CN115515628A - 腮腺炎和麻疹病毒免疫原及其用途 - Google Patents

腮腺炎和麻疹病毒免疫原及其用途 Download PDF

Info

Publication number
CN115515628A
CN115515628A CN202080096383.0A CN202080096383A CN115515628A CN 115515628 A CN115515628 A CN 115515628A CN 202080096383 A CN202080096383 A CN 202080096383A CN 115515628 A CN115515628 A CN 115515628A
Authority
CN
China
Prior art keywords
mev
seq
muv
residues
ectodomain
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.)
Pending
Application number
CN202080096383.0A
Other languages
English (en)
Inventor
B·格雷汉姆
G·斯图尔特-琼斯
R·J·卢米斯
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.)
US Department of Health and Human Services
Original Assignee
US Department of Health and Human Services
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
Application filed by US Department of Health and Human Services filed Critical US Department of Health and Human Services
Publication of CN115515628A publication Critical patent/CN115515628A/zh
Pending legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • 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
    • A61K39/12—Viral antigens
    • 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
    • A61K39/12—Viral antigens
    • A61K39/155—Paramyxoviridae, e.g. parainfluenza virus
    • A61K39/165—Mumps or measles virus
    • 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
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • C07K14/08—RNA viruses
    • C07K14/115—Paramyxoviridae, e.g. parainfluenza virus
    • C07K14/12—Mumps virus; Measles virus
    • 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
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/70—Multivalent vaccine
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C07K2319/735—Fusion polypeptide containing domain for protein-protein interaction containing a domain for self-assembly, e.g. a viral coat protein (includes phage display)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/18011—Paramyxoviridae
    • C12N2760/18022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/18011—Paramyxoviridae
    • C12N2760/18032—Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/18011—Paramyxoviridae
    • C12N2760/18034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/18011—Paramyxoviridae
    • C12N2760/18711—Rubulavirus, e.g. mumps virus, parainfluenza 2,4
    • C12N2760/18721—Viruses as such, e.g. new isolates, mutants or their genomic sequences
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/18011—Paramyxoviridae
    • C12N2760/18711—Rubulavirus, e.g. mumps virus, parainfluenza 2,4
    • C12N2760/18723—Virus like particles [VLP]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/18011—Paramyxoviridae
    • C12N2760/18711—Rubulavirus, e.g. mumps virus, parainfluenza 2,4
