CN100334214C - 奈瑟球菌蛋白质的杂交表达 - Google Patents

奈瑟球菌蛋白质的杂交表达 Download PDF

Info

Publication number
CN100334214C
CN100334214C CNB2003101036036A CN200310103603A CN100334214C CN 100334214 C CN100334214 C CN 100334214C CN B2003101036036 A CNB2003101036036 A CN B2003101036036A CN 200310103603 A CN200310103603 A CN 200310103603A CN 100334214 C CN100334214 C CN 100334214C
Authority
CN
China
Prior art keywords
protein
seq
orf46
bacterial strain
xhoi
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
CNB2003101036036A
Other languages
English (en)
Other versions
CN1508253A (zh
Inventor
M·B·阿科
M·科曼杜西
C·加莱蒂
V·马斯格阿尼
M·M·朱利亚尼
M·皮扎
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Chrion S P A
GSK Vaccines SRL
Original Assignee
Chrion S P A
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from GB0004695A external-priority patent/GB0004695D0/en
Priority claimed from GB0027675A external-priority patent/GB0027675D0/en
Application filed by Chrion S P A filed Critical Chrion S P A
Publication of CN1508253A publication Critical patent/CN1508253A/zh
Application granted granted Critical
Publication of CN100334214C publication Critical patent/CN100334214C/zh
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/02—Bacterial antigens
    • A61K39/095—Neisseria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/22—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Neisseriaceae (F)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55505—Inorganic adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
    • A61K2039/572—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 cytotoxic response
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
    • A61K2039/575—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 humoral response
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Immunology (AREA)
  • Analytical Chemistry (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biophysics (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Physics & Mathematics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Peptides Or Proteins (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

