터에 작동가능하게 연결된 제1유전자 컨스트럭트를 포함하는 제1발현벡터 및 gD, ICP0 및 ICP4를 포함하는 제2융합단백질을 암호화하는 폴리뉴클레오타이드가 프로모터에 작동가능하게 연결된 제2유전자 컨스트럭트를 포함하는 제2발현벡터를 포함하는 DNA 백신 조성물.35. The method of claim 34,
a first expression vector comprising a first gene construct operably linked to a promoter, and a second fusion protein comprising gD, ICPO and ICP4, wherein the polynucleotide encoding the first fusion protein comprising gB and UL39 is operably linked to the promoter Wherein the polynucleotide comprises a second expression construct comprising a second gene construct operably linked to a promoter.

제35항에 있어서,
상기 제1융합단백질 및/또는 상기 제2융합단백질은 분비 신호서열을 추가로 포함하는, DNA 백신 조성물.36. The method of claim 35,
Wherein the first fusion protein and / or the second fusion protein further comprises a secretory signal sequence.

제36항에 있어서,
상기 분비 신호서열은 tPA(tissue plasminogen activator) 신호서열, HSV gDs(단순포진 바이러스 당단백질 Ds) 신호서열 또는 성장호르몬 신호서열인, DNA 백신 조성물.37. The method of claim 36,
Wherein the secretory signal sequence is a tissue plasminogen activator (tPA) signal sequence, a HSV gDs (herpes simplex virus glycoprotein Ds) signal sequence or a growth hormone signal sequence.

제36항에 있어서,
상기 제1융합단백질 및/또는 상기 제2융합단백질은 면역 증진 펩타이드를 추가로 포함하는, DNA 백신 조성물.37. The method of claim 36,
Wherein the first fusion protein and / or the second fusion protein further comprises an immunity enhancing peptide.

제38항에 있어서,
상기 면역 증진 펩타이드는 CD28, ICOS(inducible costimulator), CTLA4(cytotoxic T lymphocyte associated protein 4), PD1(programmed cell death protein 1), BTLA(B and T lymphocyte associated protein), DR3(death receptor 3), 4-1BB, CD2, CD40, CD30, CD27, SLAM(signaling lymphocyte activation molecule), 2B4(CD244), NKG2D(natural-killer group 2, member D)/DAP12(DNAX-activating protein 12), TIM1(T-Cell immunoglobulin and mucin domain containing protein 1), TIM2, TIM3, TIGIT, CD226, CD160, LAG3(lymphocyte activation gene 3), B7-1, B7-H1, GITR(glucocorticoid-induced TNFR family related protein), Flt3 리간드(fms-like tyrosine kinase 3 ligand), 플라젤린(flagellin), HVEM(herpesvirus entry mediator), CD40 L(리간드) 또는 OX40L[ligand for CD134(OX40), CD252]의 세포질 도메인 또는 이들 중 둘 이상의 연결체인, DNA 백신 조성물.39. The method of claim 38,
The immunopotentiating peptides include CD28, inducible costimulator (ICOS), cytotoxic T lymphocyte associated protein 4 (CTLA4), programmed cell death protein 1 (PD1), B and T lymphocyte associated protein (BTLA), DR3 1BB, CD2, CD40, CD30, CD27, SLAM (signaling lymphocyte activation molecule), 2B4 (CD244), NKG2D / DNAX- activating protein 12, TIM1 immunoglobulin and mucin domain containing protein 1), TIM2, TIM3, TIGIT, CD226, CD160, lymphocyte activation gene 3, B7-1, B7-H1, glucocorticoid-induced TNFR family related protein, Flt3 ligand a cytoplasmic domain of a tyrosine kinase 3 ligand, flagellin, a herpesvirus entry mediator HVEM, CD40 L (ligand) or OX40L [ligand for CD134 (OX40), CD252] Vaccine composition.