    • C12N2760/18734—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Virology (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Communicable Diseases (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Molecular Biology (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Biochemistry (AREA)
  • Epidemiology (AREA)
  • Biophysics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Oncology (AREA)
  • Genetics & Genomics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Pulmonology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

提供了包含以融合前构象稳定的重组腮腺炎(MuV)胞外域三聚体或以融合前构象稳定的重组麻疹(MeV)胞外域三聚体的免疫原的实施方案。还提供了包含嵌合蛋白的免疫原的实施方案，该嵌合蛋白包含重组MuV或MeV F胞外域三聚体和一个或更多个MuV HN或MeV H胞外域。还公开了编码该免疫原的核酸及其产生方法。还提供了通过向受试者施用公开的免疫原诱导受试者产生免疫应答的方法。在一些实施方案中，免疫应答治疗或抑制受试者的MuV和/或MeV感染。

Description

腮腺炎和麻疹病毒免疫原及其用途
相关申请的交叉引用
本申请要求于2019年12月11日提交的美国临时申请号62/946,902的较早申请日的权益，其通过引用以其全文并入本文。
技术领域
本公开涉及用于引发和检测对腮腺炎病毒(MuV)和麻疹病毒(MeV)的免疫应答的多肽、多核苷酸、组合物和它们的使用方法。
背景技术
MeV和MuV是高传染性的副粘病毒，其可通过感染者的呼吸道飞沫或直接接触传播。由此产生的疾病可能导致儿童出现严重并发症或死亡。现有的MeV和MuV疫苗是减毒活病毒，其在1岁和最早一个月后分两次皮下接种。两剂麻疹、腮腺炎和风疹联合疫苗对麻疹的有效率为97％，并且对腮腺炎的有效率为88％。单剂麻疹、腮腺炎和风疹联合疫苗对麻疹的有效率为93％，并且对腮腺炎的有效率为78％。
尽管目前获得许可的疫苗对MeV和MuV具有有效性，但近年来两者的发病率均有所增加。促成因素包括由于疫苗犹豫导致疫苗接种率降低以及许可MMR疫苗对其提供有限保护的不同菌株的流通。
在MuV情况下，最近的研究已表明在通常发生在儿童时期的第二次MMR疫苗接种后，免疫力显著减弱。此外，目前流通的MuV毒株的基因型已经偏离标准腮腺炎疫苗中的Jeryl-Lynn毒株。为应对最近在美国和欧洲反复爆发的MuV疾病暴发，免疫实践咨询委员会建议进行第三次MMR疫苗接种以加强保护。然而，现有的免疫中和限制其有效性的第三次MMR疫苗接种。
发明内容
本文公开了包含原体的重组MuV F胞外域三聚体和重组MeV F胞外域三聚体，该原体包含一个或更多个修饰(例如氨基酸取代)用于以融合前构象稳定。进一步提供了与MuVHN胞外域或MeV H胞外域连接的重组MuV F胞外域三聚体和重组MeV F胞外域三聚体的实施方案。提供了嵌合蛋白，其包括MuV F胞外域三聚体和MeV H胞外域的组合，或MeV F胞外域三聚体和MuV HN胞外域的组合。此类蛋白的实施方案被证明在动物模型中产生优异的免疫应答，并且可用于例如引发或增强受试者对MeV和/或MuV的免疫应答。
在一些实施方案中，免疫原包含重组MuV F胞外域三聚体，其通过三聚体的原体中的一个或更多个氨基酸取代以融合前构象稳定，该氨基酸取代包括形成非天然二硫键以使MuV胞外域三聚体以融合前构象稳定的半胱氨酸取代。在一些实施方案中，重组MuV F胞外域三聚体通过位于MuV F第206和223位的三聚体的原体中的半胱氨酸取代之间的非天然二硫键以融合前构象稳定。在一些实施方案中，三聚体的原体进一步包含去除MuV F胞外域的F1/F2弗林蛋白酶切割位点的突变。在一些实施方案中，重组MuVF胞外域三聚体的原体在C-末端与三聚化结构域(例如GCN4三聚化结构域)融合。在另外的实施方案中，重组MuV F胞外域三聚体的原体与异源蛋白连接，例如MuV HN胞外域或MeV H胞外域。
在一些实施方案中，免疫原包含重组MeV F胞外域三聚体，其通过三聚体的原体中的一个或更多个氨基酸取代以融合前构象稳定，氨基酸取代包括形成非天然二硫键以使MeV胞外域三聚体以融合前构象稳定的半胱氨酸取代。在一些实施方案中，重组MeV F胞外域三聚体通过位于MeV F第165和171位的三聚体的原体中的半胱氨酸取代之间的非天然二硫键以融合前构象稳定。在一些实施方案中，三聚体的原体进一步包含去除MeV F胞外域的F1/F2弗林蛋白酶切割位点的突变。在一些实施方案中，重组MeV F胞外域三聚体的原体在C-末端与三聚化结构域(例如GCN4三聚化结构域)融合。在另外的实施方案中，重组MeV F胞外域三聚体的原体与异源蛋白连接，例如MuV HN胞外域或MeV H胞外域。
在一些实施方案中，提供了包含融合蛋白三聚体的免疫原，每个融合蛋白在N-末端至C-末端方向上包含：三聚化结构域和一个或更多个MuV HN胞外域或MeV H胞外域。
在一些实施方案中，免疫原包含MeV H胞外域头部的二聚体。
还提供了编码公开的蛋白的核酸分子，以及包括该核酸分子的载体，和它们的产生方法。
还提供了包括公开的免疫原的免疫原性组合物，其适合施用于受试者，并且还可以包含在单位剂型中。免疫原还可以包含运载体(carrier)以促进向免疫系统的呈递。
公开了诱导受试者产生免疫应答的方法，以及通过向受试者施用有效量的公开的免疫原、核酸分子或载体来抑制或预防受试者的MuV或MeV感染的方法。
本公开的前述和其他特征和优点将从以下参照附图进行的几个实施方案的详细描述中变得更加明显。
附图说明
图1A-1E。二硫化物稳定的融合前腮腺炎F三聚体的基于结构的设计。(图1A)通过系统筛选二硫键和GCN4附接位置的融合前腮腺炎F糖蛋白三聚体的基于结构的设计，显示通过负染色EM确定的融合前相对于融合后F构象的产率和百分比。(图1B)V206C/A223C与476-GCN4的组合以高产率观察到均质的融合前F三聚体(上图)和融合后显示在下图中。(图1C)S200凝胶过滤分析显示融合前F(PreF)、糖基化和去糖基化、HN和融合前F-HN的单分散性。(图1D)显示了具有原体的融合前腮腺炎F三聚体(SEQ ID NO:11)分辨率为