以单个杂交蛋白形式表达两种或多种奈瑟球菌的蛋白质(如A和B)，其可以式NH2-A-B-COOH简单地表示。

Description

奈瑟球菌蛋白质的杂交表达
本案是申请号为01808738.8，名为奈瑟球菌蛋白质的杂交表达的分案申请。本文将所引用的全部文献都纳入作为参考。
技术领域
本发明涉及蛋白质表达的领域。具体说，本发明涉及奈瑟球菌(如淋病奈瑟球菌或较佳地为脑膜炎奈瑟球菌)的蛋白质的异源表达。
背景技术
国际专利申请WO 99/24578、WO99/36544、WO99/57280和WO 00/22430公开了脑膜炎奈瑟球菌(Neisseria meningitidis)和淋病奈瑟球菌(Neisseriagonorrhoeae)的蛋白质。这些蛋白质一般是以N-未端GST-融合体或C-未端His-标记融合体在大肠杆菌中表达的(即异源表达)，虽然也公开了其它表达***(包括在天然的奈瑟球菌中的表达)。
本发明的目的是提供这些蛋白质的异源表达的其它或改进方法。这些方法通常影响表达的水平、纯化的简易程度、表达的细胞内定位和/或表达的蛋白质的免疫学特性。
发明的公开
在本发明中，以单杂交蛋白表达本发明的两种或多种(如3、4、5、6或更多)蛋白质。较佳地，不使用非奈瑟球菌的融合配体(如GST或聚-His)。
这有两个优点。其一，可能不稳定或本身表达很差的蛋白质可以通过加入合适的克服该问题的杂交配体予以帮助。其二，简化工业生产，即制备两种分别有用的蛋白质只需要一次表达和纯化。
因此，本发明提供了同时异源表达本发明的两种或多种蛋白质的方法，其中所述的本发明的两种或多种蛋白质是融合的(即，它们是作为单多肽链翻译的)。
该方法通常包括如下步骤：获得编码本发明第一种蛋白质的第一核酸：获得编码本发明第二种蛋白质的第二核酸；连接第一和第二核酸。将得到的核酸***表达载体中，或已作为表达载体的一部分。
当仅连接两种蛋白质时，可以式NH2-A-B-COOH简单地表示杂交蛋白。A和B各选自任何奈瑟球菌蛋白质，尤其是那些以SEQ#1-4326表示的。该方法非常适合表达蛋白质orf1、orf4、orf25、orf40、Orf46/46.1、orf83、233、287、292L、564、687、741、907、919、953、961和983。
由式NH2-A-B-COOH在下表中以‘X′表示的42个杂交体是优选的：
    ↓AB→     ORF46.1     87   741     919     953     961     983
ORF46.1 × × × × × ×
    287     ×   ×     ×     ×     ×     ×
    741     ×     ×     ×     ×     ×     ×
    919     ×     ×   ×     ×     ×     ×
    953     ×     ×   ×     ×     ×     ×
    961     ×     ×   ×     ×     ×     ×
    983     ×     ×   ×     ×     ×     ×
因此，优选的以杂交体表达的蛋白质是ORF46.1，287，741，919，953，961和983。它们可以全长形式或多-甘氨酸缺失(ΔG)形式使用(如ΔG-287、ΔGTbp2、ΔG741、ΔG983等)、或以截短形式使用(如Δ1-287、Δ2-287等)，或以结构域缺失的形式使用(如287B、287C、287BC、ORF461-433、ORF46433-608、ORF46、961c等)等。
特别优选的是：(a)含919和287的杂交蛋白；(b)含953和287的杂交蛋白；(c)含287和ORF46.1的杂交蛋白；(d)含ORF1和ORF46.1的杂交蛋白；(e)含919和ORF46.1的杂交蛋白；(f)含ORF46.1和919的杂交蛋白；(g)含ORF46.1、287和919的杂交蛋白；(h)含919和519的杂交蛋白；和(i)含ORF97和225的杂交蛋白。
附图中显示了其它优选例，它们包括ΔG287-919、ΔG287-953、ΔG287-961、ΔG983-ORF46.1、ΔG983-741、ΔG983-961、ΔG983-961C、ΔG741-961、ΔG741-961C、ΔG741-983、ΔG741-ORF46.1、ORF46.1-741、ORF46.1-961、ORF46.1-961C、961-ORF46.1、961-741、961-983、961C-ORF46.1、961C-741、961C-983、961CL-ORF46.1、961CL-741和961CL-983。
当使用287时，其优先位于杂交体的C-未端；如果在N-未端使用它，则优先使用287的ΔG形式(如与ORF46.1、919、953或961杂交的杂交体的N-未端)。
当使用287时，其优先为菌株2996或菌株394/98的。
当使用961时，其优先在N-未端。可使用961的结构域形式。
WO 99/66741公开了ORF46、287、919和953的多态形式的排序。本发明可以使用这些多态形式中的任一形式。
较佳地，本发明杂交蛋白中的组成蛋白质(A和B)来源于是同一菌株。
杂交体中的融合蛋白可以是直接相连的，或者是通过接头肽相连的，如通过聚-甘氨酸接头(即，Gn，其中n＝3、4、5、6、7、8、9、10或更多)或通过协助克隆的短肽序列相连。显然不宜将ΔG蛋白质连接于聚-甘氨酸接头的C-未端。
融合蛋白可以缺失天然前导肽或可以包含N-未端融合配体的前导肽序列。
宿主
较佳地，利用异源宿主。异源宿主可以是原核的或真核的。优先为大肠杆菌，但其它合适的宿主包括枯草芽孢杆菌(Bacillus subtilis)、霍乱弧菌(Vibriocholerae)、伤寒沙门氏菌(Salmonella typhi)、鼠伤寒沙门氏菌(salmonellatyphimurium)、脑膜炎奈瑟球菌、淋病奈瑟球菌、乳糖奈瑟球菌(Neisserialactamica)、灰色奈瑟球菌(Neisseria cinerea)、分枝杆菌(如结核分枝杆菌(M.tuberculosis)、酵母等。
载体、宿主等
如上所述的方法，本发明提供了(a)用于这些方法的核酸和载体；(b)含所述载体的宿主细胞；(c)可用这些方法表达的或可表达的蛋白质；(d)包含这些蛋白质的组合物，其可能适合作为疫苗、或例如诊断剂或免疫原性的组合物；(e)用作药物(如疫苗)或诊断剂的组合物；(f)这些组合物在制备以下物质中的用途(1)用于治疗或预防奈瑟球菌引起的感染的药物(2)检测奈瑟球菌或由奈瑟球菌引起的抗体存在与否的诊断剂，和/或(3)能产生抗奈瑟球菌抗体的药物；和(g)治疗患者的方法，其包括对该患者施用治疗有效量的这些组合物。
序列
本发明还提供了具有以下实施例中所列出的任何的蛋白质或核酸。本发明还提供了具有与这些序列是序列相同性的蛋白质和核酸。如上所述，“序列相同性”的程度最好大于50％(如60％、70％、80％、90％、95％、99％或更大)。
本文的命名
本文参考在WO 99/24578、WO 99/36544和WO 99/57280中公开的2166个蛋白质序列，并将它们编号为如下的SEQ#：
申请 蛋白质序列  本文的SEQ#
WO 99/24578 偶SEQ ID 2-892  SEQ#1-446
WO 99/36544 偶SEQ ID 2-90  SEQ#447-491
WO 99/57280 偶SEQ ID 2-3020偶SEQ ID 3040-3114SEQ ID 3115-3241  SEQ#492-2001SEQ#2002-2039SEQ#2040-2166
除了这种SEQ#编号外，本文还使用了WO 99/24578、WO 99/36544和WO99/27280中的命名约定(如WO99/24578和WO 99/36544中用的‘ORF4′、‘ORF40′、‘OFR40-1′等；WO 99/57280中用的‘m919′、‘g919′和‘a919′等)。
在本文中将Tettelin等[Science(2000)287：1809-1815]中的从NMB0001到NMB2160的2160个蛋白质称为SEQ#2167-4326[参见WO 00/66791]。
本文采用的术语“本发明的蛋白质”指包含以下的蛋白质：
(a)SEQ#1-4326中的一个序列；或
(b)与SEQ#1-4326中的一个序列相同的序列；或
(c)SEQ#1-4326中的一个序列的片段。
(b)中的“序列相同性”的程度最好大于50％(如60％、70％、80％、90％、95％、99％或更大)。其包括突变体和等位基因变体[如，参见WO 00/66741]。相同性最好用Smith-Waterman同源性搜寻算法确定，如在MPSRCH程序(OxfordMolecular)中执行的，采用参数“缺口罚分(gap open penalty)”为12，“缺口延伸罚分(gap extension penalty)”为1进行缺口仿射搜索。通常，将两种蛋白质之间50％或更高的相同性视为功能等效的指示。
(c)中的“片段”应包含SEQ#1-4326中一个序列的至少n个连续的氨基酸，且根据具体的序列n为7或更高(如8、10、12、14、16、18、20、25、30、35、40、50、60、70、80、90、100或更高)。较佳地，片段包含SEQ#1-4326中一个序列的表位。优选的片段是在WO 00/71574和WO 01/04316中公开的那些。
本发明优选的蛋白质是在脑膜炎奈瑟球菌血清群B中发现的。
根据本发明使用的优选蛋白质是血清群B脑膜炎奈瑟球菌菌株2996或菌株394/98(新西兰菌株)。除非特别指出，本文所述的蛋白质是脑膜炎奈瑟球菌菌株2996的蛋白质。但是，应该理解通常本发明并不受菌株的限制。参考具体的蛋白质(如‘287′、‘919′等)可以包括任何菌株的该蛋白质。
应该理解术语“核酸”包括DNA和RNA，以及它们的类似物，如含有修饰骨架的那些类似物，还包括肽核酸(PNA)等。
附图简述
图1-26显示本发明的杂交蛋白。
本发明的进行模式
实施例1-ORF46的杂交体
脑膜炎奈瑟球菌(血清群B，菌株2996)的完整ORF46蛋白质具有如下序列：
1    LGISRKISLI  LSILAVCLPM  HAHASDLAND SFIRQVLDRQ HFEPDGKYHL
51  FGSRGELARR SGHIGLGKIQ SHQLGNLHIQ QAAIKGNIGY IVRFSDHGHE
101 VHSPPDNHAS HSDSDEAGSP VDGFSLYRIH WDGYEHHPAD GYDGPQGGGY
151 PAPKGARDIY SYDIRGVAQN IRLNLTDNRS TGQRLADRFH NAGSMLTQGV
201 GDGFKRATRY SPELDRSGNA AEAFNGTADI VKNIIGAAGE IVGAGDAVQG
251 ISEGSNIAVM HGLGLLSTEN KMARINDLAD MAQLKDYAAA AIRDWAVQNP
301 NAAQGIEAVS NIFMAAIPIK GIGAVRGKYG LGGITAHPIK RSQMGAIALP
351 KGKSAVSDNF ADAAYAKYPS PYHSRNIRSN LEQRYGKBNI TSSTVPPSNG
401 KNVKLADQRH PKTGVPFDGK GFPNFRKHVK YDTKLDIQEL SGGGIPKAKP
451 VSDAKPRWEV DRKLNRLTTR EQVEKNVQEI RNGNKNSNPS QHAQLEREIN
501 KLKSADRINF ADGMGKFTDS MNDKAFSRLV KSVKRNGFTN PVVEYVEING
551 KAYIVRGNNR VFAAEYLGRI HELKFKKVOF PVPNTSWKNP TDVLNESGNV
601 KRPRYRSK*
在前导肽下加下划线。
可以在WO 00/66741中发现其它菌株的ORF46的序列。
ORF46在其C-未端和N-未端与287、919和ORF1融合。该杂交蛋白通常是不溶解的，但产生一些良好的ELISA和杀菌结果(针对同源2996菌株)：
 蛋白质  ELISA 杀菌Ab
 Orf1-Orf46.1-His  850 256
 919-Orf46-1-His  12900 512
 919-287-Orf46-His  n.d. n.d.
 Orf46.1-287His  150 8192
 Orf46.1-919His  2800 2048
 Orf46.1-287-919His  3200 16384
为了比较，构建了ORF46.1、287(以GST融合体或ΔG287的形式)和919的“三”杂交体，并针对各种菌株(包括同源2996菌株)将其与三种抗原的简单混合物相比。FCA用作佐剂：
2996  BZ232  MC58  NGH38  F6124  BZ133
 混合物 8192  256  512  1024  ＞2048  ＞2048
 ORF46.1-287-919his 16384  256  4096  8192  8192  8192
 △G287-919-ORF46.1his 8192  64  4096  8192  8192  16384
 △G287-ORF46.1-919his 4096  128  256  8192  512  1024
同样，这些杂交体显示相当等的或更佳的免疫活性。
针对各种异源菌株，将两种蛋白质(菌株2996)的杂交体与单种蛋白质相比：
1000  MC58  F6124(MenA)
ORF46.1-His ＜4  4096  ＜4
ORF1-His 8  256  128
ORF1-ORF46.1-His 1024  512  1024
再次，这些杂交体显示相等的或更佳的免疫活性。
实施例2-ΔG287的杂交体
发现287中(Gly)6序列的缺失对蛋白质的表达有显著影响。将缺失N-未端氨基酸多达GGGGGG的蛋白质称为‘ΔG287′。在菌株MC58中，它的基本序列(前导肽有下划线)为：
    SPDVKS ADTLSKPAAP VVSEKETEAK EDAPQAGSQG QGAPSAQGSQ DMAAVSEENT
GNGGAVTADN PKNEDEVAQN DMPQNAAGTD SSTPNHTPDP NMLAGNMENQ ATDAGESSQP
ANQPDMANAA DGMQGDDPSA GGQNAGNTAA QGANQAGNNQ AAGSSDPIPA SNPAPANGGS
NFGRVDLANG VLIDGPSQNI TLTHCKGDSC SGNNFLDEEV QLKSEFEKLS DADKISNYKK
DGKNDKFVGL VADSVQMKGI NQYIIFYKPK PTSFARFRRS ARSRRSLPAE MPLIPVNQAD
TLIVDGEAVS LTGHSGNIFA PEGNYRYLTY GAEKLPGGSY ALRVQGBPAK GEMLAGAAVY
NGEVLHFHTE NGRPYRTRGR FAARVDFGSK SVDGIIDSGD DLHMGTQKFK AAIDGNGFKG
TWTENGSGDV SGKFYGPAGE EVAGKYSYRP TDAEKGGFGV FAGKKEQD*
与‘287-His′或‘287未标记的1相比，有或无His-标记的ΔG287(分别为‘ΔG287-His′和‘ΔG287K′)以很好的水平表达。
在基因变异性数据的基础上，从许多MenB菌株(尤其是从菌株2996、MC58、100和BZ232)的大肠杆菌中表达ΔG287-His的变体。结果也好-它们都具有很高的ELISA滴定度，且血清杀菌滴定度＞8192。由pET-24b表达的ΔG287K在ELISA和血清杀菌实验中有极佳滴定度。
还将聚-Gly序列的缺失应用于Tbp2(NMB0460)、741(NMB1870)和983(NMB1969)。在不编码其前导肽的序列且没有聚-Gly(即，以“ΔG形式”)，在pET载体中克隆并在大肠杆菌中表达时，观察到相同的作用-在携带聚-甘氨酸段缺失的克隆中表达很好，若在表达的蛋白中存在甘氨酸时则表达差或不表达。
将ΔG287直接融合于919、953、961(如下所示的序列)和ORF46.1的符合读框的上游：
AQ287-919
ATGGCTAGCCCCGATGTTAAATCGGCGGACACGCTGTCAAAACCGGCCGCTCCTGTTGTTGCTGAAAAAGAGACAGAG
GTAAAAGAAGATGCGCCACAGGCAGGTTCTCAAGGACAGGGCGCGCCATCCACACAAGGCAGCCAAGATATGGCGGCA
GTTTCGGCAGAAAATACAGGCAATGGCGGTGCGGCAACAACGCACAAACCCAAAAATGAAGACGAGGGACCGCAAAAT
GATATGCCGCAAAATTCCGCCGAATCCGCAAATCAAACAGGGAACAACCAACCCGCCGATTCTTCAGATTCCGCCCCC
GCGTCAAACCCTGCACCTGCGAATGGCGGTAGCAATTTTGGAAGGGTTGATTTGGCTAATGGCGTTTTGATTGATGGG
CCGTCGCAAAATATAACGTTGACCCACTGTAAAGGCGATTCTTGTAATGGTGATAATTTATTGGATGAAGAAGCACCG
TCAAAATCAGAATTTGAAAATTTAAATGAGTCTGAACGAATTGAGAAATATAAGAAAGATGGGAAAAGCGATAAATTT
ACTAATTTGGTTGCGACAGCAGTTCAAGCTAATGGAACTAACAAATATGTCATCATTTATAAAGACAAGTCCGCTTCA
TCTTCATCTGCGCGATTCAGGCGTTCTGCACGGTCGAGGAGGTCGCTTCCTGCCGAGATGCCGCTAATCCCCGTCAAT
CAGGCGGATACGCTGATTGTCGATGGGGAAGCGGTCAGCCTGACGGGGCATTCCGGCAATATCTTCGCGCCCGAAGGG
AATTACCGGTATCTGACTTACGGGGCGGAAAAATTGCCCGGCGGATCGTATGCCCTCCGTGTGCAAGGCGAACCGGCA
AAAGGCGAAATGCTTGCTGGCACGGCCGTGTACAACGGCGAAGTGCTGCATTTTCATACGGAAAACGGCCGTCCGTAC
CCGACTAGAGGCAGGTTTGCCGCAAAAGTCGATTTCGGCAGCAAATCTGTGGACGGCATTATCGACAGCGGCGATCAT
TTGCATATGGGTACGCAAAAATTCAAAGCCGCCATCGATGGAAACGGCTTTAAGGGGACTTGGACGGAAAATGGCGGC
GGGGATGTTTCCGGAAGGTTTTACGGCCCGGCCGGCGAGGAAGTGGCGGGAAAATACAGCTATCGCCCGACAGATGCG
GAAAAGGGCGGATTCGGCGTGTTTGCCGGCAAAAAAGAGCAGGATGGATCCGGAGGAGGAGGATCCCAAAGCAAGAGC
ATCCAAACCTTTCCGCAACCCGACACATCCGTCATCAACGGCCCGGACCGGCCGGTCGGCATCCCCGACCCCGCCGGA
ACGACGGTCGGCGGCGGCGGGGCCGTCTATACCGTTGTACCGCACCTGTCCCTGCCCCACTGGGCGGCGCAGGATTTC
GCCAAAAGCCTGCAATCCTTCCGCCTCGGCTGCGCCAATTTGAAAAACCGCCAAGGCTGGCAGGATGTGTGCGCCCAA
GCCTTTCAAACCCCCGTCCATTCCTTTCAGGCAAAACAGTTTTTTGAACGCTATTTCACGCCGTGGCAGGTTGCAGGC
AACGGAAGCCTTGCCGGTACGGTTACCGGCTATTACGAGCCGGTGCTGAAGGGCGACGACAGGCGGACGGCACAAGCC
CGCTTCCCGATTTACGGTATTCCCGACGATTTTATCTCCGTCCCCCTGCCTGCCGGTTTGCGGAGCGGAAAAGCCCTT
GTCCGCATCAGGCAGACGGGAAAAAACAGCGGCACAATCGACAATACCGGCGGCACACATACCGCCGACCTCTCCCGA
TTCCCCATCACCGCGCGCACAACGGCAATCAAAGGCAGGTTTGAAGGAAGCCGCTTCCTCCCCTACCACACGCGCAAC
CAAATCAACGGCGGCGCGCTTGACGGCAAAGCCCCGATACTCGGTTACGCCGAAGACCCCGTCGAACTTTTTTTTATG
CACATCCAAGGCTCGGGCCGTCTGAAAACCCCGTCCGGCAAATACATCCGCATCGGCTATGCCGACAAAAACGAACAT
CCCTACGTTTCCATCGGACGCTATATGGCGGACAAAGGCTACCTCAAGCTCGGGCAGACCTCGATGCAGGGCATCAAA
GCCTATATGCGGCAAAATCCGCAACGCCTCGCCGAAGTTTTGGGTCAAAACCCCAGCTATATCTTTTTCCGCGAGCTT
GCCGGAAGCAGCAATGACGGTCCCGTCGGCGCACTGGGCACGCCGTTGATGGGGGAATATGCCGGCGCAGTCGACCGG
CACTACATTACCTTGGGCGCGCCCTTATTTGTCGCCACCGCCCATCCGGTTACCCGCAAAGCCCTCAACCGCCTGATT
ATGGCGCAGGATACCGGCAGCGCGATTAAAGGCGCGGTGCGCGTGGATTATTTTTGGGGATACGGCGACGAAGCCGGC
GAACTTGCCGGCAAACAGAAAACCACGGGTTACGTCTGGCAGCTCCTACCCAACGGTATGAAGCCCGAATACCGCCCG
TAACTCGAG
1      MASPDVKSAD TLSKPAAPVV AEKETEVKED APQAGSQGQG APSTQGSQDM
51     AAVSAENTGN GGAATTDKPK NEDEGPQNDM PQNSAESANQ TGNNQPADSS
101    DSAPASNPAP ANGGSNFGRV DLANGVLIDG PSQNTTLTHC KGDSCNGDNL
151    LDEEAPSKSE FENLNESERI EKYKKDGKSD KFTNLVATAV QANGTNKYVI
201    IYKDKSASSS SARFRRSARS RRSLPAEMPL IPVNQADTLI VDGEAVSLTG
251    HSGNIFAPEG NYRYLTYGAE KLPGGSYALR VQGKPAKGEM LAGTAVYNGE
301    VLHFHTENGR PYPTRGRFAA KVDFGSKSVD GIIDSGDDLH MGTQKFKAAI
351    DGNGFKGTWT ENGGGDVSGR FYGPAGEEVA GKYSYRPTDA KKGGFGVFAG
401    KKEQDGSGGG GCQSKSIQTF PQPDTSVING PDRPVGIPDP AGTTVGGGGA
451    VYTVVPHLSL PHWAAQDFAK SLQSFRLGCA NLKNRQGWQD VCAQAFQTPV
501    HSFQAKQFFE RYFTPWAVAG NGSLAGTVTG YYKPVLKGDD RRTAQARFPI
551    YGIPDDFISV PLPAGLRSGK ALVRIRQTGK NSGTIDNTGG THTADLSRFP
601    ITARTTAIKG RFEGSRFLPY HTRNQINGGA LDGKAPILGY AEDPVELFFM
651    HIQGSGRLKT PSGKYIRIGY ADKNEHPYVS IGRYMADKGY LKLGQTSMQG
701    IKAYMRQNPQ RLAEVLGQNP SYIFFRELAG SSNDGPVGAL GTPLMGEYAG
751    AVDRHYITLG APLFVATAHP VTRKALNRLI MAQDTGSAEK GAVRVDYFWG
801    YGDEAGELAG KQKTTGYVWQ LLPNGMKPEY RP*
ΔG287-953
ATGGCTAGCCCCGATGTTAAATCGGCGGACACGCTGTCAAAACCGGCCGCTCCTGTTGTTGCTGAAAAAGAGACAGAG
GTAAAAGAAGATGCGCCACAGGCAGGTTCTCAAGGACAGGGCGCGCCATCCACACAAGGCAGCCAAGATATGGCGGCA
GTTTCGGCAGAAAATACAGGCAATGGCGGTGCGGCAACAACGGACAAACCCAAAAATGAAGACGAGGGACCGCAAAAT
GATATGCCGCAAAATTCCGCCGAATCCGCAAATCAAACAGGGAACAACCAACCCGCCGATTCTTCAGATTCCGCCCCC
GCGTCAAACCCTGCACCTGCGAATGGCGGTAGCAATTTTGGAAGGGTTGATTTAACTAATGGCGTTTTGATTGATGGG
CCGTCGCAAAATATAACGTTGACCCACTGTAAAGGCGATTCTTGTAATGGTGATAATTTATTGGATGAAGAAGCACCG
TCAAAATCAGAATTTGAAAATTTAAATGAGTCTGAACGAATTGAGAAATATAAGAAAGATGGGAAAAGCGATAAATTT
ACTAATTTGGTTGCGACAGCAGTTCAAGCTAATGGAACTAACAAATATGTCATCATTTATAAAGACAAGTCCGCTTCA
TCTTCATCTGCGCGATTCAGGCGTTCTGCACGGTCGAGGAGGTCGCTTCCTGCCGAGATGCCGCTAATCCCCGTCAAT
CAGGCGGATACGCTGATTGTCGATGGGGAAGCGGTCAGCCTGACGGGGCATTCCGGCAATATCTTCGCGCCCGAAGGG
AATTACCGGTATCTGACTTACGGGGCGGAAAAATTGCCCGGCGGATCGTATGCCCTCCGTGTGCAAGGCGAACCGGCA
AAAGGCGAAATGCTTGCTGGCACGGCCGTGTACAACGGCGAAGTGCTGCATTTTCATACGGAAAACGGCCGTCCGTAC
CCGACTAGAGGCAGGTTTGCCGCAAAAGTCGATTTCGGCAGCAAATCTGTGGACGGCATTATCGACAGCGGCGATGAT
TTGCATATGGGTACGCAAAAATTCAAAGCCGCCATCGATGGAAACGGCTTTAAGGGGACTTGGACGGAAAATGGCGGC
GGGGATGTTTCCGGAAGGTTTTACGGCCCGGCCGGCGAGGAAGTGGCGGGAAAATACAGCTATCGCCCGACAGATGCG
GAAAAGGGCGGATTCGGCGTGTTTGCCGGCAAAAAAGAGCAGGATGGATCCGGAGGAGGAGGAGCCACCTACAAAGTG
GACGAATATCACGCCAACGCCCGTTTCGCCATCGACCATTTCAACACCAGCACCAACGTCGGCGGTTTTTACGGTCTG
ACCGGTTCCGTCGAGTTCGACCAAGCAAAACGCGACGGTAAAATCGACATCACCATCCCCGTTGCCAACCTGCAAAGC
GGTTCGCAACACTTTACCGACCACCTGAAATCAGCCGACATCTTCGATGCCGCCCAATATCCGGACATCCGCTTTGTT
TCCACCAAATTCAACTTCAACGGCAAAAAACTGGTTTCCGTTGACGGCAACCTGACCATGCACGGCAAAACCGCCCCC
GTCAAACTCAAAGCCGAAAAATTCAACTGCTACCAAAGCCCGATGGCGAAAACCGAAGTTTGCGGCGGCGACTTCAGC
ACCACCATCGACCGCACCAAATGGGGCGTGGACTACCTCGTTAACGTTGGTATGACCAAAAGCGTCCGCATCGACATC
CAAATCGAGGCAGCCAAACAATAACTCGAG
1      MASPDVKSAD TLSKPAAPVV AEKETEVKED APQAGSQGQG APSTQGSQDM
51     AAVSAENTGN GGAATTDKPK NEDEGPQNDM PQNSAKSANQ TGNNQPADSS
101    DSAPASNPAP ANGGSNFGRV DLANGVLIDG PSQNTTLTHC KGDSCNGDNL
151    LDEEAPSKSE FENLNESERI EKYKKDGKSD KFTNLVATAV QANGTNKYVI
201    IYKDKSASSS SARFRRSARS RRSLPAEMPL IPVNQADTLI VDGEAVSLTG
251    HSGNIFAPEG NYRYLTYGAE KLPGGSYALR VQGKPAKGEM LAGTAVYNGE
301    VLHFHTENGR PYPTRGRFAA KVDFGSKSVD GIIDSGDDLH MGTQKFKAAI
351    DGNGFKGTWT ENGGGDVSGR FYGPAGEEVA GKYSYRPTDA EKGGFGVFAG
401    KKEQDGSGGG GATYKVDEYH ANARFAIDHF NTSTNVGGFY GLTGSVEFDQ
451    AKRDGKIDIT IPVANLQSGS QHFTDHLKSA DIFDAAQYPD IRFVSTKFNF
501    NGKKLVSVDG NLTMHGKTAP VKLKAEKFNC YQSPMAKTEV CGGDFSTTID
551    RTKWGVDYLV NVGMTKSVRI DIQIEAAKQ*
ΔG287-961
ATGGCTAGCCCCGATGTTAAATCGGCGGACACGCTGTCAAAACCGGCCGCTCCTGTTGTTGCTGAAAAAGAGACAGAG
GTAAAAGAAGATGCGCCACAGGCAGGTTCTCAAGGACAGGGCGCGCCATCCACACAAGGCAGCCAAGATATGGCGGCA
GTTTCGGCAGAAAATACAGGCAATGGCGGTGCGGCAACAACGGACAAACCCAAAAATGAAGACGAGGGACCGCAAAAT
GATATGCCGCAAAATTCCGCCGAATCCGCAAATCAAACAGGGAACAACCAACCCGCCGATTCTTCAGATTCCGCCCCC
GCGTCAAACCCTGCACCTGCGAATGGCGGTAGCAATTTTGGAAGGGTTGATTTGGCTAATGGCGTTTTGATTGATGGG
CCGTCGCAAAATATAACGTTGACCCACTGTAAAGGCGATTCTTGTAATGGTGATAATTTATTGGATGAAGAAGCACCG
TCAAAATCAGAATTTGAAAATTTAAATGAGTCTGAACGAATTGAGAAATATAAGAAAGATGGGAAAAGCGATAAATTT
ACTAATTTGGTTGCGACAGCAGTTCAAGCTAATGGAACTAACAAATATGTCATCATTTATAAAGACAAGTCCGCTTCA
TCTTCATCTGCGCGATTCAGGCGTTCTGCACGGTCGAGGAGGTCGCTTCCTGCCGAGATGCCGCTAATCCCCGTCAAT
CAGGCGGATACGCTGATTGTCGATGGGGAAGCGGTCAGCCTGACGGGGCATTCCGGCAATATCTTCGCGCCCGAAGGG
AATTACCGGTATCTGACTTACGGGGCGGAAAAATTGCCCGGCGGATCGTATGCCCTCCGTGTGCAAGGCGAACCGGCA
AAAGGCGAAATGCTTGCTGGCACGGCCGTGTACAACGGCGAAGTGCTGCATTTTCATACGGAAAACGGCCGTCCGTAC
CCGACTAGAGGCAGGTTTGCCGCAAAAGTCGATTTCGGCAGCAAATCTGTGGACGGCATTATCGACAGCGGCGATGAT
TTGCATATGGGTACGCAAAAATTCAAAGCCGCCATCGATGGAAACGGCTTTAAGGGGACTTGGACGGAAAATGGCGGC
GGGGATGTTTCCGGAAGGTTTTACGGCCCGGCCGGCGAGGAAGTGGCGGGAAAATACAGCTATCGCCCGACAGATGCG
GAAAAGGGCGGATTCGGCGTGTTTGCCGGCAAAAAAGAGCAGGATGGATCCGGAGGAGGAGGAGCCACAAACGACEAC
GATGTTAAAAAAGCTGCCACTGTGGCCATTGCTGCTGCCTACAACAATGGCCAAGAAATCAACGGTTTCAAAGCTGGA
GAGACCATCTACGACATTGATGAAGACGGCACAATTACCAAAAAAGACGCAACTGCAGCCGATGTTGAAGCCGACGAC
TTTAAAGGTCTGGGTCTGAAAAAAGTCGTGACTAACCTGACCAAAACCGTCAATGAAAACAAACAAAACGTCGATGCC
AAAGTAAAAGCTGCAGAATCTGAAATAGAAAAGTTAACAACCAAGTTAGCAGACACTGATGCCGCTTTAGCAGATACT
GATGCCGCTCTGGATGCAACCACCAACGCCTTGAATAAATTGGGAGAAAATATAACGACATTTGCTGAAGAGACTAAG
ACAAATATCGTAAAAATTGATGAAAAATTAGAAGCCGTGGCTGATACCGTCGACAAGCATGCCGAAGCATTCAACGAT
ATCGCCGATTCATTGGATGAAACCAACACTAAGGCAGACGAAGCCGTCAAAACCGCCAATGAAGCCAAACAGACGGCC
GAAGAAACCAAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAAACTGCAGCAGGCAAAGCCGAAGCTGCCGCTGGC
ACAGCTAATACTGCAGCCGACAAGGCCGAAGCTGTCGCTGCAAAAGTTACCGACATCAAAGCTGATATCGCTACGAAC
AAAGATAATATTGCTAAAAAAGCAAACAGTGCCGACGTGTACACCAGAGAAGAGTCTGACAGCAAATTTGTCAGAATT
GATGGTCTGAACGCTACTACCGAAAAATTGGACACACGCTTGGCTTCTGCTGAAAAATCCATTGCCGATCACGATACT
CGCCTGAACGGTTTGGATAAAACAGTGTCAGACCTGCGCAAAGAAACCCGCCAAGGCCTTGCAGAACAAGCCGCGCTC
TCCGGTCTGTTCCAACCTTACAACGTGGGTCGGTTCAATGTAACGGCTGCAGTCGGCGGCTACAAATCCGAATCGGCA
GTCGCCATCGGTACCGGCTTCCGCTTTACCGAAAACTTTGCCGCCAAAGCAGGCGTGGCAGTCGGCACTTCGTCCGGT
TCTTCCGCAGCCTACCATGTCGGCGTCAATTACGAGTGGTAACTCGAG
1      MASPDVKSAD TLSKPAAPVV AKKETEVKED APQAGSQGQG APSTQGSQDM
1      AAVSAENTGN GGAATTDKPK NEDEGPQNDM PQNSAESANQ TGNNQPADSS
101    DSAPASNPAP ANGGSNFGRV DLANGVLIDG PSQNITLTHC KGDSCNGDNL
151    LDEEAPSKSE FENLNESERI EKYKKDGKSD KFTNLVATAV QANGTNKYVI
201    IYKDKSASSS SARFRRSARS RRSLPAEMPL IPVNQADTLI VDGKAVSLTG
251    HSGNIFAPEG NYRYLTYGAK KLPGGSYALR VQGEPAKGEM LAGTAVYNGK
301    VLHFHTENGR PYPTRGRFAA KVDFGSKSVD GIIDSGDDLH NGTQKFKAAI
351    DGNGFKGTWT ENGGGDVSGR FYGPAGEEVA GKYSYRPTDA KKGGFGVFAG
401    KKEQDGSGGG GATNDDDVKK AATVAIAAAY NNGQEINGFK AGKTIYDIDE
451    DGTITKKDAT AADVKADDFK GLGLKKVVTN LTKTVNKNKQ NVDAKVKAAE
501    SEIEKLTTKL ADTDAALADT DAALDATTNA LNKLGENITT FAEETKTNIV
551    KIDEKLEAVA DTVDKHAEAF NDIADSLDET NTKADKAVKT ANEAKQTAEE
601    TKQNVDAKVK AAETAAGKAE AAAGTANTAA DKAEAVAAKV TDIKADIATN
651    KDNIAKKANS ADVYTREESD SKFVRIDGLN ATTKKLDTRL ADAKKSIADH
701    DTRLNGLDKT VSDLRKETRQ GLAEQAALSG LFQPYNVGRF NVTAAVGGYK
751    SESAVAIGTG FRFTKNFAAK AGVAVGTSSG SSAAYHVGVN YEW*
 ELISA  杀菌
 ΔG287-953-His  3834  65536
 ΔG287-961-His  108627  65536
对919和ORF46.1而言，将针对杂交蛋白产生的抗体的杀菌效力(同源菌株)与针对组分抗原(用287-GST)的简单混合物产生的抗体相比：
    与287的混合物     与ΔG287的杂交体
   919     32000     128000
   ORF46.1     128     16000
还获得了针对异源MenB菌株和针对血清型A和C的杀菌活性的数据：
  919  ORF46.1
     菌株   混合物   杂交体  混合物  杂交体
     NGH38   1024   32000  -  16384
     MC58   512   8192  -  512
     BZ232   512   512  -  -
     MenA(F6124)   512   32000  -  8192
     MenC(C11)   ＞2048   ＞2048  -  -
     MenC(BZ133)   ＞4096   64000  -  8192
因此，在N-未端与ΔG287的杂交蛋白在免疫学上优于与ΔG287-ORF46.1的简单混合物，即使针对异源菌株也特别有效。可以在pET-24b中表达ΔG287-ORF46.1。
用新西兰菌株394/98，而非2996，制备相同的杂交蛋白：
ΔG287NZ-919
ATGGCTAGCCCCGATGTCAAGTCGGCGGACACGCTGTCAAAACCTGCCGCCCCTGTTGTTTCTGAAAAAGAGACAGAG
GCAAAGGAAGATGCGCCACAGGCAGGTTCTCAAGGACAGGGCGCGCCATCCGCACAAGGCGGTCAAGATATGGCGGCG
GTTTCGGAAGAAAATACAGGCAATGGCGGTGCGGCAGCAACGGACAAACCCAAAAATGAAGACGAGGGGGCGCAAAAT
GATATGCCGCAAAATGCCGCCGATACAGATAGTTTGACACCGAATCACACCCCGGCTTCGAATATGCCGGCCGGAAAT
ATGGAAAACCAAGCACCGGATGCCGGGGAATCGGAGCAGCCGGCAAACCAACCGGATATGGCAAATACGGCCGGAAAT
ATGCAGGGTGACGATCCGTCGGCAGGCGGGGAAAATGCCGGCAATACGGCTGCCCAAGGTACAAATCAAGCCGAAAAC
AATCAAACCGCCGGTTCTCAAAATCCTGCCTCTTCAACCAATCCTAGCGCCACGAATAGCGGTGGTGATTTTGGAAGG
ACGAACGTGGGCAATTCTGTTGTGATTGACGGGCCGTCGCAAAATATAACGTTGACCCACTGTAAAGGCGATTCTTGT
AGTGGCAATAATTTCTTGGATGAAGAAGTACAGCTAAAATCAGAATTTGAAAAATTAAGTGATGCAGACAAAATAAGT
AATTACAAGAAAGATGGGAAGAATGACGGGAAGAATGATAAATTTGTCGGTTTGGTTGCCGATAGTGTGCAGATGAAG
GGAATCAATCAATATATTATCTTTTATAAACCTAAACCCACTTCATTTGCGCGATTTAGGCGTTCTGCACGGTCGAGG
CGGTCGCTTCCGGCCGAGATGCCGCTGATTCCCGTCAATCAGGCGGATACGCTGATTGTCGATGGGGAAGCGGTCAGC
CTGACGGGGCATTCCGGCAATATCTTCGCGCCCGAAGGGAATTACCGGTATCTGACTTACGGGGCGGAAAAATTGCCC
GGCGGATCGTATGCCCTCCGTGTTCAAGGCGAACCTTCAAAAGGCGAAATGCTCGCGGGCACGGCAGTGTACAACGGC
GAAGTGCTGCATTTTCATACGGAAAACGGCCGTCCGTCCCCGTCCAGAGGCAGGTTTGCCGCAAAAGTCGATTTCGGC
AGCAAATCTGTGGACGGCATTATCGACAGCGGCGATGGTTTGCATATGGGTACGCAAAAATTCAAAGCCGCCATCGAT
GGAAACGGCTTTAAGGGGACTTGGACGGAAAATGGCGGCGGGGATGTTTCCGGAAAGTTTTACGGCCCGGCCGGCGAG
GAAGTGGCGGGAAAATACAGCTATCGCCCAACAGATGCGGAAAAGGGCGGATTCGGCGTGTTTGCCGGCAAAAAAGAG
CAGGATGGATCCGGAGGAGGAGGATGCCAAAGCAAGAGCATCCAAACCTTTCCGCAACCCGACACATCCGTCATCAAC
GGCCCGGACCGGCCGGTCGGCATCCCCGACCCCGCCGGAACGACGGTCGGCGGCGGCGGGGCCGTCTATACCGTTGTA
CCGCACCTGTCCCTGCCCCACTGGGCGGCGCAGGATTTCGCCAAAAGCCTGCAATCCTTCCGCCTCGGCTGCGCCAAT
TTGAAAAACCGCCAAGGCTGGCAGGATGTGTGCCCCCAAGCCTTTCAAACCCCCGTCCATTCCTTTCAGGCAAAACAG
TTTTTTGAACGCTATTTCACGCCGTGGCAGGTTGCAGGCAACGGAAGCCTTGCCGGTACGGTTACCGGCTATTACGAG
GTCCCCCTGCCTGCCGGTTTGCGGAGCGGAAAAGCCCTTGTCCGCATCAGGCAGACGGGAAAAAACAGCGGCACAATC
GTCCCCCTGCCTGCCGGTTTGCGGAGCGGAAAAGCCCTTGTCCGCATCAGGCAGACGGGAAAAAACAGCGGCACAATC
GACAATACCGGCGGCACACATACCGCCGACCTCTCCCGATTCCCCATCACCGCGCGCACAACGGCAATCAAAGGCAGG
TTTGAAGGAAGCCGCTTCCTCCCCTACCACACGCGCAACCAAATCAACGGCGGCGCGCTTGACGGCAAAGCCCCGATA
CTCGGTTACGCCGAAGACCCCGTCGAACTTTTTTTTATGCACATCCAAGGCTCGGGCCGTCTGAAAACCCCGTCCGGC
AAATACATCCGCATCGGCTATGCCGACAAAAACGAACATCCCTACGTTTCCATCGGACGCTATATGGCGGACAAAGGC
TACCTCAAGCTCGGGCAGACCTCGATGCAGGGCATCAAAGCCTATATGCGGCAAAATCCGCAACGCCTCGCCGAAGTT
TTGGGTCAAAACCCCAGCTATATCTTTTTCCGCGAGCTTGCCGGAAGCAGCAATGACGGTCCCGTCGGCGCACTGGGC
ACGCCGTTGATGGGGGAATATGCCGGCGCAGTCGACCGGCACTACATTACCTTGGGCGCGCCCTTATTTGTCGCCACC
GCCCATCCGGTTACCCGCAAAGCCCTCAACCGCCTGATTATGGCGCAGGATACCGGCAGCGCGATTAAAGGCGCGGTG
CGCGTGGATTATTTTTGGGGATACGGCGACGAAGCCGGCGAACTTGCCGGCAAACAGAAAACCACGGGTTACGTCTGG
CAGCTCCTACCCAACGGTATGAAGCCCGAATACCGCCCGTAAAAGCTT
  1    MASPDVKSAD TLSKPAAPVV SKKETEAKED APQAGSQGQG APSAQGGQDM
 51    AAVSEENTGN GGAAATDRRK NEDEGAQNDM PQNAADTDSL TPNHTPASNM
101    PAGNMENQAP DAGESEQPAN QPDMANTADG MQGDDPSAGG XNAGNTAAQG
151    TNQAENNQTA GSQN[ASSTN PSATNSGGDF GRTNVGNSVV IDGPSQNITL
201    THCKGDSCSG NNFLLEECQL RSEFERLSDA DKISNYKKDG KNDGKNDKFV
251    GLVADSVQMK GINQYIIFYK PKPTSFARFR RSARSRRSLP AEMPLIPVNQ
301    ADTLIVDGEA VSLTGHSGNI FAPRGNYRYL TYGAKKLPGG SYALRVQGEP
351    SKGEMLAGTA VYNGKVLHFE TENGRPSPSR GRFAAKVOFG SKSVDGIIDS
401    GDGLHMGTQK FKAAIDGNGF KGTWTENGGG DVSGKFYGPA GEEVAGKYSY
451    RPTDAEKGGF GVFAGKKEQD GSGGGGCQSK SIQTFPQPDT SVINGPDRPV
501    GIPDPAGTTV GGGGAVYYVV PHLSLPHMAA QDFAKSLQSF RLGCANLKNR
551    QGWQDVCAQA FQRPVHSFQA KQFFEHYFTP WQVAGNGSLA GTVTGYYBPV
601    LKGDDRRTAQ APFPIYGIPD DFISVPLPAG LRSGKALVRI RQTGKNSGTI
651    DNTGGTHTAD LSRFPITART TAIKGRFEGS RFLPYHTRNQ INGGALDGKA
701    PLLGYAEDPV ELFFMHIQGS GRLKTPSGKY IRIGYADKNE HPYVSIGRYM
751    ADKGYLKLGQ TSMQGIKAYM RQNPQRLAEV LGQNPSYIFF RRLAGSSNDG
801    PVGALGRPLM GEYAGAVDEH YITLGAPLFV ATAHPVTRKA LNTLIMQQDT
851    GSAIKGAVRV DYFWGYGDEA GELAGKQKTT GYVWQLLPNG MKPKYPP*
ΔG287NZ-953
ATGGCTAGCCCCGATGTCAAGTCGGCGGACACGCTGTCAAAACCTGCCGCCCCTGTTGTTTCTGAAAAAGAGACAGAG
GCAAAGGAAGATGCGCCACAGGCAGGTTCTCAAGGACAGGGCGCGCCATCCGCACAAGGCGGTCAAGATATGGCGGCG
GTTTCGGAAGAAAATACAGGCAATGGCGGTGCGGCAGCAACGGACAAACCCAAAAATGAAGACGAGGGGGCGCAAAAT
GATATGCCGCAAAATGCCGCCGATACAGATAGTTTGACACCGAATCACACCCCGGCTTCGAATATGCCGGCCGGAAAT
ATGGAAAACCAAGCACCGGATGCCGGGGAATCGGAGCAGCCGGCAAACCAACCGGATATGGCAAATACGGCGGACGGA
ATGCAGGGTGACGATCCGTCGGCAGGCGGGGAAAATGCCGGCAATACGGCTGCCCAAGGTACAAATCAAGCCGAAAAC
AATCAAACCGCCGGTTCTCAAAATCCTGCCTCTTCAACCAATCCTAGCGCCACGAATAGCGGTGGTGATTTTGGAAGG
ACGAACGTGGGCAATTCTGTTGTGATTGACGGGCCGTCGCAAAATATAACGTTGACCCACTGTAAAGGCGATTCTTGT
AGTGGCAATAATTTCTTGGATGAAGAAGTACAGCTAAAATCAGAATTTGAAAAATTAAGTGATGCAGACAAAATAAGT
AATTACAAGAAAGATGGGAAGAATGACGGGAAGAATGATAAATTTGTCGGTTTGGTTGCCGATAGTGTGCAGATGAAG
GGAATCAATCAATATATTATCTTTTATAAACCTAAACCCACTTCATTTGCGCGATTTAGGCGTTCTGCACGGTCGAGG
CGGTCGCTTCCGGCCGAGATGCCGCTGATTCCCGTCAATCAGGCGGATACGCTGATTGTCGATGGGGAAGCGGTCAGC
CTGACGGGGCATTCCGGCAATATCTTCGCGCCCGAAGGGAATTACCGGTATCTGACTTACGGGGCGGAAAAATTGCCC
GGCGGATCGTATGCCCTCCGTGTTCAAGGCGAACCTTCAAAAGGCGAAATGCTCGCGGGCACGGCAGTGTACAACGGC
GAAGTGCTGCATTTTCATACGGAAAACGGCCGTCCGTCCCCGTCCAGAGGCAGGTTTGCCGCAAAAGTCGATTTCGGC
AGCAAATCTGTGGACGGCATTATCGACAGCGGCGATGGTTTGCATATGGGTACGCAAAAATTCAAAGCCGCCATCGAT
GGAAACGGCTTTAAGGGGACTTGGACGGAAAATGGCGGCGGGGATGTTTCCGGAAAGTTTTACGGCCCGGCCGGCGAG
GAAGTGGCGGGAAAATACAGCTATCGCCCAACAGATGCGGAAAAGGGCGGATTCGGCGTGTTTGCCGGCAAAAAAGAG
CAGGATGGATCCGGAGGAGGAGGAGCCACCTACAAAGTGGACGAATATCACGCCAACGCCCGTTTCGCCATCGACCAT
TTCAACACCAGCACCAACGTCGGCGGTTTTTACGGTCTGACCGGTTCCGTCGAGTTCGACCAAGCAAAACGCGACGGT
AAAATCGACATCACCATCCCCGTTGCCAACCTGCAAAGCGGTTCGCAACACTTTACCGACCACCTGAAATCAGCCGAC
ATCTTCGATGCCGCCCAATATCCGGACATCCGCTTTGTTTCCACCAAATTCAACTTCAACGGCAAAAAACTGGTTTCC
GTTGACGGCAACCTGACCATGCACGGCAAAACCGCCCCCGTCAAACTCAAAGCCGAAAAARRCAACRGCRACCAAAGC
CCGARGGCGAAAACCGAAGRRRGCGGCGGCGACTTCAGCACCACCATCGACCGCACCAAATGGGGCGTGGACTACCTC
GTTAACGTTGGTATGACCAAAAGCGTCCGCATCGACATCCAAATCGAGGCAGCCAAACAATAAAAGCTT
1 MASPDVKSAD TLSKPAAPVV SEKETEAKKD APQAGSQGQG QPSAQGGQDM
 51    AAVSEENTGN GGAAATDKPK NEDEGAQNDM PQNAADTDSL TPNHTPASNM
101    PAGNMENQAP DAGESEQPAN QPDMANTADG MQGDDPSAGG ENAGNTAAQG
151    TNQAENNQTA GSQNPASSTN PSATNSGGDF GRTNVGNSVV IDGPSQNITL
201    THCKGDSCSG NNFLDEEVQL KSEFEKLSDA DKISNYKKDG RNDGKNDKFV
251    GLVADSVQMK GINQYIIFYK PKPTSFARFR RSARSRRSLP AEMPLIPVNQ
301    ADTLIVDGEA VSLTGHSGNI FAPEGNYRYL TYGAEKLPGG SYALRVQGEP
351    SKGEMLAGTA VYNGEVLHFH TENGRPSPSR GRFAAKVDFG SKSVDGIIDS
401    GDGLHMGTQK FKAAIDGNGF KGTWTKNGGG DVSGKFYGPA GEEVAGKYSY
451    RPTDAEKGGF GVFAGKKEQD GSGGGGATYK VDEYHANARF AIDHFNTSTN
501    VGGFYGLTGS VEFDQAKRDG KIDITIPVAN LQSGSQHFTD HLKSADIFDA
551    AQYPDIRFVS TKFNFNGKKL VSVDGNLTMH GKTAPVKLKA EKFNCYQSPM
601    AKTEVCGGDF STTIDRTKWG VDYLVNVGMT KSVRIDIQIE AAKQ*
ΔG287NZ-961
ATGGCTAGCCCCGATGTCAAGTCGGCGGACACGCTGTCAAAACCTGCCGCCCCTGTTGTTTCTGAAAAAGAGACAGAG
GCAAAGGAAGATGCGCCACAGGCAGGTTCTCAAGGACAGGGCGCGCCATCCGCACAAGGCGGTCAAGATATGGCGGCG
GTTTCGGAAGAAAATACAGGCAATGGCGGTGCGGCAGCAACGGACAAACCCAAAAATGAAGACGAGGGGGCGCAAAAT
GATATGCCGCAAAATGCCGCCGATACAGATAGTTTGACACCGAATCACACCCCGGCTTCGAATATGCCGGCCGGAAAT
ATGGAAAACCAAGCACCGGATGCCGGGGAATCGGAGCAGCCGGCAAACCAACCGGATATGGCAAATACGGCGGACGGA
ATGCAGGGTGACGATCCGTCGGCAGGCGGGGAAAATGCCGGCAATACGGCTGCCCAAGGTACAAATCAAGCCGAAAAC
AATCAAACCGCCGGTTCTCAAAATCCTGCCTCTTCAACCAATCCTAGCGCCACGAATAGCGGTGGTGATTTTGGAAGG
ACGAACGTGGGCAATTCTGTTGTGATTGACGGGCCGTCGCAAAATATAACGTTGACCCACTGTAAAGGCGATTCTTGT
AGTGGCAATAATTTCTTGGATGAAGAAGTACAGCTAAAATCAGAATTTGAAAAATTAAGTGATGCAGACAAAATAAGT
AATTACAAGAAAGATGGGAAGAATGACGGGAAGAATGATAAATTTGTCGGTTTGGTTGCCGATAGTGTGCAGATGAAG
GGAATCAATCAATATATTATCTTTTATAAACCTAAACCCACTTCATTTGCGCGATTTAGGCGTTCTGCACGGTCGAGG
CGGTCGCTTCCGGCCGAGATGCCGCTGATTCCCGTCAATCAGGCGGATACGCTGATTGTCGATGGGGAAGCGGTCAGC
CTGACGGGGCATTCCGGCAATATCTTCGCGCCCGAAGGGAATTACCGGTATCTGACTTACGGGGCGGAAAATTGCCCC
GGCGGATCGTATGCCCTCCGTGTTCAAGGCGAACCTTCAAAAGGCGAAATGCTCGCGGGCACGGCAGTGTACAACGGC
GAAGTGCTGCATTTTCATACGGAAAACGGCCGTCCGTCCCCGTCCAGAGGCAGGTTTGCCGCAAAAGTCGATTTCGGC
AGCAAATCTGTGGACGGCATTATCGACAGCGGCGATGGTTTGCATATGGGTACGCAAAAATTCAAAGCCGCCATCGAT
GGAAACGGCTTTAAGGGGACTTGGACGGAAAATGGCGGCGGGGATGTTTCCGGAAAGTTTTACGGCCCGGCCGGCGAG
GAAGTGGCGGGAAAATACAGCTATCGCCCAACAGATGCGGAAAAGGGCGGATTCGGCGTGTTTGCCGGCAAAAAAGAG
CAGGATGGATCCGGAGGAGGAGGAGCCACAAACGACGACGATGTTAAAAAAGCTGCCACTGTGGCCATTGCTGCTGCC
TACAACAATGGCCAAGAAATCAACGGTTTCAAAGCTGGAGAGACCATCTACGACATTGATGAAGACGGCACAATTACC
AAAAAAGACGCAACTGCAGCCGATGTTGAAGCCGACGACTTTAAAGGTCTGGGTCTGAAAAAAGTCGTGACTAACCTG
ACCAAAACCGTCAATGAAAACAAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAATCTGAAATAGAAAAGTTAACA
ACCAAGTTAGCAGACACTGATGCCGCTTTAGCAGATACTGATGCCGCTCTGGATGCAACCACCAACGCCTTGAATAAA
TTGGGAGAAAATATAACGACATTTGCTGAAGAGACTAAGACAAATATCGTAAAAATTGATGAAAAATTAGAAGCCGTG
GCTGATACCGTCGACAAGCATGCCGAAGCATTCAACGATATCGCCGATTCATTGGATGAAACCAACACTAAGGCAGAC
GAAGCCGTCAAAACCGCCAATGAAGCCAAACAGACGGCCGAAGAAACCAAACAAAACGTCGATGCCAAAGTAAAAGCT
GCAGAAACTGCAGCAGGCAAAGCCGAAGCTGCCGCTGGCACAGCTAATACTGCAGCCGACAAGGCCGAAGCTGTCGCT
GCAAAAGTTACCGACATCAAAGCTGATATCGCTACGAACAAAGATAATATTGCTAAAAAAGCAAACAGTGCCGACGTG
TACACCAGAGAAGAGTCTGACAGCAAATTTGTCAGAATTGATGGTCTGAACGCTACTACCGAAAAATTGGACACACGC
TTGGCTTCTGCTGAAAAATCCATTGCCGATCACGATACTCGCCTGAACGGTTTGGATAAAACAGTGTCAGACCTGCGC
AAAGAAACCCGCCAAGGCCTTGCAGAACAAGCCGCGCTCTCCGGTCTGTTCCAACCTTACAACGTGGGTCGGTTCAAT
GTAACGGCTGCAGTCGGCGGCTACAAATCCGAATCGGCAGTCGCCATCGGTACCGGCTTCCGCTTTACCGAAAACTTT
GCCGCCAAAGCCAGGCGTGCAGTCGGCACTTCGTCCGGTTCTTCCGCAGCCTACCATGTCGGCGTCAATTACGAGTGG
TAAAAGCTT
  1    MASPDVKSAD TLSKPAAPVV SEKETEAKED APQAGSQGQG APSAQGGQDM
 51    AAVSEENTGN GGAAATDKPK NEDEGAQNDM PQNAADTDSL TRNHTPASNM
101    PAGNMENQAP DAGESEQPAN QPDMDNTADG KQGDDPSAGG KNAGNTAAQG
151    TNQAKNNQTA GSQNPASSTN PSATNSGGDF GRTNVGNSVV IDGPSPNITL
2 01   THCKGDSCSG NNFLDEEVQL KSEFRKLSDA DKISNYKKDG KNDGKNDKFV
251    GLVADSVQMK GINQYIIFYX PKPTSFARFR RSARSRRSLP AEMPLIPVNQ
301    ADTLIVDGEA VSLTGHSGNI FAPRGNYRYL TYGAEKLPGG SYALRVQGEP
351    SKGEMLAGTA VYNGRVLHFH TENGRPSPSR GRFAAKVDFG SKSVDGIIDS
401    GDGLHNGTQK FKAAIDGNGF KGTWTRNGGG DVSGKFYGPA GRRVNGKYSY
451    RPTDAEKGGF GVFAGKKKQD GSGGGGATND DDVKKAATVA IAAAYNNGQE
501    INGFKAGETI YDIDEDGTIT KKDATAADVE ADDFKGLGLK KVVTNLTKTV
551    NEMLQMVDAL VKAAESSIKK LTTKLADTDA ALADTDAALD ATTNALNKLG
601    ENITTFAEET KTNIVKIDKK LKAVADTVDK EAKAFNDIAD SLDETNTKAD
651    EAVKTANEAK QTAEETKQNV DAKVKAAETA AGKAEAAAGT ANTAADKAEA
701    VAAKVTDIKA DIATNKDNIA KKANSADVYY RKRSDSKFVR IDGLNATTEK
151    LDTRLASAEF SIADHDTRLN GLDITVSDLR KETRQGLAEQ AALSGLFQPY
801    NVGRFNVTAA VGGYKSESAV AIGTGFRFTE NFAAKAGVAV GTSSGSSAAY
851    HVGVNYEW*                                           实施例3-ΔG983的杂交体
蛋白质983具有如下序列：
983                                          →ΔG983
1     MRTTPTFPTK TFKPTAMALA VATTLSACLG GGGGGTSAPD FNAGGTGIGS
51   NSRATTAKSA AVSYAGIKNE MCKDRSMLCA GRDDVAVTDR DAKINAPPPN
101  LHTGDFPNPN DAYKNLINLK PAIEAGYTGR GVEVGIVDTG ESVGSISFPE
151  LYGRKEHGYN ENYKNYTAYM RKEAPEDGGG KDIEASFDDE AVIETEAKPT
201  DIRHVKEIGH IDLVSHIIGG RSVDGRPAGG IAPDATLHIM NTNDETKNEM
251  MVAAIRNAWV KLGERGVRIV NNSFGTTSRA GTADIFQIAN SEEQYRQALL
301  DYSGGDKTDE GIRLMQQSDY GNLSYHIRNK NMLFIFSTGN DAQAQPNTYA
351  LLPFYEKDAQ KGIITVAGVD RSGEKFKREM YGEPGTEPLE YGSNHCGITA
401  MWCLSAPYEA SVRFTRTNPI QIAGTSFSAP IVTGTAALLL QKYPWMSNDN
451  LRTTLLTTAQ DIGAVGVDSK FGWGLLDAGK AMNGPASFPF GDFTADTKGT
501  SDIAYSFRND ISGTGGLIKK GGSQLQLHGN NTYTGKTIIE GGSLVLYGNN
551  KSDMRVETKG ALIYNGAASG GSLNSDGIVY LADTDQSGAN ETVHIKGSLQ
601  LDGKGTLYTR LGKLLKVDGT AIIGGKLYMS ARGKGAGYLN STGRRVFFLS
651  AAKIGQDYSF FTNIETDGGL LASLDSVEKT AGSHGDTLSY YVRRGNAART
701  ASAAAHSAPA GLKHAVEQGG SNLENLMVEL DASESSATPE TVETAAADRT
751  DMPGIRPYGA TFRAAAAVQH ANAADGVRIF NSLAATVYAD STAAHADMQG
801  RRLKAVSDGL DHNGTGLRVI AQTQQDGGTW EQGGVEGKMR GSTQTVGEIA
851  KTGENTTAAA TLGMGRSTWS ENSANAKTDS ISLFAGIRHD AGDIGYLKGL
901  FSYGRYKNSI SRSTGADEHA EGSVNGTLNQ LGALGGVNVP FAATGDLTVE
951  GGLRYDLLKQ DAFAEKGSAL GWSGNSLTEG TLVGLAGLKL SQPLSDKAVL
1001 FATAGVERDL NGRDYTVTGG FTGATAATGK TGARNMPHTR LVAGLGADVE
1051 FGNGWNGLAR YSYAGSKQYG NHSGRVGVGY RF*
因此ΔG983具有如下基本序列：
                                      TSAPD FNAGGTGIGS
NSRATTAKSA AVSYAGIKNE NCKDRSMLCA GRDDVAVTDR DAKINAPPPN
LHTGDFPNPN DAYKNLINLK PAIEAGYTGR GVEVGIVDTG ESVGSISFPE
LYGRKEHGYN ENYKNYTAYM RKEAPEDGGG KDIEASFDDE AVIETEAKPT
DIRHVKEIGH IDLVSHIIGG RSVDGRPAGG IAPDATLHIM NTNDETKNEM
MVAAIRNAWV KLGERGVRIV NNSFGTTSRA GTADLFQIAN SEEQYRQALL
DYSGGDKTDE GIRLMQQSDY GNLSYHIRNK NMLFIFSTGN DAQAQPNTYA
LLPFYEKDAQ KGIITVAGVD RSGEKFKREM YGEPGTEPLE YGSNHCGITA
MWCLSAPYEA SVRFTRTNPI QIAGTSFSAP IVTGTAALLL QKYPWMSNDN
LRTTLLTTAQ DIGAVGVDSK FGWGLLDAGK ANNGPASFPF GDFTADTKGT
SDIAYSFRND ISGTGGLIKK GGSQLQLHGN NTYTGKTIIE GGSLVLYGNN
KSDMRVETKG ALIYNGAASG GSLNSDGIYY LADTDQSGAN ETVHIKGSLQ
LDGKGTLYTR LGKLLDVDGT AIIGGKLYMS ARGKGAGYLN STGRRVPFLS
AAKIGQDYSF FTNIETDGGL LASLDSVEKT AGSEGDTLSY YVRRGNAART
ASAAAHSAPA GLKHAVEQGG SNLENLMVEL DASESSATPE TVETAAADRT
DMPGIRPYGA TFRAAAAVQH ANAADGVRIF NSLAATVYAD STAAHADMQG
RRLKAVSDGL DHNGTGLRVI AQTQQDGGTN EQGGVEGKMR GSTQTVGIAA
KTGENTTAAA TLGMGRSTWS ENSANAKTDS ISLFAGIRHD AGDIGYLKGL
FSYGRYKNSI SRSTGADEHA EGSVNGTLMQ LGALGGVNVP FAATGDLTVE
GGLRYDLLKQ DAFAEKGSAL GWSGNSLTEG TLVGLAGLKL SQPLSDKAVL
FATAGVKRDL NGRDYTVTGG FTGATAATGK TGARNNPHTR LVAGLGADVE
FGNGWNGLAR YSYAGSKQYG NHSGRVGVGY RF*
将ΔG983作为在其C-未端与ORF46.1、741、961或961c的杂交体表达：
AG983-ORF46.1
ATGATTCTGCGCCCGACTTCAATGCAGGCGGTACCGGTATCGGCACGCAACAGCAGAGCAACAACAGCGAAATCAGCA
GCAGTATCTTACQCCQQTATCAGAACGAAATQTCAAAGACAGAAGCATQCTCTCTGTQCCQQTCGGGATGACQTTQCG
GTTACACGACAGGGATQCCAAAATCAATGCCCCCCCCCCCGATCTGCATACCQGAGACTTTCCAAACCCAAATGCQCA
TACAAGAATTTGATCAACCTCAACCTQCAATTGAAQCAQQCTATACAQGACCQCQQQGTAGQTAQQTATATCGTCGAC
ACAGGCGAATCCQTCQQCAQCATATCCTTTCCCQAACTGTATQQCAGAAAAGAACACQQCTATAACGAAAATTACAAA
AACTATACQQCGTATATQCQQAAQQAAGCQCCTGAAGACQQAQQCOOTAAAGACATTGAAQCTTCTTTCGACGATGAG
GCCGTTATAGAGACTGAAGCAAAGCCQACGGATATCCQCCACGTAAAAGAAATCQQACACATGGATTTGGTCTCCCAT
ATTATTGGCGGGCGTTCCGTGGACGGCAGACCTGCAGGCGGTATTGCGCCCGATGCGACGCTACACATAATGAATACG
AATGATGAAACCAAGAACGAAATGATGGTTGCAGCCATCCGCAATGCATGGGTCAAGCTGGGCGAACGTGGCGTGCGC
ATCGTCAATAACAGTTTTGGAACAACATCGAGGGCAGGCACTGCCGACCTTTTCCAAATAGCCAATTCGGAGGAGCAG
TACCGCCAAGCGTTGCTCGACTATTCCGGCGGTGATAAAACAGACGAGGGTATCCGCCTGATGCAACAGAGCGATTAC
GGCAACCTGTCCTACCACATCCGTAATAAAAACATGCTTTTCATCTTTTCGACAGGCAATGACGCACAAGCTCAGCCC
AACGCATATGCCCTATTGCCATTTTATGAAAAAGACGCTCAAAAAGGCATTATCACAGTCGCAGGCGTAGACGCCAGT
GGAGAAAAGTTCAAACGGGAAATGTATGGAGAACCGGGTACAGAACCGCTTGAGTATGGCTCCAACCATTGCGGAATT
ACTGCCATGTGGTGCCTGTCGGCACCCTATGAAGCAAGCGTCCGTTTCACCCGTACAAACCCGATTCAAATTGCCGGA
ACATCCTTTTCCGCACCCATCGTAACCGGCACGGCGGCTCTGCTGCTGCAGAAATACCCGTGGATGAGCAACGACAAC
CTGCGTACCACGTTGCTGACGACGGCTCAGGACATCGGTGCAGTCGGCGTGGACAGCAAGTTCGGCTGGGGACTGCTG
GATGCGGGTAAGGCCATGAACGGACCCGCGTCCTTTCCGTTCGGCGACTTTACCGCCGATACGAAAGGTACATCCGAT
ATTGCCTACTCCTTCCGTAACGACATTTCAGGCACGGGCGGCCTGATCAAAAAAGGCGGCAGCCAACTGCAACTGCAC