Figure GDA0003943442610000021
的晶体结构，其中经历>
Figure GDA0003943442610000022
构象变化以转变成以黑色显示的融合后构象的残基。GCN4三聚(TD)基序以虚线显示，其与F残基476连接。放大插图突出显示突变残基以稳定三聚体融合前F结构。(图1E)融合前腮腺炎三聚体的单一原体显示D1-3亚区和每个原体的六个N-连接聚糖的位置。
图2A-2H。腮腺炎和PIV5融合前F的结构比较以及腮腺炎F和HN基因型变异分析。(图2A)腮腺炎融合前F三聚体和相关副粘病毒PIV5融合前F三聚体的结构叠加显示出相似的总体拓扑结构
Figure GDA0003943442610000023
，但具有49％的序列同一性。(图2B)腮腺炎preF的顶端环以“闭合帽”组件缔合，而在PIV5中，这些环展开(上图)，并且稳定腮腺炎preF顶端的残基是T178、T179和N181(下图)。(图2C)腮腺炎F糖蛋白被糖基化，每个原体有6个聚糖，每个三聚体共有18个聚糖，其中在构象可移动区域(黑色)中有6个聚糖以及在构象可变性较小的区域中有12个聚糖，从而为三聚体提供了大量的聚糖屏蔽。(图2D)来自腮腺炎基因型A-J的融合糖蛋白的系统发育分析。(图2E)来自腮腺炎基因型A-J的血凝素-神经氨酸酶糖蛋白的系统发育分析。(图2F和2G)Jeryl Lynn疫苗株(基因型A，SEQ ID NO:98)与来自腮腺炎基因型C(SEQ ID NO:102)、D(SEQ ID NO:103)、F(SEQ ID NO:104)、G(SEQ ID NO:105)和H(SEQID NO:106)的序列比对。(图2H)腮腺炎基因型变异在融合前F(左图)和HN(右图)上的结构图，显示A-J基因型变异(上图)和基因型G与A(Jeryl Lynn(JL))(下图)，显示残基差异和聚糖差异。
图3A和3B。腮腺炎F和preF-HN免疫原变体的设计和免疫方案。(图3A)在0、3和10周的时间用Poly(I:C)佐剂以10μg/剂量对CB6F1/J小鼠进行3次免疫，并在第2、5、12和16周进行血液取样。(图3B)腮腺炎F和preF-HN蛋白的负染色表征确证了免疫原的构象和分子身份以及在小鼠免疫1、2或3次后引发的融合后F(左)、融合前F(中)和融合前F-HN(右)免疫原的腮腺炎中和滴度(如箭头所示)，以及基因型G病毒、Jeryl Lynn基因型A病毒和基因型H病毒的PRN滴度。
图4A-4E。融合前腮腺炎F-HN免疫原的设计，针对三种不同腮腺炎基因型病毒的中和滴度的特异性和持久性的血清分析。(图4A)腮腺炎preF-HN融合蛋白的负染色表征，显示了preF头部、GCN4三聚化结构域和三个HN头部基团的位置。(图4B)使用OctetBioinferometry，使用腮腺炎融合前F探针和(图4C)HN探针，免疫小鼠的结合滴度。(图4D)用三剂10μg融合前F蛋白和Poly(I:C)免疫小鼠的持久性分析，每月进行血清取样并针对基因型G病毒、Jeryl Lynn病毒和基因型H病毒进行PRNT分析，持续6个月。(图4E)用三剂10μg融合前F-HN蛋白和Poly(I:C)免疫小鼠的持久性分析，每月进行血清取样并针对基因型G病毒、Jeryl Lynn病毒和基因型H病毒进行PRNT分析，持续6个月。
图5A-5C。融合前麻疹F的基于结构的设计。(图5A)稳定融合前构象的融合前稳定的麻疹F糖蛋白突变的基于结构的设计。(图5B)包括所示的非天然二硫键和C-末端GCN4三聚化结构域的麻疹F变体的负染色EM分析，以及(图5C)融合前(MeV F R165C/M171C-486-GCN4，SEQ ID NO:38)和融合后(具有-486-GCN4的天然MeV F序列)麻疹F糖蛋白的的免疫原性。
图6A-6C。腮腺炎pre-F-麻疹H嵌合免疫原的设计。(图6A)麻疹H二聚体(左上)、麻疹H三聚体(中上)、腮腺炎HN三聚体(右上)和具有麻疹H的腮腺炎pre-F(MuV F 206C/223C-476+GCN4/Fd+MeV-H,SEQ ID NO:28)(下)的负染色表征。(图6B)麻疹斑减少中和滴度测定(PRNT)。(图6C)腮腺炎(基因型G)PRNT测定。
序列表
随附序列表中列出的核酸和氨基酸序列使用标准字母缩写显示核苷酸碱基并使用三字母代码显示氨基酸，如37C.F.R.1.822所定义。仅显示每个核酸序列的一条链，但互补链应理解为包括在对显示链的任何引用中。序列表以ASCII文本文件的形式提交，文件命名为“Sequence.txt”(～456kb)，创建于2020年12月11日，其通过引用并入本文。
结构坐标
以融合前构象稳定的MuV F胞外域三聚体的晶体结构的原子坐标记载于2019年12月11日提交的美国临时申请号62/946,902的表1中，其通过引用以其全文并入本文，并以ASCII文本文件的形式提交，文件命名为“Table_1.txt”(～555KB)，其创建于2019年12月9日。
具体实施方式
I.术语总结
除非另有说明，否则根据常规用法使用技术术语。分子生物学中常用术语的定义可见于Benjamin Lewin,Genes X,由Jones&Bartlett出版社出版,2009；和Meyers等人(编辑),The Encyclopedia of Cell Biology and Molecular Medicine,由Wiley-VCH出版，共16卷,2008；以及其他类似参考文献。如本文所用，单数形式“一种(a、an)”和“该(the)”既指单数也指复数，除非上下文另有说明。例如，术语“一种抗原”包括单个或多个抗原并且可以被认为等同于短语“至少一种抗原”。如本文所用，术语“包含”意指“包括”。还应理解，除非另有说明，否则对于核酸或多肽给出的任何和所有碱基大小或氨基酸大小以及所有分子量或分子质量值均是近似的，并且提供用于描述目的。尽管可以使用与本文所述的那些相似或等同的许多方法和材料，但以下描述了特别合适的方法和材料。如有冲突，以本说明书(包括术语解释)为准。此外，材料、方法和实例仅是说明性的而不是限制性的。为了便于查看各种实施方案，提供以下术语解释：
佐剂：用于增强抗原性的溶媒。在一些实施方案中，佐剂包括其上吸附抗原的矿物质(铝剂(alum，明矾剂)、氢氧化铝或磷酸盐)的悬浮液；或油包水乳液，例如，其中抗原溶液在矿物油(弗氏不完全佐剂)中乳化，有时包括杀死的分枝杆菌(弗氏完全佐剂)以进一步增强抗原性(抑制抗原降解和/或导致巨噬细胞流入)。在一些实施方案中，在公开的免疫原性组合物中使用的佐剂是卵磷脂和卡波姆均聚物的组合(例如可获自AdvancedBioAdjuvants的ADJUPLEXTM佐剂，另见Wegmann,Clin Vaccine Immunol,22(9):1004-1012,2015)。用于公开的免疫原性组合物的另外的佐剂包括QS21纯化植物提取物、Matrix M、AS01、MF59和ALFQ佐剂。免疫刺激性寡核苷酸(例如包括CpG基序的那些)也可以用作佐剂。佐剂包括生物分子(“生物佐剂”)，例如共刺激分子。示例性佐剂包括IL-2、RANTES、GM-CSF、TNF-α、IFN-γ、G-CSF、LFA-3、CD72、B7-1、B7-2、OX-40L、4-1BBL、免疫刺激复合物(ISCOM)基质和Toll样受体(TLR)激动剂，例如TLR-9激动剂、Poly I:C或PolyICLC。(参见，例如，Singh(编辑)Vaccine Adjuvants and Delivery Systems.Wiley-Interscience,2007)。
施用：通过选择的途径将组合物引入受试者。施用可以是局部的或全身的。例如，如果选择的途径是鼻内，则通过将组合物引入受试者的鼻道中来施用组合物(例如包括公开的重组MuV F胞外域三聚体或重组MeV F胞外域三聚体的组合物)。示例性施用途径包括但不限于口服、注射(例如皮下、肌内、皮内、腹腔和静脉内)、舌下、直肠、经皮(例如，外用)、鼻内、阴道和吸入途径。
氨基酸取代：用一种或更多种不同的氨基酸取代多肽中的氨基酸。在蛋白序列的背景下，氨基酸取代也称为突变。
抗体：特异性结合并识别分析物(抗原)例如MuV或MeV F蛋白、其抗原性片段或抗原的二聚体或多聚体的免疫球蛋白、抗原结合片段或其衍生物。术语“抗体”在本文中以最广泛的含义使用并且涵盖各种抗体结构，包括但不限于单克隆抗体、多克隆抗体、多特异性抗体(例如双特异性抗体)和抗体片段，只要它们表现出期望的抗原结合活性。抗体的非限制性实例包括，例如，完整的免疫球蛋白和其保留对抗原的结合亲和力的变体和片段。抗体片段的实例包括但不限于Fv、Fab、Fab'、Fab'-SH、F(ab')2；双体；线性抗体；单链抗体分子(例如scFv)；以及由抗体片段形成的多特异性抗体。抗体片段包括通过修饰全抗体产生的或使用重组DNA方法从头合成的那些抗原结合片段(参见，例如，Kontermann和Dubel(编辑),Antibody Engineering,Vols.1-2,2nd Ed.,Springer Press,2010)。
运载体(Carrier)：可以与抗原连接的免疫原性分子。当与运载体连接时，抗原可能变得更具有免疫原性。选择运载体以增加抗原的免疫原性和/或引发对诊断、分析和/或治疗有益的针对运载体的抗体。有用的运载体包括聚合物运载体，其可以是天然的(例如，来自细菌或病毒的蛋白)、半合成或合成材料，其含有一个或更多个可附接反应物部分的官能团。
保守变体：“保守”氨基酸取代是那些基本上不影响或降低蛋白质功能(例如当施用于受试者时蛋白质诱导免疫应答的能力)的取代。术语保守变异还包括使用取代的氨基酸代替未取代的亲本氨基酸。此外，改变、添加或缺失编码序列中的单个氨基酸或小百分比的氨基酸(例如小于5％，在一些实施方案中小于1％)的单独取代、缺失或添加是保守变异，其中改变导致氨基酸被化学上相似的氨基酸取代。
以下六组是被认为是彼此保守取代的氨基酸实例：
1)丙氨酸(A)、丝氨酸(S)、苏氨酸(T)；
2)天冬氨酸(D)、谷氨酸(E)；
3)天冬酰胺(N)、谷氨酰胺(Q)；
4)精氨酸(R)、赖氨酸(K)；
5)异亮氨酸(I)、亮氨酸(L)、蛋氨酸(M)、缬氨酸(V)；和
6)苯丙氨酸(F)、酪氨酸(Y)、色氨酸(W)。
非保守取代是降低重组MuV或MeV F胞外域三聚体的活性或功能(例如当施用于受试者时诱导免疫应答的能力)的那些。例如，如果氨基酸残基对于蛋白质的功能是必需的，那么即使是保守取代也可能破坏该活性。因此，保守取代不改变目的蛋白的基本功能。
对照：参考标准。在一些实施方案中，对照是从健康患者获得的阴性对照样本。在其他实施方案中，对照是从被诊断患有MuV或MeV感染的患者获得的阳性对照样本。在还其他实施方案中，对照是历史对照或标准参考值或值范围(例如先前检测的对照样本，例如具有已知预后或结局的一组MuV或MeV患者，或代表基线或正常值的一组样本)。
检测样本与对照之间的差异可以是增加或相反地减少。该差异可以是定性差异或定量差异，例如统计学上显著性差异。在一些实例中，差异是相对于对照增加或减少至少约5％，例如至少约10％、至少约20％、至少约30％、至少约40％、至少约50％、至少约60％、至少约70％、至少约80％、至少约90％、至少约100％、至少约150％、至少约200％、至少约250％、至少约300％、至少约350％、至少约400％、至少约500％或大于500％。
简并变体：在本公开的上下文中，“简并变体”是指编码多肽的多核苷酸，该多肽包括由于遗传密码简并的序列。有20种天然氨基酸，其中大部分由多于一个密码子指定。因此，所有编码肽的简并核苷酸序列均被包括，只要该核苷酸序列编码的肽的氨基酸序列未改变。
有效量：足以引发受试者产生期望的应答(例如免疫应答)的试剂(例如免疫原)的量。应当理解，为了获得针对目的抗原的保护性免疫应答，可能需要多次施用公开的免疫原，和/或施用公开的免疫原作为初免加强方案中的“初免”，其中加强免疫原可以不同于初免免疫原。因此，公开的免疫原的有效量可以是足以引发受试者产生免疫应答的免疫原的量，随后可以用相同或不同的免疫原加强免疫应答以引发保护性免疫应答。
在一个实例中，期望的应答是抑制或减少或预防MuV感染。不需要完全消除或减少或预防MuV感染以使该方法有效。例如，与合适的对照相比，施用有效量的试剂可以使MuV感染(例如，通过细胞感染，或通过MuV感染的受试者的数量或百分比来测量)降低期望量，例如降低至少50％、至少60％、至少70％、至少80％、至少90％、至少95％、至少98％或甚至至少100％(消除或预防可检测的MuV感染)。
在一个实例中，期望的应答是抑制或减少或预防MeV感染。不需要完全消除或减少或预防MeV感染以使该方法有效。例如，与合适的对照相比，施用有效量的试剂可以使MeV感染(例如，通过细胞感染，或通过MeV感染的受试者的数量或百分比来测量)降低期望量，例如降低至少50％、至少60％、至少70％、至少80％、至少90％、至少95％、至少98％或甚至至少100％(消除或预防可检测的MeV感染)。