GGCAACAACACCTATACGGGCAAAACCATTATCGAAGGCGGTTCGCTGGTGTTGTACGGCAACAACAAATCGGATATG
CGCGTCGAAACCAAAGGTGCGCTGATTTATAACGGGGCGGCATCCGGCGGCAGCCTGAACAGCGACGGCATTGTCTAT
CTGGCAGATACCGACCAATCCGGCGCAAACGAAACCGTACACATCAAAGGCAGTCTGCAGCTGGACGGCAAAGGTACG
CTGTACACACGTTTGGGCAAACTGCTGAAAGTGGACGGTACGGCGATTATCGGCGGCAAGCTGTACATGTCGGCACGC
GGCAAGGGGGCAGGCTATCTCAACAGTACCGGACGACGTGTTCCCTTCCTGAGTGCCGCCAAAATCGGGCAGGATTAT
TCTTTCTTCACAAACATCGAAACCGACGGCGGCCTGCTGGCTTCCCTCGACAGCGTCGAAAAAACAGCGGGCAQTGAA
GGCGACACGCTGTCCTATTATGTCCGTCGCGGCAATGCGGCACGGACTGCTTCGGCAGCGGCACATTCCGCGCCCGCC
GGTCTGAAACACGCCGTAGAACAGGGCGGCAGCAATCTGGAAAACCTGATGGTCGAACTGGATGCCTCCGAATCATCC
GCAACACCCGAGACGGTTGAAACTGCGGCAGCCGACCGCACAGATATGCCGGGCATCCGCCCCTACGGCGCAACTTTC
CGCGCAGCGGCAGCCCTACAGCATGCGAATGCCGCCGACGGTGTACGCATCTTCAACAGTCTCGCCGCTACCGTCTAT
GCCGACAGTACCGCCGCCCATGCCGATATGCAGGGACGCCGCCTGAAAGCCGTATCGGACGGGTTGGACCACAACGGC
ACGGGTCTGCGCGTCATCGCGCAAACCCAACAGGACGGTGGAACGTGGGAACAGGGCGGTGTTGAAGGCAAAATGCGC
GGCAGTACCCAAACCGTCGGCATTGCCGCGAAAACCGGCGAAAATACGACAGCAGCCGCCACACTGGGCATGGGACGC
AGCACATGGAGCGAAAACAGTGCAAATGCAAAAACCGACAGCATTAGTCTGTTTGCAGGCATACGGCACGATGCGGGC
GATATCGGCTATCTCAAAGGCCTGTTCTCCTACGGACGCTACAAAAACAGCATCAGCCGCAGCACCGGTGCGGACGAA
CATGCGGAAGGCAGCGTCAACGGCACGCTGATGCAGCTGGGCGCACTGGGCGGTGTCAACGTTCCGTTTGCCGCAACG
GGAGATTTGACGGTCGAAGGCGGTCTGCGCTACGACCTGCTCAAACAGGATGCATTCGCCGAAAAAGGCAGTGCTTTG
GGCTGGAGCGGCAACAGCCTCACTGAAGGCACGCTGGTCGGACTCGCGGGTCTGAAGCTGTCGCAACCCTTGAGCGAT
AAAGCCGTCCTGTTTGCAACGGCGGGCGTGGAACGCGACCTGAACGGACGCGACTACACGGTAACGGGCGGCTTTACC
GGCGCGACTGCAGCAACCGGCAAGACGGGGGCACGCAATATGCCGCACACCCGTCTGGTTGCCGGCCTGGGCGCGGAT
GTCGAATTCGGCAACGGCTGGAACGGCTTGGCACGTTACAGCTACGCCGGTTCCAAACAGTACGGCAACCACAGCGGA
CGAGTCGGCGTAGGCTACCGGTTCCTCGACGGTGGCGGAGGCACTGGATCCTCAGATTTGGCAAACGATTCTTTTATC
CGGCAGGTTCTCGACCGTCAGCATTTCGAACCCGACGGGAAATACCACCTATTCGGCAGCAGGGGGGAACTTGCCGAG
CGCAGCGGCCATATCGGATTGGGAAAAATACAAAGCCATCAGTTGGGCAACCTGATGATTCAACAGGCGGCCATTAAA
GGAAATATCGGCTACATTGTCCGCTTTTCCGATCACQQQCACGAAGTCCATTCCCCCTTCGACAACCATGCCTCACAT
TCCGATTCTGATGAAGCCQQTAGTCCCGTTGACGGATTTAGCCTTTACCGCATCCATTQQQACQQATACGAACACCAT
CCCGCCGACGGCTATGACQGGCCACAQQQCQQCQQCTATCCCQCTCCCAAAQQCQCQAQQQATATATACAQCTACGAC
ATAAAAQQCGTTGCCCAAAATATCCGCCTCAACCTGACCGACAACCQCAGCACCQGACAACQQCTTQCCGACCQTTTC
CACAATGCCGGTAGTATGCTGACGCAAGGAGTAGGCGACGGQATTCAAACGCQCCACCCGATACAGCCCGAGCTGGAC
AGCTCGGGCAATGCCGCCGAAGCCTTCAACGGCACTGCAQATATCGTTAAAAACATCATCGGCGCQGCAGGAGAAATT
GTCGGCGCAGQCGATQCCGTGCAQQQCATAAGCQAAQQCTCAAACATTQCTTQCATQCACQGCTTQQQTCTGCTTTCC
ACCCGAAAAACAACATQQCQCGCATCAACGATTTGGCAGATATCGCAACTCAAAGACTATQCCGCAGCAQCCATCCGC
GATTGGGCAGTCCAAAACCCCAATGCCGCACAAGGCATAQAAQCCQTCAGCAATATCTTTATQQCAQCCQTCCCCATC
AAAGGGATTGGAGCTGTTCQQQQAAAATACGGCTTQQQCQQCATCACGGCACATCCTATCAAGCQQTCGCAGATGGGC
GCGATCGCATTGCCGAAAGGGAAATCCGCCQTCAQCGACAATTTTQCCGATQCGGCATACGCCAAATACCCGTCCCCT
TACCATTCCCGAAATATCCGTTCAAACTTQGAGCAQCQTTACGGCAAAQAAAACATCACCTCCTCAACCGTQCCQCCG
TCAAACGGCAAAAATGTCAAACTGGCAGACCAACGCCACCCGAAGACAQGCGTACCGTTTGACQQTAAAGGGTTTCCG
AATTTTGAGAAGCACGTGAAATATGATACGCTCGAQCACCACCACCACCACCACTGAA
  1    MTSAPDENAG GTGIGSNSRA TTAKSAAVSY AGIKNKNCKD RSMLCAGRDD
  51   VAVTDRDAKI NAPPPNLHTG DFPNPNDAYK NLINLKPAIR AGYTGRGVEV
101    GIVDTGRSVG SISFPRLYGR KRHGYNENYK NYTAYMRKBA PRDQQGKDIE
151    ASFDDEAVIB TEAKPTDIRH VKKIGHIDLV SHIIGGRSVD GRPAQQIAPD
201    ATLHIMNTND STKNRMMVAA IRNAWVKLGB RGVRIVNNSF GTTSRAGTAD
251    IFQIANSEEQ YRQALLDYSG GDKTDEGIRL MQQSDYGNLS YHIRNKNMLF
301    IFSTGNDAQA QPNTYALLPF YEKDAQKGII TVAGVDRSGE KFKREMYGEP
351    GTEPLEYGSN HCGITAMWCL SAPYEASVRF TRTNPIQIAG TSFSAPIVTG
401    TAALLLQKYP WMSNDNLRTT LLTTAQDIGA VGVDSKPGWG LLDAGKAMNG
451    PASFPFGDFT ADTKQTSDIA YSFRNDISQT QQLIKKQGSQ LQLHQNNTTT
501    GKTIIRQQSL VLYQNNKSDM RVBTKGALYY NGAASQQSIN SDGIVYLADT
551    DQSGANSTVR IKGSLQLDCK GTLYTRLGKL LKVDGTAIIG GKLYMSARGK
601    GAGYINSTGR RVPFISAAKI GQDYSFFINI BTDGGLLASL DSVRKTAGSE
651    GDTLSYYVRR GNAARTASAA AHSAPAGLKH AVSQGGSNLR NLMVHLDASE
701    SSATPETVET AAADRTDMPG IRPYGATFRA AAAVQHANAA DGVRIFNSLA
751    ATVYADSTAA HADMQGRRLK AVSDGLDHNG TGLRVIAQTQ QDQQTWRQGG
801    VEGKMRGSTQ TVGIAAKTGE NTTAAATLGM GRSTWSENSA NAKTDSISLF
851    AGIRHDAGDI GYLKGLFSYG RYKNSISRST GADEHAEGSV NGTLMQLGAL
901    GGVNVPFAAT GDLTVEGGLR YDLLKQDAFA KKGSALGWSG NSLTEGTLVG
951    LAGLKLSQPL SDKAVLFATA GVERDLNGRD YTVTGGFTGA TAATGKTGAR
1001   NMPHTRLVAG LGADVEFGNG WNGLARYSYA GSKQYGNHSG RVGVGYRFLD
1051   GGGGTGSSDL ANDSFIRQVL DRQHFEPDGK YHLFGSRGEL AERSGHIGLG
1101   KIQSHQLGNL MIQQAAIKGN IGYIVRFSDH GHEVHSPFDN HASHSDSDEA
1151   GSPVDGFSLY RIHWDGYEHH PADGYDGPQG GGYPAPKGAR DIYSYDIKGV
1201   AQNIRLNLTD NRSTGQRLAD RFHNAGSNLT QGVGDGFKRA TRYSPKLDRS
1251   GNAAEAFNGT ADIVKNIIGA AGEIVGAGDA VQGISEGSNI AVMHGLGLLS
1301   TENKMARIND LADMAQLKDY AAAAIRDWAV QNPNAAQGIE AVSNIFMAAT
1351   PIKGIGAVRG KYGLGGITAH PIKRSQMGAI ALPKGKSAVS DNFADAAYAK
1401   YPSPYHSRNI RSNLEQRYGK ENITSSTVPP SNGKNVKLAD QRHPKTGVPF
1451   DGKGFPNFEK HVKYDTLEHH HHHH*
ΔG983-741
ATGACTTCTGCGCCCGACTTCAATGCAGGCGGTACCQGTATCGGCAQCAACAGCAGAGCAACAACAQCGAAATCAGCA
GCAGTATCTTACGCCGGTATCAAGAACGAAATGTGCAAAGACAGAAQCATQCTCTGTQCCGGTCQQQATGACGTTGCG
GTTACAGAGAQQQATGCCAAAATCAATQCCCCCCCCCCGAATCTQCATACCQQAGACTTTCCAAACCCAAATGACGCA
TACAAGAATTTGATCAACCTCAAACCTGCAATTGAAQCAQGCTATACAQGACGCQQQQTAGAQQTAQQTATCGTCGAC
ACAGGCGAATCCGTCGGCAGCATATCCTTTCCCGAACTGTATQQCAGAAAAGAACACQQCTATAACGAAAATTACAAA
AACTATACGGCGTATATGCGGAAGGAAGCGCCTGAAGACGGAGGCQGTAAAGACATTGAAGCTTCTTTCGACGATGAG
GCCCGTTATAGAGACTGAAQCAAAQCCGACQQATATCCQCCACQTAAAAGAAATCGACACATCGATTTGGTCTCCCAT
ATTATTQGCGGGCGTTCCQTGGACGGCAGACCTGCAGGCQQTATTGCGCCCQATGCGACGCTACACATAATGAATACG
AATGATGAAACCAAGAACGAAATGATQCTTGCAGCCATCCGCAATGCATQQQTCAAQCTQQGCGAACGTQQCGTQCGC
ATCGTCAATAACAGTTTTGGAACAACATCGAGGGCAGGCACTGCCGACCTTTTCCAAATAGCCAATTCQGAGGAGCAG
TACCQCCAAGCGTTQCTCGACTATTCCQQCQGTGATAAAACAGACGAGQCTATCCQCCTGATQCAACAGAGCGATTAC
GGCAACCTGTCCTACCACATCCGTAATAAAAAGATGCTTTTCATCTTTTCGACAGGCAATGACQCACAAGCTCAGCCC
AACACATATGCCCTATTGCCATTTTATGAAAAAGACGCTCAAAAAQQCATTATCACAQTCGCAGGCGTAGACCGCAGT
GGAGAAAAGTTCAAACGGGAAATGTATGGAGAACCGGGTACAGAACCQCTTGAGTATQGCTCCAACCATTGCGGAATT
ACTGCCATGTGGTQCCTGTCGGCACCCTATGAAGCAAGCGTCCGTTTCACCCGTACAAACCCGATTCAAATTGCCGGA
ACATCCTTTTCCGCACCCATCGTAACCGGCACQGCGGCTCTGCTGCTQCAGAAATACCCQTGGATGAGCAACGACAAC
CTGCGTACCACGTTGCTGACGACQQCTCAGGACATCGGTGCAGTCGGCQTGGACAGCAAGTTCQGCTQQQQACTGCTG
GATGCGGGTAAQQCCATGAACGGACCCGCGTCCTTTCCGTTCGQCGACTTTACCGCCQATACGAAAGGTACATCCGAT
ATTGCCTACTCCTTCCGTAACQACATTTCAGGCACQQQCQGCCTGATCAAAAAAQQCQQCAGCCAACTQCAACTGCAC
GGCAACAACACCTATACQQQCAAAACCATTATCGAAGGCGGTTCQCTGGTGTTQTACQQCAACAACAAATCGGATATG
CGCGTCGAAACCAAAGGTGCGCTGATTTATAACGQQQCGGCATCCGGCGQCAGCCTGAACAGCGACQGCATTGTCTAT
CTGGCAGATACCGACCAATCCGGCGCAAACGAAACCGTACACATCAAAGGCAGTCTGCAGCTGGACQGCAAAQQTACG
CTGTACACACGTTTGGGCAAACTGCTGAAAGTQGACOGTACQQCGATTATCGGCGGCAAQCTQTACATGTCGGCACGC
GGCAAGGGGGCAGGCTATCTCAACAGTACCGGACGACGTGTTCCCTTCCTGAGTGCCGCCAAAATCGGGCAGGATTAT
TCTTTCTTCACAAACATCGAAACCGACQQCQGCCTGCTQGCTTCCCTCGACAGCGTCGAAAAAACAGCQGGCAGTGAA
GGCGACACGCTGTCCTATTATGTCCGTCGCGGCAATGCGGCACGGACTGCTTCGGCAGCGGCACATTCCQCGCCCGCC
GGTCTGAAACACGCCGTAGAACAGGGCGGCAGCAATCTGGAAAACCTGATQQTCGAACTQGATQCCTCCQAATCATCC
OCAACACCCGAGACGGTTGAAACTQCGQCAGCCGACCGCACAGATATGCCQQQCATCCGCCCCTACQQCGCAACTTTC
CGCGCAQCQQCAGCCGTACAGCATGCGAATGCCGCCGACQQTGTACGCATCTTCAACAGTCTCQCCGCTACCGTCTAT
GCCGACAGTACCGCCGCCCATGCCGATATGCAQQQACGCCQCCTGAAAQCCQTATCQQACGGGTTGGACCACAACGGC
ACQQQTCTGCGCGTCATCGCGCAAACCCAACAGGACGGTGGAACQTQQGAACAGQQCQGTGTTGAAGGCAAAATGCGC
GGCAGTACCCAAACCGTCGGCATTGCCQCGAAAACCGGCGAAAATACGACAGCAGCCGCCACACTGGGCATGGGTCGC
AGCACATGGAGCGAAAACAGTGCAAATGCAAAAACCGACAGCATTAGTCTGTTTGCAGGCATACGGCACGATGCQQQC
GATATCGGCTATCTCAAAGGCCTGTTCTCCTACGGACGCTACAAAAACAGCATCAGCCGCAGCACCGGTGCGGACGAA
CATGCGGAAGGCAGCGTCAACGGCACGCTGATQCAGCTQQQCQCACTQQQCQQTGTCAACGTTCCQTTTGCCGCAACG
GGAGATTTGACQQTCQAAQQCGGTCTGCQCTACQACCTQCTCAAACAQQATQCATTCQCCGAAAAAGGCAGTGCTTTG
GGCTGGAGCGGCAACAQCCTCACTGAAQQCACQCTGGTCGGACTCGCGQQTCTGAAGCTQTCGCAACCCTTGAGCGAT
AAAGCCGTCCTGTTTGCAACGGCQQQCGTGGAACQCQACCTGAACQGACQCGACTACACQGTAACGGGCGGCTTTACC
GGCGCGACTGCAGCAACCGGCAAGACGGGGGCACGCAATATGCCQCACACCCGTCTQQTTGCCGGCCTGGGCGCGGAT
GTCGAATTCGGCAACGGCTGGAACGGCTTGGCACGTTACAGCTAGCCGGTTCCAAACAGTACGGCAACCACAGCCGGA
CGAGTCGGCGTAGGCTACCGGTTCCTCGAGGGATCCGGAGGGGGTGGTGTCGCCGCCGACATCGGTGCQQQGCTTGCC
GATGCACTAACCGCACCGCTCGACCATAAAGACAAAGGTTTGCAQTCTTTGACGCTGGATCAGTCCGTCAGGAAAAAC
GAGAAACTGAAGCTGGCGGCACAAGGTGCGGAAAAAACTTATGGAAACGGTGACAGCCTCAATACGGGCAAATTGAAG
AACGACAAGGTCAQCCGTTTCGACTTTATCCQCCAAATCGAAQTQGACGGGCAGCTCATTACCTTGGAGAGTGGAGAG
TTCCAAGTATACAAACAAAGCCATTCCQCCTTAACCGCCTTTCAGACCGAGCAAATACAAGATTCGGAGCATTCCGQG
AAGATGGTTGCGAAACGCCAGTTCAGAATCGGCGACATAGCGGGCGAACATACATCTTTTGACAAGCTTCCCGAAGGC
GGCAGGGCGACATATCGCGGGACGGCGTTCGGTTCAGACGATGCCQQCGGAAAACTGACCTACACCATAQATTTCGCC
GCCAAGCAGGGAAACGGCAAAATCGAACATTTGAAATCGCCAGAAACTCAATQTCQACCTQQCCGCCGCGATATCAAG
CCGGATGGAAAACGCCATQCCQTCATCAGCGGTTCCGTCCTTTACAACCAAGCCGAGAAAGGCAGTTACTCCCTCGGT
ATCTTTGGCAAAAGCCCAGGAAGTTGCCGGCAGCGCGGAAGAACCGTAAACGGCACGCCATCGGCCTT
GCCGCCAAGCAACTCGAGCACCACCACCACCACCATGA
1     MTSAPDFNAG GTGIGSNSRA TTAKSAAVSY AGIKNEMCKD RSMLCAGRDD
51    VAVTDRDAKI NAPPPNLHTG NFPNPNDAYK NLINLKPAIE AGYTGRGVEV
101   GIVDTGESVG SISFPELYGR KKHGYNENYK NYTAYMRKEA PEDGGGKDIE
151   ASFDDEAVIE TEAKPTDIRH VKEIGHIDLV SHIIGRRSVD GRPAGGIAPD
201   ATTLHIMTND ETKNEMMVAA IRNAWVKLGE RGVRIVNNSF GTTSRAGTAD
251   LFFQLASEEQ YRQALLDYSG GDKTEGIRLK QQSDYGNLDS YHIRNKNMLF
301   IFSTGNDAQA QPNTYALLPF YEKDAQKGII TVAGVDRSGE KFKREMYGEP
351   GTEPLEYGSN HCGITAMWCL SAPYEASVRF TRTNPIQIAG TSFSAPIVTG
401   TAALLLQKYP WMSNDNLRTT LLTTAQDIGA VGVDSKFGWG LLDAGKAMNG
451   PASFPFGDFT ADTKGTSDIA YSFRNDISGT GGLIKKGGSQ LQLHGNNTYT
501   GKTIIEGGSL VLYGNNKSDM RVETKGALIY NGAASGGSLN SDGIVYIADT
551   DQSGANETVH IKGSLQLDGK GTLYTRLGKL LKVDGTAIIG GKLYMSARGK
601   GAGYLNSTGR RVPFLSAAKI GQDYSFFTNT ETDGGLLASL DSVEKTAGSE
651   GDTLSYYVRR GNAARTASAA AHSAPGLKHD DVKQGGSNLE NLMVELDASE
701   SSATPETVET AAADRTMPGD IRPYGAFRAS AAAVQHANAA DGVRIFNSLA
751   ATVYADSTAA HADMQGRRLK AVSDGLDHNG TGLRVIAQTQ QDGGTWEQGG
801   VEGKMRGSTQ TVGIAAKTGR NTTAAATLGM GRSTWSRRSA NAKTDSISLF
851   AGLRHDAGDI GYLGLFSYGR YKNSISRSTA GADHAEGGSV NGTLMQLGAL
901   GGVNPFATGD LTVRGGLRYD LLKQDAFAEK GSALGWSGNS LTEGTLDAVG
951   LAGLKLSQPL SDKAVLFATA GVERDLNGRD YTVTGGFTGA TAATGKTGAR
1001  NMPETRLVAG LGADVRFGNG WNGLARYSYA GSKQYGNHSG RVGVGGYFLE
1051  GSGGGGVAAD IGAGLADALT APLDHKDKGL QSLTLDQSVR KNEKIKLAAQ
1101  GAEKTYGNGD SINTGKIKND KVSRFDFIRQ IEVDGQLITL ESGEFQVYKQ
1151  SHSALTAFQT EQIQDSEHSG KMVAKRQFRI GDIAGRHTSF DKLPEGGRAT
1201  YRGTAFGSDD AGGKLTYTID FAAKQGNGKI ERLKSPKLNV DLAAADIKPD
1251  GKRHAVISGS VLYNQAEKGS YSLGIFGGKA QEDVGSAEVK TVNGIRRHRGL
1301  AAKQLEHHHH HH*
ΔG983-961
ATGACTTCTGCGCCCGACTTCAATGCAGGCGGTACCGGTATCGGCAGCAACAGCAGAGCAACAACAGCGAAATCAGCA
GCAGTATCTTACGCCGGTATCAAGAACGAAATGTGCAAAGACAGAAGCATGCTCTGTGCCGGTCGGGATGACGTTGCG
GTTACAGACAGGGATGCCAAAATCAATGCCCCCCCCCCGAATCTGCATACCGGAGACTTTCCAAACCCAAATGACQCA
TACAAGAATTTGATCAACCTCAAACCTGCAATTGAAGCAGGCTATACAGGACGCGGGGTAGAGGTAGGTATCGTCGAC
ACAGGCGAATCCGTCGGCAGCATATCCTTTCCCGAACTGTATGGCAGAAAAGAACACGGCTATAACGAAAATTACAAA
AACTATACGGCGTATATGCGGAAGGAAGCGCCTGAAGACGGAGGCGGTAAAGACATTGAAGCTTCTTTCGACGATGAG
GCCGTTATAGAGACTGAAGCAAAGCCGACGGATATCCGCCACGTAAAAGAAATCGGACACATCGATTTGGTCTCCCAT
ATTATTGGCGGGCGTTCCGTGGACGGCAGACCTGCAGGCGGTATTGCGCCCGATGCGACGGTACACATAATGAATACG
AATGATGAAACCAAGAACGAAATGATGGTTGCAGCCATCCQCAATGCATGGGTCAAGCTGGGCGAACGTGGCGTGCGC
ATCGTCAATAACAGTTTTGGAACAACATCGAGGGCAGGCACTGCCGACCTTTTCCAAATAGCCAATTCGGAGGAGCAG
TACCGCCAAGCGTTGCTCGACTATTCCGGCGGTGATAAAACAGACGAGGGTATCCGCCTGATGCAACAGAGCGATTAC
GGCAACCTGTCCTACCACATCCGTAATAAAAACATGCTTTTCATCTTTTCGACAGGCAATGACGCACAAGCTCAGCCC
AACACATATGCCCTATTGCCATTTTATGAAAAAGACGCTCAAAAAGGCATTATCACAGTCGCAGGCGTAGACCGCAGT
GGAGAAAAGTTCAAACGGGAAATGTAGGAGAACCTGGGTACAGAACCCGCTTGAGTATGQGTCCAACATTGCGGAATT
ACTGCCATGTGGTGCCTGTCGGCACCCTATGAAGCAAGCGTCCGTTTCACCCGTACAAACCCGATTCAAATTGCCGGA
ACATCCTTTTCCGCACCCATCGTAACCGGCACGGCGGCTCTGCTGCTGCAGAAATACCCGTGGATGAGCAACGACAAC
CTGCGTACCACGTTGCTGACGACGGCTCAGGACATCGGTGCAGTCGGCGTGGACAGCAAGTTCGGCTGGGGACTGCTG
GATGCGGGTAAGGCCATGAACGGACCCGCGTCCTTTCCGTTCGGCGACTTTACCGCCGATACGAAAGGTACATCCGAT
ATTGCCTACTCCTTCCGTAACGATTTCAGGCACGGGCGGCCTGATCAAAAAGGCGGCAGCCAACTGAAACTGCATCAC
GGCAACAACACCTATACGGGCAACCATATCGAAGGCGGTTCAGTTCGCTGCTGGTTGCGGCAAAAFCAAACCGATATG
CGCGTCGAAACCAAAGGTGCGCTGATTTATAACGGGGCGGCATCCGGCGGCAGCCTGAACAGCGACGGCATTGTCTAT
CTGGCAGATACCGACCAATCCGGCGCAAACGAAACCGTACACATCAAAGGCAGTCTGCAGCTGGACGGCAAAGGTACG
CTGTACACACGTTTGGGCAAACTGCTGAAAGTGGACGGTACGGCGATTATCGGCGGCAAGCTGTACATGTCGGCACGC
GGCAAGGGGGCAGGCTATCTCAACAGTACCGGACGACGTGTTCCCTTCCTGAGTGCCGCCAAAATCGGGCAGGATTAT
TCTTTCTTCACAAACATCGAAACCGACGGCGGCCTGCTGGCTTCCCTCGACAGCGTCGAAAAAACAGCGGGCAGTGAA
GGCGACACGCTGTCCTATTATGTCCGTCGCGGCAATGCGGCACGGACTGCTTCGGCAGCGGCACATTCCGCGCCCGCC
GGTCTGAAACACGCCGTAGAACAGGGCGGCAGCAATCTGGAAAACCTGATGGTCGAACTGGATGCCTCCCAATCAGCC
CATGCGGAAGGCAGCGTCAACGGCACGCTGATGCAGCTGGGCGCACTGGGCGGTGTCAACGTTCCGTTTGCCGCAACG
GGAGATTTGACGGTCGAAGGCGGTCTQCGCTACGACCTGCTCAAACAGGATGCATTCGCCGAAAAAGGCAGTGCTTTG
GGCTGGAGCGGCAACAGCCTCACTGAAGGCACGCTGGTCGGACTCGCGGGTCTGAAGCTGTCGCAACCCTTGAGCGAT
AAAGCCGTCCTGTTTGCAACGGCGGGCGTGGAACGCGACCTGAACGGACGCGACTACACQQTAACGGGCGGCTTTACC
GGCGCGACTGCAGCAACCGGCAAGACGGGGGCACGCAATATGCCGCACACCCGTCTGGTTQCCGGCCTGGGCGCGGAT
GTCGAATTCGGCAACGGCTGGAACGGCTTGGCACGTTACAGCTACGCCQGTTCCAAACAGTACGQCAACCACAQCGGA
GCTGCCACTGTGGCCATGCTGCTGCCTCAACAATGGCCAAGAAATCAACGGTTTCAAACGCTGGAGAGACCATCTACA
GACATTGATGAAGACGGCACAATTACCAAAAAAGACGCAACTGCAGCCGATGTTGAAGCCGACGACTTTAAAGGTCTG
GGTCTGAAAAAAGTCGTGACTAACCTGACCAAAACCGTCAATGAAAACAAACAAAACGTCGATGCCAAAGTAAAAGCT
GCAGAATCTGAAATAGAAAAGTTAACAACCAAGTTAGCAGACACTGATGCCGCTTTAGCAGATACTGATGCCGCTCTG
GATGCAACCACCAACGCCTTGAATAAATTGGGAGAAAATATAACGACATTTGCTGAAGAGACTAAGACAAATATCGTA
AAAATTGAAAAATTAGAAGCCGTGGCTGTATACCGTCGACAAGCACTGCCGAAGCDATTCAACGATATCGCCGATTCA
TTGGATGAAACCAACACTAAGGCAGACGAAGCCGTCAAAACCGCCAATGAAGCCAAACAGACGGCCGAAGAAACCAAA
CAAAACGTCGATGCCAAAGTAAAAGCTGCAGAAACTGCAGCAGGCAAAGCCGAAGCTGCCGCTGGCACAGCTAATACT
GCAGCCGACAAGGCCGAAGCTGTCGCTGCAAAAGTTACCGACATCAAAGCTGATATCGCTACGAACAAAGATAATATT
GCTAAAAAAGCAAACAGTGCCGACGTGTACACCAGAGAAGAGTCTGACAGCAAATTTGTCAGAATTGATGGTCTGAAC
GCTACTACCGAAAAATTGGACACACGCTTGGCTTCTGCTGAAAAATCCATTGCCGATCACGATACTCGCCTGAACGGT
TTGGATAAAACAGTGTCAGACCTGCGCAAAGAAACCCGCCAAGGCCTTGCAGAACAAGCCGCGCTCTCCGGTCTGTTC
CAACCTTACAACGTGGGGTCGGTTCAATGTAACGGCGCAGTCGGCGGCTACAAATCCGAATCGGCAGTCGCCATCGGT
ACCGGCTTCCGCTTTACCGAAAACTTTGCCGCCAAAGCAGGCGTGGCAGTCGGCACTTCGTCCGGTTCTTCCGCAGCC
TACCATGTCGGCGTCAATTACGAGTGGCTCGAGCACCACCACCACCACCACTGA
1    MAPAGGIGSN  GTGSRATTRA  TTAKSAAVSY  AGDIKNMCKD  RSMKCAGRDD
51   VAVTDRDAKI  NAPPPNLETG  DFPNPNDAYK  NLINDLKAIE  AGYTGRGVEV
101  GIVDTGESVG  SISFPELYGR  KEHGYNKNYT  NYTAYMRKRA  PEDGGCKDIE
151  ASFDDEAVIE  TEKPTDIERH  VVEIGIDDLV  SHIIGGRSVD  GRPAGGIAPD
201  ATLHIMNDND  ETKRMTMVAA  IRNAWVKKGE  RGVRIVNNSF  GTTSRAGTAD
251  LFLANSEESQ  YRQAYLLYSG  GRQALLDYSG  GDKTDEGIRL  KQQSDYGNLF
301  IFSTGNDAQA  QPNTYALLPF  YEKDAQKGII  TVAGVIDSGE  KFKRKNYGEP
351  GTEPLEYGSN  HCGITAMWCL  SAPYEASVRF  TRTNPIQIAG  TSFSAPIVTG
401  TAALLLQKYP  WMSNDNLRTT  LLTTAQDIGA  VGVDSKFGWG  LLDAGKAMNG
451  PASFPFGDFT  ADTKGTSDIA  YSFRNDISGT  GGLIKKGGSQ  LQLHGNNTYT
501  GKTIIEGGSL  VLYGNNKSDM  RVETKGALIY  NGAASGGSLN  SDGIVYLADT
551  DQSGANETVH  IKGSLQLDGK  GTLYTRLGKL  LKVDGTAIIG  GKLYMSARGE
601  GAGYLNSTGR  RVPFLSAAKI  GQDYSFFTNI  ETDGGLLASL  DSVEKTAGSS
651  GDTLSYYVRR  GNAARTASAA  AESAPAGLKH  AVEQQGSNLE  NLMVELDASE
701  SSATPETVET  AAADRTDMPG  IRPYGATFRA  AAAVQEANAA  DGVRIFNSLA
751  ATVYADSTAA  HADMQGRRLK  AVSDGLDHNG  TGLRVIAQTQ  QDGGTWRQGG
801  VEGKMRGSTQ  TVGIAAKTGE  NTTAAATLGM  GRSTWSENSA  NAKTDSISLF
851  AGIRHDAGDI  GYLKGLFSYG  RYKNSISRST  GADEHAEGSV  NGTLMQLGAL
901  GGVNVPFAAT  GDLTVEGGLR  YDLLKQDAFA  KKGSALGWSG  NSLTEGTLVG
951  LAGLKLSQPL  SDKAVLFATA  GVERDLNGRD  YTVTGGFTGA  TAATGKTGAR
1001 NMPHTRLVAG  LGADVEFGNG  WNGLARYSYA  GSKQYGNESG  RVGVGYRFLE
1051 GGGGTGSATN  DDDVKKAATV  ALAAAYNNGQ  EINGFKAGET  IYDIDKDGTI
1101 TKKDATAADV  EADDFKGLGL  KKVVTNLTKT  VNENKQNVDA  KVKAAESEIE
1151 KLTTKLADTD  AALADTDAAL  DATTNALNKL  GENITTFAEE  TKTNIVKIDE
1201 KLEAVADTVD  KHAEAFNDIA  DSLDETNTKA  DEAVKTANEA  KQTAEETKQN
1251 VDAKVKAAET  AAGKAKAAAG  TANTAADKAE  AVAAKVTDIK  ADIATNKDNI
1301 AKKANSADVY  TREESDSKFV  RIDGLNATTE  KLDTRLASAE  KSIADHDTRL
1351 NGLDKTVSDL  RKETRQGLAE  QAALSGLFQP  YNVGRFNVTA  AVGGYKSESA
1401 VAIGTGFRFT  ENFAAKAGVA  VGTSSGSSAA  YHVGVNYEWL  EHHHHHH*
ΔG983-963C
ATGACTTCTGCGCCCGACTTCAATGCAGGCGGTACCGGTATCGGCAGCAACAGCAGAGCAACAACAGCGAAATCAGCA
GCAGTATCTTACGCCGGTATCAAGAACGAAATGTGCAAAGACAGAAGCATGCTCTGTGCCGGTCGGGATGACGTTGCG
GTTACAGACAGGGATGCCAAAATCAATGCCCCCCCCCCGAATCTGCATACCGGAGACTTTCCAAACCCAAATGACGCA
TACAAGAATTTGATCAACCTCAAACCTGCAATTGAAGCAGGCTATACAGGACGCGGGGTAGAGGTAGGTATCGTCGAC
ACAGGCGAATCCGTCGGCAGCATATCCTTCCCGAACTGTCATGGCAGAAAAGAACACGGCTATAACGAAAATTACAAA
AACTATACGGCGTATATGCGGAAGGAAGCGCCTGAAGACGGAGGCGGTAAAGACATTGAAGCTTCTTTCGACGATGAG
GCCGTTATAGAGACTGAAGCAAAGCCGACGGATATCCGCCACGTAAAAGAAATCGGACACATCGATTTGGTCTCCCAT
ATTATTGGCGGGCGTTCCGTGGACGGCAGACCTGCAGGCGGTATTGCGCCCGATGCGACGCTACACATAATGAATACG
AATGATGAAACCAAGAACGAAATGATGGTTGCAGCCATCCGCAATGCATGGGTCAAGCTGGGCCAACGTGGCGTGCGC
ATCGTCAATAACAGTTTTGGAACAACATCGAGGGCAGGCACTGCCGACCTTTTCCAAATAGCCAATTCGGAGGAGCAG
TACCGCCAAGCGTTGCTCGACTATTCCGGCGGTGATAAAACAGACGAGGGTATCCGCCTGATGCAACAGAGCGATTAC
GGCAACCTGTCCTACCACATCCGTAATAAAAACATGCTTTTCATCTTTTCGACAGGCAATGACGCACAAGCTCAGCCC
AACACATATGCCCTATTGCCATTTTATGAAAAAGACGCTCAAAAAGGCATTATCACAGTCGCAGGCGTAGACCGCAGT
GGAGAAAAGTTCAAACQGGAAATGTATGGAGAACCGGGTACAGAACCGCTTGWGTATGGCTCCAACCATTGCGGAATT
ACTGCCATGTGGTGCCTGTCGGCACCCTATGAAGCAAGCGTCCGTTTCACCCGTACAAACCCGATTCAAATTGCCGGA
ACATCCTTTTCCGCACCCATCGTAACCGGCACGGCGGCTCTGCTGCTGCAGAAATACCCGTGGATGAGCAACGACAAC
CTGCGTACCACGTTGCTGACGACGGCTCAGGACATCGGTGCAGTCGGCGTGGACAGCAAGTTCGGCTGGGGACTGCTG
GATGCGGGTAAGGCCATGAACGGACCCGCGTCCTTTCCGTTCGGCGACTTTACCGCCGATACGAAAGGTACATCCGAT
ATTGCCTACTCCTTCCGTAACGACATTTCAGGCACGGGCGGCCTGATCAAAAAAGGCGGCAGCCAACTGCAACTGCAC
GGCAACAACACCTATACGGGCAAAACCATTATCGAAGGCGGTTCGCTGGTGTTGTACGGCAACAACAAATCGGATATG
CGCGTCGAAACCAAAGGTGCGCTGATTTATAACGGGCGGCATCCCGGCGGCAGCCTGAACAGCGACGGCATTGTCTAT
CTGGCAGATACCGACCAATCCGGCGCAAACGAAACCGTACACATCAAAGGCAGTCTGCAGCTGGACGGCAAAGGTACG
CTGTACACACGTTTGGGCAAACTGCTGAAAGTGGACGGTACGGCGATTATCGGCGGCAAGCTGTACATGTCGGCACGC
GGCAAGGGGGCAGGCTATCTCAACAGTACCGGACGACGTGTTCCCTTCCTGAGTGCCGCCAAAATCGGGCAGGATTAT
TCTTTCTTCACAAACATCGAAACCGACGGCGGCCTGCTGGCTTCCCTCGACAGCGTCGAAAAAACAGCGGGCAGTGAA
GGCGACACGCTGTCCTATTATGTCCGTCGCGGCAATGCGGCACGGACTGCTTCGGCAGCGGCACATTCCGCGCCCGCC
GGTCTGAAACACGCCGTAGAACAGGGCGGCAGCAATCTGGAAAACCTGATGGTCGAACTGGATGCCTCCGAATCATCC
GCAACACCCGAGACGGTTGAAACTGCGGCAGCCGACCGCACAGATATGCCGGGCATCCGCCCCTACGGCGCAACTTTC
CGCGCAGCGGCAGCCGTACAGCATGCGAATGCCGCCGACGGTGTACGCATCTTCAACAGTCTCGCCGCTACCGTCTAT
GCCGACAGTACCGCCGCCCATGCCGATATGCAGGGACGCCGCCTGAAAGCCGTATCGGACGGGTTGGACCACAACGGC
ACGGGTCTGCGCGTCATCGCGCAAACCCAACAGGACGGTGGAACGTGGGAACAGGGCGGTCTTGAAGGCAAAATGCGC
GGCAGTACCCAAACCGTCGGCATTGCCGCGAAAACCGGCGAAAATACGACAGCAGCCGCCACACTGGGCATGGGACGC
AGCACATGGAGCGAAAACAGTGCAAATGCAAAAACCGACAGCATTAGTCTGTTTGCAGGCATACGGCACGATGCGGGC
GATATCGGCTATCTCAAAGGCCTGTTCTCCTACGGACGCTACAAAAACAGCATCAGCCGCAGCACCGGTGCGGACGAA
CATGCGGAAGGCAGCGTCAACGGCACGCTGATGCAGCTGGGCGCACTGGGCGGTGTCAACGTTCCGTTTGCCGCAACG
GGAGATTTGACGGTCGAAGGCGGTCTGCGCTACGACCTGCTCAAACAGGATGCATTCGCCGAAAAAGGCAGTGCTTTG
GGCTGGAGCGGCAACAGCCTCACTGAAGGCACGCTGGTCGGACTCGCGGGTCTGAAGCTGTCGCAACCCTTGAGCGAT
AAAGCCGTCCTGTTTGCAACGGCGGGCGTGGAACGCGACCTGAACGGACGCGACTACACGGTAACGGGCGGCTTTACC
GGCGCGACTGCAGCAACCGGCAAGACGGGGGCACGCAATATGCCGCACACCCGTCTGGTTGCCGGCCTGGGCGCGGAT
GTCGAATTCGGCAACGGCTGGAACGGCTTGGCACGTTACAGCTACGCCGGTTCCAAACAGTACGGCAACCACAGCGGA
CGAGTCGGCGTAGGCTACCGGTTCCTCGAGGGTGGCGGAGGCACTGGATCCGCCACAAACGACGACGATGTTAAAAAA
GCTGCCACTGTGGCCATTGCTGCTGCCTACAACAATGGCCAAGAAATCAACGGTTTCAAAGCTGGAGAGACCATCTAC
GACATTGATGAAGACGGCACAATTACCAAAAAAGACGCAACTGCAGCCGATGTTGAAGCCGACGACTTTAAAGGTCTG
GGTCTGAAAAAAGTCGTGACTAACCTGACCAAAACCGTCAATGAAAACAAACAAAACGTCGATGCCAAAGTAAAAGCT
GCAGAATCTGAAATAGAAAAGTTAACAACCAAGTTAGCAGACACTGATGCCGCTTTAGCAGATACTGATGCCGCTCTG
GATGCAACCACCAACGCCTTGAATAAATTGGGAGAAAATATAACGACATTTGCTGAAGAGACTAAGACAAATATCGTA
AAAATTGATGAAAAATTAGAAGCCGTGGCTGATACCGTCGACAAGCATGCCGAAGCATTCAACGATATCGCCGATTCA
TTGGATGAAACCAACACTAAGGCAGACGAAGCCGTCAAAACCGCCAATGAAGCCAAACAGACGGCCGAAGAAACCAAA
CAAAACGTCGATGCCAAAGTAAAAGCTGCAGAAACTGCAGCAGGCAAAGCCGAAGCTGCCGCTGGCACAGCTAATACT
GCAGCCGACAAGGCCGAAGCTGTCGCTGCAAAAGTTACCGACATCAAAGCTGATATCGCTACGAACAAAGATAATATT
GCTAAAAAAGCAAACAGTGCCGACGTGTACACCAGAGAAGAGTCTGACAGCAAATTTGTCAGAATTGATGGTCTGAAC
GCTACTACCGAAAAATTGGACACACGCTTGGCTTCTGCTGAAAATCCATTGCCGATTCACGATACTCGCCTGAACGGT
TTGGATAAAACAGTGTCAGACCTGCGCAAAGAAACCCGCCAAGGCCTTGCAGAACAAGCCGCGCTCTCCGGTCTGTTC
CAACCTTACAACGTGGTCTCGAGCACCACCACCACCACCACTGA
1     MTSAPDFNAG GTGIGSNSRA TTAKSAVSY AGIKNEMCKD RSMLCAGRDD
51    VAVTDRDAKI NAPPPNLRTG DFPNPNDAYK NLINLKPAIK AGYTGRGVEV
101   GRVDTGESVG SISFPELYGR KKHGYNKNYK NYTAYMRKEA PEDGGGKDIB
151   ASFDNEAVIB TBAKPTDLRH VKEIGHIDLV SHIIGGRSVD GRPAGGIAPD
201   ATLHDMNIND ETKNEMMVAA IRNAWVKLGE RGVRIVMNSF GTTSRAGTAD
251   LFQIANSKEQ YRQALLDYSG GDKTDEGIRL MQQSDYGRLB YHIRNKNMLF
301   IFSTGNDAQA QPNTYALLPF YEKDAQKGII TVAGVDBBGK KFKRBMYGEP
351   GTEPLEYGSN HCGITAKWCL SAPYBASVRF TRTNPIQIAG TSFSAPIVTG
401   TAALLLQKYP WMSNDNLRTT LLTTAQDIGA VGVDSKFGWG LLDAGKAMNG
451   PASFPFGDFT ADTKGTSDLA YSFRNDISGT GGLIKKGGSQ LQLHGNNTYT
501   GKTIIEGGSL VLYGNNKSDM RVRTKGALTY NGAASGGSIN SDGIVYLADT
551   DQSGANETVH IKGSLQLDGK GTLYTRLGKL LKVDGTAIIG GKLYMSARGK
601   GAGYLNSTGR RVPFLSAAKI GQDYSFFINI KIDGGLLASL DSVRKTAGSE
651   GDTLSYYVRR GNAARTASAA AHSAPAGLKH AVBQGGSNLK NLMVKLDASE
701   SSATPETVKT AAADRTDNPG IRPYGATPRA AAAVQHANAA DGVRIFNSLA
751   ATVYADSTAA HADMQGRRLK AVSDGLDHNG TGLRVLAQTQ QDGGTWRQGG
801   VEGKMRGSTQ TVGIAAKTGK NTTAAATLGM GRSTMSRNSA NAKTDSISLF
851   AGLRHDAGDI GYLKGLFSYG RYKNSISRST GADKHABGSV NGTLMQLGAL
901   GGVNVPFAAT GDLTVBGGLR YDLLKQDAFA KKGSALGWSG NSLTBGTLVG
951   LAGLKLSQPL SDKAVLFATA GVBRDLNGRD YTVTGGFTGA TAATGKTGAR
1001  NMPHTRLVAG LGADVRFGNG WNGLARYSYA GSKQYGHHSG RVGVGYRFLB
1051  GGGGTGSATN DDDVKKAATV AIAAAYNNGQ BINGFKAGST TYDIDBDGTI
1101  TKKDATAADV EADDFKGLGL KKVVTNLTKT VNBNKQNVDA KVKAAESEIB
1151  KLTTKLADTD AALADTDAAL DATTNALNKL GBNITTFAKE TKTNIVKIDE
1201KLEAVADTVD KBAEAFNDLA DSLDETNTKA DEAVKTANEA KQTAEBTKQN
1251VDAKVKAAET AAGKAEAAAG TANTAADKAB AVAAKVTDIK ADIATNKDNI
1301AKKANSADVY TREESDSKFV RIDGLNATTB KLDTRLASAE KSIADHDTRL
1351NGLDKTVSDL RKETRQGLAE QAALSGLFQP YNVGLEHHHH HH*
实施例4-ΔG741的杂交体
蛋白质741具有如下序列：
  1 VNRTAFCCLS LTTALILTAC SSGGGGVAAD IGAGLADALT APLDHKDKGL
 51QSLTLDQSVR KNKKLKLAAQ GAEKTYGNGD SLNTGKLKND KVSRFDFIRQ
101IEVDGQLITL ESGEFQVYKQ SHSALTAFQT EQIQDSKHSG KMVAKRQFRI
151GDIAGEHTSF DKLPEGGRAT YRGTAFGSDD AGGKLTYTID FAAKQGNGKI
201EHLKSPELNV DLAAADIKPD GKRHAVISGS VLYNQAEKGS YSLGIFGGKA
251QEVAGSAEVK TVNGIRHIGL AAKQ*
因此，ΔG741具有如下基本序列：
VAAD IGAGLADALT APLDHKDKGL
QSLTLDQSVR KNEKLKLAAQ GAEKTYGNGD SLNTGKLKND KVSRFDFIRQ
IEVDGQLITL ESGEFQVYKQ SHSALTAFQT BQIQDSKHSG KMVAKRQFRI
GDIAGEHTSF DKLPBGGRAT YRGTAFGSDD AGGKLTYTID FAAKQGNGKI
EHLKSPELNV DLAAADIKPD GKRHAVISGS VLYNQAEKGS YSLGIFGGKA
QEVAGSAEVK TVNGIRHIGL AAKQ*
将ΔG741直接融合于蛋白质961、961c、983和ORF46.1的符合读框的上游：
ΔG7741-961
ATGGTCGCCGCCGACATCGGTGCGGGGCTTGCCGATGCACTAACCGCACCGCTCGACCATAAAGACAAAGGTTTGCAG
TCTTTGACGCTGGATCAGTCCGTCAGGAAAAACGAGAAACTGAAGCTGGCGGCACAAGGTGCGGAAAAAACTTATGGA
AACGGTGACAGCCTCAATACGGGCAAATTGAAGAACGACAAGGTCAGCCGTTTCGACTTTATCCGCCAAATCGAAGTG
GACGGGCAGCTCATTACCTTGGAGAGTGCAGAGTTCCAAGTATACAAACAAAGCCATTCCGCCTTAACCGCCTTTCAG
ACCGAGCAAATACAAGATTCGGAGCATTCCGGGAAGATGGTTGCGAAACGCCAGTTCAGAATCGGCGACATAGCGGGC
GAACATACATCTTTTGACAAGCTTCCCGAAGGCGGCAGGGCGACATATCGCGGGACGGCGTTCGGTTCAGACGATGCC
GGCGGAAAACTGACCTACACCATAGATTTCGCCGCCAAGCAGGGAAACGGCAAAATCGAACATTTGAAATCGCCAGAA
CTCAATGTCGACCTGGCCGCCGCCGATATCAAGCCGGATGGAAAACGCCATGCCGTCATCAGCGGTTCCGTCCTTTAC
AACCAAGCCGAGAAAGGCAGTTACTCCCTCGGTATCTTTGGCGGAAAAGCCCAGGAAGTTGCCGGCAGCGCGGAAGTG
AAAACCGTAAACGGCATACGCCATATCGGCCTTGCCGCCAAGCAACTCGAGGGTGGCGGAGGCACTGGATCCGCCACA
AACGACGACGATGTTAAAAAAGCTGCCACTGTGGCCATTGCTGCTGCCTACAACAATGGCCAAGAAATCAACGGTTTC
AAAGCTGGAGAGACCATCTACGACATTGATGAAGACGGCACAATTACCAAAAAAGACGCAACTGCAGCCGATGTTGAA
GCCGACGACTTTAAAGGTCTGGGTCTGAAAAAAGTCGTGACTAACCTGACCAAAACCGTCAATGAAAACAAACAAAAC
GTCGATGCCAAAGTAAAAGCTGCAGAATCTGAAATAGAAAAGTTAACAACCAAGTTAGCAGACACTGATGCCGCTTTA
GCAGATACTGATGCCGCTCTGGATGCAACCACCAACGCCTTGAATAAATTGGGAGAAAATATAACGACATTTGCTGAA
GAGACTAAGACAAATATCGTAAAAATTGATGAAAAATTAGAAGCCGTGGCTGATACCGTCGACAAGCATGCCGAAGCA
TTCAACGATATCGCCGATTCATTGGATGAAACCAACACTAAGGCAGACGAAGCCGTCAAAACCGCCAATGAAGCCAAA
CAGACGGCCGAAGAAACCAAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAAACTGCAGCAGGCAAAGCCGAAGCT
GCCGCTGGCACAGCTAATACTGCAGCCGACAAGGCCGAAGCTGTCGCTGCAAAAGTTACCGACATCAAAGCTGATATC
GCTACGAACAAAGATAATATTGCTAAAAAAGCAAACAGTGCCGACGTGTACACCAGAGAAGAGTCTGACAGCAAATTT
GTCAGAATTGATGGTCTGAACGCTACTACCGAAAAATTGGACACACGCTTGGCTTCTGCTGAAAAATCCATTGCCGAT
CACGATACTCGCCTGAACGGTTTGGATAAAACAGTGTCAGACCTGCGCAAAGAAACCCGCCAAGGCCTTGCAGAACAA
GCCGCGCTCTCCGGTCTGTTCCAACCTTACAACGTGGGTCGGTTCAATGTAACGGCTGCAGTCGGCGGCTACAAATCC
GAATCGGCAGTCGCCATCGGTACCGGCTTCCGCTTTACCGAAAACTTTGCCGCCAAAGCAGGCGTGGCAGTCGGCACT
TCGTCCGGTTCTTCCGCAGCCTACCATGTCGGCGTCAATTACGAGTGGCTCGAGCACCACCACCACCACCACCACTGA
1    MVAADIGAGL ADALTAPLDH KDKGLQSLTL DQSVRKNEKL KLAAQGAEKT
51   YGNGDSLNTG KLKNDKVSRF DFIRQIEVDG QLITLESGRF QVYKQSHSAL
101  TAFQTEQIQD SKHSGKMVAK RQFRIGDIAG ERTSFDKLPE GGRATYRGTA
151  FGSDDAGGKL TYTIDFAAKQ GNGKIKHLKS PKLNVDLAAA DIKPDGKRHA
201  VISGSVLYNQ AKKGSYSLGI FGGKAQEVAG SAKVKTVDGI RHIGLAAKQL
251  EGGGGTGSAT NDDDVKKAAT VAIAAAYNNG QEINGFKAGE TIYDIDBDGT
301  ITKKDATAAD VEADDFKGLG LKKVVTNLTK TVNENKQNVD AKVKAABSEI
351  EKLTTKLADT DAALADTDAA LDATTNALNK LGENITTFAE ETKTNIVKID
401  EKLEAVADTV DKHAEAFNDI ADSLDETNTK ADEAVKTANE AKQTAEETKQ
451  NVDAKVKAAE TAAGKAEAAA EAVAAKVTDI KADIATNKDN
501  IAKKANSADV YTREESDSKF VRIDGLNATT EKLDTRLASA EKSIADHDTR
551  LNGLDKTVSD LRKETRQGLA EQAALSGLFQ PYNVGRFNVT AAVGGYKSES
601  AVAIGTGFRF TENFAAKAGV AVGTSSGSSA AYHVGVNYKW LEHHHHHH*
ΔG741-961c
ATGGTCGCCGCCGACATCGGTGCGGGGCTTGCCGATGCACTAACCGCACCGCTCGACCATAAAGACAAAGGTTTGCAG
TCTTTGACGCTGGATCAGTCCGTCAGGAAAAACGAGAAACTGAAGCTGGCGGCACAAGGTGCGGAAAAAACTTATGGA
AACGGTGACAGCCTCAATACGGGCAAATTGAAGAACGACAAGGTCAGCCGTTTCGACTTTATCCGCCAAATCGAAGTG
GACGGGCAGCTCATTACCTTGGAGAGTGGAGAGTTCCAAGTATACAAACAAAGCCATTCCGCCTTAACCGCCTTTCAG
ACCGAGCAAATACAAGATTCGGAGCATTCCGGGAAGATGGTTGCGAAACGCCAGTTCAGAATCGGCGACATAGCGGGC
GAACATACATCTTTTGACAAGCTTCCCGAAGGCGGCAGGGCGACATATCGCGGGACGGCGTTCGGTTCAGACGATGCC
GGCGGAAAACTGACCTACACCATAGATTTCGCCGCCAAGCAGGGAAACGGCAAAATCGAACATTTGAAATCGCCAGAA
CTCAATGTCGACCTGGCCGCCGCCGATATCAAGCCGGATGGAAAACGCCATGCCGTCATCAGCGGTTCCGTCCTTTAC
AACCAAGCCGAGAAAGGCAGTTACTCCCTCGGTATCTTTGGCGGAAAAGCCCAGGAAGTTGCCGGCAGCGCGGAAGTG
AAAACCGTAAACGGCATACGCCATATCGGCCTTGCCGCCAAGCAACTCGAGGGTGGCGGAGGCACTGGATCCGCCACA
AACGACGACGATGTTAAAAAAGCTGCCACTGTGGCCATTGCTGCTGCCTACAACAATGGCCAAGAAATCAACGGTTTC
AAAGCTGGAGAGACCATCTACGACATTGATGAAGACGGCACAATTACCAAAAAAGACGCAACTGCAGCCGATGTTGAA
GCCGACGACTTTAAAGGTCTGGGTCTGAAAAAAGTCGTGACTAACCTGACCAAAACCGTCAATGAAAACAAACAAAAC
GTCGATGCCAAAGTAAAAGCTGCAGAATCTGAAATAGAAAAGTTAACAACCAAGTTAGCAGACACTGATGCCGCTTTA
GCAGATACTGATGCCGCTCTGGATGCAACCACCAACGCTTGAATAAATTGGGAGAAAATATAACGAACATTTGCTGAA
GAGACTAAGACAAATATCGTAAAAATTGATGAAAAATTAGAAGCCGTGGCTGATACCGTCGACAAGCATGCCGAAGCA
TTCAACGATATCGCCGATTCATTGGATGAAACCAACACTAAGGCAGACGAAGCCGTCAAAACCGCCAATGAAGCCAAA
CAGACGGCCGAAGAAACCAAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAAACTGCAGCAGGCAAAGCCGAAGCT
GCCGCTGGCACAGCTAATACTGCAGCCGACAAGGCCGAAGCTGTCGCTGCAAAAGTTACCGACATCAAAGCTGATATC
GCTACGAACAAAGATAATATTGCTAAAAAAGCAAACAGTGCCGACGTGTACACCAGAGAAGAGTCTGACAGCAAATTT
GTCAGAATTGATGGTCTGAACGCTACTACCGAAAAATTGGACACACGCTTGGCTTCTGCTGAAAAATCCATTGCCGAT
CACGATACTCGCCTGAACGGTTTGGATAAAACAGTGTCAGACCTGCGCAAAGAAACCCGCCAAGGCCTTGCAGAACAA
GCCGGCGCTCTCCGGTCTGTTCCAACCTTACAACGTGGGTCTCGAGCACCACCACCACCACCACTGA
1   MVAADIGAGL ADALTAPLDH KDKGLQSLTL DQSVRKNKKL KLAAQGAEKT
51  YGNGDSLNTG KLKNDKVSRF DFIRQIEVDG QLITLKSGKF QVYKQSHSAL
101 TAFQTEQIQD SEHSGKMVAK RQFRIGDIAG KHTSFDKLPE GGRATYRGTY
151 FGSDDAGGKL TYTIDFAAKQ GNGKIKHLKS PKINVDLAAA DIKPDGKRHA
201 VISGSVLYNQ AEKGSYSLGI FGGKAQEVAG SAEVKTVNGI RHIGLAAKQL
251 EGGGGTGSAT NDDDVKKAAT VAIAAAYNNG QEINGFKAGE TIYDIDEDGT
301 ITKKDATAAD VEADDFKGLG LKKVVTNLTK TVNENKQNVD AKVKAAESEI
351 EKLTTKLADT DAALADTDAA LDATTNALNK LGENTTTFAE ETKTNIVKID
401 EKLEAVADTV DKHAEAFNDI ADSLDETNTK ADEAVKTANE AKQTAKETKQ
451 NVDAKVKAAE TAAGKAEAAA GTANTAADKA EAVAAKVTDI KADIATNKDN
501 IAKKANSADV YTREESDSKF VRIDGLNATT EKLDTRLASA EKSIADHDTR
551 LNGLDKTVSD LRKETRQGLA EQAALFQGLA PYNVGLEHHH HHH*
ΔG741-983
ATGGTCGCCGCCGACATCGGTGCGGGGCTTGCCGATGCACTAACCGCACCGCTCGACCATAAAGAGAAAGGTTTGCAG
TCTTTGACGCTGGATCAGTCCGTCAGGAAAAACGAGAAACTGAAGCTGGCGGCACAAGGTGCGGAAAAAACTTATGGA
AACGGTGACAGCCTCAATACGGGCAAATTGAAGAACGACAAGGTCAGCCGTTTCGACTTTATCCGCCAAATCGAAGTG
GACGGGCAGCTCATTACCTTGGAGAGTGGAGAGTTCCAAGTATACAAACAAAGCCATTCCGCCTTAACCGCCTTTCAG
ACCGAGCAAATACAAGATTCGGAGCATTCCGGGAAGATGGTTGCGAAACGCCAGTTCAGAATCGGCGACATACGGGGC
GAACATACATCTTTTGACAAGCTTCCCGAAGGCGGCAGGGCGACATATCGCGGGACGGCGTTCGGTTCAGACGATGCC
GGCGGAAAACTGACCTACACCATAGATTTCGCCGCCAAGCAGGGAAACGGCAAAATCGAACATTTGAAATCGCCAGAA
CTCAATGTCGACCTGGCCGCCGCCGATATCAAGCCGGATGGAAAACGCCATGCCGTCATCAGCGGTTCCGTCCTTTAC
AACCAAGCCGAGAAAGGCAGTTACTCCCTCGGTATCTTTGGCGGAAAAGCCCAGGAAGTTGCCGGCAGCGCGGAAGTG
AAAACCGTAAACGGCATACGCCATATCGGCCTTGCCGCCAAGCAACTCGAGGGATCCGGCGGAGGCGGCACTTCTGCG
CCCGACTTCAATGCAGGCGGTACCGGTATCGGCAGCAACAGCAGAGCAACAACAGCGAAATCAGCAGCAGTATCTTAC
GCCGGTATCAAGAACGAAATGTGCAAAGACAGAAGCATGCTCTGTGCCGGTCGGGATGACGTTGCGGTTACAGACAGG
GATGCCAAAATCAATGCCCCCCCCCCGAATCTGCATACCGGAGACTTTCCAAACCCAAATGACGCATACAAGAATTTG
ATCAACCTCAAACCTGCAATTGAAGCAGGCTATACAGGACGCGGGGTAGAGGTAGGTATCGTCGACACAGGCGAATCC
CTCGGCAGCATATCCTTTCCCGAACTGTATGGCAGAAAAGAACACGGCTATAACGAAAATTACAAAAACTATACGGCG
TATATGCGGAAGGAAGCGCCTGAAGACGGAGGCGGTAAAGACATTGAAGCTTCTTTCGACGATGAGGCCGTTATAGAG
ACTGAAGCAAAGCCGACGGATATCCGCCACGTAAAAGAAATCGGACACATCGATTTGGTCTCCCATATTATTGGCGGG
CGTTCCGTGGACGGCAGACCTGCAGGCGGTATTGCGCCCGATGCGACGCTACACATAATGAATACGAATGATGAAACC
AAGAACGAAATGATGGTTGCAGCCATCCGCAATGCATGGGTCAAGCTGGGCGAACGTGGCGTGCGCATCGTCAATAAC
AGTTTTGGAACAACATCGAGGGCAGGCACTGCCGACCTTTTCCAAATAGCCAATTCGGAGGAGCAGTACCGCCAAGCG
TTGCTCGACTATTCCGGCGGTGATAAAACAGACGAGGGTATCCGCCTGATGCAACAGAGCGATTACGGCAACCTGTCC
TACCACATCCGTAATAAAAACATGCTTTTCATCTTTTCGACAGGCAATGACGCACAAGCTCAGCCCAACACATATGCC
CTATTGCCATTTTATGAAAAAGACGCTCAAAAAGGCATTATCACAGTCGCAGGCGTAGACCGCAGTGGAGAAAAGTTC
AAACGGGAAATGTATGGAGAACCGGGTACAGAACCGCTTGAGTATGGCTCCAACCATTGCGGAATTACTGCCATGTGG
TGCCTGTCGGCACCCTATGAAGCAAGCGTCCGTTTCACCCGTACAAACCCGATTCAAATTGCCGGAACATCCTTTTCC
GCACCCATCGTAACCGGCACGGCGGCTCTGCTGCTGCAGAAATACCCGTGGATGAGCAACGACAACCTGCGTACCACG
TTGCTGACGACGGCTCAGGACATCGGTGCAGTCGGCGTGGACAGCAAGTTCGGCTGGGGACTGCTGGATGCGGGTAAG
GCCATGAACGGACCCGCGTCCTTTCCCGTTCGCGACTTTACCGCCGATACGAAAGGTACATCCGATATTGCCTACTCC
TTCCGTAACGACATTTCAGGCACGGGCGGCCTGATCAAAAAAGGGGGCAGCCAACTGCAACTGCACGGCAACAACACC
TATACGGGCAAAACCATTATCGAAGGCGGTTCGCTGGTGTTGTACGGCAACAACAAATCGGATATGCGCGTCGAAACC
AAAGGTGCGCTGATTTATAACGGGGCGGCATCCGGCGGCAGCCTGAACAGCGACGGCATTGTCTATCTGGCAGATACC
GACCAATCCGGCGCAAACGAAACCGTACACATCAAAGGCAGTCTGCAGCTGGACGGCAAAGGTACGCTGTACACACGT
TTGGGCAAACTGCTGAAAGTGGACGGTACGGCGATTATCGGCGGCAAGCTGTACATGTCGGCACGCGGCAAGGGGGCA
GGCTATCTCAACAGTACCGGACGACGTGTTCCCTTCCTGAGTGCCGCCAAAATCGGGCAGGATTATTCTTTCTTCACA
AACATCGAAACCGACGGCGGCCTGCTGGGCTTCCCTCGACAGCGTCGAAAAAACAGCGGCAGTGAAGGCGACACGCTG
TCCTATTATGTCCGTCGCGGCAATGCGGCACGGACTGCTTCGGCAGCGGCACATTCCGCGCCCGCCGGTCTGAAACAC
GCCGTAGAACAGGGCGGCAGCAATCTGGAAAACCTGATGGTCGAACTGGATGCCTCCGAATCATCCGCAACACCCGAG
ACGGTTGAAACTGCGGCAGCCGACCGCACAGATATGCCGGGCATCCGCCCCTACGGCGCAACTTTCCGCGCAGCGGCA
GCCGTACAGCATGCGAATGCCGCCGACGGTGTACGCATCTTCAACAGTCTCGCCGCTACCGTCTATGCCGACAGTACC
GCCGCCCATGCCGATATGCAGGGACGCCGCCTGAAAGCCGTATCGGACGGGTGGACCACAACGGCACGGGTGCTGCGC
GTCATCGCGCAAACCCAACAGGACGGTGGAACGTGGGAACAGGGCGGTGTTGAAGGCAAAATGCGCGGCAGTACCCAA
ACCGTCGGCATTGCCGCGAAAACCGGCGAAAATACGACAGCAGCCGCCACACTGGGCATGGGACGCAGCACATGGAGC
GAAAACAGTGCAAATGCAAAAACCGACAGCATTAGTCTGTTTGCAGGCATACGGCACGATGCGGGCGATATCGGCTAT
CTCAAAGGCCTGTTCTCCTACGGACGCTACAAAAACAGCATCAGCCGCAGCACCGGTGCGGACGAACATGCGGAAGGC
AGCGTCAACGGCACGCTGATGCAGCTGGGCGCACTGGGCGGTGTCAACGTTCCGTTTGCCGCAACGGGAGATTTGACG
GTVHAAGGCGGTCTGCGCTACGACCTGCTCAAACAGGATGCATTCGCCGAAAAAGGCAGTGCTTTGGGCTGGAGCGGC
AACAGCCTCACTGAAGGCACGCATGGTCGGACTCGCGGGTCTGAAGCTGTCGCAACCTTGAGCGATAAAGCCGTCCTG
TTTGCAACGGCGGGCGTGGAACGCGACCTGAACGGACGCGACTACACGGTAACGGGCGGCTTTACCGGCGCGACTGCA
GCAACCGGCAAGACGGGGGCACGCAATATGCCGCACACCCGTCTGGTTGCCGGCCTGGGCGCGGATGTCGAATTCGGC
GCACGTTACAGCTACGCCGGTTCCAAACAGTACGGCAACCACAGCGGACGAGTCGGCGTA
GGCTACCGGTTCCTCGGAGCACCACCACCACCACCACTGA
1    MVAADIGAGL ADALTAPLDH KDKGLQSLTL DQSVRKNEKL KLAAQGAEKT
51   YGNGDSLNTG KLKNDKVSRF DFIRQIEVDG QLITLESGEF QVYKQSHSAL
101  TAFQTEQIQD SEHSGKMVAK RQFRIGDIAG KHTSFDKLPE GGRATYRGTA
151  FGSDDNGGKL TYTIDFAAKQ GNGKIEHLKS PKLNVDLAAA DIKPDGKRHA
201  VISGSVLYNQ AEKGSYSLGI FGGKAQEVAG SAEVKTVNGI RHIGLAAKQL
251  EGSGGGGTSA PDFNAGGTGI GSNSRATTAK SAAVSYAGIK NEHCKDRSHL
301  CNGRDDVAVT DRDAKINAPP PNLHTGDFPN PNDAYKNLIN LKPAIEAGYT
351  GRGVEVGIVD TGESVGSISF PELYGRKEHG YNENYKNYTA YMRKBAPKDG
401  GGKDIEASFD DEAVIETKAK PTDIREVKEI GHIDLVSHII GGRSVDGRPA
451  GGIAPDATLE IMNTNDETKN EMMVAAIRNA WVKLGERGVR IVNNSFGTTS
501  RAGTADLFQI ANSEEQYRQA LLDYSGGDKT DEGIRLMQQS DYGNLSYHIR
551  NKNMLFIFST GNDAQAQPNT YALLPFYEKD AQKGIITVAG VDRSGEKFKR
601  KMYGEPGIER LEYGSNHCGI TAMWCISAPY EASVRFTRTN PIQIAGTSFS
651  APIVTGTAAL LLQKYPWMSN DNLRTTLLTT AQDIGAVGVD SKFGWGLLDA
701  GKAMNGPASF PFGDFTADTK GTSDLAYSFR NDISGTGGLI KKGGSQLQLH
751  GNNTYTGKTI IEGGSLVLYG NNKSDMRVET KGALIYNGAA SGGSINSDGI
801  VYLADTDQSG ANETVHIKGS LQLDGKGTLY TRLGKLLKVD GTAIIGGKLY
851  MSARGKGAGY LNSTGRRVPF  LSAAKIGQDY SFFTNIETDG GLLASLDSVE
901  KTAGSEGDTL SYYVRRGNAA RTASAAAHSA PAGLKHAVEQ GGSNLENLMV
951  ELDASESSAT PETVETAAAD RTDMPGIRPY GATFRAAAAV QHANAADGVR
1001 IFNSLAATVY ADSTAAHADM QGRRLKAVSD GLDHNGTGLR VIAQTQQDGG
1051 TWEQGGVEGK MRGSTQTVGI AAKTGENTTA AATLGMGRST WSENSANAKT
1101 DSISLFAGIR  HDAGDIGYLK GLFSYGRYKN SISRSTGADE HAEGSVNGTL
1151 MQLGALGGVN VPFAATGDLT VEGGLRYDLL KQDAFAEKGS ALGWSGNSLT
1201 EGTLVGLAGL KLSQPLSDKA VLFATAGVER DLNGRDYTVT GGFTGATAAT
1251 GKTGARNMPH TRLVAGLGAD VEFGNGNNGL ARYSYAGSKQ YGNHSGRVGV
1301 GYRFLEHHHH HH*
ΔG741-ORF46.1
ATGGTCGCCGCCGACATCGGTGCGGGGCTTGCCGATGCACTAACCGCACCGCTCGACCATAAAGACAAAGGTTTGCAG
TCTTTGACGCTGGATCAGTCCGTCAGGAAAAACGAGAAACTGAAGCTGGCGGCACAAGGTGCGGAAAAAACTTATGGA
AACGGTGACAGCCTCAATACGGGCAAATTGAAGAACGACAAGGTCAGCCGTTTCGACTTTATCCGCCAAATCGAAGTG
GACGGGCAGCTCATTACCTTGGAGAGTGGAGAGTTCCAAGTATACAAACAAAGCCATTCCGCCTTAACCGCCTTTCAG
ACCGAGCAAATACAAGATTCGGAGCATTCCGGGAAGATGGTTGCGAAACDCCAGTTCAGAATCGGCGACATAGCGGGC
GAACATACATCTTTTGACAAGCTTCCCGAAGGCGGCAGGGCGACATATCGCGGGACGGCGTTCGGTTCAGACGATGCC
GGCGGAAAACTGACCTACACCATAGATTTCGCCGCCAAGCAGGGAAACGGCAAAACGAACATTTGGAAATCGCCAGAA
CTCAATGTCGACCTGCCGCCGATATCAAGCCGGATGGAAAACGCCATGCCGTCAATGCTCAGCGGTTCCGTCCTTTAC
AACCAAGCCGAGAAAGGCAGTTACTCCCTCGGTATCTTTGGCGGAAAAGCCCAGGAAGTTGCCGGCAGCGCGGAAGTC
AAAACCGTAAACGGCATACGCCATATCGGCCTTGCCGCCAAGCAACTCGACGGTGGCGGAGGCACTGGATCCTCAGAT
TTGGCAAACGATTCTTTTATCCQGCAGGTTCTCGACCGTCAGCATTTCGAACCCGACGGGAAATACCACCTATTCGGC
AGCAGGGGGGAACTTGCCGAGCGCAGCGGCCATATCGGATTGGGAAAAATACAAAGCCATCAGTTGGGCAACCTGATG
ATTCAACAGGCGGCCATTAAAGGAAATATCGGCTACATTGTCCGCTTTTCCGATCACGGGCACGAAGCCCATTCCCCC
TTCGACAACCATGCCTCACATTCCGATTCTGATGAAGCCGGTAGTCCCGTTGACGGATTTAGCCTTTACCGCATCCAT
TGGGACGGATACGAACACCATCCCGCCGACGGCTATGACGGGCCACAGGGCGGCCGCTATCCCGCTCCCAAAGGCGCG
AGGGATATATACAGCTACGACATAAAAGGCGTTGCCCAAAATATCCGCCTCAACCTGACCGACAACCGCAGCACCGGA
CAACGGCTTGCCGACCGTTTCCACAATGCCGGTAGTAPGCTGACGCAAGGAGTAGGCGACGGATTCAAACGCGCCACC
CGATACAGCCCCGAGCTGGACAGATCGGGCAATGCCGCCGAAGCCTTCAACGGCACTGCAGATATCGTTAAAAACATC
ATCGGCGCGGCAGGAGAAATTGTCGGCGCAGGCGATGCCGTGCAGGGCATAAGCGAAGGCTCAAACATTGCTGTCATG
CATGCCGCAGCAGCCATCCGCGATTGGGCAGTCCAAAACCCCAATGCCGCACAAGGCATAGAAGCCGTCAGCAATATC
TATGCCGCAGCAGCCATCCGCGATTGGGCAGTCCAAAACCCCAATGCCGCACAAGGCATAGAAGCCGTCAGCAATATC
TTTATGGCAGCCATCCCCATCAAAGGGATTGGAGCTGTTCGGGGAAAATACGGCTTGGGCGGCATCACGGCACATCCT
ATCAAGCGGTCGCAGATGGGCGCGATCGCATTGCCGQAAAGGGAAATCGCCGTCAGCGACAATTTTGCCGATGCGGCA
TACGCCAAATACCCGTCCCCTTACCATTCCCGAAATATCCGTTCAAACTTGGAGCAGCGTTACGGCAAAGAAAACATC
ACCTCCTCAACCGTGCCGCCGTCAAACGGCAAAAATGTCAAACTGGCAGACCAACGCCACCCGAAGACAGGCGTACCG
TTTGACGGTAAAGGGTTTCCGAATTTTGQAGAAGCACGTGAAATATGATACGCTCGAGCACCACCACCACCACCACTG
1      MVAADIGAGL ADALTAPLDH KDKGLQSLTL DQSVRKNEKL KLAAQGAKKT
51     YGNGDSLNTG KLKNDKVSRF DFIRQIKVDG QLITLZSGKF QVYKQSHSAL
101    TAFQTEQIQD SEHSGKNVAK RQFRIGDIAG RHTSFDKLPE GGRATYRGTA
151    FGSDDAGGKL TYTIDFAAKQ GNGKIEHLKS PELNVDLAAA DIKPDGKRHA
201    VISGSVLYNQ AEKGSYSLGI PGGKAQRVAG SAKVKTVNGI RHIGLAAKQL
251    DGGGGTGSSD LANDSFIRQV LDRQHFRPDG KYHLFGSRGK LAKRSGHIGL
301    GKIQSHQLGN LMIQQAAIKG NIGYIVRFSD HGHEVESPFD NHASHSDSDE
351    AGSPVDGFSL YRIHWDGYEH HPADGYDGPQ GGGYPAPKGA RDTYSYDIKG
401    VAQNIRLNLT DNRSTGQRLA DRFHNAGSML TQGVGDGFRR ATRYSPRLDR
451    SGNAAEAFNG TADIVKNIIG AAGEIVGAGD AVQGISEGSN IAVMHGLGLL
501    STENKMARIN DLADMAQLKD YAAAAIRDWA VQNPNAAQGI RAVSNIFNAA
551    IPIKGIGAVR GKYGLGGITA HPIKRSQMGA IALPKGKSAV SDNFADAAYA
601    KYPSPYHSRN IRSNLBQRYG KENIISSTVP PSNGKNVKLA DQRHPXTGVP
651    FDGKGFPNFE KHVKYDTLEE HHHHH*
实施例5-287的杂交体
以全长、具有C-未端His-标记或无其前导肽但具有C-未端His-标记的287的表达水平非常低。用N-未端GST-融合实现了较好的表达。将GST用作N-未端融合配体的替代方法，将287置于蛋白质919的C-未端(‘919-287′)、蛋白质953的C-端(‘953-287′)和蛋白质ORF46.1的C-未端(‘ORF46.1-287′)。在这两种方法中，前导肽都是缺失的，且杂交体是直接的符合读杠的融合。
为了制备953-287杂交体，通过设计正向引物自各序列前导区的下游，除去两种蛋白质的前导肽；在953反向引物中除去终止密码子序列，但在287反向引物中包含终止密码子序列。对953基因而言，用于扩增的5′和3′引物分别包括NdeI和BamHI限制酶切位点，而对287基因的扩增而言，5′和3′引物分别包括BamHI和XhoI限制酶切位点。用这种方法，用NdeI-BamHI(克隆第一个基因)且随后用BamHI-XhoI(克隆第二个基因)，可以实现pET21b+中两个基因的顺序定向克隆。
通过将编码287的成熟部分的序列克隆入pET21b+中的919-His克隆的3′-端的XhoI位点，可以得到919-287杂交体。设计用于扩增287基因的引物，从而在PCR片段的5′-引入SalI限制酶切位点并在3′-引入XhoI位点。因为由SalI和XhoI限制酶产生的粘性末端是相容的，因此可以将由SalI-XhoI消化的287PCR产物***到由XhoI切割的pET21b-919克隆中。
类似地得到ORF46.1-287杂交体。
将针对杂交蛋白产生的抗体的杀菌效力(同源菌株)与针对组分抗原的简单混合物产生的抗体相比：
    与287的混合物    与287的杂交体
 919     32000    16000
 953     8192    8192
 ORF46.1     128    8192
对919-287和953-287而言，还获得了针对异源MenB菌株和针对血清型A和C的杀菌活性的数据：
 919   953 ORF46.1
  菌株  混合物  杂交体   混合物  杂交体 混合物   杂交体
  MC58  512  1024   512  1024 -   1024
  NGH38  1024  2048   2048  4096 -   4096
  BZ232  512  128   1024  16 -   -
  MenA(F6124)  512  2048   2048  32 -   1024
  MenC(CU) ＞2048  n.d.   ＞2048  n.d. -   n.d.
  MenC(BZ133) ＞4096  ＞8192   ＞4096  ＜16 -   2048
还构建了ORF46.1和919的杂交体。在N-未端用919得到了最佳结果(高4倍的滴定度)。
还测试了杂交体919-519His、ORF97-225His和225-ORF97His。它们的ELISA滴定度和杀菌抗体应答结果中等。
两种蛋白质A和B的杂交体可以是NH2-A-B-COOH或NH2-B-A-COOH，用ΔG287还制得了在N-未端与287“反向”杂交的杂交体。使用一系列菌株，包括同源菌株2996。FCA用作佐剂：
287&919 287&953 287&ORF46.1
菌株 ΔG287-919 919-287 ΔG287-953 953-287 ΔG287-46.1 46.1-287
2996 128000 16000 65536 8192 16384 8192
BZ232 256 128 128 ＜4 ＜4 ＜4
1000 2048 ＜4 ＜4 ＜4 ＜4 ＜4
MC58 8192 1024 16384 1024 512 128
NGH38 32000 2048 ＞2048 4096 16384 4096
394/98 4096 32  256 128 128 16
MenA(F6124) 32000 2048 ＞2048 32 8192 1024
MenC(BZ133) 64000 ＞8192 ＞8192 ＜16 8192 2048
通常在N-未端用287观察到较好的杀菌滴定度。
当融合于蛋白质961[如上所示的NH2-ΔG287-961-COOH-序列]时，得到的蛋白质是不溶解的，纯化必须将其变性和复性。在复性后，发现约50％蛋白质仍不溶解。比较可溶和不溶蛋白质，用可溶蛋白质得到更好的杀菌滴定度(FCA作为佐剂)：
2996 BZ232 MC58 NGH38 F6124 BZ133
可溶的 65536 128 4096 ＞2048 ＞2048 4096
不溶的 8192 ＜4 ＜4 16 n.d. n.d.
但用明矾佐剂替代，可以改善不溶形式的蛋白质的滴定度：
不溶的 32768  128  4096 ＞2048 ＞2048  2048
还在杂交蛋白中使用961c(见上)。由于961及其结构域变体指导有效的表达，它们非常适合作为杂交蛋白的N-未端部分。
实施例23-其它杂交体
附图中显示本发明的其它杂交蛋白，它们具有如下所示的序列。当与单独蛋白质相比时，这此杂交蛋白是有利的：
ORF46.1-741
ATGTCAGATTTGGCAAACGATTCTTTTATCCGGCAGGTTCTCGACCGTCAGCATTTCGAACCCGACGGGAAATACCAC
CTATTCGGCAGCAGGGGGGAACTTGCCGAGCGCAGCGGCCATATCGGATTGGGAAAAATACAAAGCCATCAGTTGGGC
AACCTGATGATTCAACAGGCGGCCATTAAAGGAAATATCGGCTACATTGTCCGCTTTTCCGATCACGGGCACGAAGTC
CATTCCCCCTTCGACAACCATGCCTCACATTCCGATTCTGATGAAGCCGGTAGTCCCGTTGACGGATTTAGCCTTTAC
CGCATCCATTGGGACGGATACGAACACCATCCCGCCGACGGCTATGACGGGCCACAGGGCGGCGGCTATCCCGCTCCC
AAAGGCGCGAGGGATATATACAGCTACGACATAAAAGGCGTTGCCCAAAATATCCGCCTCAACCTGACCGACAACCGC
AGCACCGGACAACGGCTTGCCGACCGTTTCCACAATGCCGGTAGTATGCTGACGCAAGGAGTAGGCGACGGATTCAAA
CGCGCCACCCGATACAGCCCCGAGCTGGACAGATCGGGCAATGCCGCCGAAGCCTTCAACGGCACTGCAGATATCGTT
AAAAACATCATCGGCGCGGCAGGAGAAATTGTCGGCGCAGGCGATGCCGTGCAGGGCATAAGCGAAGGCTCAAACATT
GCTGTCATGCACGGCTTGGGTCTGCTTTCCACCGAAAACAAGATGGCGCGCATCAACGATTTGGCAGATATGGCGCAA
CTCAAAGACTATGCCGCAGCAGCCATCCGCGATTGGGCACTCCAAAACCCCAATGCCGCACAAGGCATAGAAGCCGTC
AGCAATATCTTTATGGCAGCCATCCCCATCAAAGGGATTGGAGCTGTTCGGGGAAAATACGGCTTGGGCGGCATCACG
GCACATCCTATCAAGCGGTCGCAGATGGGCGCGATCGCATTGCCGAAAGGGAAATCCGCCGTCAGCGACAATTTTGCC
GATGCGGCATACGCCAAATACCCGTCCCCTTACCATTCCCGAAATATCCGTTCAAACTTGGAGCAGCGTTACGGCAAA
GAAAACATCACCTCCTCAACCGTGCCGCCGTCAAACGGCAAAAATGTCAAACTGGCAGACCAACGCCACCCGAAGACA
GGCGTACCGTTTGACGGTAAAGGGTTTCCGAATTTTGAGAAGCACGTGAAATATGATACGGGATCCGGAGGGGGTGGT
GTCGCCGCCGACATCGGTGCGGGGCTTGCCGATGCACTAACCGCACCGCTCGACCATAAAGACAAAGGTTTGCAGTCT
TTGACGCTGGATCAGTCCGTCAGGAAAAACGAGAAACTGAAGCTGGCGGCACAAGGTGCGGAAAAAACTTATGGAAAC
GGTGACAGCCTCAATACGGGCAAATTGAAGAACGACAAGGTCAGCCGTTTCGACTTTATCCGCCAAATCGAAGTGGAC
GGGCAGCTCATTACCTTGGAGAGTGGAGAGTTCCAAGTATACAAACAAAGCCATTCCGCCTTAACCGCCTTTCAGACC
GAGCAAATACAAGAGATTCGGAGCATTCCGGAAGATGGTTGCGAAACGCCAGTTCAGAATCGCGACATAGCGGGCGAA
CATACATCTTTTGACAAGCTTCCCGAAGGCGGCAGGGCGACATATCGCGGGACGGCGTTCGGTTCAGACGATGCCGGC
GGAAAACTGACCTACACCATAGATTTCGCCGCCAAGCAGGGAAACGGCAAAATCGAACATTTGAAATCGCCAGAACTC
AATGTCGACCTGGCCGCCGCCGATATCAAGCCGGATGGAAAACGCCATGGCCGTCATCAGCGGTCCGTCCTTTACAAC
CAAGCCGAGAAAGGCAGTTACTCCCTCGGTATCTTTGGCGGAAAAGCCCAGGAAGTTGCCGGCAGCGCGGAAGTGAAA
ACCGTAAACGGCATACGCCATATCGGCCTTGCCGCCAAGCAACTCGAGCACCACCACCACCACCACTTGA
1      MSDLANDSFI RQVLDRQHFE PDGKYHLFGS RGELAERSGH IGLGKIQSHQ
51     IGNLMIQQAA IKGNIGYIVR FSDHGHEVHS PFDNHASHSD SDEAGSPVDG
101    FSLYRIHWDG YEHHPADGYD GPQGGGYPAP KGARDIYSYD IKGVAQNIRL
151    NLTDNRSTGQ RLADRFHNAG SMLTQGVGDG FKRATRYSPE LDRSQNAAEA
201    FNGTADIVKN IIGAAGEIVG AGDAVQGISE GSNIAVMHGL GLLSTENKMA
251    RINDLADMAQ LKDYAAAAIR DWAVQNPNAA QGIRAVSNIF MAAIPIKGIG
301    AVRGKYGLGG ITAHPIKRSQ MGAIALPKGK SAVSDNFADA AYAKYPSPYH
351    SRNIRSNLEQ RYGKENITSS TVPPSNGKNV KLADQRHPKT GVPFDGKGFP
401    NFEKHVKYDT GSGGGGVAAD IGAGLADALT APLDHKDKGL QSLTLDQSVR
451    KNKKLKLAAQ GAKKTYGNGD SLNTGKLKND KVSRVDFORQ IEVDGQLITL
501    ESGEFQVYKQ SHSALTAFQT EQIQDSEHSG KMVAKRQFRI GDIAGEHTSF
551    DKLPEGGRAT YRGTAFGSDD AGGKLTYTID FAAKQGNGKI EHLKSPELNV
601    DLAADIKPD  GKRHAVISGS VLYNQAEKGS YSLGIFGGKA QEVAGSAEVK
651    TVNGIRHIGL AAKQLEHHHH HH*
ORF46.1-961
ATGTCAGATTTGGCAAACGATTCTTTTATCCGGCAGGTTCTCGACCGTCAGCATTTCGAACCCGACGGGAAATACCAC
CTATTCGGCAGCAGGGGGGAACTTGCCGAGCGCAGCGGCCATATCGGATTGGGAAAAATACAAAGCCATCAGTTGGGCAACCTGATGATTCAACAGGCGGCCATTAAAGGAAATATCGCCTACATTGTCCGCTTTTCCGATCACGGGCACGAAGTG
CATTCCCCCTTCGACAACCATGCCTCACATTCCGATTCTGATGAAGCCGGTAGTCCCGTTGACGGATTTAGCCTTTAC
CGCATCCTATTGGGACGGATACGAACACCATCCCGCCGACGGCTATGACGGGCACAGGGCGGCGGCTATCCCGCTCCC
AAAGGCGCGAGGGATATATACAGCTACGACATAAAAGGCGTTGCCCAAAATATCCGCCTCAACCTGACCGACAACCGC
AGCACCGGACAACGGCTTGCCGACCGTTTCCACAATGCCGGTAGTATGCTGACGCAAGGAGTAGGCGACGGATTCAAA
CGCGCCACCCGATACAGCCCCGAGCTGGACAGATCGGGCAATGCCGCCGAAGCCTTCAACGGCACTGCAGATATCGTT
AAAAACATCATCGGCGCGGCAGGAGAAATTGTCGGCGCAGGCGATGCCGTGCAGGGCATAAGCGAAGGCTCAAACATT
GCTGTCATGCACGGCTTGGGTCTGCTTTCCACCGAAAACAAGATGGCGCGCATCAACGATTTGGCAGATATGGCGCAA
CTCAAAGACTATGCCGCAGCAGCCATCCGCGATTGGGCAGTCCAAAACCCCAATGCCGCACAAGGCATAGAAGCCGTC
AGCAATATCTTTATGGCAGCCATCCCCATCAAAGGGATTGGAGCTGTTCGGGGAAAATACGGCTTGGGCGGCATCACG
GCACATCCTATCAAGCGGTCGCAGATGGGCGCGATCGCATTGCCGAAAGGGAAATCCGCCGTCAGCGACAATTTTGCC
GATGCGGCATACGCCAAATACCCGTCCCCTTACCATTCCCGAAATATCCGTTCAAACTTGGAGCAGCGTTACGGCAAA
GAAACATCACCTCCTCAACCGTGCCGCCGTCAAACGGCAAAAATGTCAAACTGGCAGACCAACGCCACCCGAAAGACA
GGCGTACCGTTTGACGGTAAAGGGTTTCCGAATTTTGAGAAGCACGTGAAATATGATACGGGATCCGGAGGAGGAGGA
GCCACAAACGACGACGATGTTAAAAAAGCTGCCACTGTGGCCATTGCTGCTGCCTACAACAATGGCCAAGAAATCAAC
GGTTTCAAAGCTGGAGAGACCATCTACGACATTGATGAAGACGGCACAATTACCAAAAAAGACGCAACTGCAGCCGAT
GTTGAAGCCGACGACTTTAAAGGTCTGGGTCTGAAAAAAGTCGTGACTAACCTGACCAAAACCGTCAATGAAAACAAA
CAAAACGTCGATGCCAAAGTAAAAGCTGCAGAATCTGAAATAGAAAAGTTAACAACCAAGTTAGCAGACACTGATGCC
GCTTTAGCAGATACTGATGCCGCTCTGCATCGAACCACCAACGCCTTGAATAAATTGGGAGAAAATATAACGACATTT
GCTGAAGAGACTAAGACAAATATCGTAAAAATTGATGAAAAATTAGAAGCCGTGGCTGATACCGTCGACAAGCATGCC
GAAGCATTCAACGATATCGCCGATTCATTGGATGAAACCAACACTAAGGCAGACGAAGCCGTCAAAACCGCCAATGAA
GCCAAACAGACGGCCGAAGAAACCAAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAAACTGCAGCAGGCAAAGCC
GAAGCTGCCGCTGGCACAGCTAATACTGCAGCCGACAAGGCCGAAGCTGTCGCTGCAAAAGTTACCGACATCAAAGCT
GATATCGCTACGAACAAAGATAATATTGCTAAAAAAGCAAACAGTGCCGACGTGTACACCAGAGAAGAGTCTGACAGC
AAATTTGTCAGAATTGATGGTCTGAACGCTACTACCGAAAAATTGGACACACGTTGGCTTCTGCTGAAAAATCCCATT
GCCGGATCACGATACTCGCCTGAACGGTTTGGATAAAACAGTGTCAGACCTGCGCAAAGAAACCCGCCAAGGCCTGCA
GAACAAGCCGCGCTCTCCGGTCTGTTCCAACCTTACAACGTGGGTCGGTTCAATGTAACGGCTGCAGTCGGCGGCTAC
AAATCCGAATCGGCAGTCGCCATCGGTACCGGCTTCCGCTTTACCGAAAACTTTGCCGCCAAAGCAGGCGTGGCAGTC
GGCACTTCGTCCGGTTCTTCCGCAGCCTACCATGTCGGCGTCAATTACGAGTGGCTCGAGCACCACCACCACCACCAC
TGA
1    MSDLANDSFI RQVLDRQHFE PDGKYHLFGS RGELAERSGH IGLGKIQSHQ
51   LGNLMIQQAA IKGNIGYIVR FSDHGHEVHS PFDNHASHSD SDEAGSPVDG
101  FSLYRIHWDG YEHHPADGYD GPQGGGYPAP KGARDIYSYD IKGVAQNIRL
151  NLTDNRSTGQ RLADRFHNAG SMLTQGVGDG FKRATRYSPE LDRSPNAAEA
201  FNGTADIVKN IIGAAGEIVG AGDAVQGISE GSNIAVNHGL GLLSTENKMA
251  RINKLADMAQ LKDYAAAAIR DWAVQNPNAA QGIEAVSNIF MAAIPIKGIG
301  AVRGKYGLGG IPAHPIKRSQ MGAIALPKGK SAVSDNFADA AYAKYPSPYH
351  SRNIRSNLEQ RYGKKNITSS TVPPSNGKNV KLADQRHPKT GVPFDGKGFP
401  NFEKHVKYDT GSGGGGATND DDVKKAATVA LAAAYNNGQE INGFKAGETI
451  YDIDEDGTIT KKDATAADVE ADDFKGLGLK KVVTNLTKTV NENKQNVDAK
501  VKAAESEIEK LTTKLADTDA ALADTDAALD ATTNALNKLG ENITTFAEET
551  KTNIVKIDEK LEAVADTVDK HAEAFNDIAD SLDETNTKAD EAVKTANEAK
601  QTAEETKQNV DAKVKAAETA AGKABAAAGT ANTAADKAEA VAAKVTDIKA
651  DIATNKDNIA KKANSADVYT REESDSKFVR IDGLNATTEK LDTRLASAEK
701  SIADHDTRLN GLDKTVSDLR KETRQGLAEQ AALSGLFQPY NVGRFNVTAA
751  VGGYKSESAV AIGTGFRFTE NFAAKAGVAV GTSSGSSAAY HVGVNYEWLE
801  HHHHHH*
ORF46.1-961c
ATGTCAGATTTGGCAAACGATTCTTTTATCCGGCAGGTTCTCGACCGTCAGCATTTCGAACCCGACGGGAAATACCAC
CTATTCGGCAGCAGGGGGGAACTTGCCGAGCGCAGCGGCCATATCGGATTGGGAAAAATACAAAGCCATCAGTTGGGC
AACCTGATGATTCAACAGGCGGCCATTAAAGGAAATATCGGCTACATTGTCCGCTTTTCCGATCACGGGCACGAAGTC
CATTCCCCCTTCGACAACCATGCCTCACATTCCGATTCTGATGAAGCCGGTAGTCCCGTTGACGGATTTAGCCTTTAC
CGCATCCATTGGGACGGATACGAACACCATCCCGCCGACGGCTATGACGGGCCACAGGGCGGCGGCTATCCCGCTCCC
AAAGGCGCGAGGGATATATACAGCTACGACATAAAAGGCGTTGCCCAAAATATCCGCCTCAACCTGACCGACAACCGC
AGCACCGGACAACGGCTTGCCGACCGTTTCCACAATGCCGGTAGTATGCTGACGCAAGGAGTAGGCGACGGATTCAAA
CGCGCCACCCGATACAGCCCCGAGCTGGACAGATCGGGCAATGCCGCCGAAGCCTTCAACGGCACTGCAGATATCGTT
AAAAACATCATCGGCGCGGCAGGAGAAATTGTCGGCGCAGGCGATGCCGTGCAGGGCATAAGCGAAGGCTCAAACATT
GCTGTCATGCACGGCTTGGGTCTGCTTTCCACCGAAAACAAGATGGCGCGCATCAACGATTTGGCAGATATGGCGCAA
CTCAAAGACTATGCCGCAGCAGCCATCCGCCATTGGGCAGTCCAAAACCCCAATGCCGCACAAGGCATAGAAGCCGTC
AGCAATATCTTTATGGCAGCCATCCCCATCAAAGGGATTGGAGCTGTTCGGGGAAAATACGGCTTGGGCGGCATCACG
GCACATCCTATCAAGCGGTCGCAGATGGGCGCGATCGCATTGCCGAAAGGGAAATCCGCCGTCAGCGACAATTTTGCC
GATGCGGCATACGCCAAATACCCGTCCCCTTACCATTCCCGAAATATCCGTTCAAACTTGGAGCAGCGTTACGGCAAA
GAAAACATCACCTCCTCAACCGTGCCGCCGTCAAACGGCAAAAATGTCAAACTGGCAGACCAACGCCACCCGAAGACA
GGCGTACCGTTTGACGGTAAAGGGTTTCCGAATTTTGAGAAGCACGTGAAATATGATACGGGATCCGGAGGAGGAGGA
GCCACAAACGACGACGATGTTAAAAAAGCTGCCACTGTGGCCATTGCTGCTGCCTACAACAATGGCCAAGAAATCAAC
GCTTTCAAAGCTGGAGAGACCATCTACGACATTGATGAAGACGGCACAATTACCAAAAAAGACGCAACTGCAGCCGAT
GTTGAAGCCGACGACTTTAAAGGTCTGGGTCTGAAAAAAGTCGTGACTAACCTGACCAAAACCGTCAATGAAAACAAA
CAAACGTCGATGCCAAAGTAAAGCTGCAGAGAATCTGAAATAGAAAAGTTAACAACCAAGTTAGCAGACACTGATGCC
GCTTTAGCAGATACTGATGCCGCTCTGGATGCAACCACCAACGCCTTGAATAAATTGGGAGAAAATATAACGACATTT
GCTGAAGAGACTAAGACAAATATCGTAAAAATTGATGAAAAATTAGAAGCCGTGGCTGATACCGTCGACAAGCATGCC
GAAGCATTCAACGATATCGCCGATTCATTGGATGAAACCAACACTAAGGCAGACGAAGCCGTCAAAACCGCCAATGAA
GCCAAACAGACGGCCGAAGAAACCAAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAAACTGCAGCAGGCAAAGCC
GAAGCTGCCGCTGGCACAGCTAATACTGCAGCCGACAAGGCCGAAGCTGTCGCTGCAAAAGTTACCGACATCAAAGCT
GATATCGCTACGAACAAAGATAATATTGCTAAAAAAGCAAACAGTGCCGACGTGTACACCAGAGAAGAGTCTGACAGC
AAATTTGTCAGAATTGATGGTCTGAACGCTACTACCGAAAAATTGGACACACGCTTGGCTTCTGCTGAAAAATCCATT
GCCGATCACGATACTCGCCTGAACGGTTTGGATAAAACAGTGTCAGACCTGCGCAAAGAAACCCGCCAAGGCCTTGCA
GAACAAGCCGCGCTCTCCGGTCTGTTCCAACCTTACAACGTGGGTCTCGAGCACCACCACCACCACCACTGA
1    MSDLANDSFI RQVLDRQHFE PDGKYHLFGS RGELAERSGH IGLGKIQSHQ
51   LGNLMIQQAA IKGNIGYIVR FSDHGHEVHS PFDNHASHSD SDRAGSPVDG
101  FSLYRIHWDG YEHHPADGYD GPQGGGYPAP KGARDIYSYD IKGVAQNIRL
151  NLTDNRSTGQ RLADRFEHNG SMLTQGVGDG FKRATRYSPE LDRSGNAAEA
201  FNGTADIVKN IIGAAGBIVG AGDAVQGISE GSNIAVMRGL GLLSTENKMA
251  RINDLADMAQ LKDYAAAAIR DWAVQNPNAA QGIKAVSHIF NAAIPIKGIG
301  AVRGKYGLGG ITAHPIKRSQ NGAIALPKGK SAVSDNFADA AYAKYPSPYH
351  SRNIRSNLBQ RYGKKNITSS TVPPSNGKNV KLADQRHPKT GVPFDGKGFP
401  NFEKHVKYDT GSGGGGATND DDVKKAATVA IAAAYNNGQE INGFKNGETI
451  YDIDEDGTIT KKDATAADVE ADDFKGLGLK KVVTNLTKTV NENKQNVKAK
501  VKAAESSIEK LTTKLADTDA ALAKTDAALD ATTNALNKLG ENITTFAEET
551  KTNIVKIDEK LEAVADTVDK HAEAFNDIAD SLDSTNTKAD BAVKTANEAK
601  QTAEBTKQNV DAKVKAAETA AGKASAAAGT ANTAADKAEA VAAKVTDIKA
651  DIATNKDNIA KKANSADVYT RERSDSKFVR IDGLNATTEK LDTRLASAEK
701  SIADHDTRLN GLDKTVSDLR KETRQGLABQ AALSGLFQPY NVGLEHHHHH
751  H*
961-ORF46.1
ATGGCCACAAACGACGACGATGTTAAAAAAGCTGCCACTGTGGCCATTGCTGCTGCCTACAACAATGGCCAAGAAATC
AACGGTTTCAAAGCTGGAGAGACCATCTACCACATTCATGAAGACGGCACAATTACCAAAAAAGACGCAACTGCAGCC
GATGTTGAAGCCGACGACTTTAAAGGTCTGGGTCTGAAAAAAGTCGTGACTAACCTGACCAAAACCGTCAATGAAAAC
AAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAATCTGAAATAGAAAAGTTAACCACCAAGTTAGCAGACACTGAT
GCCGCTTTAGCAGATACTGATGCCGCTCTGGATGCAACCACCAACGCCTTGAATAAATTGGGAGAAAATATAACGACA
TTTGCTGAAGAGACTAAGACAAATATCGTAAAAATTGATGAAAAATTAGAAGCCGTGGCTGATACCGTCGACAAGCAT
GCCGAAGCATTCAACGATATCGCCGATTCATTGGATGAAACCAACACTAAGGCAGACGAAGCCGTCAAAACCGCCAAT
GAAGCCAAACAGACGGCCGAAGAAACCAAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAAACTGCAGCAGGCAAA
GCCGAAGCTGCCGCTGGCACAGCTAATACTGCAGCCGACAAGGCCGAAGCTGTCGCTGCAAAAGTTACCGACATCAAA
GCTGATATCGCTACGAACAAAGATAATATTGCTAAAAAAGCAAACAGTGCCGACGTGTACACCAGAGAAGAGTCTGAC
AGCAAATTTGTCAGAATTGATGGTCTGAACGCTACTACCGAAAAATTGGACACACGCTTGGCTTCTGCTGAAAAATCC
ATTGCCGATCACGATACTCGCCTGAACGGTTTGGATAAAACAGTGTCAGACCTGCGCAAAGAAACCCGCCAAGGCCTT
GCAGAACAAGCCGCGCTCTCCGGTCTCTTCCAACCTTACAACGTGGGTCGGTTCAATGTAACGGCTGCAGTCGGCGGC
TACAAATCCGAATCGGCAGTCGCCATCGGTACCGGCTTCCGCTTTACCGAAAACTTTGCCGCCAAAGCAGGCGTGGCA
GTCGGCACTTCGTCCGGTTCTTCCGCAGCCTACCATGTCGGCGTCAATTACGAGTGGGGATCCGGAGGAGGAGGATCA
GATTTGGCAAACGATTCTTTTATCCGGCAGGTTCTCGACCGTCAGCATTTCGAACCCGACGGGAAATACCACCTATTC
GGCAGCAGGGGGGAACTTGCCGAGCGCAGCGGCCATATCGGATTGGGAAAAATACAAAGCCATCAGTTGGGCAACCTG
ATGATTCAACAGGCGGCCATTAAAGGAAATATCGGCTACATTGTCCGCTTTTCCGATCACGGGCACGAAGTCCATTCC
CCCTTCGACAACCATGCCTCACATTCCGATTCTGATGAAGCCGGTAGTCCCGTTGACGGATTTAGCCTTTACCGCATC
CATTGGGACGGATACGAACACCATCCCGCCGACGGCTATGACGGGCCACAGGGCGGCGGCTATCCCGCTCCCAAAGGC
GCGAGGGATATATACAGCTACGACATAAAAGGCGTTGCCCAAAATATCCGCCTCAACCTGACCGACAACCGCAGCACC
GCACAACGGCTTGCCGACCGTTTCCACAATGCCGGTAGTATGCTGACGCAAGGAGTAGGCGACGGATTCAAACGCGCC
ACCCGATACAGCCCCGAGCTGGACAGATCGGGCAATGCCGCCGAAGCCTTCAACGGCACTGCAGATATCGTTAAAAAC
ATCATCGGCGCGGCAGGAGAAATTCTCGGCGCAGGCGATGCCGTGCAGGGCATAAGCGAAGGCTCAAACATTGCTGTC
ATGCACGGCTTGGGTCTGCTTTCCACCGAAAACAAGATGGCGCGCATCAACGATTTGGCAGATATGGCGCAACTCAAA
GACTATGCCGCAGCAGCCATCCGCGATTGGGCAGTCCAAAACCCCAATGCCGCACAAGGCATAGAAGCCGTCAGCAAT
ATCTTTATGGCAGCCATCCCCATCAAAGGGATTGGAGCTGTTCGGGGAAAATACGGCTTGGGCGGCATCACGGCACAT
CCTATCAAGCGGTCGCAGATGGGCGCGATCGCATTGCCGAAAGGGAAATCCGCCGTCAGCGACAATTTTGCCGATGCG
GCATACGCCAAATACCCGTCCCCTTACCATTCCCGAAATATCCGTTCAAACTTGGAGCAGCGTTACGGCAAAGAAAAC
ATCACCTCCTCAACCGTGCCGCCGTCAAACGGCAAAAATGTCAAACTGGCAGACCAACGCCACCCGAAGACAGGCGTA
CCGTTTGACGGTAAAGGGTTTCCGAATTTTGAGAAGCACGTGAAATATGATACGCTCGAGCACCACCACCACCACCAC
TGA
1    MATNKKKVKK AATVAIAAAY NNGQEINGFK AGETIYDIDE DGTITKKDAT
51   AADVEADDFK GLGLKKVVTN LTKTVNKNKQ NVKAKVKAAE SEIEKLTTKL
101  ADTDAALADT DAALDATTNA LNKLGKNITT FAEETKTNIV KIDEKLEAVA
151  DTVDKHAEAF NDIADSLDST NTKADEAVKT ANEAKQTAEK TKQNVDAKVK
201  AAETAAGKAK AAAGTANTAA DKAEAVAAKV TDIKADIATN KDNIAKKANS
251  ADVYTREESD SKFVRIDGLN ATTEKLDTRL ASARKSLADH DTRLNGLDKT
301  VSDLRKETRQ GLAEQAALSG LFQPYNVGRF NVTAAVGGYK SESAVAIGTG
351  FRFTENFAAK AGVAVGTSSG SSAAYHVGVN YKWGSGGGGS DLANDSFIRQ
401  VLDRQHFEPD GKYHLFGSRG ELAERSGHIG LGKIQSHQLG NLMIQQAAIK
451  GNIGYIVRFS DHGHEVHSPF DNHASHSDSD EAGSPVDGFS LYRIHWDGYE
501  HHPADGYDGP QGGGYPAPKG ARDIYSYDIK GVAQNIRLNL TDNRSTGQRL
551  ADRFHNAGSN LTQGVGDGFK RATRYSPELD RSGNAAEAFN GTADIVKNII
601  GAAGEIVGAG DAVQGISEGS NIAVNHGIGL LSTRNKMARI NDLADMAQLK
651  DYAAAAIRDW AVQNPNAAQG IRAVSNIFMA AIPIKGIGAV RGKYGLGGIT
701  AHPIKRSQMG AIALPKGKSA VSDNFADAAY AKYPSPYESR NIRSNLBQRY
751  GKENITSSTV PPSNGKNVKL ADQREPKTGV PFDGKGFPNF EKHVKYDTLE
801  HHHHHH*
961-741
ATGGCCACAAACGACGACGATGTTAAAAAAGCTGCCACTGTGGCCATTGCTGCTGCCTACAACAATGGCCAAGAAATC
AACGGTTTCAAAGCTGGAGAGACCATCTACGACATTGATGAAGACGGCACAATTACCAAAAAAGACGCAACTGCAGCC
GATGTTGAAGCCGACGACTTTAAAGGTCTGGGTCTGAAAAAACTCGTGACTAACCTGACCAAAACCGTCAATGAAAAC
AAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAATCTGAAATAGAAAAGTTAACAACCAAGTTAGCAGACACTGAT
GCCGCTTTAGCAGATACTGATGCCGCTCTGGATGCAACCACCAACGCCTTGAATAAATTGGGAGAAAATATAACGACA
TTTGCTGAAGAGACTAAGACAAATATCCTAAAAATTGATGAAAAATTAGAAGCCGTGGCTGATACCGTCGACAAGCAT
GCCGAAGCATTCAACGATATCGCCGATTCATTGGATGAAACCAACACTAAGGCAGACGAAGCCGTCAAAACCGCCAAT
GAAGCCAAACAGACGGCCGAAGAAACCAAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAAACTGCAGCAGGCAAA
GCCGAAGCTGCCGCTGGCACAGCTAATACTGCAGCCGACAAGGCCGAAGCTGTCGCTGCAAAAGTTACCGACATCAAA
GCTGATATCGCTACGAACAAAGATAATATTGCTAAAAAAGCAAACAGTGCCGACGTGTACACCAGAGAAGAGTCTGAC
AGCAAATTTGTCAGAATTGATGGTCTGAACGCTACTACCGAAAAATTGGACACACGCTTGGCTTCTGCTGAAAAATCC
ATTGCCGATCACGATACTCGCCTGAACGGTTTGGATAAAACAGTGTCAGACCTGCGCAAAGAAACCCGCCAAGGGCTT
GCAGAACAAGCCGCGCTCTCCGGTCTGTTCCAACCTTACAACGTGGGTCGGTTCAATGTAACGGCTGCAGTCGGCGGC
TACAAATCCGAATCGGCAGTCGCCATCGGTACCGGCTTCCGCTTCACCGAAAACTTTGCCGCCAAAGCAGGCGTGGCA
GTCGGCACTTCGGTCCGGTTCTTCTTCCGCACCTACCATGTCGGCGTCAATTACGAGTGGGGATCCGGGGGTGGTGTC
GCCGCCGACATCGGTGCGGGGCTTGCCGATGCACTAACCGCACCGCTCGACCATAAAGACAAAGGTTTGCAGTCTTTG
ACGCTGGATCAGTCCGTCAGGAAAAACGAGAAACTGAAGCTGGCGGCACAAGGTGCGGAAAAAACTTATGGAAACGGT
GACAGCCTCAATACGGGCAAATTGAAGAACGACAAGGTCAGCCGTTTCGACTTTATCCGCCAAATCGAAGTGGACGGG
CAGCTCATTACCTTGGAGAGTGGAGAGTTCCAAGTATACAAACAAAGCCATTCCGCCTTAACCGCCTTTCAGACCGAG
CAAATACAAGATTCGGAGCATTCCGGGAAGATGGTTGCGAAACGCCAGTTCAGAATCGGCGACATAGCGGGCGAACAT
ACATCTTTTGACAAGCTTCCCGAAGGCGGCAGGGCGACATATCGCGGGACGGCGTTCGGTTCAGACGATGCCGGCGGA
AAACTGACCTACACCATAGATTTCGCCGCCAAGCAGGGAAACGGCAAAATCGAACATTTGAAATCGCCAGAACTCAAT
GTCGACCTGGCCGCCGCCGATATCAAGCCGGATGGAAAACGCCATGCCGTCATCAGCGGTTCCGTCCTTTACAACCAA
GCCGAGAAAGGCAGTTACTCCCTCGGTATCTTTGGCGGAAAAGCCCAGGAAGTTGCCGGCAGCGCGGAAGTGAAAACC
GTAAACGGCATACGCCATATCGGCCTTGCCGCCAAGCAACTCGAGCACCACCACCACCACCACTGA
1   MATNDDDVKK AATVAIAAAY NNGQEINGFK AGETIYDIDE DGTITKKDAT
51  AADVEADDFK GLGLKKVVTN LTKTVNKNKQ NVDAKVKAAE SEIEKLTTKL
101 ADTDAALADT DAALDATTNA LNKLGENITT FAEETKTNIV KIDEKLEAVA
151 DTVDKHAEAF NDIADSLDET NTKADKAVKT ANBAKQTAEE TKQNVDAKVK
201 AAETAAGKAE AAAGTANTAA DKAEAVAAKV TDIKADIATN KDNIAKKANS
251 ADVYTRKKSD SKFVRIDGLN ATTEKLDTRL ASAKKSIADH DTRLNGLDKT
301 VSDLRKBTRQ GLAEQAALSG LFQPYNVGRF NVTAAVGGYK SESAVAIGTG
351 FRFTENFAAK AGVAVGTSSG SSAAYHVGVN YBVGSGGGGV AADIGAGLAD
401 ALTAPLDHKD KGLQSLTLDQ SVRKNEKLKL AAQGAKKTYG NGDSLNTGKL
451 KNDKVSRFDF IRQIBVDGQL ITLESQKFQV YKQSHSALTA FQTBQIQDSE
501 HSQKMVAKRQ FRIGDIAGEH TSFDKLPEGG RATYRGTAFG SDDAGGKLTY
551 TIDFAAKQGN GKIEHLKSPB LNVDLAAADI KPDGKRHAVI SGSVLYNQAE
601 KGSYSLGIFG GKAQEVAGSA EVKTVNGIRH IGLAAKQLEH HHHH*
961-983
ATGGCCACAAACGACGACGATGTTAAAAAAGCTGCCACTGTGGCCATTGCTGCTGCCTACAACAATGGCCAAGAAATC
AACGGTTTCAAAGCTGGAGAGACCATCTACCGACATTGATGAAGACGGCACAATTACCAAAAAGACGCAACTGCAGCC
GATGTTGAAGCCCGACGACTTTAAAGGTCTGGGTCTGAAAAAGTCGTGACTAACCTGACCAAAACCGTCAATGAAAAC
AAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAATCTGAAATAGAAAAGTTAACAACCAAGTTAGCAGACACTGAT
GCCGCTTTAGCAGATACTGATGCCGCTCTGGATCCAACCACCAACGCCTTGAATAAATTGGGAGAAAATATAACGACA
TTTGCTGAAGAGACTAGAGACAAATATCGTAAAAATTGATGAAAAATTAGAAGCCGTGGCTGATACGTCGACAAGCAT
GCCGAAGCATTCAACGATATCGCCGATTCATTGGATGAAACCAACACTAAGGCAGACGAAGCCGTCAAAACCGCCAAT
GAAGCCAAACAGACGGCCGAAGAAACCAAACAAAACGTCGATGCGAAAGTAAAAGCTGCAGAAACTGCAGCAGGCAAA
GCCGAAGCTGCCGCTGGCACAGCTAATACTGCAGCCGACAAGGCCGAAGCTCTCGCTGCAAAAGTTACCGACATCAAA
GCTGATATCGCTACGAACAAAGATAATATTGCTAAAAAAGCAAACAGTGCCGACGTGTACACCAGAGAAGAGTCTGAC
AQCAAATTTGTCAGAATTGATGGTCTGAACGCTACTACCGAAAAATTGGACACACGCTTGGCTTCTGCTGAAAAATCC
ATTGCCGATCACGATACTCGCCTGAACGGTTTGGATAAAACAGTGTCAGACCTGCGCAAAGAAACCCGCCAAGGCCTT
GCAGAACAAGCCGCGCTCTCCGGTCTGTTCCAACCTTACAACGTGGGTCGGTTCAATGTAACGGCTGCAGTCGGCGGC
TACAAATCCGAATCGGCAGTCGCCATCGGTACCGGCTTCCGCTTTACCGAAAACTTTGCCGCCAAAGCAGGCGTGGCA
GTCGGCACTTCGTCCGGTTCTTCCGCAGCCTACCATGTCGGCGTCAATTACGAGTGGGGATCCGGCGGAGGCGGCACT
TCTGCGCCCGACTTCAATGCAGGCGGTACCGGTATCGGCAGCAACAGCAGAGCAACAACAGCGAAATCAGCAGCAGTA
TCTTACGCCGGTATCAAGAACGAAATGTGCAAAGACAGAAGCATGCTCTGTGCCGGTCGGGATGACGTTGCGGTTACA
GACAGGGATGCCAAAATCAATGCCCCCCCCCCGAATCTGCATACCGGAGACTTTCCAAACCCAAATGACGCATACAAG
AATTTGATCAACCTCAAACCTGCAATTGAAGCAGGCTATACAGGACGCGGGGTAGAGGTAGGTATCGTCGACACAGGC
GAATCCGTCGGCAGCATATCCTTTCCCGAACTGTATGGCAGAAAAGAACACGGCTATAACGAAAATTACAAAAACTAT
ACGGCGTATATGCGGAAGGAAGCGCCTGAAGACGGAGGCGGTAAAGACATTGAAGCTTCTTTCGACGATGAGGCCGTT
ATAGAGACTGAAGCAAAGCCGACGGATATCCGCCACGTAAAAGAAATCGGACACATCGATTTGGTCTCCCATATTATT
GGCGGGCGTTCCGTGGACGGCAGACCTGCAGGCGGTATTGCGCCCGATQCGACGCTACACATAATGAATACGAATGAT
GAAACCAAGAACGAAATGATGGTTGCAGCCATCCGCAATGCATGGGTCAAGCTGGGCGAACGTGGCGTGCGCATCGTC
AATAACAGTTTTGGAACAACATCGAGGGCAGGCACTGCCGACCTTTTCCAAATAGCCAATTCGGAGGAGCAGTACCGC
CAAGCGTTGCTCGACTATTCCGGCGGTGATAAAACAGACGAGGGTATCCGCCTGATGCAACAGAGCGATTACGGCAAC
CTGTCCTACCACATCCGTAATAAAAACATGCTTTTCATCTTTTCGACAGGCAATGACGCACAAGCTCAGCCCAACACA
TATGCCCTATTGCCATTTTATGAAAAAGACGCTCAAAAAGGCATTATCACAGTCGCAGGCGTAGACCGCAGTGGAGAA
AAGTTCAAACGGGAAATGTATGGAGAACCGGGTACAGAACCGCTTGAGTATGGCTCCAACCATTGCGGAATTACTGCC
ATGTGGTGCCTGTCGGCACCDTATGAAGCAAGCGTCCGTTTCACCCGTACAAACCCGATTCAAATTGCCGGAACATCC
TTTTCCGCACCCATCGTAACCGGCACGGCGGCTCTGCTGCTGCAGAAATACCCGTGGATGAGCAACGACAACCTGCGT
ACCACGTTGCTGACGACGGCTCAGGACATCGGAGCAGTCGGCGTGGACAGCAAGTTCGGCTGGGGACTGCTGGATGCG
GGTAAGGCCATGAACGGACCCGCGTCCTTTCCGTTCGGCGACTTTACCCCCGATACGAAAGGTACATCCGATATTGCC
TACTCCTTCCGTAACGACATTTCAGGCACGGGCGGCCTGATCAAAAAAGGCGGCAGCCAACTGCAACTGCACGGCAAC
AACACCTATACGGGCAAAACCATTATCGAAGGCGGTTCGCTGGTGTTGTACGGCAACAACAAATCGGATATGCGCGTC
GAAACCAAAGGTGCGCTGATTTATAACGGGGCGGCATCCGGCGGCAGCCTGAACAGCGACGGCATTGTCTATCTGGCA
GATACCGACCAATCCGGCGCAAACGAAACCGTACACATCAAAGGCAGTCTGCAGCTGGACGGCAAAGGTACGCTGTAC
ACACGTTTGGGCAAACTGCTGAAAGTGGACGGTACGGCGATTATCGGCGGCAAGCTGTACATGTCGGCACGCGGCAAG
GGGGCAGGCTATCTCAACAGTACCGGACGACGTGTTCCCTTCCTGAGTGCCGCCAAAATCGGGCAGGATTATTCTTTC
TTCACAAACATCGAAACCGACGGCGGCCTGCTGGCTTCCCTCGACAGCGTCGAAAAAACAGCGGGCAGTGAAGGCGAC
ACGCTGTCCTATTATGTCCGTCGCGGCAATGCGGCACGGACTGCTTCGGCAGCGGCACATTCCGCGCCCGCCGGTCTG
AAACACGCCGTAGAACAGGGCGGCAGCAATCTGGAAAACCTGATGGTCGAACTGGATGCCTCCGAATCATCCGCAACA
CCCGACACGGTTGAAACTGCGGCAGCCGACCGCACAGATATGCCGGGCATCCGCCCCTACGGCGCAACTTTCCGCGCA
GCGGCAGCCGTACAGCATGCGAATGCCGCCGACGGTGTACGCATCTTCAACAGTCTCGCCGCTACCGTCTATGCCGAC
AGTZCCGCCGCCCATGCCGATATGCAGGGAGGCCGCCTGAAAGCCGTATCGGACGGGTTGGACCACAACGGCACGGGT
CTGCGCGTCATCGCGCAAACCCAACAGGACGGTGGAACGTGGGAACAGGGCGGTGTTGAAGGCAAAATGCGCGGCAGT
ACCCAAACCGTCGGCATTGCCGCGAAAACCGGCGAAAATACGACAGCAGCCGCCACACTGGGCATGGGACGCAGCACA
TGGAGCGAAAACAGTGCAAATGCAAAAACCGACAGCATTAGTCTGTTTGCAGGCATACGGCACGATGCGGGCGATATC
GGCTATCTCAAAGGCCTGTTCTCCTACGGACGCTACAAAAACAGCATCAGCCGCAGCACCGGTGCGGACGAACATGCG
GAAGGCAGCGACAACGGCACGCTGATGCAGCTGGGCGCACTGGGCGGTGTCAACGTTCCGTTTGCCGCAACGGGAGAT
TTGACGGTCGAAGGCGGTCTGCGCTACGACCTGCTCAAACAGGATGCATTCGCCGAAAAAGGCAGTGCTTTGGGCTGG
AGCGGCAACAGCCTCACTGAAGGCACGCTGGTCGGACTCGCGGGTCTGAAGCTGTCGCAACCCTTGAGCGATAAAGCC
GTCCTGTTTGCAACGGCGGGCGTGGAACGCGACCTGAACGGACGCGACTACACGGTAACGGGCGGCTTTACCGGCGCG
ACTGCAGCAACCGGCAAGACGGGGGCACGCAATATGCCGCACACCCGTCTGGTTGCCGGCCTGGGCGCGGATGTCGAA
TTCGGCAACGGCTGGAACGGCTTGGCACGTTACAGCTACGCCGGTTCCAAACAGTACGGCAACCACAGCGGACGAGTC
GGCGTAGGCTACCGGTTCCTCGAGCACCACCACCACCACCACTGA
1    MATNDDDVKK AATVAIAAAY NNQQEINGFK AGBTIYDIDB DGTITXKDAT
51   AADVEADDFK GLGLKKVVTN LTKTVNKNKQ NVDAKVKAAE SEIEKLTTKL
101  ADTDAALADT DAALDATTNA LNKLGKNITT FABBIKINIV KIDKKLBAVA
151  DTVDKHAEAF NDIADSLDET NTKADEAVKT ANEAKQTAEE TKQNVDAKVK
201  AAETAAGKAE AAAGTANTAA DKAEAVAAKV TDIKADIATN KINIAKKANS
251  ADVYTRKKSD SKFVRIDGLN ATTEKLDTRL ASAKKSIADH DTRLNGLDKT
301  VSDLRKETRQ GLABQAALSG LFQPYNVGRF HVTAAVGGYK SESAVAIGTG
351  FRFTKNFAAK AGVAVGTSSG SSAAYHVGVN YENGSGGGGT SAPDFNAGGT
401  GIGSNSRATT AKSAAVSYAG IKNENCKDRS MLCAGRDDVA VTDRDAKINA
451  PPPNLHNGDF PNPNDAYKNL LNLKPAIRAG YTGRGVBVGI VDTGKSVGSI
501  SFPELYGRKE HGYNENYKNY TAYMRKBAPB DGGGKDIKAS FDDBAVIBTE
551  AKPTDIRHVK BIGHIDLVSH IIGGRSVDGR PAGGIAPDAT LHDANTNDET
601  KNEMMVAAIR MAWVKLGEFQ VRIVNNSFGT TSRAGTADLF QLANSEEQYR
651  QALLDYSGGD KTDBGIRLMQ QSDYGNLSYH IPNKMMLFIF STGNDAQAQP
701  NTYALLPFYB KDAQKGIITV AGVDRSGEKF KFEMYGBPGT EPLKYGSNHC
751  GITAMWCLSA FYEASVRFTR TNPIQIAGTS FSAPTVTGTA ALLLQKYPWM
801  SNDNLRTTLL TTAQDIGAVQ VDSKFGWGLL DAGKAMNGPA SFPFGDFTAD
851  TKGTSDLAYS FRNDISGTGG LIKKGGSQLQ LHGNNTYTGK TIIBGGBLVL
901  YGNNKSDMRV ETKGALIYNG AASGGSLNSD GIVYLADTDQ SGANETVHIK
951  GSLQLDGKGT LYTRLGKLLK VDGTAIIQQK LYMSARGKQA GYLNSTGRRV
1001 PFLSAAKIGQ DYSFFTNIET DGGLLASLDS VKKTAGSEGD TLSYYVRRGN
1051 AARTASAAAH SAPAGLKHAV EQGGSNLENL MVELPASESS ATPBTVETAA
1101 ADRTDMPGIR PYGATFRAAA AVQHANAADG VRIFNSLAAT VYADSTAAHA
1151 DMQGRRLKAV SDGLDHNGTG LRVIAQTQQD QQTWBQGGVE GKMRGSTQTV
1201 GLAAKTGRNT TAAATLGMGR STWSENSANA KTDSISLFAG IRHDAGDIGY
1251 LKGLFSYGRY KNSISRSTGA DKHABGSVNG TLMQLGALGG VNVPFAATGD
1301 LTVEGGLRYD LLKQDAFAEK GBALGWSGNS LTBGTLVGLA GLKLSQPLSD
1351 KAVLFATAGV KRDLNGRDYT VTGGFTGATA ATGKTGARNM PHTRLVAGLG
1401 ADVEFGNGWN GLARYSYAGS KQYGNHSGRV GVGYRFLKHH HHHH*
961c-ORF46.1
ATGGCCACAAACGACGACGATGTTAAAAAAGCTGCCACTGTGGCCATTGCTGCTGCCTACAACAATGGCCAAGAAATC
AACGGTTTCAAAGCTGGAGAGACCATCTACGACATTGATGAAGACGGCACAATTACCAAAAAAGACGCAACTGCAGCC
GATGTTGAAGCCGACGACTTTAAAGGTCTGGGTCTGAAAAAAGTCGTGACTAACCTGACCAAAACCQTCAATGAAAAC
AAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAATCTGAAATAGAAAAGTTAACAACCAAGTTAGCAGACACIGAT
GCCGCTTTAGCAGATACTGATGCCGCTCTGGATGCAACCACCAACGCCTTGAATAAATTGGGAGAAAATATAACGACA
TTTGCTGAAGAGACTAAGACAAATATCCTAAAAATTGATGAAAAATTAGAAGCCGTGGCTGATACCGTCGACAAGCAT
GCCGAAGCATTCAACGATATCGCCGATTCATTGGGATGAAACCAACACTAAGGCAGACGAAGCCGTCAAAACCGCCAT
GAAGCCAAACAGACGGCCGAAGAAACCAAACAAAACGTCGATGCCAAAGTAAAAGCTQCAGAAACTGCAGCAGGCAAA
GCCGAAGCTGCCGCTGGCACAGCTAATACTGCAGCCGACAAGGCCGAAGCTGTCGCTGCAAAAGTTACCGACATCAAA
GCTGATATCGCTACGAACAAAGATAATATTGCTAAAAAAQCAAACAGTGCCGACGTGTACACGGAGAAGAGTCTGACC
AGCAAATTTGTCAGAATTGATGGTCTGAACGCTACTACCGAAAAATTGGACACACGCTTGGCTTCTQCTQAAAAATCC
ATTGCCGATCACGATACTCGCCTGAACGGTTTGGATAAAACAGTGTCAGACCTGCGCAAAGAAACCCGCCAAGGCCTT
GCAGAACAAGCCGCGCTCTCCGGTCTGTTCCAACCTTACAACGTGGGTGGATCCGGAGGAGGAGCATCAGATTTGGCA
AACGATTCTTTTATCCGGCAGGTTCTCGACCGTCAGCATTTCGAACCCGACGGGAAATACCACCTATTCGGCAGCAGG
GGGGAACTTGCCGAGCGCAGCGGCCATATCGGATTGGGAAAAATACAAAGCCATCAGTTGGGCAACCTGATGATTCAA
CAGGCGGCCATTAAAGGAAATATCGGCTACATTGTCCGCTTTTCCGATCACGGGCACGAAGTCCATTCCCCCTTCQAC
AACCATGCCTCACATTCCGATTCTGATGAAGCCGGTAGTCCCGTTGACGGATTTAQCCTTTACCGCATCCATTGGGAC
GGATACGAACACCATCCCGCCGACGGCTATGACGGGCCACAGGGCGGCGGCTATCCCGCTCCCAAAGGCGCGAGGGAT
ATATACAGCTACGACATAAAAGGCGTTGCCCAAAATATCCGCCTCAACCTGACCGACAACCGCAGCACCGGACAACGG
CTTGCCGACCGTTTCCACAATGCCGGTAGTATGCTGACGCAAGGAGTAGGCGACGGATTCAAACGCGCCACCCGATAC
AGCCCCGAGCTGGACAGATCGGGCAATGCCGCCGAAGCCTTCAACGGCACTGCAGATATCGTTAAAAACATCATCGGC
GCGGCAGGAGAAATTGTCGGCGCAGGCGATGCCGTGCAGGGCATAAGCGAAGGCTCAAAGATTGCTGTCATGCACGGC
TTGGGTCTGCGGGCCACCGAAAACAAGATGGCGCGCATCAACGATTTGGCAGATATGGCGCAACTCAAAGACTATGCC
GCAGCAGCCATCCGCGATTGGGCAGTCCAAAACCCCAATGCCGCACAAGGCATAGAAGCCGTCAGCAATATCTTTATG
GCAGCCATCCCCATCAAAGGGATTGGAGCTGTTCGGGGAAAATACGGCTTGGGCGGCATCACGGCACATCCTATCAAG
CGGTCGCAGATGGGCGCGATCGCATTGCCGAAAGGGAAATCCGCCGTCAGCGACAATTTTGCCGATGCGGCATACGCC
AAATACCCGTCCCCTTACCATTCCCGAAATATCCGTTCAAACTTGGAGCAGCGTTACGGCAAAGAAAACATCACCTCC
TCAACCGTGCCGCCGTCAAACGGCAAAAATGTCAAACTGGCAGACCAACGCCACCCGAAGACAGGCGTACCGTTTGAC
GGTAAAGGGTTTCCGAATTTTGAGAAGCACGTGAAATATGATACGCTCGAGCACCACCACCACCACCACTGA
1   MATNDDDVKK AATVAIAAAY NNGQEINGFK AGETIYDIDE DGTITKKDAT
51  AADVBADDFK GLGLKKVVTN LTKTVNKNKQ NVDAKVKAAE SEIKKLTTKL
101 ADTDAALADT DAALDATTNA LNKLGBNITT FAKBTKTNIV KIDKKLBAVA
151 DTVDKHAEAF NDIADSLDET NTKADEAVKT AEEAKQTAEE TAQNVDAKVK
201 AAETAAGKAE AAAGTANTAA DKAEAVAAKV TAIKADIATN KINIAKKANS
251 ADVYTREESP SKFVRIDGLN ATTEKLDTRL ASAEKSIADE DTRLNGLDKT