在一个实例中，期望的应答是抑制或减少或预防MuV和MeV感染。不需要完全消除或减少或预防MuV和MeV感染以使该方法有效。例如，与合适的对照相比，施用有效量的试剂可以使MuV和MeV感染(例如，通过细胞感染，或通过MuV和/或MeV感染的受试者的数量或百分比来衡量)降低期望量，例如降低至少50％、至少60％、至少70％、至少80％、至少90％、至少95％、至少98％或甚至至少100％(消除或预防可检测的MuV和/或MeV感染)。
表达：核酸序列的转录或翻译。例如，当基因的DNA被转录成RNA或RNA片段时，基因被表达，在一些实例中，该RNA或RNA片段经加工成mRNA。当基因的mRNA被翻译成氨基酸序列，例如蛋白或蛋白片段时，基因也可以被表达。在特定的实例中，异源基因在转录成RNA时被表达。在另一个实例中，异源基因在其RNA被翻译成氨基酸序列时被表达。术语“表达”在本文中用于表示转录或翻译。表达的调节可以包括对转录、翻译、RNA转运和加工、中间分子(例如mRNA)的降解的控制，或通过产生特定蛋白分子后其激活、失活、区室化或降解的控制。
表达控制序列：调节与其可操作地连接的异源核酸序列的表达的核酸序列。当表达控制序列控制和调节核酸序列的转录和适当时的翻译时，表达控制序列与核酸序列有效地连接。因此，表达控制序列可以包括适当的启动子、增强子、转录终止子、蛋白质编码基因前面的起始密码子(ATG)、内含子的剪接信号、维持该基因的正确阅读框以允许正确翻译mRNA，和终止密码子。术语“控制序列”旨在至少包括其存在可以影响表达的组分，并且还可以包括其存在是有利的另外的组分，例如前导序列和融合配偶体序列。表达控制序列可以包括启动子。
启动子是足以指导转录的最小序列。还包括那些足以使启动子依赖性基因表达可控为具有细胞类型特异性、组织特异性或可被外部信号或试剂诱导的启动子元件；此类元件可能位于基因的5'或3'区域。组成型和诱导型启动子均被包括(参见例如，Bitter等人，Methods in Enzymology 153:516-544,1987)。例如，当在细菌系统中克隆时，可以使用诱导型启动子，例如λ噬菌体的pL、plac、ptrp、ptac(ptrp-lac杂合启动子)等。在一个实施方案中，当在哺乳动物细胞系统中克隆时，可以使用来自哺乳动物细胞基因组的启动子(例如金属硫蛋白启动子)或来自哺乳动物病毒(例如逆转录病毒长末端重复序列；腺病毒晚期启动子；牛痘病毒7.5K启动子)。通过重组DNA或合成技术产生的启动子也可用于提供核酸序列的转录。
表达载体：包含重组多核苷酸的载体，该重组多核苷酸包含与待表达的核苷酸序列可操作地连接的表达控制序列。表达载体包含足够的用于表达的顺式作用元件；用于表达的其他元件可以由宿主细胞或在体外表达系统中提供。表达载体包括本领域已知的所有那些，例如掺入重组多核苷酸的粘粒、质粒(例如，裸露的或包含在脂质体中的)和病毒(例如，慢病毒、逆转录病毒、腺病毒和腺相关病毒)。
GCN4三聚化结构域：来自GCN4蛋白的三聚化结构域，其包含天然形成三聚体结构的亮氨酸拉链氨基酸序列。GCN4三聚化结构域的实施方案描述于，例如，Harbury等人(1993Science 262:1401-1407)。在一些实例中，GCN4三聚化结构域可以包括在公开的重组蛋白的氨基酸序列中，使得重组蛋白将三聚化。与公开的实施方案一起使用的GCN4三聚化结构域序列的非限制性实例以IEDKIEEILSKIYHIENEIIARIKKLIGEAP(SEQ ID NO:33)提供。
异源的：起源于不同的遗传来源。
宿主细胞：可以在其中繁殖载体并表达其核酸的细胞。该细胞可以是原核的或真核的。该术语还包括主题宿主细胞的任何后代。应当理解，所有后代可能与亲代细胞不相同，因为在复制过程中可能发生突变。然而，当使用术语“宿主细胞”时，包括此类后代。
免疫应答：免疫系统细胞(例如B细胞、T细胞或单核细胞)对刺激的应答。在一个实施方案中，应答对特定抗原具有特异性(“抗原特异性应答”)。在一个实施方案中，免疫应答是T细胞应答，例如CD4+应答或CD8+应答。在另一个实施方案中，应答是B细胞应答，并导致产生特异性抗体。
免疫原：可以引发动物产生免疫应答的化合物、组合物或物质(例如，重组MuV或MeV F胞外域三聚体)，包括注射或吸收至动物的组合物。向受试者施用免疫原可以导致针对目的病原体的保护性免疫。
免疫原性组合物：包含公开的免疫原的组合物，当施用于受试者时，该免疫原诱导针对MuV或MeV的可测量的CTL应答，或诱导针对MuV或MeV的可测量的B细胞应答(例如产生抗体)。它还涉及编码公开的重组MuV或MeV F胞外域三聚体的原体的分离的核酸分子和载体，其可用于表达该原体(并因此用于引发针对重组MuV或MeV F胞外域三聚体的免疫应答)。对于体内使用，免疫原性组合物通常包括在药学上可接受的运载体中的重组MuV或MeVF胞外域三聚体或编码重组MuV或MeV F胞外域三聚体的原体的核酸分子，并且还可以包括其他试剂，例如佐剂。
抑制或治疗疾病：抑制疾病或病况的完全发展，例如，在具有患疾病(例如MuV感染或MeV感染)风险的受试者中。“治疗”是指在疾病或病理状况开始发展后改善其体征或症状的治疗干预。提及疾病或病理状况的术语“改善”是指治疗的任何可观察到的有益效果。抑制疾病可以包括预防或降低疾病的风险，例如预防或降低病毒感染的风险。有益效果可以例如通过以下进行证明：易感受试者中疾病临床症状的延迟发作、疾病的一些或所有临床症状的严重程度降低、疾病进展较慢、病毒载量降低、受试者的整体健康或健康情况改善，或特定疾病特有的其他参数。“预防性”治疗是对未表现出疾病体征或仅表现出早期体征的受试者进行的治疗，目的是降低发展病理的风险。
分离的：“分离的”生物组分已与其他生物组分(例如该组分天然存在于其中的其他生物组分)基本上分离或纯化，例如其他染色体和染色体外DNA、RNA和蛋白。已“分离的”蛋白、肽、核酸和病毒包括通过标准纯化方法纯化的那些。分离的不需要绝对纯度，并且可以包括至少50％分离的，例如至少75％、80％、90％、95％、98％、99％或甚至99.9％分离的蛋白、肽、核酸或病毒分子。
接头和连接的：可用于将两个分子连接成一个连续分子的双功能分子。肽接头的非限制性实例包括甘氨酸-丝氨酸肽接头。除非上下文另有说明，否则提及“连接”第一多肽和第二多肽，或提及“连接”在一起的两个多肽，或提及具有与第二多肽“连接”的第一多肽，是指通过肽键共价连接(例如通过肽接头)，使得第一多肽和第二多肽形成连续多肽链。如果涉及肽接头，则第一多肽和第二多肽的共价键可以连接至肽接头的N-末端和C-末端。通常，此类连接是使用分子生物学技术以遗传操作编码通过肽接头连接至第二多肽的第一多肽的DNA来实现的。