301 VSDLRKBTRQ QLAEQAALSQ LFQPYNVGGS GGGGSDLAND SFIRQVLDRQ
351 HFEPDGKYHL FGSRGELAER SQHIGLGKIQ SHQLGNLNIQ GAAIXGNIGY
401 TVRFSDHQHK VHSPFDNHAS VHSPFDNHAS HSDSDEAGSP VDGFSLYRIH
451 GYDGPQGGGY PAPKGAPDIY SYDIKGVAQN IRLNLTDNRS TGQRLADRFH
501 NAGSMLTQGV QDGFKRATRY SPELDRSGNA SPELDRSGNA VKNIIGAAGE
551 TVGAGDAVQG ISEGSNIAVN BGLGLLSTBN KMARINDLAD MAQLKDYAAA
601 AIRDWAVQNP NAAQGIEAVS NIFMAAIPIK GIGAVRGKYG LGGITAHPIK
651 RSQMGAIALP KQKSAVSINF ADAAYAKYPS FYHSRNIRSN LBQRYGKENI
701 TSSTVPPSNG KNVKLADQRH PKTGVPFDGK GFPNFEKHVK YDTLEHHHHH
751 H*
961c-741
ATGGCCACAAACGACGACGATGTTAAAAAAGCTGCCACTGTGGCCATTGCTGCTGCCTACAACAATGGCCAAGAAATC
AACGGTTTCAAAGCTGGAGAGACCATCTACGACATTGATGAAGACGGCACAATTACCAAAAAAGACGCAACTQCAGCC
GATGTTGAAGCCGACGACTTTAAAAGGTCTGGGTCTGAAAAAAGTCGTGACTAACCTGACCAAAACCGTCAATGAAAC
AAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAATCTGAAATAGAAAAGTTAACAACCAAGTTAGCAGACACTGAT
GCCGCTTTAGCAGATACTGATGCCCGCGTCTGGATGCAACCACCAACGCTTGAATAAATTGGGAGAAATATAACGACA
TTTGCTGAAGAGACTAAGACAAATATCGTAAAAATTGATGAAAAATTAGAAGCCGTGGCTGATACCGTCGACAAGCAT
GCCGAAGCATTCAACGATATCGCCGATTCATTGGATGAAACCAACACTAAGGCAGACGAAGCCGTCAAAACCGCCAAT
GAAGCCAAACAGACGGCCGAAAGAAACCAAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAAACTGCACAGGCAAA
GCCGAAGCTGCCGCTGGCACAGCTAATACTGCAGCCGACAAGGCCGAAGCTGTCGCTGCAAAAGTTACCGACATCAAA
GCTGATATCGCTACGAACAAAGATAATATTGCTAAAAAAGCAAACAGTGCCGACGTGTACACCAGAGAAGAGTCTGAC
AGCAAATTTGTCAGAATTGATGGTCTGAACGCTACTACCGAAAAATTGGACACACGCTTGGCTTCTGCTGAAAAATCC
ATTGCCGATCACGATACTCGCCTGAACGGTTTGGATAAAACAGTGTCAGACCTGCGCAAAGAAACCCGCCAAGGCCTT
GCAGAACAAGCCGCGCTCTCCGGTCTGTTCCAACCTTACAACGTGGGTGGATCCGGAGGGGGTGGTGTCGCCGCCGAC
ATCGGTGCGGGGCTTGCCGATGCACTAACCGCACCGCTCGACCATAAAGACAAAGGTTTGCAGTCTTTGACGCTGGAT
CAGTCCGTCAGGAAAAACGAGAAACTGAAGCTGGCGGCACAAGGTGCGGAAAAAACTTATGGAAACGGTGACAGCCTC
AATACGGGCAAATTGAAGAACGACAAGGTCAGCCGTTTCGACTTTATCCGCCAAATCGAAGTGGACGGGCAGCTCATT
ACCTTGGAGAGTGGAGAGTTCCAAGTATACAAACAAAGCCATTCCGCCTTAACCGCCTTTCAGACCGAGCAAATACAA
GATTCGGAGCATTCCGGGAAGATGGTTGCGAAACGCCAGTTCAGAATCGGCGACATAGCGGGCGAACATACATCTTTT
GACAAGCTTCCCGAAGGCGGCAGGGCGACATATCGCGGGACGGCGTTCGGTTCAGACGATGCCGGCGGAAAACTGACC
TACACCATAGATTTCGCCGCCAAGCAGGGAAACGGCAAAATCGAACATTTGAAATCGCCAGAACTCAATGTCGACCTG
GCCGCCGCCGATATCAAGCCGGATGGAAACGCCCATGCCGTCATCAGCGGTTCCGTCCTTTACAACCAAGCCGAGAAA
GGCAGTTACTCCCTCGGTATCTTTGGCGGAAAAGCCCAGGAAGTTGCCGGCAGCGCGGAAGTGAAAACCGTAAACGGC
ATACGCCTATCGGCCTTGCCGCCAAGCAAGCAACTCGAGCACCACCACCACCACTGA
1   MATNDDDVKK AATVAIAAAY NNQQBINGFK AGETIYDIDE DGTITKKDAT
51  AADVEADDFK GLGLKKVVTN LTKTVNENKQ NVDAKVKAAE SEIEKLTTKL
101 ADTDAALADT DAALDATTNA LNKLGENITT FAEETKTNIV KIDEKLEAVA
151 DTVDKHAEAF NDIADSLDET NTKADEAVKT ANEAKQTABB TKQNVDAKVK
201 AABTAAGDAE AAAGTANTAA DKAEAVAAKV TDIKADIATN KDNIAKKANS
251 ADVTTRBBSD SKFVRIDGLN ATTKKLDTRL ASAEESIADH DTRLNLNDKT
301 VSDLRKETRQ GLABQAALSG LFQPYHVGGS GGGGVAADIG AGLADALTAP
351 LDHKDKGLQS LTLDQSVRKN KKLKLAAQGA KKTYGNGDSL NTGKLKNDKV
401 SRFDFIRQIE VDGQLITLRS GRFQVYKQSH SALTAFQTRQ IQDSEHSGKM
451  VAKRQFRIGD IAGEHTSFDK LREGGRARYB GTAFGSDDAG GKLTYTIDFA
501  AKQGNGKIEH LKSPELNVDL AAADIKPDGK RHAVISGSVL YNQAEKGSYS
551  LGIFGGKAQE VAGSAEVKTV NGIRHIGLAA KQLBHHHHHH *
961c-983
ATGGCCACAAACGACGACGATGTTAAAAAAGCTGCCACTGTGGCCATTGCTGCTGCCTACAACAATGGCCAAGAAATC
AACGGTTTCAAAGCTGGAGAGACCATCTACGACATTGATGAAGACGGCACAATTACCAAAAAAGACGCAACTGCAGCC
GATGTTGAAGCCGACGACTTTAAAGGTCTGGGTCTGAAAAAAGTCGTGACTAACCTGACCAAAACCGTCAATGAAAAC
AAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAATCTGAAATAGAAAAGTTAACAACCAAGTTAGCAGACACTGAT
GCCGCTTTAGCAGATACTGATGCCGCTCTGGATGCAACCACCAACGCCTTGAATAAATTGGGAGAAAATATAACGACA
TTTGCTGAAGAGACTAAGACAAATATCGTAAAAATTGATGAAAAATTAGAAGCCGTGGCTGATACCGTCGACAAGCAT
GCCGAAGCATTCAACGATATCGCCGATTCATTGGATGAAACCAACACTAAGGCAGACGAAGCCGTCAAAACCGCCAAT
GAAGCCAAACAGACGGCCGAAGAAACCAAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAAACTGCAGCAGGCAAA
GCCGAAGCTGCCGCTGGCACAGCTAATACTGCAGCCGACAAGGCCGAAGCTGTCGCTGCAAAAGTTACCGACATCAAA
GCTGATATCGCTACGAACAAAGATAATATTGCTAAAAAAGCAAACAGTGCCACGTGTACACCGAGAGAAGAGTCTGAC
AGCAAATTTGTCAGAATTGATGGTCTGAACGCTACTACCGAAAAATTGGACACACGCTTGGCTTCTGCTGAAAAATCC
ATTGCCGATCACGATACTCGCCTGAACGGTTTGGATAAAACAGTGTCAGACCTGCGCAAAGAAACCCGCCAAGGCCTT
GCAGAACAAGCCGCGCTCTCCGGTCTGTTCCAACCTTACAACGTGGGTGGATCCGGCGGAGGCGGCACTTCTGCGCCC
GACTTCAATGCAGGCGGTACCGGTATCGGCAGCAACAGCAGAGCAACAACAGCGAAATCAGCAGCAGTATCTTACGCC
GGTATCAAGAACGAAATGTGCAAAGACAGAAGCATGCTCTGTGCCGGTCGGGATGACGTTGCGGTTACAGACAGGGAT
GCCAAAATCAATGCCCCCCCCCCGAATCTGCATACCGGAGACTTTCCAAACCCAAATGACGCATACAAGAATTTGATC
AACCTCAAACCTGCAATTGAAGCAGGCTATACAGGACGCGGGGTAGAGGTAGGTATCGTCGACACAGGCGAATCCGTC
GGCAGCATATCCTTTCCCGAACTGTATGGCAGAAAAGAACACGGCTATAACGAAAATTACAAAAACTATACGGCGTAT
ATGCGGAAGGAAGCGCCTGAAGACGGAGGCGGTAAAGACATTGAAGCTTCTTTCGACGATGAGGCCGTTATAGAGACT
GAAGCAAAGCCGACGGATATCCGCCACGTAAAAGAAATCGGACACATCGATTTGGTCTCCCATATTATTGGCGGGCGT
TCCGTGGACGGCAGACCTGCAGGCGGTATTGCGCCCGATGCGACGCTACACATAATGAATACGAATGATGAAACCAAG
AACGAAATGATGGTTGCAGCCATCCGCAAGCATGGGTCAAGCATGGGCGAACGTGGCGTGCGCATCGTCAATAACAGT
TTTGGAACAACATCGAGGGCAGGCACTGCCGACCTTTTCCAAATAGCCAATTCGGAGGAGCAGTACCGCCAAGCGTTG
CTCGACTATTCCGGCGGTGATAAAACAGACGAGGGTATCCGCCTGATGCAACAGAGCGATTACGGCAACCTGTCCTAC
CACATCCGTAATAAAAACATGCTTTTCATCTTTTCGACAGGCAATGACGCACAAGCTCAGCCCAACACATATGCCCTA
TTGCCATTTTATGAAAAAGACGCTCAAAAAGGCATTATCACAGTAGCAGGCGTAGACCGCAGTGGAGAAAAGTTCAAA
CGGGAAATGTATGGAGAACCGGGTACAGAACCGCTTGAGTATGGCTCCAACCATTGCGGAATTACTGCCATGTGGTGC
CTGTCGGCACCCTATGAAGCAAGCGTCCGTTTCACCCGTACAAACCCGATTCAAATTGCCGGAACATCCTTTTCCGCA
CCCATCGTAACCGGCACGGCGGCTCTGCTGCTGCAGAAATACCCGTGGATGAGCAACGACAACCTGCGTACCACGTTG
CTGACGACGGCTCAGGACATCGGTGCAGTCGGCGTGGACAGCAAGTTCGGCTGGGGACTGCTGGATGCGGGTAAGGCC
ATGAACGGACCCGCGTCCTTTCCGTTCGGCGACTTTACCGCCGATACGAAAGGTACATCCGATATTGCCTACTCCTTC
CGTAACGACATTTTCAGCACGGGCGGCCTGATCAAAAAAGGCGGCAGCCAACTGCAACTGCACGGCAACAACACCTAY
ACGGGCAAAACCATTATCGAAGGCGGTTCGCTGGTGTTGTACGGCAACAACAAATCGGATATGCGCGTCGAAACCAAA
GGTGCGCTGATTTATAAVGGGGCGGVATCCGGCGGCAGCCTGAACAGCGACGGCATTGTCTATCTGGCAGATACCGAC
CAATCCGGCGCAAACGAAACCGTACACATCAAAGGCAGTCTGCAGCTGGACGGCAAAGGTACGCTGTACACACGTTTG
GGCAAACTGCTGAAAGTGGACGGTACGGCGATTATCGGCGGCAAGCTGTACATGTCGGCACGCGGCAAGGGGGCAGGC
TATCTCAACAGTACCGGACGACGTGTTCCCTTCCTGAGTGCCGCCAAAATCGGGCAGGATTATTCTTTCTTCACAAAC
ATCGAAACCGACGGCGGCCTGCTGGCTTCCCTCGACAGCGTCGAAAAAACAGCGGGCAGTGAAGGCGACACGCTGTCC
TATTATGTCCGTCGCGGCAATGCGGCACGGACTGCTTCGGCAGCGGCACATTCCGCGCCCGCCGGTCTGAAACACGCC
GTAGAACAGGGCGGCAGCAATCTGGAAAACCTGATGGTCGAACTGGATGCCCTCCGAATCATCCGAACACCCGAGACG
GTTGAAACTGCGGCAGCGACCGCACAGATATGCCGGGCATCCGCCCCTACGGCGCAACTTCTCCGCGCAGCGGCAGCC
GTACAGCATGCGAATGCCGCCGACGGTGTACGCATCTTCAACAGTCTCGCCGCTACCGTCTATGCCGACAGTACCGCC
GCCCATGCCGATATGCAGGGACGCCGCCTGAAAGCCGTATCGGACGGGTTGGACCACAACGGCACGGGTCTGCGCGTC
ATCGCGCAAACCCAACAGGACGGTGGAACGTGGGAACAGGGCGGTGTTGAAGGCAAAATGCGCGGCAGTACCCAAACC
GTCGGCATTGCCGCGAAAACCGGCGAAAATACGACAGCAGCCGCCACACTGGGCATGGGACGCAGCACATGGAGCGAA
AACAGTGCAAATGCAAAAACCGACAGCATTAGTCTGTTTGCAGGCATACGGCACGATGCGGGCGATATCGGCTATCTC
AAAGGCCTGTTCTCCTACGGACGCTACAAAAACAGCATCAGCCGCAGCACCQQTGCGGACGAACATGCGGAAGGCAGC
GTCAACGGCACGCTGATGCAGCTGGGCGCACTGGGCGGTGTCAACGTTCCGTTTGCCGCAACGGGAGATTTGACGGTC
GAAGGCGGTCTGCGCTACGACCTGCTCAAACAGGATGCATTCGCCGAAAAAGGCAGTGCTTTGGGCTGGAGCGGCAAC
AGCCTCACTGAAGGCACGCTGGTCGGACTCGCGGGTCTGAAGCTGTCGCAACCCTTGAGCGATAAAGCCGTCCTGTTT
GCAACGGCGGGCGTGGAACGCGACCTGAACGGACGCGACTACACGGTAACGGGCGGCTTTACCGGCGCGACTGCAGCA
ACCGGCAAGACGGGGGCACGCAATATGCCGCACACCCGTCTGGTTGCCGGCCTGGGCGCGGATGTCGAATTCGGCAAC
GGCTGGAACGGCTTGGCACGTTACAGCTACGCCGGTTCCAAACAGTACGGCAACCACAGCGGACGAGTCGGCGTAGGC
TACCGGTCCTCGAGCACCACCACCACCACCACTGA
1    MATNDDDCKK AATVAIAAAY NNGQBINGFK AGETIYDIDK DGTITKKDAT
51   AADVEADDFK GLGLKKCCTN LTKTVNEKNK NVKAKVKAAE SEIEKLTTKL
101  ADTDAALADT DAALDATTNA LNKLGENITT FAEETKTNIV KIDEKLKAVA
151  DTVDKHARAF NDIADSLDRT NTKADRAVKT ANBAKQTAKB TKQNVDAKVK
201  AAETAAGKAR AAAGRABTAA DKAKAVAAAV TDIKADIATN KTNIAKKANS
251  ADVYTRKESK SKFVRIDGLN ATTEKLDTRL ASAEKSIADH DTRLNGLDKT
301    VSDLRKETRQ GLARQAALSG LFQPYNVGGS GGGGTSAPDF NAGGTGIGSN
351    SRATTAKSAA VSYAGIKNEM CKDRSMLCAG RDDVAVTDRD AKINAPPPNL
401    HTGDFPNPND AYKNLINLKP AIEAGYTGRG VEVGIVDTGE SVGSISFPEL
451    YGRKEEGYNE NYKNYTAYMR KEAPEDGGGK DIEASFDDEA VIETEAKPTD
501    IRHVKEIGHI DLVSEIIGGR SVDGRPAGGI APDATLHIMN TNDETKNEMM
551    VAAIRNAWVK LGERGVRIVN NSFGTTSRAG TADLFQIANS EEQYRQALLD
601    YSGGDKTDEG IRLMQQSDYG MLSYHIRNKN MLFIFSTGND AQAQPNTYAL
651    LPFYEKDAQK GIITVAGVDR SGEKPKREMY GEPGTEPLEY GSNHCGITAM
701    WCLSAPYEAS VRFTRTNPIQ IAGTSFSAPI VTGTAALLLQ KYPWMSNDNL
751    RTTLLTTAQD IGAVGVDSKF GWGLLDAGKA MNGPASFPFG DFTADTKGTS
801    DIAYSFFNDI SGTGGLIKKG GSQLQLHGNN TYTGKTIIEG GSLVLYGNNK
851    SDMRVETKGA LIYNGAASGG SLNSDGIVYL ADTDQSGANE TVHIKGSLQL
901    DGKGTLYTRL GKLLKVDGTA IIGGKLYMSA RGKGAGYLNS TGRRVPFLSA
951    AKIGQDYSFF TNIETDGGLL ASLDSVEKTA GSEGDTLSYY VRRGNAARTA
1001   SAAAHSAPAG LKHAVEQGGS NLENLMVKLD ASESSATPET VETAAADRTD
1051   MPGIRPYGAT FRAAAAVQEA NAADGVRIFN SLAATVYADS TAAHADMQGR
1101   RLKAVSDGLD HNGTGLFVIA QTQQDGGTWE QGGVEGKHRG STQTVGIAAK
1151   TGENTTAAAT LGMGRSTWSE NSANAKTDSI SLFAGIREDA GDIGYLKGLF
1201   SYGRYKNSIS RSTGADRHAE GSVNGTLMQL GALGGVNVPF AATGDLTVEG
1251   GLRYDLLKQD AFAEKGSALG WSGNSLTEGT LVGLAGLKLS QPLSDKAVLF
1301   ATAGVERDLN GRDYTVTGGF TGATAATGKT GARNMPHTRL VAGLGADVEF
1351   GNGWNGLARY SYAGSKQYGN HSGRVGVGYR FLEHHHHHH*
961cL-ORF46.1
ATGAAACACTTTCCATCCAAAGTACTGACCACAGCCATCCTTGCCACTTTCTGTAGCGGCGCACTGGCAGCCACAAAC
GACGACGATGTTAAAAAAGCTGCCACTGTGGCCATTGCTGCTGCCTACAACAATGGCCAAGAAATCAACGGTTTCAAA
GCTGGAGAGACCATCTACGACATTGATGAAGACGGCACAATTACCAAAAAAGACGCAACTGCAGCCGATGTTGAAGCC
GACGACTTTAAAGGTCTGGGTCTGAAAAAAGTCGTGACTAACCTGACCAAAACCGTCAATGAAAACAAACAAAACGTC
GATGCCAAAGTAAAAGCTGCAGAATCTGAAATAGAAAAGTTAACAACCAAGTTAGCAGACACTGATGCCGCTTTAGCA
GATACTGATGCCCCTCTGGATGCAACCACCAACGCCTTGAATAAATTGGGAGAAAATATAACGACATTTGCTGAAGAG
ACTAAGACAAATATCGTAAAAATTGATGAAAAATTAGAAGCCGTGGCTGATACCGTCGACAAGCATGCCGAAGCATTC
AACGATATCGCCGATTCATTGGATGAAACCAACACTAAGGCAGACGAAGCCGTCAAAACCGCCAATGAAGCCAAACAG
ACGGCCGAAGAAACCAAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAAACTGCAGCAGGCAAAGCCGAAGCTGCC
GGTGGCACAGCTAATACTGCAGCCGACAAGGCCGAAGCTGTCGCTGCAAAAGTTACCGACATCAAAGCTGATATCGCT
ACGAACAAAGATAATATTGCTAAAAAAGCAAACAGTGCCGACGTGTACACCAGAGAAGAGTCTQACAGCAAATTTGTC
AGAATTGATGGTCTGAACGCTACTACCGAAAAATTGGACACACGCTTGGCTTCTGCTGAAAAATCCATTGCCGATCAC
GATACTCGCCTGAACGGTTTGGATAAAACAGTGTCAGACCTGCGCAAAGAAACCCGCCAAGGCCTTGCAGAACAAGCC
GCGCTCTCCGGTCTGTTCCAACCTTACAACGTGGGTGGATCCGGAGGAGGAGGATCAGATTTGGCAAACGATTCTTTT
ATCCGGCAGGTTCTCGACCGTCAGCATTTCGAACCCGACGGGAAATACCACCTATTCGGCAGCAGGGGGGAACTTGCC
GAGCGCAGCGGCCATATCGGATTGGGAAAAATACAAAGCCATCAGTTGGGCAACCTGATGATTCAACAGGCGGCCATT
AAAGGAAATATCGGCTACATTGTCCGCTTTTCCGATCACGGGCACGAAGTCCATTCCCCCTTCGACAACCATGCCTCA
CATTCCGATTCTGATGAAGCCGCTAGTCCCGTTGACGGATTTAGCCTTTACCGCATCCATTGGGACGGATACGAACAC
CATCCCGCCGACGGCTATGACGGGCCACAGGGCGGCGGCTATDCCCGTCCCAAAGGCGCGAGGGATATATACAGCTAC
GACATAAAAGGCGTTGCCCAAAATATCCGCCTCAACCTGACCGACAACCGCAGCACCGGACAACGGCTTGCCGACCGT
TTCCACAATGCCGGTAGTATGCTGACGCAAGGAGTAGGCGACGGATTCAAACGCGCCACCCGATACAGCCCCGAGCTG
GACAGATCGGGCAATGCCGCCGAAGCCTTCAACGGCACTGCAGATATCGTTAAAAACATCATCGGCGCGGCAGGAGAA
ATTGTCGGCGCAGGCGATGCCGTGCAGGGCATAAGCGAAGGCTCAAACATTGCTGTCATGCACGGCTTGGGTCTGCTT
TCCACCGAAAACAAGATGGCGCGCATCAACGATTTGGCAGATATGGCGCAACTCAAAGACTATGCCGCAGCAGCCATC
CGCGATTGGGCAGTCCAAAACCCCAATGCCGCACAAGGCATAGAAGCCGTCAGCAATATCTTTATGGCAGCCATCCCC
ATCAAAGGGATTGGAGCTGTTCGGGGAAAATACGGCTTGGGCGGCATCACGGCACATCCTATCAAGCGGTCGCAGATG
GGCGCGATCGCATTGCCGAAAGGGAAATCCGCCGTCAGCGACAATTTTGCCGATGCGGCATACGCCAAATACCCGTCC
CCTTACCATTCCCGAAATATCCGTTCAAACTTGGAGCAGCGTTACGGCAAAGAAAACATCACCTCCTCAACCGTGCCG
CCGTCAAACGGCAAAAATGTCAAACTGGCAGACCAACGCCACCCGAAGACAGGCGTACCGTTTGACGGTAAAGGGTTT
CCGAATTTTGAGAAGCACGTGAAATATGATACGTAACTCGAG
1      MKHFPSKVLT TAILATPCSG ALAATNDDDV KKAATVAIAA AYNNGQEING
51     FKAGETIYDI DKDGTITKKD ATAADVSADD FKGLGLKKVV TNLTKTVNEN
101    KQNVDAKVKA AESEIEKLTT KLADTDAALA DTDAALDATT NALNKLGKNI
151    TTFAEETKTN IVKIDEKLEA VADTVDKHAE AFNDIADSLD EINTKADEAV
201    KTANEAKQTA EETKQNVDAK VKAARTAAGK AEAAAGTANT AADKAEAVAA
251    KVTDIKAKIA TNKDNIAKKA KSADVYTRRR SDSKFVRIDG LNATTRKIDT
301    RLASAERSIA DHDTELNGLD KTVSDLRKET RQGLABQAAL SGLFQFYNVG
351    GSGGGGSDLA NDSFIRQVLD RQHFEPDGKY ELFGSRQELA ERSGHIGLGK
401    IQSHQLGNLM IQQAAIKGNI GYIVRFSDEG EEVESPFDNH ASHSDSDRAG
451    SPVDGFSLYR IHWDGYEEEP ADGYDGPQGG GYPAPKGARD IYSYDIEGVA
501    QNIRLNLTDN RSTGQRLADR FHNAGSMLTQ GVGDGFKRAT RYSPELDRSG
551    NAAEAFNGTA DIVKNIIGAA GEIVGAGDAV QGISRGSNIA VMHGLGLLST
601    ENKMARINDL ADMAQLKDYA AAAIRDWAVQ NPNAAQGIEA VSNIFMAAIP
651    IKGIGAVRGK YGLGGITAHP IKRSQMGAIA LPKGKSAVSD NFADAAYAKY
701    PSPYHSRNIR SNLEQRYGKE NITSSTVPPS NGKNVKLADQ RHPKTGVPFD
751    GKGFPNFEKH VKYDT*
961cL-741
ATGAAACACTTTCCATCCAAAGTACTGACCACAGCCATCCTTGCCACTTTCTGTAGCGGCGCACTGGCAGCCACAAAC
GACGACGATGTTAAAAAAGCTGCCACTGGTGGCATTGCTGCTGCCTACAACAATGGCCAAGAAATCAACGGTTTCAAA
GCTGGAGAGACCATCTACGACATTGATGAAGACGGCACAATTACCAAAAAAGACGCAACTGCAGCCGATGTTGAAGCC
GACGACTTTAAAGGTCTGGGTCTGAAAAAAGTCGTGACTAACCTGACCAAAACCGTCAATGAAAACAAACAAAACGTC
GATGCCAAAGTAAAAGCTGCAGAATCTGAAATAGAAAAGTTAACAACCAAGTTAGCAGACACTGATGCCGCTTTAGCA
GATACTGATGCCGCTCTGGATGCAACCACCAACGCCTTGAATAAATTGGGAGAAAATATAACGACATTTGCTGAAGAG
ACTAAGACAAATATCGTAAAAATTGATGAAAAATTAGAAGCCGTGGCTGATACCGTCGACAAGCATGCCGAAGCATTC
AACGATATCGCCGATTCATTGGATGAAACCAACACTAAGGCAGACGAAGCCGTCAAAACCGCCAATGAAGCCAAACAG
ACGGCCGAAGAAACCAAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAAACTGCAGCAGGCAAAGCCGAAGCTGCC
GCTGGCACAGCTAATACTGCAGCCGACAAGGCCGAAGCTGTCGCTGCAAAAGTTACCGACATCAAAGCTGATATCGCT
ACGAACAAAGATAATATTGCTAAAAAAGCAAACAGTGCCGACGTGTACACCAGAGAAGAGTCTGACAGCAAATTTGTC
AGAATTGATGGTCTGAACGCTACTACCGAAAAATTGGACACACGCTTGGCTTCTGCTGAAAAATCCATTGCCGATCAC
GATACTCGCCTGAACGGTTTGGATAAAACAGTGTCAGACCTGCGCAAAGAAACCCGCCAAGGCCTTGCAGAACAAGCC
GCGCTCTCCGGTCTGTTCCAACCTTACAACGTGGGTGGATCCGGAGGGGGTGGTGTCGCCGCCGACATCGGTGCGGGG
CTTGCCGATGCACTAACCGCACCGCTCGACCATAAAGACAAAGGTTTGCACTCTTTGACGCTGCATCAGTCCGTCAGG
AAAAACGAGAAACTGAAGCTGGCGGCACAAGGTGCGGAAAAAACTTATGGAAACGGTGACAGCCTCAATACGGGCAAA
TTGAAGAACGACAAGGTCAGCCGTTTCGACTTTATCCGCCAAATCGAAGTGGACGGGCAGCTCATTACCTTGGAGAGT
GGAGAGTTCCAAGTATACAAACAAAGCCATTCCGCCTTAACCGCCTTTCAGACCGAGCAAATACAAGATTCGGAGCAT
TCCGGGAAGATGGTTGCGAAACGCCAGTTCAGAATCGGCGACATAGCGGGCGAACATACATCTTTTGACAAGCTTCCC
GAAGGCGGCAGGGCGACATATCGCGGGACGGCGTTCGGTTCAGACGATGCCGGCGGAAAACTGACCTACACCATAGAT
TTCGCCGCCAAGCAGGGAAACGGCAAAATCGAACATTTGAAATCGCCAGAACTCAATGTCGACCTGGCCGCCGCCGAT
ATCAAGCCGGATGGAAAACGCCATGCCCTCATCAGCGGTTCCGTCCTTTACAACCAAGCCGAGAAAGGCAGTTACTCC
CTCGGTATCTTTGGCGGAAAAGCCCAGGAAGTTGCCGGCAGCGCGGAAGTGAAAACCGTAAACGGCATACGCCATATC
GGCCTTGCCGCCAAGCAACTCGAGCACCACCACCACCACCACTGA
1      MKHFPSKVLT TAILATFCSG ALAATNDDDV KKAATVAIAA AYNNGQEING
51     FKAGETIYDI DEDGTITKKD ATAADVEADD FKGLGLKKVV TNLTKTVNEN
101    KQNVDAKVKA AESEIEKLII KLADTDAALA DTDAALDATT NALNKLGENI
151    TTFAEETKTN IVKIDKKLEA VADTVDKHAR AFNDIADSLD ETNTKADEAV
201    KTANEAKQTA EETKQNVDAK VKAAETAAGK AEAAAGTANT AADKAEAVAA
251    KVTDIKADIA TNKDNIAKKA NSADVYTREE SDSKFVRIDG LNATTEKLDT
301    RLASAEKSIA DHDTRLNGLD KTVSDLRKET RQGLAEQAAL SGLFQPYNVG
351    GSGGGGVAAD IGAGLADALT APLDHKDKGL QSLTLDQSVR KNEKLKLAAQ
401    GAEKTYGNGD SLNTGKLKND KVSRFDFIRQ IEVDGQLITL RSGEFQVYKQ
451    SHSALTAFQT EQIQDSEHSG KMVAKRQFRI GDLAGKHTSF DKLPRGGRAT501    YRGTAFGSDD AGGKLTYTID FAAKQGNGKI EHLKSPELNV DLAAADIKPD
551    GKRHAVISGS VLYNQAEKGS YSLGIFGGKA QKVAGSAEVK TVNGIRHIGL
601    AAKQLEHHHH HH*
961cL-983
ATGAAACACTTTCCATCCAAAGTACTGACCACAGCCATCCTTGCCACTTTCTGTAGCGGCGCACTGGCAGCCACAAAC
GACGACGATGTTAAAAAAGCTGCCACTGTGGCCATTGCTGCTGCCTACAACAATGGCCAAGAAATCAACGGTTTCAAA
GCTGGAGAGACCATCTACGACATTGATGAAGACGGCACAATTACCAAAAAAGACGCAACTGCAGCCGATGTTGAAGCC
GACGACTTTAAAGGTCTGGGTCTGAAAAAAGTCGTGACTAACCTGACCAAAACCGTCAATGAAAACAAACAAAACGTC
GATGCCAAAGTAAAAGCTGCAGAATCTGAAATAGAAAAGTTAACAACCAAGTTAGCAGACACTGATGCCGCTTTAGCA
GATGCTGATGCCGCTCTGGATGCAACCACCAACGCCTTGAATAAATTGGGAGAAAATATAACGACATTTGCTGAAGAG
ACTAAGACAAATATCGTAAAAATTGATGAAAAATTAGAAGCCGTGGCTGATACCGTCGACAAGCATGCCGAAGCATTC
AACGATATCGCCGATTCATTGGATGAAACCAACACTAAGGCAGACGAAGCCGTCAAAACCCCCAATGAAGCCAAACAG
ACGGCCGAAGAAACCAAACAAAACGTCGATGCCAAAGTAAAAGCTGCAGAAACTGCAGCAGGCAAAGCCGAAGCTGCC
GCTGGCACAGCTAATACTGCAGCCGACAAGGCCGAAGCTGTCGCTGCAAAAGTTACCGACATCAAAGCTGATATCGCT
ACGAACAAAGATAATATTGCTAAAAAAGCAAACAGTGCCGACGTGTACACCAGAGAAGAGTCTGACAGCAAATTTGTC
AGAATTGATGGTCTGAACGCTACTACCGAAAAATTGGACACACGCTTGGCTTCTGCTGAAAAATCCATTGCCGATCAC
GATACTCGCCTGAACGGTTTGGATAAAACAGTGTCAGACCTGCGCAAAGAAACCCGCCAAGGCCTTGCAGAACAAGCC
GCGCTCTCCGGTCTGTTCCAACCTTACAACGTGGGTGGATCCGGCGGAGGCGGCACTTCTGCGCCCGACTTCAATGCA
GGCGGTACCGGTATCGGCAGCAACAGCAGAGCAACAACAGCGAAATCAGCAGCAGTATCTTACGCCGGTATCAAGAAC
GAAATGTGCAAAGACAGAAGCATGCTCTGTGCCGGTCGGGATGACGTTGCGGTTACAGACAGGGATGCCAAAATCAAT
GCCCCCCCCCCGAATCTGCATACCGGAGACTTTCCAAACCCAAATGACGCATACAAGAATTTGATCAACCTCAAACCT
GCAATTGAAGCAGGCTATACAGGACGCGGGGTAGAGGTAGGTATCGTCGACACAGGCGAATCCGTCGGCAGCATATCC
TTTCCCGAACTGTATGGCAGAAAAGAACACGGCTATAACGAAAATTACAAAAACTATACGGCGTATATGCGGAAGGAA
GCGCCTGAAGACGGAGGCGGTAAAGACATTGAAGCTTCTTTCGACGATGAGGCCGTTATAGAGACTGAAGCAAAGCCG
ACGGATATCCGCCACGTAAAAGAAATCGGACACATCGATTTGGTCTCCCATATTATTGGCGGGCGTTCCGTGGACGGC
AGACCTGCAGGCGGTATTGCGCCCGATGCGACGCTACACATAATCAATACGAATGATGAAACCAAGAACGAAATGATG
GTTGCAGCCATCCGCAATGCATGGGTCAAGCTGGGCGAACGTGGCGTGCGCATCGTCAATAACAGTTTTGGAACAACA
TCGAGGGCAGGCACTGCCGACCTTTTCCAAATAGCCAATTCGGAGGAGCAGTACCGCCAAGCGTTGCTCGACTATTCC
GGCGGTGATAAAACAGACGAGGGTATCCGCCTGATGCAACAGAGCGATTACGGCAACCTGTCCTACCACATCCGTAAT
AAAAACATGCTTTTCATCTTTTCGACAGGCAATGACGCACAAGCTCAGCCCAACACATATGCCCTATTGCCATTTTAT
GAAAAAGACGCTCAAAAAGGCATTATCACAGTCGCAGGCGTAGACCGCAGTGGAGAAAAGTTCAAACGGGAAATGTAT
GGAGAACCGGGTACAGAACCGCTTGAGTATCCCTCCAACCATTGCGGAATTACTGCCATGTGGTGCCTGTCGGCACCC
TATGAAGCAAGCGTCCGTTTCACCCGTACAAACCCGATTCAAATTGCCGGAACATCCTTTTCCGCACCCATCGTAACC
GGCACGGCGGCTCTGCTGCTGCAGAAATACCCGTGGATGAGCAACGACAACCTGCGTACCACGTTGCTGACGACGGCT
CAGGACATCGGTGCAGTCGGCGTGGACAGCAAGTTCGGCTGGGGACTGCTGGATGCGGGTAAGGCCATGAACGGACCC
GCGTCCTTTCCGTTCGGCGACTTTACCGCCGATACGAAAGGTACATCCGATATTGCCTACTCCTTCCGTAACGACATT
TCAGGCACGGGCGGCCTGATCAAAAAAGGCGGCAGCCAACTGCAACTGCACGGCAACAACACCTATACGGGCAAAACC
ATTATCGAAGGCGGTTCGCTGGTGTTGTACGGCAACAACAAATCGGATATGCGCGTCGAAACCAAAGGTGCGCTGATT
TATAACGGGGCGGCATCCGGCGGCAGCCTGAACAGCGACGGCATTGTCTATCTGGCAGATACCGACCAATCCGGCGCA
AACGAAACCGTACACATCAAAGGCAGTCTGCAGCTGGACGGCAAAGGTACGCTGTACACACGTTTGGGCAAACTGCTG
AAAGTGGACGGTACGGCGATTATCGGCGGCAAGCTGTACATGTCGGCACGCGGCAAGGGGGCAGGCTATCTCAACAGT
ACCGGACGACGTGTTCCCTTCCTGAGTGCCGCCAAAATCGGGCAGGATTATTCTTTCTTCACAAACATCGAAACCGAC
GGCGGCCTGCTGGCTTCCCTCGACAGCGTCGAAAAAACAGCGGGCAGTGAAGGCGACACGCTGTCCTATTATGTCCGT
CGCGGCAATGCGGCACGGACTGCTTCGGCAGCGGCACATTCCGCGCCCGCCGGTCTGAAACACGCCGTAGAACAGGGC
GGCAGCAATCTGGAAAACCTGATGGTCGAACTGGATGCCTCCGAATCATCCGCAACACCCGAGACGGTTGAAACTGCG
GCAGCCGACCGCACAGATATGCCGGGCATCCGCCCCTACGGCGCAACTTTCCGCGCAGCGGCAGCCGTACAGCATGCG
AATGCCGCCGACGGTGTACGCATCTTCAACAGTCTCGCCGCTACCGTCTATGCCGACAGTACCGCCGCCCATGCCGAT
ATGCAGGGACGCCGCCTGAAAGCCGTATCGGACGGGTTGGACCACAACGGCACGGGTCTGCGCGTCATCGCGCAAACC
CAACAGGACGGTGGAACGTGGGAACAGGGCGGTGTTGAAGGCAAAATGCGCGGCAGTACCCAAACCGTCGGCATTGCC
GCGAAAACCGGCGAAAATACGACAGCAGCCGCCACACTGGGCATGGGACGCAGCACATGGAGCGAAAACAGTGCAAAT
GCAAAAACCGACAGCATTAGTCTGTTTGCAGGCATACGGCACGATGCGGGCGATATCGGCTATCTCAAAGGCCTGTTC
TCCTACGGACGCTACAAAAACAGCATCAGCCGCAGCACCGGTGCGGACGAACATGCGGAAGGCAGCGTCAACGGCACG
CTGATGCAGCTGGGCGCACTGGGCGGTGTCAACGTTCCGTTTGCCGCAACGGGAGATTTGACGGTCGAAGGCGGTCTG
CGCTACGACCTGCTCAAACAGGATGCATTCGCCGAAAAAGGCAGTGCTTTGGGCTGGAGCGGCAACAGCCTCACTGAA
GGCACGCTGGTCGGACTCGCGGGTCTGAAGCTGTCGCAACCCTTGAGCGATAAAGCCGTCCTGTTTGCAACGGCGGGC
GTGGAACGCGACCTGAACGGACGCGACTACACGGTAACGGGCGGCTTTACCGGCGCGACTGCAGCAACCGGCAACACG
GGGGCACGCAATATGCCGCACACCCGTCTGGTTGCCGGCCTGGGCGCGGATGTCGAATTCGGCAACGGCTGGAACGGC
TTGGCACGTTACAGCTACGCCGGTTCCAAACAGTACGGCAACCACAGCGGACGACTCGGCGTAGGCTACCGGTTCTGA
CTCGAG
1    MKHFPSKVLT TAILATFCSG ALAATNDDDV KKAATVAIAA AYNNGQEING
51   FKAGBTIYDI DKDGTITKKD ATAADVRADD FKGLGLKKVV TNLTKTVNEN
101  KQNVDAKVKA ARSEIBKLTT KLADTDAALA DTDAALDATT NALNKLGRNI
151  TTFAEETKTN IVKIDEKLRA VADTVTRHAB AFNDIADSLD BTNTKADEAV
201  KTANRAKQTA RBTKQNVDAK VKAAETAAGK ABAAAGTANT AADKABAVAA
25l  KVTDIKADIA TNKINIAKKA NBADVYTRBB BDSKFVRIDG LNATTRKLDT
301  RLASAEKSIA DHDTRLNGLD KTVSDLRKET RQGLQBQAAL SGLFQPYNVG
351  GSGGGGTSAP DFNAGGTGIG SNSRATTAKS AAVSYAGIKN KMCKDRSMLC
401  AGRDDVAVTD RDARINAPPP NLHTGDFPNP NDAYKNLINL KPAIBAGYTG
451  RGVEVGIVDT GRSVGSISFP ELYGRKRHGY NKNYKNYTAY MRKRAPRDGG
501  GKDIEASFDD EAVIBTBAKP TDRRHVKBIG HIDLVSHIIG GRSVDGRPAG
551  GIAPDATLHT MNTNDFTKNB MMVAAIRNAW VKIGRRGVRI VNNSFGTTSR
601  AGTADLFQIA NSBBQYRQAL LDYSGGDKTD EGIRLMQQSD YGNLSYEIRN
651  KNMLFIFSTG NDAQAQPNTY ALLPFYEKDA QKGIITVAGV DRSGRKFKRB
701  MYGEPGTEPL EYGSNBCGIT AMWCLSAPYB ASVRFTRTNP IQIAGTSFSA
751  PIVTGTAALL LQKYPWMSND NLRTTLLTTA QDIQAVGVDS KFGWGLLDAG
801  KAMNGPASFP FGDFTADTKG TSDIAYSFRN DISGTGGLIK KGGSQLQLHG
851  NNTYTGKTII EGGSLVLYGN NKSDNRVETR GAIIYNGAAS GGSINSDGIV
901  YLADTDQSGA NETVHIKGSL QLDGKGTLYT RIGKLLKVDG TAIIGGKLYM
95l  SARGKGAGYL NSTGRRVPPL SAAKIGQDYS FFTNIBTDGG LLASLDSVRR
1001 TAGSEGLYLS YYVRRGNAAR TASAAAESAP AGLKEQVBAG GSNLENLMVE
1051 LDASESSATP ETVETAAADR TDNPGIRPYG ATFRAAAAVQ EANAADGVRI
1101 FNSLAATVYA DSTAAHAIMQ GRRLKAVSDG LDEVGTGLRV IAQTQQGGGT
1151 LDASESSATP ETVETAAADR TDMPGIRPYG ATFRAAAAVQ SRNSANAKTD
1201 SISLFAGIRE DAGDIGYLKG LFSYGRYKNS ISRSTGADRE AEGSVNGTLM
1251 QLGALGGVNV PFAATGDLTV EGGLRYDLLR QDAFAEKGSA LGWSGNSLTE
1301 GTLVGLAGLK LSQPLSDKAV LFATAGVERD LNGRDYTVTG GFTGATAATG
1351 KTGARNMPHT RLVAGLGADV EFGNGWNGLA RYSYAGSKQY GNHBGRVGVG
1401 YRF*
可以理解本发明仅以实施例的方式进行描述，在本发明的范围和精神内还可进行改变。例如，设想可以使用其它菌株的蛋白质[如，参见WO00/66741，ORF4、ORF40、ORF46、225、235、287、519、726、919和953的多态序列]。
实验详述
克隆策略和寡核苷酸设计
用以脑膜炎奈瑟球菌B MC58的基因组序列为基础设计的寡核苷酸，通过PCR扩增编码感兴趣的抗原的基因。除非特别指出，通常将菌株2996的基因组DNA用作PCR反应的模板，将扩增的片段克隆入表达载体pET21b+(Novagen)，从而以C-未端His标记的产物形式表达该蛋白，或将其克隆入pET-24b+(Novagen)以‘未标记的′形式(如ΔG287K)表达该蛋白。
不用融合配体和用其自身前导肽(如果存在时)表达蛋白质，进行开放读框(ATG到终止密码子)的扩增。
当蛋白质以‘未标记的′的形式表达时，通过从预定的前导序列设计5′-端扩增引物下游除去前导肽。
用于PCR的引物的解链温度取决于整个引物中杂交核苷酸的数量和类型，并用以下公式确定：
Tm1＝4(G+C)+2(A+T)              (除去尾部的)
Tm2＝64.9+0.41(％GC)-600/N      (完整的引物)
对整个低聚物而言，所选寡核苷酸的解链温度通常为65-70℃，仅针对杂交区域，解链温度为50-60℃。
用Perkin Elmer 394 DNA/RNA合成仪合成寡核苷酸，将其从柱上洗脱到2.0ml NH4OH中，在56℃培育5小时以去保护。加入0.3M乙酸钠和2体积的乙醇沉淀寡聚物。将样品离心，将沉淀物重悬浮于水中。
 序列  限制酶切位点
 fu(961)-  Fwd  CGC GGATCC-GGAGGGGGTGGTGTCG  BamHI
 741(MC58)-His
 Rev  CCCG CTCGAG-TTGCTTGGCGGCAAGGC  XhoI
 fu(961)-983-His  Fwd  CGC GGATCC-GGCGGAGGCCGCACTT  BamHI
 Rev  CCCG CTCGAG-GAACCGGTAGCCTACG  XhoI
fu(961)-Orf46.1-His  Fwd  CGC GGATCCGGTGGTGGTGGT-TCAGATTTGGCAAACGATTC  BamHI
 Rev  CCCG CTCGAG-CGTATCATATTTCACGTGC  XhoI
fu(961c-L)-741(MC58)  Fwd  CGC GGATCC-GGAGGGGGTGGTGTCG  BamHI
 Rev  CCCG CTCGAG-TTATTGCTTGGCGGCAAG  XhoI
fu(961c-L)-983  Fwd  CGC GGATCC-GGCGGAGGCGGCACTT  BamHI
 Rev  CCCG CTCGAG-TCAGAACCGGTAGCCTAC  XhoI
fu(961c-L)-Orf46.1  Fwd  CGC GGATCCGGTGGTGGTGGT-TCAGATTTGGCAAACGATTC  BamHI
 Rev  CCCG CTCGAG-TTACGTATCATATTTCACGTGC  XhoI
fu-(ΔG287)-919-His  Fwd  CGC GGATCCGGTGGTGGTGGT-CAAAGCAAGAGCATCCAAACC  BamHI
 Rev  CCC AAGCTT-TTCGGGCGGTATTCGGGCTTC  HindIII
fu-(ΔG287)-953-His  Fwd  CGC GGATCCGGTGGTGGTGGT-GCCACCTACAAAGTGGAC  BamHI
 Rev  GCCC AAGCTT-TTGTTTGGCTGCCTCGAT  HindIII
fu-(ΔG287)-961-His  Fwd  CGC GGATCCGGTGGTGGTGGT-ACAAGCGACGACG  BamHI
 Rev  GCCC AAGCTT-CCACTCGTAATTGACGCC  HindIII
fu-(ΔG287)-Orf46.1-His  Fwd  CGC GGATCCGGTGGTGGTGGT-TCAGATTTGGCAAACGATTC  BamHI
 Rev  CCC AAGCTT-CGTATCATATTTCACGTGC  HindIII
fu-(ΔG287-919)-Orf46.1-His  Fwd  CCC AAGCTTGGTGGTGGTGGTGGT-TCAGATTTGGCAAACGATTC  HindIII
 Rev  CCC GCTCGAG-CGTATCATATTTCACGTGC  XhoI
fu-(ΔG287-Orf46.1)-919-His  Fwd  CCC AAGCTTGGTGGTGGTGGTGGT-CAAAGCAAGAGCATCCAAACC  HindIII
 Rev  CCC GCTCGAG-CGGGCGGTATTCGGGCTT  XhoI
fuΔG287(394.98)-…  Fwd  CGCGGATCC GCTAGC-CCCGATGTTAAATCGGC  NheI
 Rev  CGG GGATCC-ATCCTGCTCTTTTTTGCCGG  BamHI
fu Orf1-(Orf46.1)-His  Fwd  CGCGGATCC GCTAGC-GGACACACTTATTTCGGCATC  NheI
 Rev  CGCGGATCC-CCAGCGGTAGCCTAATTTGAT
fu(OrF1)-Orf46.1-His  Fwd  CGC GGATCCGGTGGTGGTGGT-TCAGATTTGGCAAACGATTC  BamHI
 Rev  CCC AAGCTT-CGTATCATATTTCACGTGC  HindIII
fu(919)-Orf46.1-His  Fwd1  GCGGC GTCGACGGTGGCGGAGGCACTGGATCCTCAG  SalI
 Fwd2  GGAGGCACTGGATCCTCAGATTTGGCAAACGATTC
 Rev  CCC GCTCGAG-CGTATCATATTTCACGTGC  XhoI
Fu(orf46)-287-His  Fwd  CGG GGATCCGGGGGCGGCGGTGGCG  BamHI
 Rev  CCC AAGCTTATCCTGCTCTTTTTTGCCGGC  HindIII
Fu(orf46)-919-His  Fwd  CGC GGATCCGGTGGTGGTGGTCAAAGCAAGAGCATCCAAACC  BamHI
 Rev  CCC AAGCTTCGGGCGGTATTCGGGCTTC  HindIII
 Fu(orf46-919)-287-His  Fwd  CCCC AAGCTTGGGGGCGGCGGTGGCG  HindIII
 Rev  CCCG CTCGAGATCCTGCTCTTTTTTGCCGGC  XhoI
 Fu(orf46-287)-919-His  Fwd  CCC AAGCTTGGTGGTGGTGGTGGTCAAAGCAAGAGCATCCAAACC  HindIII
 Rev  CCCG CTCGAGCGGGCGGTATTCGGGCTT  XhoI
 (ΔG741)-961c-His  Fwd1Fwd2  GGAGGCACTGGATCCGCAGCCACAAACGACGACGAGCGGC CTCGAG-GGTGGCGGAGGCACTGGATCCGCAG  XhoI
 Rev  CCCG CTCGAG-ACCCAGCTTGTAAGGTTG  XhoI
 (ΔG741)-961-His  Fwd1Fwd2  GGAGGCACTGGATCCGCAGCCACAAACGACGACGAGCGGC CTCGAG-GGTGGCGGAGGCACTGGATCCGCAG  XhoI
 Rev  CCCG CTCGAG-CCACTCGTAATTGACGCC  XhoI
 (ΔG741)-983-His  Fwd  GCGGC CTCGAG-GGATCCGGCGGAGGCGGCACTTCTGCG  XhoI
 Rev  CCCG CTCGAG-GAACCGGTAGCCTACG  XhoI
 (ΔG741)-orf46.1-His  Fwd1Fwd2  GGAGGCACTGGATCCTCAGATTTGGCAAACGATTCGCGGC GTCGACGGTGGCGGAGGCACTGGATCCTCAGA  SalI
 Rev  CCCG CTCGAG-CGTATCATATTTCACGTGC  XhoI
 (ΔG98.3)-741(MC58)-His  Fwd  GCGGC CTCGAG-GGATCCGGAGGGGGTGGTGTCGCC  XhoI
 Rev  CCCG CTCGAG-TTGCTTGGCGGCAAG  XhoI
 (ΔG983)-961c-His  Fwd1Fwd2  GGAGGCACTGGATCCGCAGCCACAAACGACGACGAGCGGC CTCGAG-GGTGGCGGAGGCACTGGATCCGCAG  XhoI
 Rev  CCCG CTCGAG-ACCCAGCTTGTAAGGTTG  XhoI
 (ΔG983)-961-His  Fwd1Fwd2  GGAGGCACTGGATCCGCAGCCACAAACGACGACGAGCGGC CTCGAG-GGTGGCGGAGGCACTGGATCCGCAG  XhoI
 Rev  CCCG CTCGAG-CCACTCGTAATTGACGCC  XhoI
 (ΔG983)-Orf46.1-His  Fwd1Fwd2  GGAGGCACTGGATCCTCAGATTTGGCAAACGATTCGCGGC GTCGACGGTGGCGGAGGCACTGGATCCTCAGA  SalI
 Rev  CCCG CTCGAG-CGTATCATATTTCACGTGC  XhoI
*将该引物用作反向引物，将所有287的C未端融合于His-标记。
§与287-His反向引物联用的正向引物。
NB-所有PCR反应使用菌株2996，除非特别指出(如菌株MC58)。
在所有以ATG起始不跟随唯一NheI位点的构建物中，ATG密码子是用于克隆的NdeI位点的一部分。在5′端用NheI作为克隆位点制备的构建物(如所有那些在N-未端包含287的)另有两个融合于抗原的编码序列的密码子(GCTAGC)。
染色体DNA模板的制备
在100ml GC培养基中使脑膜炎奈瑟球菌前株2996、MC58、394.98、1000和BZ232(及其它)生长至指数期，离心收获，并重悬浮于5ml缓冲液(20％w/v蔗糖、50mM Tris-HCl、50mM EDTA、pH8)中。在冰上培育10分钟后，加入10ml裂解液(50mM NaCl、1％十二烷基肌氨酸钠(Na-Sarkosyl)、50μg/ml蛋白酶K)裂解细菌，悬浮液在37℃培育2小时。进行2次苯酚提取(平衡至pH8)和一次CHCl3/异戊醇(24∶1)提取。加入0.3M乙酸钠和2体积乙醇沉降DNA，并离心收集。用70％(v/v)乙醇洗涤沉淀物1次，并重溶解于4.0ml TE缓冲液(10mM Tris-HCl、1mM EDTA、pH 8.0)。读取OD260测定DNA浓度。
PCR扩增
标准的PCR过程：进行如下在40μM各寡核苷酸引物、400-800μM dNTP溶液、1×PCR缓冲液(包含1.5mM MgCl2)、2.5单位TaqI DNA聚合酶(用Perkin-Elmer AmpliTaQ，Boerhingher Mannheim ExpandTM长模板)存在时，将200ng 2996、MC581000、或BZ232菌株的基因组DNA或10ng重组克隆的质粒DNA制备物用作模板。
将整个混合物在95℃初步培育3分钟后，每份样品进行2-步的扩增：用除去引物(Tm1)的限制酶尾部的杂交温度进行前5轮。然后按全长低聚物(Tm2)计算的杂交温度进行30轮。根据要扩增的Orf的长度，在68℃或72℃进行的延伸时间各不相同。对Orf1而言，自3分钟开始的延伸时间每轮递增15秒。以在72℃10分钟延伸步骤完成循环。
将扩增的DNA直接加到1％琼脂糖凝胶上。按制造商的说明，用Qiagen凝胶提取试剂盒纯化相应于正确大小条带的DNA片段。
PCR片段和克隆载体的消化
用适合的限制酶消化相应于扩增片段的纯化DNA，从而克隆入pET-21b+、pET22b+、或pET-24b+。用QIAquick PCR纯化试剂盒(按制造商的说明)纯化消化的片段，用H2O或10mM Tris(pH 8.5)洗脱。用适合的限制酶消化质粒载体，加到1.0％琼脂糖凝胶上，用Qiagen AIQquick凝胶提取试剂盒纯化相应于消化的载体的条带。
克隆
将预先消化和纯化的、相应于各基因的片段连接到pET21b+、pET22b+或pET-24b+中。将在连接缓冲液(由制造商提供)中的T4 DNA连接酶用于摩尔比为3∶1的片段/载体。
通过在冰上培育连接酶反应溶液和细菌40分钟，然后在37℃培育3分钟，将重组质粒转化入感受态大肠杆菌DH5或HB101中。
然后添加800μl LB肉汤，并在37℃培育20分钟。在Eppendorf微型离心机中以最高速度离心这些细胞，并重悬浮于约200μl的上清液中，并涂布在LB氨苄青霉素(100mg/ml)琼脂上。
在4.0ml LB肉汤+100μg/ml氨苄青霉素中培育随机选择的集落过夜，进行重组集落的筛选。使细胞沉淀，并按制造商的说明用Qiagen QIAprep SpinMiniprep试剂盒提取质粒DNA。用适合的限制酶消化约1μg的各微型制备物，将消化物加到1-1.5％琼脂糖凝胶(取决于预计的***大小)与分子量标记(1kbDNA Ladder，GIBCO)平行。根据***的大小选择阳性克隆。
表达
各基因克隆入表达载体后，将重组质粒转化入适合表达重组蛋白的大肠杆菌菌株中。如上所述，用1μl各构建物转化大肠杆菌BL21-DE3。将单重组集落接种入2ml LB+Amp(100μg/ml)中，在37℃培育过夜，然后在100ml烧瓶中用20ml LB+Amp(100μg/ml)以1∶30稀释，使OD600在0.1-0.2之间。将烧瓶置于旋转式水浴摇床中于30℃或37℃培育，直到OD600显示适合诱导表达的指数生长期(0.4-0.8OD)。加入1.0mM IPTG诱导蛋白质表达。在30℃或37℃培育3小时后，测定OD600并检测表达。用微型离心机离心1.0ml各样品，将沉淀物重悬浮于PBS中，用SDS-PAGE和考马斯蓝染色分析。
His-标记的蛋白质的纯化
从菌株2996和MC58中克隆了287的各种形式。用C-未端His标记的融合体进行构建，其包括成熟形式(aa18-427)、含缺失(Δ1、Δ2、Δ3和Δ4)的构建物和由B或C结构域组成的克隆。对以His-融合体纯化的各克隆而言，划线接种单集落，并且在37℃LB/Amp(100μg/ml)琼脂板上培育过夜。将从该平板上分离的集落接种到20ml LB/Amp(100μg/ml)的液体培养基中，并在37℃振荡生长过夜。以1∶30将过夜培养物稀释到1.0L LB/Amp(100μg/ml)液体培养基中，让其在最佳温度(30或37℃)生长，直到OD550达到0.6-0.8。添加IPTG(终浓度为1.0mM)诱导重组蛋白的表达，再培育培养物3小时。于4℃，以8000g离心15分钟收获细菌。将细菌沉淀物重悬浮于7.5ml(i)冷缓冲液A(300mMNaCl、50mM磷酸盐缓冲液、10mM咪唑、pH 8.0)，用于可溶性蛋白质；或(ii)缓冲液B(10mM Tirs-HCl、100mM磷酸盐缓冲液，pH 8.8和任选地8M尿素)，用于不溶性蛋白质。以可溶形式纯化的蛋白质包括287-His、Δ1、Δ2、Δ3和Δ4287-His、Δ4287MC58-His、287c-His和287cMC58-His。蛋白质287bMC58-His是不溶的并相应地纯化。用Branson Sonifier 450，在冰上以40W、30秒超声处理破坏细胞4次，并于4℃以13000xg离心30分钟。对于不溶蛋白质，将沉淀物重悬浮于2.0ml缓冲液C(6M盐酸胍、100mM磷酸盐缓冲液、10mM Tris-HCl、pH 7.5)，并用Dounce匀浆器处理10次。以13000g离心匀浆30分钟并保留上清液。将可溶的和不溶制备物的上清液与150μl Ni2+-树脂(预先用缓冲液A或缓冲液B平衡)混合，并在室温温和振荡培育30分钟。树脂是按制造商说明制备的Chelating Sepharose Fast Flow(Pharmcia)。于4℃，以700g离心分批制备物5分钟，弃去上清液。用10ml缓冲液A或B洗涤树脂2次(分批)10分钟，重悬浮于1.0ml缓冲液A或B中，加到一次性柱上。用(i)缓冲液A(4℃)或(ii)缓冲液B(室温)持续洗涤树脂，直到流出物的OD280达到0.02-0.01。再用(i)冷缓冲液C(300mM NaCl、50mM磷酸盐缓冲液、20mM咪唑、pH 8.0)或(ii)缓冲液D(10mM Tris-HCl、100mM磷酸盐缓冲液、pH 6.3和任选地8M尿素)进一步洗涤树脂，直到流出物的OD280达到0.02-0.01。加入700μl(i)冷洗脱缓冲液A(300mM NaCl、50mM磷酸盐缓冲液、250mM咪唑、pH 8.0)或(ii)洗脱缓冲液B(10mM Tris-HCl、100mM磷酸盐缓冲液、pH 4.5和任选地8M尿素)洗脱His-融合蛋白，收集组分直到OD280显示获得了所有重组蛋白。用SDS-PAGE分析20μl量的各洗脱组分。用Bradford试验法计算蛋白质浓度。
变性的His-融合蛋白的复性
需要变性以稳定287bMC8，因此在免疫接种前需进行复性步骤。将甘油加到上述得到的变性组分中，使终浓度为10％v/v。用透析缓冲液I(10％v/v甘油，0.5M精氨酸、50mM磷酸盐缓冲液、5.0mM还原的谷胱甘肽、0.5mM氧化的谷胱甘肽、2.0M尿素，pH 8.8)将蛋白质稀释至200μg/ml，用相同的缓冲液在4℃透析12-14小时。于4℃，用缓冲液II(10％v/v甘油，0.5M精氨酸、50mM磷酸盐缓冲液、5.0mM还原的谷胱甘肽、0.5mM氧化的谷胱甘肽、pH 8.8)再进行透析12-14小时。用以下公式计算蛋白质的浓度：
蛋白质(mg/ml)＝(1.55×OD280)-(0.76×OD260)
免疫接种
在第0、21和35天，用抗原免疫接种Balb/C小鼠，在第49天分析血清。
血清分析-ELISA
将不包囊的MenB M7和包囊的菌株置于巧克力琼脂板上，在37℃、5％CO2中培育过夜。用无菌Dracon刷从琼脂板上收集细菌菌落，并接种到含有0.25％葡萄糖的Mueller-Hinton肉汤(Difco)中。每30分钟监测一次细菌的生长，随后测定OD620。让细菌生长直到OD达到0.4-0.5。将培养物以4000rpm离心10分钟。弃去上清液，用PBS洗涤细菌2次，重悬浮于含有0.025％甲醛的PBS中，并在37℃培育1小时，然后在4℃搅拌培育过夜。在96孔Greiner板的各孔中添加100μl细菌细胞，并在4℃培育过夜。然后用PBT洗涤缓冲液(0.1％Tween-20的PBS溶液)冲洗这些孔3次。在各孔中加入200μl饱和缓冲液(2.7％聚乙烯吡咯烷酮10的水溶液)，将这些平板在37℃培育2小时。用PBT冲洗这些孔3次。在各孔中加入200μl稀释的血清(稀释缓冲液：1％BSA、0.1％Tween-20、0.1％NaN3的PBS溶液)，将这些平板在37℃培育2小时。用PBT冲洗这些孔3次。在各孔中加入100μl HRP-缀合的兔抗-小鼠(Dako)血清(用稀释缓冲液以1∶2000稀释)，将这些平板置于37℃培育90分钟。用PBT缓冲液洗涤这些孔3次。在各孔中加入100μl HRP的底物缓冲液(25ml柠檬酸缓冲液pH 5，10mg邻-苯二胺(phenildiamine)和10μl H2O2)，并将平板在室温中静置20分钟。在各孔中加入100μl 12.5％H2SO4，随后测定OD490。计算ELISA滴定度，即高于预先免疫血清稀释度的、OD490值为0.4的血清稀释度。当OD490值为0.4的血清稀释度高于1∶400时，将ELISA视为阳性。
血清分析-FACS扫描细菌结合分析
将不包囊的MenB M7菌株置于巧克力琼脂板上，在37℃、5％CO2中培育过夜。用无菌Dracon刷从琼脂板上收集菌落，并接种到4支装有0.25％葡萄糖的8ml Mueller-Hinton肉汤(Difco)试管中。每30分钟监测细菌的生长，然后测定OD620。让细菌生长直到OD达到0.35-0.5。将培养物以4000rpm离心10分钟。弃去上清液，用封阻缓冲液(1％BSA的PBS溶液，0.4％NaN3)重悬浮沉淀物，并以4000rpm离心5分钟。将细胞重悬浮于封阻缓冲液，使OD620达到0.05。在96孔Costar板的各孔中添加100μl细菌细胞。在各孔中加入100μl稀释的(1∶100、1∶200、1∶400)血清(以封阻缓冲液配制)，并将这些平板在4℃培育2小时。以4000rpm离心细胞5分钟，吸出上清液，在各孔中加入200μl/孔封阻缓冲液洗少涤细胞。在各孔中加入100μl R-Phicoerytrin缀合的F(ab)2山羊抗-小鼠(以1∶100稀释)，将平板置于4℃培育1小时。以4000rpm离心5分钟以沉淀细胞，添加200μl/孔封阻缓冲洗涤细胞。吸出上清液，细胞重悬浮于200μl/孔PBS、0.25％甲醛。将样品转移到FACScan管中并读数。FACScan(LaserPower 15mW)的条件设定为：FL2开；FSC-H阈值：92；FSC PMT电压：E01；SSCPMT：474；Amp.Gains 6.1；FL-2 PMT：586；补偿值：0。
血清分析-杀菌试验
于37℃，在5％CO2、在巧克力琼脂板(以冷冻原液起始)上培育脑膜炎奈瑟球菌菌株2996过夜。收集菌落，并将其接种到7ml含有0.25％葡萄糖的Mueller-Hinton肉汤中，使OD620达到0.05-0.08。在37℃振荡培育约1.5小时，直到OD620达到0.23-0.24。用50mM磷酸盐缓冲液(pH7.2，含10mM MgCl2、10mMCaCl2和0.5％(w/v)BSA(分析缓冲液))以105CFU/ml工作稀释度稀释细菌。最终反应混合物的总体积为50μl，其中25μl连续2倍稀释的测试血清，12.5μl工作稀释度的细菌，12.5μl幼兔补体(终浓度25％)。
对照包括：用补体血清培育的细菌、用细菌培育并在56℃加热30分钟补充灭活的免疫血清。在加入幼兔补体后，用斜置方法立即将10μl对照置于Mueller-Hinton琼脂板上(0时间)。37℃旋转培育96孔板1小时。将每份样品的7μl涂布在Mueller-Hinton琼脂板上作为斑点，而用斜置方法将10μl对照涂布在Mueller-Hinton琼脂板上(1时间)。37℃培育琼脂板18小时，计算相应于0时间和1时间的菌落数量。
血清分析-Western印迹法
将纯化的蛋白质(500ng/泳道)、外膜小泡(5μg)和MenB菌株2996衍生的全细胞提取物(25μg)加到12％SDS-聚丙烯酰胺凝胶上，并转移到硝基纤维素膜上。用转化缓冲液(0.3％Tris碱，1.14％甘氨酸、20％(v/v)甲醇)在4℃、150mA进行转化2小时。于4℃，在饱和缓冲液(10％脱脂乳、0.1％Triton X100的PBS溶液)中培育过夜使膜饱和。用冲洗缓冲液(3％脱脂乳、0.1％Triton X100的PBS溶液)冲洗膜2次，并在37℃与用洗涤缓冲液以1∶200稀释的小鼠血清一起培育2小时。冲洗膜2次，与1∶2000稀释的辣根过氧化物酶标记的抗-小鼠Ig一起培育90分钟。用0.1％Triton X100的PBS溶液冲洗膜2次，用Opti-4CN底物试剂盒(Bio-Rad)显色。加入水终止反应。
如下制备OMV：于37℃、5％CO2，在5个GC板上让脑膜炎奈瑟球菌2996生长过夜，用接种环收获，并重悬浮于10ml 20mM Tris-HCl pH 7.5、2mM EDTA中。在56℃热灭活45分钟，在冰上超声处理破碎细胞5分钟(50％负载循环(dutycycle)、50％输出，Branson超声仪3mm微型针头)。以5000g离心10分钟除去未被破坏的细胞，回收含有完整细胞包膜组分的上清液，于4℃以50000g进一步离心过夜。将含有膜的沉淀物重悬浮于2％二烷基肌氨酸钠、20mM Tris-HClpH7.5、2mM EDTA中，在室温培育20分钟以溶解内膜。以10000g离心上清液10分钟除去聚集体，以50000g进一步离心上清液3小时。用PBS冲洗含有外膜的沉淀物，并重悬浮于相同的缓冲液中。用BSA作为标准，由D.C.Bio-Rad蛋白分析(改进的Lowry方法)测定蛋白质的浓度。
如下制备全细胞提取物：使脑膜炎奈瑟球菌在GC板上培育过夜，用接种环收获，并重悬浮于1ml 20mM Tris-HCl中。在56℃热灭活30分钟。

Claims (18)

1.式NH2-A-B-COOH的杂交蛋白，其特征在于，所述的A和B是不同的奈瑟球菌蛋白，选自orf1、orf4、orf25、orf40、ORF461-433、ORF46433-608、ORF46、Orf46.1、orf83、97、225、233、287、ΔG287、Δ1-287、Δ2-287、Δ3-287、Δ4-287、287B、287C、287BC、292L、519、564、687、741、ΔG741、907、919、953、961c、961cL和ΔG983，其中蛋白质的氨基酸序列如下限定：(a)“orf1”定义为WO99/24578的SEQ ID NO：650，“orf4”定义为WO99/24578的SEQ IDNO：218，“orf25”定义为WO99/24578的SEQ ID NO：684，“orf40”定义为WO99/36544的SEQ ID NO：4，“orf46.1”定义为SEQ ID NO：18的氨基酸1058-1466，“orf83”定义为WO99/24578的SEQ ID NO：314，“233”定义为WO99/57280的SEQ ID NO：862，“ΔG287”定义为SEQ ID NO：2，“292L”定义为WO99/57280的SEQ ID NO：1220，“564”定义为WO99/57280的SEQ IDNO：1670，“687”定义为WO99/57280的SEQ ID NO：2282，“ΔG741”定义为SEQ ID NO：26，“907”定义为WO99/57280的SEQ ID NO：2732，“919”定义为SEQ ID NO：4的氨基酸412-832，“953”定义为SEQID NO：6的氨基酸412-579，“961c”定义为SEQ ID NO：48的氨基酸2-327，和“ΔG983”定义为SEQ ID NO：16。
2.权利要求1所述的蛋白质，其特征在于，所述的A和B各选自：ORF46.1、287、741、919、953、961和983。
3.如权利要求1所述的蛋白质，其特征在于，所述的A和/或B具有聚-甘氨酸缺失。
4.如权利要求3所述的蛋白质，其特征在于，所述的A和/或B是ΔG287、ΔG741或ΔG983。
5.如权利要求1所述的蛋白质，其特征在于，所述的A和/或B是Δ1-287、Δ2-287、Δ3-287或Δ4-287。
6.如权利要求1所述的蛋白质，其特征在于，所述的A和/或B是287B、287C、287BC、ORF461-433、ORF46433-608、ORF46或961c。
7.如权利要求1所述的蛋白质，其特征在于，所述的A和B是：(a)919和287；(b)953和287；(c)287和ORF46.1；(d)ORF1和ORF46.1；(e)919和ORF46.1；(f)ORF46.1和919；(g)919和519；或(h)ORF97和225。
8.如权利要求1所述的蛋白质，其特征在于，所述的蛋白质是ΔG287-919、ΔG287-953、ΔG287-961、ΔG983-ORF46.1、ΔG983-741、ΔG983-961、ΔG983-961C、ΔG741-961、ΔG741-961C、ΔG741-983、ΔG741-ORF46.1、ORF46.1-741、ORF46.1-961、ORF46.1-961C、961-ORF46.1、961-741、961-983、961C-ORF46.1、961C-741、961C-983、961CL-ORF46.1、961CL-741或961CL-983。
9.如权利要求1所述的蛋白质，其特征在于，所述的A或B是287。
10.如权利要求9所述的蛋白质，其特征在于，所述的B是287。
11.如权利要求9所述的蛋白质，其特征在于，所述的A是ΔG287。
12.如权利要求11所述的蛋白质，其特征在于，所述的B是ORF46、919、953或961。
13.如权利要求9所述的蛋白质，其特征在于，所述的287来自菌株2996或394/98。
14.如权利要求2所述的蛋白质，其特征在于，所述的A是961。
15.如权利要求1所述的蛋白质，其特征在于，所述的A和B来自相同的菌株。
16.如权利要求1所述的蛋白质，其特征在于，所述的A和B直接相连。
17.如权利要求1所述的蛋白质，其特征在于，所述的A和B通过接头肽相连。
18.如权利要求17所述的蛋白质，其特征在于，所述的接头肽是聚-甘氨酸接头，但条件是B不是聚甘氨酸缺失的蛋白。
CNB2003101036036A 2000-02-28 2001-02-28 奈瑟球菌蛋白质的杂交表达 Expired - Fee Related CN100334214C (zh)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
GB0004695.3 2000-02-28
GB0004695A GB0004695D0 (en) 2000-02-28 2000-02-28 Protein expression
GB0027675.8 2000-11-13
GB0027675A GB0027675D0 (en) 2000-11-13 2000-11-13 Protein Expression
PCT/IB2001/000420 WO2001064920A2 (en) 2000-02-28 2001-02-28 Hybrid expression of neisserial proteins