天然蛋白、序列或二硫键：未经例如选择性突变修饰的多肽、序列或二硫键。例如，选择性突变以将抗原的抗原性集中到靶标表位，或将二硫键引入天然蛋白中不存在的蛋白中。天然蛋白或天然序列也称为野生型蛋白或野生型序列。非天然二硫键是天然蛋白中不存在的二硫键，例如，由于通过基因工程将一个或更多个半胱氨酸残基引入蛋白中而在蛋白中形成的二硫键。
麻疹：由麻疹病毒引起的感染性疾病。症状通常在暴露于感染者后10-12天出现并持续7-10天。初始症状通常包括发烧、咳嗽、流鼻涕和眼睛发炎。症状开始后两至三天，口腔内可能形成称为Koplik斑的小白点。红色扁平皮疹通常从面部开始，然后扩散至身体其他部位，通常在症状开始至三到五天开始。常见的并发症包括腹泻、中耳感染和肺炎。这些发生的部分原因是麻疹诱导的免疫抑制。不太常见的癫痫发作、失明或脑部炎症可能发生。
麻疹病毒：非区段负链RNA病毒，属于副粘病毒科麻疹病毒属，可引起麻疹疾病。麻疹病毒基因组RNA含有6个相连的转录单元，它们编码以下八种蛋白的开放阅读框：核蛋白(N)、磷蛋白(P)、C蛋白、V蛋白、基质(M)蛋白、融合(F)蛋白、血凝素(H)蛋白和大(L)蛋白。至少有7种已知的MeV基因型，称为基因型A、B、C、D、F、G和H，目前在全球流行。
MeV融合(F)蛋白：MeV的包膜糖蛋白，其促进病毒与细胞膜的融合。在自然界中，来自MeV的F蛋白最初被合成为长度为约550个氨基酸的单一多肽前体，称为F0。F0包括指导定位至内质网的N-末端信号肽，其中信号肽被蛋白水解切割。其余F0残基寡聚化形成三聚体，并且可以通过细胞蛋白酶进行蛋白水解加工以产生两个二硫键连接的片段F1和F2。在MeV F中，切割位点大致位于残基113/114之间。这些片段中较小的F2源自F0前体的N-末端部分(大致残基24-113)。这些片段中较大的F1包括F0前体的C-末端部分(大致残基114-550)，包括细胞外/腔区(大致残基110-486)，以及跨膜和胞质区(大致残基487-550)。MeV F蛋白的胞外部分是MeV F胞外域，其包括F2蛋白和F1胞外域。
MeV F蛋白在MeV毒株中表现出明显的序列保守性。鉴于这种保守性，本领域普通技术人员可以容易地比较不同MeV F蛋白的氨基酸位置。除非上下文另有说明，否则MeV F氨基酸的编号参考SEQ ID NO:36(NCBI参考序列P35973.1，其通过引用并入本文)进行：
mglkvnvsaifmavlltlqtptgQIHWGNLSKIGVVGIGSASYKVMTRSSHQSLVIKLMPNITLLNNCTRVEIAEYRRLLRTVLEPIRDALNAMTQNIRPVQSVASSRRHKRFAGVVLAGAALGVATAAQITAGIALHQSMLNSQAIDNLRASLETTNQAIEAIRQAGQEMILAVQGVQDYINNELIPSMNQLSCDLIGQKLGLKLLRYYTEILSLFGPSLRDPISAEISIQALSYALGGDINKVLEKLGYSGGDLLGILESRGIKARITHVDTESYLIVLSIAYPTLSEIKGVIVHRLEGVSYNIGSQEWYTTVPKYVATQGYLISNFDESSCTFMPEGTVCSQNALYPMSPLLQECLRGSTKSCARTLVSGSFGNRFILSQGNLIANCASILCKCYTTGTIINQDPDKILTYIAADHCPVVEVNGVTIQVGSRRYPDAVYLHRIDLGPPILLERLDVGTNLGNAIAKLEDAKELLESSDQILRSMKGLSSTCIVYILIAVCLGGLIGIPALICCCRGRCNKKGEQVGMSRPGLKPDLTGTSKSYVRSL
三个MeV F原体在成熟F蛋白中寡聚化，其采用亚稳态的融合前构象(prefusionconformation)，当与靶细胞膜接触时，该融合前构象被触发经历构象变化为融合后构象。这种构象变化暴露疏水序列，称为融合肽，其位于F1胞外域的N-末端，并且其与宿主细胞膜缔合并促进病毒或感染细胞的膜与靶细胞膜融合。
“以融合前构象稳定”的MeV F胞外域三聚体包含与由相应天然MeV F序列形成的MeV F胞外域三聚体相比，提供与相应天然MeV F序列相比融合前构象保留增加的一个或更多个氨基酸取代、缺失或插入。融合前构象的“稳定化”可以是，例如，能量稳定化(例如，相对于融合后开放构象降低融合前构象的能量)和/或动力学稳定化(例如，降低从融合前构象到融合后构象的转变速率)。此外，与相应天然MeV F序列相比，MeV F胞外域三聚体以融合前构象的稳定性可以包括对变性的抗性增加。本文提供了确定MeV F胞外域三聚体是否处于融合前构象的方法，包括(但不限于)负染色电子显微镜和使用融合前构象特异性抗体的抗体结合测定。关于MeV F蛋白的术语“pre-F”描述了一种分子，该分子是通过一个或更多个氨基酸取代以融合前构象稳定的三聚体I类融合蛋白。
MeV F融合前特异性抗体：与以融合前构象的MeV F蛋白特异性结合，但不与以融合后构象的MeV F蛋白特异性结合的抗体。
MeV血凝素(H)蛋白：MeV包膜糖蛋白，其是一种II型膜蛋白并且促进MeV与宿主细胞膜的附接。全长H蛋白具有N-末端胞质尾和跨膜结构域(CT和TM，大致氨基酸1-58)，以及包括茎部(大致氨基酸59-179)和头部(大致氨基酸180-617)区域的胞外域(大致氨基酸59-617)。示例性MeV H蛋白序列在本文中以SEQ ID NO:49(NCBI参考序列AAA56644.1，其通过引用并入本文)提供：
MspqrdrinafykdnphpkgsrivinrehlmidrpyvllavlfvmflsligllaiagirlhraaiytaeihkslstnldvtnsiehqvkdvltplfkiigdevglrtpqrftdlvkfisdkikflnpdreydfrdltwcinpperikldydqycadvaaeelmnalvnstlletrttnqFLAVSKGNCSGPTTIRGQFSNMSLSLLDLYLGRGYNVSSIVTMTSQGMYGGTYLVEKPNLSSKRSELSQLSMYRVFEVGVIRNPGLGAPVFHMTNYLEQPVSNDLSNCMVALGELKLAALCHGEDSITIPYQGSGKGVSFQLVKLGVWKSPTDMQSWVPLSTDDPVIDRLYLSSHRGVIADNQAKWAVPTTRTDDKLRMETCFQQACKGKIQALCENPEWAPLKDNRIPSYGVLSVDLSLTVELKIKIASGFGPLITHGSGMDLYKSNHNNVYWLTIPPMKNLALGVINTLEWIPRFKVSPYLFTVPIKEAGEDCHAPTYLPAEVDGDVKLSSNLVILPGQDLQYVLATYDTSRVEHAVVYYVYSPSRSFSYFYPFRLPIKGVPIELQVECFTWDQKLWCRHFCVLADSESGGHITHSGMVGMGVSCTVTREDGTNRR
如本文所用，MeV H残基定位参照如SEQ ID NO:49所示的序列进行。
腮腺炎：由腮腺炎病毒引起的感染性疾病。腮腺炎的特征在于唾液腺(通常是腮腺)炎症。腮腺炎病毒感染可能出现重度并发症，例如脑膜炎、脑炎、胰腺炎、卵巢炎(女性)、睾丸炎(男性)和听力损失。
腮腺炎病毒(MuV)：非区段负链RNA病毒，属于副粘病毒科副粘病毒亚科风疹病毒属，可引起腮腺炎疾病。腮腺炎病毒基因组RNA含有7个衔接的转录单元，它们编码核蛋白(N)、磷蛋白(P)、V蛋白、I蛋白、基质(M)蛋白、融合(F)蛋白、小疏水性(SH)蛋白、血凝素-神经氨酸酶(HN)蛋白和大(L)蛋白的开放阅读框蛋白。腮腺炎病毒基因组的示意图在图6中显示。由于通过插入鸟嘌呤核苷酸进行RNA编辑，P基因(也称为“V/P/I基因”)产生对应于V、P和I蛋白的三个mRNA转录本。具体地，P基因的忠实转录产生V蛋白，插入两个鸟嘌呤核苷酸产生编码P蛋白的mRNA，以及插入四个鸟嘌呤残基产生编码I蛋白的mRNA。SH基因是MuV不同基因型中变异最大的基因，因此通常用作基因分型的基础。MuV有12种已知基因型，称为基因型A、B、C、D、F、G、H、I、J、K、L和N，目前在全球流行。目前大多数MuV疫苗基于基因型A(Jeryl Lynn)、基因型B(Urabe-AM9)或未确定的基因型(Leningrad-Zagreb)病毒。
MuV融合(F)蛋白：MuV的包膜糖蛋白，其促进病毒余细胞膜的融合。在自然界中，来自MuV的F蛋白最初被合成为长度为约538个氨基酸的单一多肽前体，称为F0。F0包括指导定位至内质网的N-末端信号肽，其中信号肽被蛋白水解切割。其余F0残基寡聚化形成三聚体，并且可以通过细胞蛋白酶进行蛋白水解加工以产生两个二硫键连接的片段F1和F2。在MuV F中，切割位点大致位于残基103/104之间。这些片段中较小的F2源自F0前体的N-末端部分(大致残基20-103)。这些片段中较大的F1包括F0前体的C-末端部分(大致残基104-538)，包括细胞外/腔区(大致残基110-483)，以及跨膜和胞质区(大致残基484-538)。MuV F蛋白的胞外部分是MuV F胞外域，其包括F2蛋白和F1胞外域。
MuV F蛋白在MuV毒株中表现出明显的序列保守性。鉴于这种保守性，本领域普通技术人员可以容易地比较不同MuV F蛋白的氨基酸位置。除非上下文另有说明，否则MuV F氨基酸的编号参考SEQ ID NO:1(NCBI参考序列P09458.1，其通过引用并入本文)进行：
mkafsvtclgfavfsssicVNINILQQIGYIKQQVRQLSYYSQSSSSYIVVKLLPNIQPTDNSCEFKSVTQYNKTLSNLLLPIAENINNIASPSPGSRRHKRFAGIAIGIAALGVATAAQVTAAVSLVQAQTNARAIAAMKNSIQATNRAIFEVKEGTQQLAIAVQAIQDHINTIMNTQLNNMSCQILDNQLATYLGLYLTELTTVFQPQLINPALSPISIQALRSLLGSMTPAVVQATLSTSISAAEILSAGLMEGQIVSVLLDEMQMIVKINIPTIVTQSNALVIDFYSISSFINNQESIIQLPDRILEIGNEQWSYPAKNCKLTRHHIFCQYNEAERLSLESKLCLAGNISACVFSPIAGSYMRRFVALDGTIVANCRSLTCLCKSPSYPIYQPDHHAVTTIDLTTCQTLSLDGLDFSIVSLSNITYAENLTISLSQTINTQPIDISTELSKVNASLQNAVKYIKESNHQLQSVSVNSKIGAIIVAALVLSILSIIISLLFCCWAYIATKEIRRINFKTNHINTISSSVDDLIRY
三个MuV F原体在成熟F蛋白中寡聚化，其采用亚稳态的融合前构象，当与靶细胞膜接触时，该融合前构象被触发经历构象变化为融合后构象。这种构象变化暴露疏水序列，称为融合肽，其位于F1胞外域的N-末端，并且其与宿主细胞膜缔合并促进病毒或感染细胞膜与靶细胞膜融合。
“以融合前构象稳定”的MuV F胞外域三聚体包含与由相应天然MuV F序列形成的MuV F胞外域三聚体相比，提供与相应天然MuV F序列相比融合前构象保留增加的一个或更多个氨基酸取代、缺失或插入。融合前构象的“稳定化”可以是，例如，能量稳定化(例如，相对于融合后开放构象降低融合前构象的能量)和/或动力学稳定化(例如，降低从融合前构象到融合后构象的转变速率)。此外，与相应天然MuV F序列相比，MuV F胞外域三聚体以融合前构象的稳定性可以包括对变性的抗性增加。本文提供了确定MuV F胞外域三聚体是否处于融合前构象的方法，包括(但不限于)负染色电子显微镜和使用融合前构象特异性抗体的抗体结合测定。关于MuV F蛋白的术语“pre-F”描述了一种分子，该分子是通过一个或更多个氨基酸取代以融合前构象稳定的三聚体I类融合蛋白。
MuV血凝素-神经氨酸酶(HN)蛋白：MuV包膜糖蛋白，其是一种II型膜蛋白并且促进MuV与宿主细胞膜的附接。全长MuV HN蛋白具有N-末端胞质尾和跨膜结构域(CT和TM，大致氨基酸1-53)和包括茎部(大致氨基酸54-130)和头部区(大致氨基酸131-582)的胞外域(大致氨基酸54-582)。示例性MuV HN蛋白序列在本文中以SEQ ID NO:50(NCBI参考序列AQT03695.1，其通过引用并入本文)提供：
mepskfftisdsatfapgpvsnaadkktfrtcfrilvlsvqavtlilvivtlgelvrmindqglsnqlssitdkiresatmiasavgvmnqvihgvtvslplqiegnqnqllatlaticasqkqvsncstNIPLVNDLRFINGINKFIIEDYATHDFSIGHPLNMPSFIPTATSPNGCTRIPSFSLGKTHWCYTHNVINANCKDHTSSNQYVSMGILVQTASGYPMFKTLKIQYLSDGLNRKSCSIATVPDGCAMYCYVSTQLETDDYAGSSPPTQKLTLLFYNDTVTERTISPSGLEGNWATLVPGVGSGIYFENKLIFPAYGGVLPNSTLGVKSAREFFRPVNPYNPCSGPQQDLDQRALRSYFPSYFSNRRIQSAFLVCAWNQILVTNCELVVPSSNQTMMGAEGRVLLINNRLLYYQRSTSWWPYELLYEISFTFTNSGPSSVNMSWIPIYSFTRPGSGNCSGENVCPTACVSGVYLDPWPLTPYSHQSGINRNFYFTGALLNSSTTRVNPTLYVSALNNLKVLAPYGTQGLFASYTTTTCFQDTGDASVYCVYIMELASNIVGEFQILPVLTRLTIT