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
CNB018087388A Division CN100339482C (zh) 2000-02-28 2001-02-28 奈瑟球菌蛋白质的杂交表达

Publications (2)

Publication Number Publication Date
CN1508253A CN1508253A (zh) 2004-06-30
CN100334214C true CN100334214C (zh) 2007-08-29

Family

ID=26243750

Family Applications (7)

Application Number Title Priority Date Filing Date
CNB2003101028453A Expired - Fee Related CN100473663C (zh) 2000-02-28 2001-02-28 奈瑟球菌蛋白质的异源表达
CNB2003101036036A Expired - Fee Related CN100334214C (zh) 2000-02-28 2001-02-28 奈瑟球菌蛋白质的杂交表达
CNB018087388A Expired - Fee Related CN100339482C (zh) 2000-02-28 2001-02-28 奈瑟球菌蛋白质的杂交表达
CN200510116305XA Expired - Fee Related CN1800385B (zh) 2000-02-28 2001-02-28 奈瑟球菌蛋白质的异源表达
CN2007101391568A Expired - Fee Related CN101139590B (zh) 2000-02-28 2001-02-28 奈瑟球菌蛋白质的杂交表达
CNB018087507A Expired - Fee Related CN1201011C (zh) 2000-02-28 2001-02-28 奈瑟球菌蛋白质的异源表达
CN2010101656630A Pending CN101906413A (zh) 2000-02-28 2001-02-28 奈瑟球菌蛋白质的异源表达

Family Applications Before (1)

Application Number Title Priority Date Filing Date
CNB2003101028453A Expired - Fee Related CN100473663C (zh) 2000-02-28 2001-02-28 奈瑟球菌蛋白质的异源表达

Family Applications After (5)

Application Number Title Priority Date Filing Date
CNB018087388A Expired - Fee Related CN100339482C (zh) 2000-02-28 2001-02-28 奈瑟球菌蛋白质的杂交表达
CN200510116305XA Expired - Fee Related CN1800385B (zh) 2000-02-28 2001-02-28 奈瑟球菌蛋白质的异源表达
CN2007101391568A Expired - Fee Related CN101139590B (zh) 2000-02-28 2001-02-28 奈瑟球菌蛋白质的杂交表达
CNB018087507A Expired - Fee Related CN1201011C (zh) 2000-02-28 2001-02-28 奈瑟球菌蛋白质的异源表达
CN2010101656630A Pending CN101906413A (zh) 2000-02-28 2001-02-28 奈瑟球菌蛋白质的异源表达

Country Status (20)

Country Link
US (9) US20040092711A1 (zh)
EP (6) EP1947187B9 (zh)
JP (8) JP4763210B2 (zh)
CN (7) CN100473663C (zh)
AT (3) ATE387502T1 (zh)
AU (4) AU2001239488B2 (zh)
BR (2) BR0108711A (zh)
CA (5) CA2400562C (zh)
CY (6) CY1106532T1 (zh)
DE (3) DE60126249T2 (zh)
DK (4) DK1947187T5 (zh)
ES (5) ES2360746T3 (zh)
HK (4) HK1064120A1 (zh)
LU (1) LU92240I2 (zh)
MX (2) MXPA02008313A (zh)
NL (1) NL300605I2 (zh)
NZ (2) NZ521531A (zh)
PT (4) PT1790660E (zh)
RU (2) RU2304617C2 (zh)
WO (2) WO2001064920A2 (zh)

Families Citing this family (86)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1860192A1 (en) 1996-09-17 2007-11-28 Novartis Vaccines and Diagnostics, Inc. Compositions and methods for treating intracellular diseases
CA2317815A1 (en) * 1998-01-14 1999-07-22 Chiron S.P.A. Neisseria meningitidis antigens
ES2304065T3 (es) 1998-05-01 2008-09-01 Novartis Vaccines And Diagnostics, Inc. Antigenos y composiciones de neisseria meningitidis.
US20070026021A1 (en) * 1998-05-01 2007-02-01 Chiron S.R.I. Neisseria meningitidis antigens and compositions
ES2228454T3 (es) * 1999-02-26 2005-04-16 Chiron S.R.L. Mejora de la actividad bacteriana de antigenos de neisseria con oligonucleotidos que contienen motivos cg.
NZ581940A (en) 1999-04-30 2011-07-29 Novartis Vaccines & Diagnostic Conserved neisserial antigens
GB9911683D0 (en) * 1999-05-19 1999-07-21 Chiron Spa Antigenic peptides
AU783894B2 (en) * 1999-05-19 2005-12-22 Novartis Vaccines And Diagnostics S.R.L. Combination neisserial compositions
RU2281956C2 (ru) 1999-10-29 2006-08-20 Чирон С.Р.Л. Антигенные пептиды neisseria
GB9928196D0 (en) 1999-11-29 2000-01-26 Chiron Spa Combinations of B, C and other antigens
DK2289545T3 (en) * 2000-01-17 2016-09-05 Glaxosmithkline Biologicals Sa Supplemented OMV vaccine against meningococcus
CN100473663C (zh) 2000-02-28 2009-04-01 启龙股份公司 奈瑟球菌蛋白质的异源表达
ATE455793T1 (de) * 2001-04-17 2010-02-15 Novartis Vaccines & Diagnostic Monoklonale antikörper gegen molekulare mimetika von meningokokken-b-epitopen
GB0115176D0 (en) 2001-06-20 2001-08-15 Chiron Spa Capular polysaccharide solubilisation and combination vaccines
US6928463B1 (en) * 2001-07-06 2005-08-09 Nortel Networks Limited Broadband content delivery via personal content tunnel
GB0118249D0 (en) 2001-07-26 2001-09-19 Chiron Spa Histidine vaccines
GB0121591D0 (en) 2001-09-06 2001-10-24 Chiron Spa Hybrid and tandem expression of neisserial proteins
JP4592284B2 (ja) * 2001-07-27 2010-12-01 カイロン ソチエタ ア レスポンサビリタ リミタータ 髄膜炎菌付着因子
AU2014201962B2 (en) * 2001-09-06 2016-06-09 Novartis Vaccines And Diagnostics S.R.L. Hybrid and tandem expression of Neisserial proteins
AR045702A1 (es) 2001-10-03 2005-11-09 Chiron Corp Composiciones de adyuvantes.
US7838015B2 (en) 2001-10-03 2010-11-23 Novartis Vaccines And Diagnostics, Inc. Adjuvanted meningococcus compositions
EP1438323A4 (en) * 2001-10-03 2007-08-01 Novartis Vaccines & Diagnostic ADJUVOUS MENINGOCOCCUS COMPOSITIONS
MX339524B (es) * 2001-10-11 2016-05-30 Wyeth Corp Composiciones inmunogenicas novedosas para la prevencion y tratamiento de enfermedad meningococica.
JP2006500963A (ja) 2002-08-02 2006-01-12 グラクソスミスクライン バイオロジカルズ ソシエテ アノニム ワクチン組成物
US7785608B2 (en) 2002-08-30 2010-08-31 Wyeth Holdings Corporation Immunogenic compositions for the prevention and treatment of meningococcal disease
AU2003274511B2 (en) * 2002-10-11 2009-06-04 Glaxosmithkline Biologicals S.A. Polypeptide-vaccines for broad protection against hypervirulent meningococcal lineages
WO2004039417A2 (en) 2002-11-01 2004-05-13 Glaxosmithkline Biologicals S.A. Drying process
DK2279746T3 (da) 2002-11-15 2013-11-25 Novartis Vaccines & Diagnostic Overfladeproteiner i neisseria meningitidis
GB0227346D0 (en) * 2002-11-22 2002-12-31 Chiron Spa 741
WO2004067030A2 (en) 2003-01-30 2004-08-12 Chiron Srl Injectable vaccines against multiple meningococcal serogroups
WO2005019265A1 (en) 2003-08-13 2005-03-03 Chiron Corporation Improved method of purifying tfpi and tfpi analogs
GB0323103D0 (en) 2003-10-02 2003-11-05 Chiron Srl De-acetylated saccharides
ES2397923T3 (es) 2003-10-02 2013-03-12 Novartis Ag Vacunas líquidas para múltiples serogrupos meningocócicos
GB0408977D0 (en) * 2004-04-22 2004-05-26 Chiron Srl Immunising against meningococcal serogroup Y using proteins
GB0409748D0 (en) 2004-04-30 2004-06-09 Chiron Srl Lactoferrin cleavage
GB0419408D0 (en) * 2004-09-01 2004-10-06 Chiron Srl 741 chimeric polypeptides
GB0424092D0 (en) 2004-10-29 2004-12-01 Chiron Srl Immunogenic bacterial vesicles with outer membrane proteins
MX291624B (es) 2005-02-18 2011-11-04 Novartis Vaccines & Diagnostic Inmunogenos de escherichia coli uropatogenica.
AU2006214064B2 (en) 2005-02-18 2012-04-26 J. Craig Venter Institute, Inc. Proteins and nucleic acids from meningitis/sepsis-associated escherichia coli
CN100475965C (zh) * 2005-07-22 2009-04-08 上海高科联合生物技术研发有限公司 一种大肠杆菌高效外分泌表达溶葡萄球菌酶的方法
GB0524066D0 (en) 2005-11-25 2006-01-04 Chiron Srl 741 ii
WO2007066226A2 (en) * 2005-12-06 2007-06-14 Universita Degli Studi Di Padova Methods and compositions relating to adhesins as adjuvants
JP2010500399A (ja) 2006-08-16 2010-01-07 ノバルティス アーゲー 尿路病原性大腸菌由来の免疫原
AR064642A1 (es) 2006-12-22 2009-04-15 Wyeth Corp Polinucleotido vector que lo comprende celula recombinante que comprende el vector polipeptido , anticuerpo , composicion que comprende el polinucleotido , vector , celula recombinante polipeptido o anticuerpo , uso de la composicion y metodo para preparar la composicion misma y preparar una composi
GB0700562D0 (en) 2007-01-11 2007-02-21 Novartis Vaccines & Diagnostic Modified Saccharides
GB0713880D0 (en) 2007-07-17 2007-08-29 Novartis Ag Conjugate purification
PT2200642E (pt) 2007-10-19 2012-05-30 Novartis Ag Formulações de vacinas meningocócicas
CA2716212A1 (en) 2008-02-21 2009-08-27 Novartis Ag Meningococcal fhbp polypeptides
PT2268618E (pt) 2008-03-03 2015-10-12 Novartis Ag Compostos e composições moduladores da actividade tlr
US20100105875A1 (en) * 2008-06-09 2010-04-29 Maria Scarselli Antibodies against neisserial factor H binding protein
JP5487463B2 (ja) * 2008-08-08 2014-05-07 独立行政法人産業技術総合研究所 非拡散植物ウイルスベクター
CA2756522C (en) 2009-03-24 2018-06-26 Novartis Ag Adjuvanting meningococcal factor h binding protein
WO2010144734A1 (en) 2009-06-10 2010-12-16 Novartis Ag Benzonaphthyridine-containing vaccines
ES2596653T3 (es) 2009-06-16 2017-01-11 Glaxosmithkline Biologicals Sa Ensayos bactericidas de opsonización y dependientes de anticuerpo mediado por el complemento de alto rendimiento
JP2013502918A (ja) 2009-08-27 2013-01-31 ノバルティス アーゲー 髄膜炎菌ｆＨＢＰ配列を含むハイブリッドポリペプチド
AU2010290896B2 (en) 2009-09-02 2014-07-03 Glaxosmithkline Biologicals S.A. Immunogenic compositions including TLR activity modulators
TWI445708B (zh) 2009-09-02 2014-07-21 Irm Llc 作為ｔｌｒ活性調節劑之化合物及組合物
US20130022639A1 (en) 2009-09-30 2013-01-24 Novartis Ag Expression of meningococcal fhbp polypeptides
EP2493499A1 (en) 2009-10-27 2012-09-05 Novartis AG Modified meningococcal fhbp polypeptides
WO2011057148A1 (en) 2009-11-05 2011-05-12 Irm Llc Compounds and compositions as tlr-7 activity modulators
AU2010339921B2 (en) 2009-12-15 2016-08-11 Glaxosmithkline Biologicals S.A. Homogeneous suspension of immunopotentiating compounds and uses thereof
WO2011119759A1 (en) 2010-03-23 2011-09-29 Irm Llc Compounds (cystein based lipopeptides) and compositions as tlr2 agonists used for treating infections, inflammations, respiratory diseases etc.
US10478483B2 (en) 2010-06-25 2019-11-19 Glaxosmithkline Biologicals Sa Combinations of meningococcal factor H binding proteins
RU2580620C2 (ru) 2010-08-23 2016-04-10 ВАЙЕТ ЭлЭлСи СТАБИЛЬНЫЕ КОМПОЗИЦИИ АНТИГЕНОВ Neisseria meningitidis rLP2086
PE20140173A1 (es) 2010-09-10 2014-02-20 Wyeth Llc Variantes no lipidadas de antigenos orf2086 de neisseria meningitidis
AU2011300418B2 (en) 2010-09-10 2016-05-12 Glaxosmithkline Biologicals Sa Meningococcus overexpressing NadA and/or NHBA and outer membrane vesicles derived therefrom
GB201102090D0 (en) * 2011-02-08 2011-03-23 Univ Sheffield Antigenic polypeptide
SG11201400199XA (en) 2011-08-31 2014-03-28 Children S Hospital & Res Ct Oakland Engineered sequences to facilitate expression of antigens in neisseria and methods of use
MY198910A (en) 2012-03-09 2023-10-02 Pfizer Neisseria meningitidis compositions and methods thereof
SA115360586B1 (ar) 2012-03-09 2017-04-12 فايزر انك تركيبات لعلاج الالتهاب السحائي البكتيري وطرق لتحضيرها
JP2015521595A (ja) 2012-06-14 2015-07-30 ノバルティス アーゲー 血清群ｘ髄膜炎菌のためのワクチン
CN104736563A (zh) 2012-07-27 2015-06-24 国家健康与医学研究院 Cd147作为受体用于脑膜炎球菌至血管内皮的菌毛介导的粘附
WO2014084432A1 (ko) * 2012-11-30 2014-06-05 경상대학교 산학협력단 헬리코박터 파일로리 발현 벡터
EP2964665B1 (en) 2013-03-08 2018-08-01 Pfizer Inc Immunogenic fusion polypeptides
MX369534B (es) 2013-09-08 2019-11-11 Pfizer Composiciones de neisseria meningitidis y sus metodos.
SG11201606478YA (en) 2014-02-28 2016-09-29 Glaxosmithkline Biolog Sa Modified meningococcal fhbp polypeptides
CN107249626A (zh) 2015-02-19 2017-10-13 辉瑞大药厂 脑膜炎奈瑟球菌组合物及其方法
GB201522153D0 (en) 2015-12-15 2016-01-27 Univ Southampton Meningococcal infection and modified neisseria lactamica
AU2017321039B2 (en) 2016-09-02 2021-03-18 Glaxosmithkline Biologicals Sa Vaccines for Neisseria gonorrhoeae
EP3312192B1 (en) * 2016-10-24 2023-02-22 BiOMVis Srl Immunogenic compositions containing bacterial outer membrane vesicles
SG11201906519RA (en) 2017-01-31 2019-08-27 Pfizer Neisseria meningitidis compositions and methods thereof
CN110516550B (zh) * 2019-07-26 2022-07-05 电子科技大学 一种基于fpga的车道线实时检测方法
TW202245836A (zh) 2021-02-19 2022-12-01 美商賽諾菲巴斯德公司 重組ｂ型腦膜炎球菌疫苗
GB202208089D0 (en) 2022-06-01 2022-07-13 Glaxosmithkline Biologicals Sa Immunogenic composition
GB202208093D0 (en) 2022-06-01 2022-07-13 Glaxosmithkline Biologicals Sa Immunogenic composition
WO2024030931A1 (en) 2022-08-03 2024-02-08 Sanofi Pasteur Inc. Adjuvanted immunogenic composition against neisseria meningitidis b

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992016643A1 (en) * 1991-03-14 1992-10-01 Imclone Systems Incorporated Recombinant hybrid porin epitopes
WO1999036544A2 (en) * 1998-01-14 1999-07-22 Chiron S.P.A. Neisseria meningitidis antigens

Family Cites Families (129)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4239749A (en) * 1979-09-27 1980-12-16 United States Of America Neisseria gonorrhoeae vaccine
US5288641A (en) * 1984-06-04 1994-02-22 Arch Development Corporation Herpes Simplex virus as a vector
CA1282721C (en) 1984-06-04 1991-04-09 Bernard Roizman Herpes simplex virus as a vector
EP0273116A3 (en) 1986-10-09 1990-05-02 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. Gonococcal and meningococcal polypeptides, vaccines and diagnostics
GB8702816D0 (en) 1987-02-07 1987-03-11 Al Sumidaie A M K Obtaining retrovirus-containing fraction
US5270176A (en) * 1987-11-20 1993-12-14 Hoechst Aktiengesellschaft Method for the selective cleavage of fusion proteins with lysostaphin
CA1340506C (en) * 1987-11-24 1999-04-20 Nicholas H. Carbonetti Production of gonorrheal pi proteins and vaccines
US5591624A (en) * 1988-03-21 1997-01-07 Chiron Viagene, Inc. Retroviral packaging cell lines
CN1038306A (zh) 1988-03-21 1989-12-27 维吉恩公司 重组反转录病毒
US5422120A (en) * 1988-05-30 1995-06-06 Depotech Corporation Heterovesicular liposomes
AP129A (en) 1988-06-03 1991-04-17 Smithkline Biologicals S A Expression of retrovirus gag protein eukaryotic cells
AU640118B2 (en) 1988-12-19 1993-08-19 De Staat Der Nederlanden Vertegenwoordigd Door De Minister Van Welzijn, Volksgezonheid En Cultuur Meningococcal class 1 outer-membrane protein vaccine
NL8803111A (nl) 1988-12-19 1990-07-16 Nederlanden Staat Multivalent meningococcen klasse i buitenmembraaneiwit vaccin.
US5703055A (en) * 1989-03-21 1997-12-30 Wisconsin Alumni Research Foundation Generation of antibodies through lipid mediated DNA delivery
DE69032284T2 (de) 1989-03-21 1998-10-08 Vical, Inc., San Diego, Calif. Expression von exogenen polynukleotidsequenzen in wirbeltieren
GB8919607D0 (en) 1989-08-30 1989-10-11 Wellcome Found Novel entities for cancer therapy
CU22302A1 (es) * 1990-09-07 1995-01-31 Cigb Secuencia nucleotidica codificante para una proteina de la membrana externa de neisseria meningitidis y uso de dicha proteina en preparados vacunales
IE912559A1 (en) 1990-07-19 1992-01-29 Merck & Co Inc The class ii protein of the outer membrane of neisseria¹meningitidis, and vaccines containing same
WO1992005266A2 (en) 1990-09-21 1992-04-02 Viagene, Inc. Packaging cells
US5965424A (en) * 1991-01-11 1999-10-12 Boehringer Mannheim Gmbh Methods for making neisseria or hemophilus IgA protease and DNA encoding the proteases
FR2681786A1 (fr) 1991-09-27 1993-04-02 Centre Nat Rech Scient Vecteurs recombinants d'origine virale, leur procede d'obtention et leur utilisation pour l'expression de polypeptides dans des cellules musculaires.
IL103059A0 (en) 1991-09-30 1993-02-21 Boehringer Ingelheim Int Conjugates for introducing nucleic acid into higher eucaryotic cells
NZ244306A (en) 1991-09-30 1995-07-26 Boehringer Ingelheim Int Composition for introducing nucleic acid complexes into eucaryotic cells, complex containing nucleic acid and endosomolytic agent, peptide with endosomolytic domain and nucleic acid binding domain and preparation
US6100380A (en) * 1991-10-28 2000-08-08 Cytran, Inc. Immunomodulating peptides and methods of use
EP0625049A4 (en) 1992-01-23 1995-07-12 Vical Inc EX VIVO GENTRANSFER.
FR2692592B1 (fr) * 1992-06-19 1995-03-31 Pasteur Merieux Serums Vacc Fragments d'ADN codant pour les sous-unités du récepteur de la transferrine de Neisseria meningitidis et procédés les exprimant.
ATE307200T1 (de) * 1992-07-07 2005-11-15 Fuso Pharmaceutical Ind Sonde zur diagnose einer ansteckenden krankheit verursacht durch staphylococcus epidermidis
US6592873B1 (en) 1992-10-30 2003-07-15 Iowa State University Research Foundation, Inc. Polynucleic acids isolated from a porcine reproductive and respiratory syndrome virus (PRRSV) and proteins encoded by the polynucleic acids
US5785974A (en) * 1993-01-23 1998-07-28 Immunologia Y Genetica Aplicada, S.A. Synthetic peptides and vaccines against parvovirus
US5439808A (en) * 1993-07-23 1995-08-08 North American Vaccine, Inc. Method for the high level expression, purification and refolding of the outer membrane group B porin proteins from Neisseria meningitidis
AU7408494A (en) * 1993-07-30 1995-02-28 University Of North Carolina At Chapel Hill, The Production of gonorrheal pi proteins and vaccines in e. coli and salmonella
US5914254A (en) 1993-08-02 1999-06-22 Celtrix Pharmaceuticals, Inc. Expression of fusion polypeptides transported out of the cytoplasm without leader sequences
US5510264A (en) 1993-09-28 1996-04-23 Insight Biotech Inc. Antibodies which bind meningitis related homologous antigenic sequences
NZ276305A (en) 1993-11-16 1997-10-24 Depotech Corp Controlled release vesicle compositions
JP4303315B2 (ja) 1994-05-09 2009-07-29 オックスフォード バイオメディカ（ユーケー）リミテッド 非交差性レトロウイルスベクター
FR2720408B1 (fr) 1994-05-31 1996-08-14 Pasteur Merieux Serums Vacc Fragments Tbp2 de Neisseria meningitidis.
US6245337B1 (en) 1994-08-25 2001-06-12 Washington University Haemophilus adherence and penetration proteins
IL117483A (en) 1995-03-17 2008-03-20 Bernard Brodeur MENINGITIDIS NEISSERIA shell protein is resistant to proteinase K.
US5646259A (en) 1995-03-24 1997-07-08 St. Louis University DNA encoding haemophilus adhesion proteins
US6265567B1 (en) 1995-04-07 2001-07-24 University Of North Carolina At Chapel Hill Isolated FrpB nucleic acid molecule
WO1996040718A1 (en) * 1995-06-07 1996-12-19 Connaught Laboratories, Inc. Expression of lipoproteins
DE19534579C2 (de) 1995-09-18 2000-06-08 Max Planck Gesellschaft Nucleinsäure-Moleküle codierend Proteine, die die Adhäsion von Neisseria-Zellen an humane Zellen vermitteln
PT854729E (pt) 1995-09-18 2004-08-31 Us Army Med Res Mat Com Usamrm Metodos melhorados para a producao de vacinas de subunidade multivalentes de proteossomas complexados nao covalentemente
FR2739624B1 (fr) 1995-10-10 1997-12-05 Pasteur Merieux Serums Vacc Nouvelle sous-unite tbp2 de neisseria meningitidis
ZA9610456B (en) * 1995-12-20 1997-06-20 Novo Nordisk As N-terminally extended proteins expressed in yeast
EP0877816A1 (en) * 1996-02-01 1998-11-18 North American Vaccine, Inc. Expression of group b neisseria meningitidis outer membrane (mb3) protein from yeast and vaccines
US6083499A (en) * 1996-04-19 2000-07-04 Mycogen Corporation Pesticidal toxins
US6472518B1 (en) 1996-10-24 2002-10-29 Centers For Disease Control And Prevention, As Represented By The Secretary, Department Of Health And Human Services Invasion associated genes from Neisseria meningitidis serogroup B
US5980898A (en) 1996-11-14 1999-11-09 The United States Of America As Represented By The U.S. Army Medical Research & Material Command Adjuvant for transcutaneous immunization
JP2001515467A (ja) 1996-12-20 2001-09-18 ボード・オヴ・リージェンツ，ザ・ユニヴァーシティ・オヴ・テキサス・システム モラクセラ・カタラーリスのｕｓｐａ１及びｕｓｐａ２抗原
US6017531A (en) 1997-06-02 2000-01-25 W. R. Grace & Co. Hydrophilic composition containing protease produced by Vibrio
US6583275B1 (en) * 1997-07-02 2003-06-24 Genome Therapeutics Corporation Nucleic acid sequences and expression system relating to Enterococcus faecium for diagnostics and therapeutics
AU9363798A (en) * 1997-11-06 1999-05-31 Chiron S.P.A. Neisserial antigens
US6914131B1 (en) * 1998-10-09 2005-07-05 Chiron S.R.L. Neisserial antigens
GB9726398D0 (en) * 1997-12-12 1998-02-11 Isis Innovation Polypeptide and coding sequences
US7635486B1 (en) * 1998-02-03 2009-12-22 The United States Of America As Represented By The Department Of Health And Human Services Recombinant lipidated PsaA protein, methods of preparation and use
WO1999041230A1 (en) * 1998-02-11 1999-08-19 Eli Lilly And Company Processes and intermediates useful to make antifolates
GB9808734D0 (en) 1998-04-23 1998-06-24 Smithkline Beecham Biolog Novel compounds
GB9808866D0 (en) 1998-04-24 1998-06-24 Smithkline Beecham Biolog Novel compounds
US6150502A (en) * 1998-04-29 2000-11-21 Genesis Research & Development Corporation Limited Polypeptides expressed in skin cells
US20070026021A1 (en) * 1998-05-01 2007-02-01 Chiron S.R.I. Neisseria meningitidis antigens and compositions
ES2304065T3 (es) 1998-05-01 2008-09-01 Novartis Vaccines And Diagnostics, Inc. Antigenos y composiciones de neisseria meningitidis.
US6615024B1 (en) * 1998-05-01 2003-09-02 Arraycomm, Inc. Method and apparatus for determining signatures for calibrating a communication station having an antenna array
US5990085A (en) * 1998-05-04 1999-11-23 Michigan State University Inhibin-HBc fusion protein
GB9810276D0 (en) 1998-05-13 1998-07-15 Smithkline Beecham Biolog Novel compounds
US6248329B1 (en) * 1998-06-01 2001-06-19 Ramaswamy Chandrashekar Parasitic helminth cuticlin nucleic acid molecules and uses thereof
AU1202200A (en) * 1998-10-09 2000-05-01 Chiron Corporation Neisseria genomic sequences and methods of their use
EP1144645A1 (en) 1999-01-15 2001-10-17 SMITHKLINE BEECHAM BIOLOGICALS s.a. Neisseria meningitidis polypeptide basb052
EP1147194A1 (en) 1999-01-22 2001-10-24 SMITHKLINE BEECHAM BIOLOGICALS s.a. Neisseria meningitidis antigenic polypeptides, corresponding polynucleotides and protective antibodies
GB9902084D0 (en) 1999-01-29 1999-03-24 Smithkline Beecham Biolog Novel compounds
ES2228454T3 (es) * 1999-02-26 2005-04-16 Chiron S.R.L. Mejora de la actividad bacteriana de antigenos de neisseria con oligonucleotidos que contienen motivos cg.
NZ581940A (en) * 1999-04-30 2011-07-29 Novartis Vaccines & Diagnostic Conserved neisserial antigens
BR0010361A (pt) 1999-04-30 2003-06-10 Chiron Corp Seq ências genÈmicas de neisseria e uso destas
GB9911683D0 (en) * 1999-05-19 1999-07-21 Chiron Spa Antigenic peptides
AU783894B2 (en) * 1999-05-19 2005-12-22 Novartis Vaccines And Diagnostics S.R.L. Combination neisserial compositions
GB9916529D0 (en) * 1999-07-14 1999-09-15 Chiron Spa Antigenic peptides
RU2281956C2 (ru) * 1999-10-29 2006-08-20 Чирон С.Р.Л. Антигенные пептиды neisseria
RU2002117308A (ru) * 1999-11-29 2004-03-10 Чирон Спа (It) 85 кДа АНТИГЕН NEISSERIA
GB9928676D0 (en) 1999-12-03 2000-02-02 Provalis Uk Ltd Pseudomonas aeruginosa antigens
US20010031268A1 (en) * 1999-12-17 2001-10-18 Baldwin Thomas John Antigen preparations
DK2289545T3 (en) * 2000-01-17 2016-09-05 Glaxosmithkline Biologicals Sa Supplemented OMV vaccine against meningococcus
KR100698561B1 (ko) 2000-01-25 2007-03-21 더 유니버서티 어브 퀸슬랜드 Ｎｅｉｓｓｅｒｉａ Ｍｅｎｉｎｇｉｔｉｄｉｓ 표면 항원ΝｈｈΑ의 보존 부위를 포함하는 단백질
CN100473663C (zh) * 2000-02-28 2009-04-01 启龙股份公司 奈瑟球菌蛋白质的异源表达
WO2001081581A2 (en) 2000-04-21 2001-11-01 Corixa Corporation Compositions and methods for the therapy and diagnosis of acne vulgaris
AU2001276619A1 (en) 2000-07-03 2002-01-14 Chiron S.P.A. Immunisation against chlamydia pneumoniae
GB0017149D0 (en) 2000-07-12 2000-08-30 Chiron Spa Helicobacter pylori mutants
GB0103170D0 (en) 2001-02-08 2001-03-28 Smithkline Beecham Biolog Vaccine composition
US20040073294A1 (en) * 2002-09-20 2004-04-15 Conor Medsystems, Inc. Method and apparatus for loading a beneficial agent into an expandable medical device
GB0103424D0 (en) * 2001-02-12 2001-03-28 Chiron Spa Gonococcus proteins
EP1409013B1 (en) 2001-07-26 2009-11-18 Novartis Vaccines and Diagnostics S.r.l. Vaccines comprising aluminium adjuvants and histidine
GB0118249D0 (en) * 2001-07-26 2001-09-19 Chiron Spa Histidine vaccines
JP4592284B2 (ja) 2001-07-27 2010-12-01 カイロン ソチエタ ア レスポンサビリタ リミタータ 髄膜炎菌付着因子
GB0121591D0 (en) * 2001-09-06 2001-10-24 Chiron Spa Hybrid and tandem expression of neisserial proteins
MX339524B (es) 2001-10-11 2016-05-30 Wyeth Corp Composiciones inmunogenicas novedosas para la prevencion y tratamiento de enfermedad meningococica.
GB0129007D0 (en) * 2001-12-04 2002-01-23 Chiron Spa Adjuvanted antigenic meningococcal compositions
JP2006500963A (ja) * 2002-08-02 2006-01-12 グラクソスミスクライン バイオロジカルズ ソシエテ アノニム ワクチン組成物
US7785608B2 (en) * 2002-08-30 2010-08-31 Wyeth Holdings Corporation Immunogenic compositions for the prevention and treatment of meningococcal disease
AU2003274511B2 (en) * 2002-10-11 2009-06-04 Glaxosmithkline Biologicals S.A. Polypeptide-vaccines for broad protection against hypervirulent meningococcal lineages
GB0227346D0 (en) * 2002-11-22 2002-12-31 Chiron Spa 741
WO2004065603A2 (en) 2003-01-15 2004-08-05 Wyeth Holdings Corporation Methods for increasing neisseria protein expression
WO2004067030A2 (en) * 2003-01-30 2004-08-12 Chiron Srl Injectable vaccines against multiple meningococcal serogroups
US20070253964A1 (en) 2003-04-16 2007-11-01 Zlotnick Gary W Novel Immunogenic Compositions for the Prevention and Treatment of Meningococcal Disease
GB0315021D0 (en) * 2003-06-26 2003-07-30 Chiron Srl Immunogenic gonococcal compositions
GB0323103D0 (en) * 2003-10-02 2003-11-05 Chiron Srl De-acetylated saccharides
ES2397923T3 (es) * 2003-10-02 2013-03-12 Novartis Ag Vacunas líquidas para múltiples serogrupos meningocócicos
GB0408977D0 (en) * 2004-04-22 2004-05-26 Chiron Srl Immunising against meningococcal serogroup Y using proteins
GB0409748D0 (en) * 2004-04-30 2004-06-09 Chiron Srl Lactoferrin cleavage
GB0410866D0 (en) * 2004-05-14 2004-06-16 Chiron Srl Haemophilius influenzae
GB0419408D0 (en) * 2004-09-01 2004-10-06 Chiron Srl 741 chimeric polypeptides
EP2682126B1 (en) 2005-01-27 2016-11-23 Children's Hospital & Research Center at Oakland GNA1870-based vesicle vaccines for broad spectrum protection against diseases caused by Neisseria meningitidis
GB0524066D0 (en) 2005-11-25 2006-01-04 Chiron Srl 741 ii
NZ572054A (en) 2006-03-22 2011-12-22 Novartis Ag Regimens for immunisation with meningococcal conjugates
TW200806315A (en) 2006-04-26 2008-02-01 Wyeth Corp Novel formulations which stabilize and inhibit precipitation of immunogenic compositions
NZ573931A (en) * 2006-06-29 2012-03-30 Craig J Venter Inst Inc Polypeptides from neisseria meningitidis
AR064642A1 (es) * 2006-12-22 2009-04-15 Wyeth Corp Polinucleotido vector que lo comprende celula recombinante que comprende el vector polipeptido , anticuerpo , composicion que comprende el polinucleotido , vector , celula recombinante polipeptido o anticuerpo , uso de la composicion y metodo para preparar la composicion misma y preparar una composi
WO2008125985A2 (en) 2007-04-11 2008-10-23 Novartis Ag Blocking interaction between pathogen factors and factor h to inhibit hemorrhagic syndromes
CA2688268A1 (en) 2007-06-04 2008-12-11 Novartis Ag Formulation of meningitis vaccines
CA2716212A1 (en) 2008-02-21 2009-08-27 Novartis Ag Meningococcal fhbp polypeptides
PT2268618E (pt) * 2008-03-03 2015-10-12 Novartis Ag Compostos e composições moduladores da actividade tlr
US9511131B2 (en) 2008-03-10 2016-12-06 Children's Hospital & Research Center Oakland Chimeric factor H binding proteins (fHBP) containing a heterologous B domain and methods of use
EP2331562A4 (en) 2008-09-03 2012-07-04 Childrens Hosp & Res Ct Oak PEPTIDES WITH AN EPITOPE OF A DOMAIN OF THE FACTOR H-BINDING PROTEIN AND METHOD OF USE
IT1394288B1 (it) 2008-09-12 2012-06-06 Novartis Vaccines & Diagnostic Immunogeni di proteine che legano il fattore h.
GB0819633D0 (en) 2008-10-25 2008-12-03 Isis Innovation Composition
CA2756522C (en) * 2009-03-24 2018-06-26 Novartis Ag Adjuvanting meningococcal factor h binding protein
ES2458355T3 (es) * 2010-09-01 2014-05-05 Novartis Ag Adsorción de inmunopotenciadores sobre sales metálicas insolubles
ES2744471T3 (es) * 2010-09-04 2020-02-25 Glaxosmithkline Biologicals Sa Ensayos de anticuerpos bactericidas para evaluar la inmunogenia y la potencia de vacunas de sacárido capsular meningocócico
AU2011300418B2 (en) * 2010-09-10 2016-05-12 Glaxosmithkline Biologicals Sa Meningococcus overexpressing NadA and/or NHBA and outer membrane vesicles derived therefrom
ES2864764T3 (es) * 2011-05-11 2021-10-14 Childrens Medical Center Composición inmunogénica que presenta múltiples antígenos, y métodos y usos de la misma
US9184727B2 (en) * 2012-06-11 2015-11-10 Phonon Corporation SAW device and method for post-seal frequency trimming

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992016643A1 (en) * 1991-03-14 1992-10-01 Imclone Systems Incorporated Recombinant hybrid porin epitopes
US5547670A (en) * 1991-03-14 1996-08-20 Imclone Systems Incorporated Recombinant hybrid porin epitopes
WO1999036544A2 (en) * 1998-01-14 1999-07-22 Chiron S.P.A. Neisseria meningitidis antigens

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Expression in Escherichia coli and immunologicalcharacterization of a hybrid class 1-P64K protein fromNesisseria Meningitidis. Guillen G et al,Biotecnologia Aplicada,Vol.13 No.4 1996 *
抗***外膜蛋白单克隆抗体的研制及应用研究 马远方，白惠玲，河南医学研究，第6卷第2期 1997 *

Also Published As

Publication number Publication date
EP1947187A1 (en) 2008-07-23
CN1800385B (zh) 2010-06-02
ES2391153T3 (es) 2012-11-22
JP2011101657A (ja) 2011-05-26
US20040092711A1 (en) 2004-05-13
CN1544462A (zh) 2004-11-10
DK1790660T3 (da) 2012-09-17
CN1201011C (zh) 2005-05-11
DE60132978D1 (de) 2008-04-10
CN100473663C (zh) 2009-04-01
ES2360746T3 (es) 2011-06-08
CA2875231A1 (en) 2001-09-07
JP2013005816A (ja) 2013-01-10
DE60126249T2 (de) 2008-05-29
CA2689666A1 (en) 2001-09-07
RU2002125882A (ru) 2004-03-10
AU2001239488B2 (en) 2006-01-19
US9150898B2 (en) 2015-10-06
CA2400562A1 (en) 2001-09-07
EP1259627A2 (en) 2002-11-27
JP4846160B2 (ja) 2011-12-28
BR0108711A (pt) 2004-06-22
CN1426471A (zh) 2003-06-25
US9267163B2 (en) 2016-02-23
BR0108713A (pt) 2004-06-22
WO2001064920A3 (en) 2002-03-14
US20140363462A1 (en) 2014-12-11
EP1790660A2 (en) 2007-05-30
PT1261723E (pt) 2008-06-05
DE60126249D1 (de) 2007-03-15
JP2011083287A (ja) 2011-04-28
JP2016128520A (ja) 2016-07-14
PT1947187E (pt) 2011-07-04
MXPA02008313A (es) 2002-12-09
EP2270030A2 (en) 2011-01-05
NZ521531A (en) 2005-12-23
EP1790660B1 (en) 2012-06-20
RU2002125880A (ru) 2004-03-10
JP4763210B2 (ja) 2011-08-31
LU92240I2 (fr) 2013-09-04
ATE503837T1 (de) 2011-04-15
ATE352631T1 (de) 2007-02-15
ATE387502T1 (de) 2008-03-15
JP5941027B2 (ja) 2016-06-29
CY1113477T1 (el) 2016-06-22
JP2014121333A (ja) 2014-07-03
EP2270031A3 (en) 2011-02-16
US20170080077A1 (en) 2017-03-23
JP5346004B2 (ja) 2013-11-20
EP2270030A3 (en) 2011-02-02
CN1800385A (zh) 2006-07-12
ES2281409T3 (es) 2007-10-01
HK1071147A1 (en) 2005-07-08
EP2270030B1 (en) 2012-05-23
ES2360746T9 (es) 2012-02-13
US8703914B2 (en) 2014-04-22
DE60132978T2 (de) 2009-02-26
DK1947187T5 (da) 2011-10-24
CN101139590B (zh) 2012-07-18
US20170080076A1 (en) 2017-03-23
AU3947801A (en) 2001-09-12
CA2400570A1 (en) 2001-09-07
DK1261723T3 (da) 2008-06-23
EP1947187B9 (en) 2011-09-21
US20130005667A1 (en) 2013-01-03
JP2014000092A (ja) 2014-01-09
AU2001239478B2 (en) 2007-05-17
CA2744921C (en) 2014-05-13
RU2299906C2 (ru) 2007-05-27
AU3948801A (en) 2001-09-12
CA2744921A1 (en) 2001-09-07
US20140294885A1 (en) 2014-10-02
WO2001064922A2 (en) 2001-09-07
JP2003525049A (ja) 2003-08-26
HK1064120A1 (en) 2005-01-21
EP1790660A3 (en) 2007-10-31
ES2386534T3 (es) 2012-08-22
NL300605I2 (zh) 2016-09-22
CY2013025I1 (el) 2015-11-04
MXPA02008314A (es) 2002-12-09
EP1947187B1 (en) 2011-03-30
PT1790660E (pt) 2012-09-17
US20100267931A1 (en) 2010-10-21
CN1508253A (zh) 2004-06-30
US20040110670A1 (en) 2004-06-10
DE60144353D1 (de) 2011-05-12
EP1261723A2 (en) 2002-12-04
HK1055993A1 (en) 2004-01-30
CA2400570C (en) 2010-04-27
CN101139590A (zh) 2008-03-12
CY1106532T1 (el) 2012-01-25
CY1107447T1 (el) 2012-12-19
US20060051840A1 (en) 2006-03-09
RU2304617C2 (ru) 2007-08-20
CY2013025I2 (el) 2015-11-04
CA2400562C (en) 2011-09-20
EP2270031A2 (en) 2011-01-05
CY1111591T1 (el) 2015-10-07
CN1426473A (zh) 2003-06-25
NZ521396A (en) 2004-06-25
EP1261723B1 (en) 2008-02-27
ES2299476T3 (es) 2008-06-01
CN101906413A (zh) 2010-12-08
US7803387B2 (en) 2010-09-28
WO2001064920A2 (en) 2001-09-07
DK1947187T3 (da) 2011-05-09
EP1259627B1 (en) 2007-01-24
HK1056197A1 (en) 2004-02-06
CN100339482C (zh) 2007-09-26
CA2689666C (en) 2015-02-24
DK2270030T3 (da) 2012-08-13
CY1113032T1 (el) 2016-04-13
US8114960B2 (en) 2012-02-14
WO2001064922A3 (en) 2001-12-06
JP2003525050A (ja) 2003-08-26
PT2270030E (pt) 2012-07-24

Similar Documents

Publication Publication Date Title
CN100334214C (zh) 奈瑟球菌蛋白质的杂交表达
Van der Ende et al. Variable expression of class 1 outer membrane protein in Neisseria meningitidis is caused by variation in the spacing between the-10 and-35 regions of the promoter
Feavers et al. Molecular analysis of the serotyping antigens of Neisseria meningitidis
AU2001239488A1 (en) Heterologous expression of neisserial proteins
AU2001239478A1 (en) Hybrid expression of neisserial proteins
CA2438080A1 (en) Gonococcal proteins and nucleic acids
Zhu et al. The opcA and ΨopcB regions in Neisseria: genes, pseudogenes, deletions, insertion elements and DNA islands
AU707691B2 (en) Nucleic acid probes and amplification oligonucleotides for neisseria species
US6548246B1 (en) Method and probes for the identification of microbial genes specifically induced during host infection
WO1997044487A1 (en) Method and probes for the identification of microbial genes specifically induced during host infection
AU2004201216A1 (en) Hybrid expression of Neisserial proteins

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
REG Reference to a national code

Ref country code: HK

Ref legal event code: DE

Ref document number: 1064120

Country of ref document: HK

C14 Grant of patent or utility model
GR01 Patent grant
REG Reference to a national code

Ref country code: HK

Ref legal event code: GR

Ref document number: 1064120

Country of ref document: HK

CF01 Termination of patent right due to non-payment of annual fee
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20070829

Termination date: 20170228