如本文所用，MuV HN残基定位参照如SEQ ID NO:50所示的序列进行。
MuV F融合前特异性抗体：与以融合前构象的MuV F蛋白特异性结合，但不与以融合后构象的MuV F蛋白特异性结合的抗体。
核酸分子：核苷酸的聚合形式，其可以包括RNA、cDNA、基因组DNA的正义链和反义链，以及上述的合成形式和混合聚合物。核苷酸是指核糖核苷酸、脱氧核苷酸或任一类型核苷酸的修饰形式。如本文所用，术语“核酸分子”与“核酸”和“多核苷酸”同义。除非另有说明，否则核酸分子的长度通常为至少10个碱基。该术语可以包括单链和双链形式的DNA。多核苷酸可以包括通过天然存在的和/或非天然存在的核苷酸键连接在一起的天然存在的和修饰的核苷酸中的一者或两者。“cDNA”是指与单链或双链形式的mRNA互补或相同的DNA。“编码”是指多核苷酸(例如基因、cDNA或mRNA)中特定核苷酸序列的固有特性，用作在生物过程中合成其他聚合物和大分子的模板，这些聚合物和大分子具有确定的核苷酸序列(即rRNA、tRNA和mRNA)或确定的氨基酸序列以及由此产生的生物学特性。
可操作地连接：当第一核酸序列与第二核酸序列处于功能关系时，第一核酸序列与第二核酸序列可操作地连接。例如，如果启动子影响编码序列的转录或表达，则启动子与编码序列可操作地连接。通常，可操作地连接的核酸序列是连续的，并且在需要连接两个蛋白编码区时，在相同的阅读框中。
药学上可接受的运载体：使用的药学上可接受的运载体是常规的。雷明顿制药科学(Remington’s Pharmaceutical Sciences)，E.W.Martin,Mack Publishing Co.,Easton,PA,第19th版,1995描述了适用于公开的免疫原的药物递送的组合物和制剂。
通常，运载体的性质将取决于所采用的特定施用方式。例如，肠胃外制剂通常包含可注射流体，其包括药学上和生理学上可接受的流体，例如水、生理盐水、平衡盐溶液、葡萄糖水溶液、甘油等作为溶媒。对于固体组合物(例如，粉剂、丸剂、片剂或胶囊形式)，常规的无毒固体运载体可以包括，例如，药物级的甘露醇、乳糖、淀粉或硬脂酸镁。除了生物中性运载体之外，待施用的药物组合物(例如免疫原性组合物)可以含有少量的无毒辅助物质，例如润湿剂或乳化剂、防腐剂和pH缓冲剂等，例如乙酸钠或脱水山梨糖醇单月桂酸酯。在特定实施方案中，适合于向受试者施用的运载体可以是无菌的，和/或悬浮的或以其他方式包含在单位剂型中，该单位剂型包含一个或更多个测量剂量的适合诱导期望免疫应答的组合物。它还可以与用于治疗目的的药物伴随用药。单位剂型可以，例如，在包含无菌内容物的密封小瓶或用于注射至受试者的注射器中，或冻干用于随后的溶解和施用，或以固体或控释剂型。
多肽：任何氨基酸链，无论长度或翻译后修饰(例如，糖基化或磷酸化)。“多肽”适用于氨基酸聚合物，包括天然存在的氨基酸聚合物和非天然存在的氨基酸聚合物，以及其中一个或更多个氨基酸残基是非天然氨基酸，例如，相应的天然氨基酸的人工化学模拟物。“残基”是指通过酰胺键或酰胺键模拟物掺入多肽中的氨基酸或氨基酸模拟物。多肽具有氨基末端(N-末端)和羧基末端(C-末端)。“多肽”与肽或蛋白可互换使用，并且在本文中用于指氨基酸残基的聚合物。
初免-加强疫苗接种：免疫疗法，包括向受试者施用第一免疫原性组合物(初免疫苗)，然后施用第二免疫原性组合物(加强疫苗)以诱导免疫应答。初免疫苗和/或加强疫苗包括表达免疫应答所针对的抗原的载体(例如病毒载体、RNA或DNA载体)。在初免疫苗之后向受试者施用加强疫苗；本文公开了施用初免疫苗和加强疫苗之间的合适时间间隔，以及此类时间范围的实例。在一些实施方案中，初免疫苗、加强疫苗或初免疫苗和加强疫苗两者另外包括佐剂。在一个非限制性实例中，初免疫苗是基于DNA的疫苗(或基于基因递送的其他疫苗)，而加强疫苗是基于蛋白亚基或蛋白纳米颗粒的疫苗。
蛋白纳米颗粒：自组装、多亚基、基于蛋白的多面体形结构。亚基各自由蛋白或多肽(例如糖基化多肽)组成，并且任选地由以下项的单个或多个特征组成：核酸、辅基、有机和无机化合物。蛋白纳米颗粒的非限制性实例包括铁蛋白纳米颗粒(参见，例如，Zhang,Y.Int.J.Mol.Sci.,12:5406-5421,2011，其通过引用并入本文)、encapsulin纳米颗粒(参见，例如，Sutter等人，Nature Struct.and Mol.Biol.,15:939-947,2008，其通过引用并入本文)、硫氧化酶还原酶(SOR)纳米颗粒(参见，例如，Urich等人，Science,311:996-1000,2006，其通过引用并入本文)、lumazine合酶纳米颗粒(参见，例如，Zhang等人，J.Mol.Biol.,306:1099-1114,2001)或丙酮酸脱氢酶纳米颗粒(参见，例如，Izard等人，PNAS 96:1240-1245,1999，其通过引用并入本文)。铁蛋白、encapsulin、SOR、lumazine合酶和丙酮酸脱氢酶是单体蛋白，它们自组装成球状蛋白复合物，在某些情况下分别由24、60、24、60和60个蛋白亚基组成。在一些实例中，铁蛋白、encapsulin、SOR、lumazine合酶或丙酮酸脱氢酶单体与重组MuV或MeV F胞外域连接并自组装成蛋白纳米颗粒，在其表面呈现重组MuV或MeV F胞外域三聚体，其可以施用于受试者以刺激对抗原的免疫应答。
重组：重组核酸分子是具有非天然存在的序列的一种分子，例如，包括一个或更多个核酸取代、缺失或插入，和/或具有通过人工组合两个否则分开的序列片段而产生的序列。这种人工组合可以通过化学合成或更常见地通过人工操作分离的核酸片段来实现，例如通过基因工程技术。
重组病毒是包括基因组的一种病毒，该基因组包括重组核酸分子。
重组蛋白是具有非天然存在的序列或具有通过人工组合两个否则分开的序列片段而产生的序列的一种蛋白。在若干实施方案中，重组蛋白由已引入宿主细胞(例如细菌或真核细胞)或重组病毒基因组的异源(例如重组)核酸编码。
序列同一性：氨基酸序列之间的相似性以序列之间的相似性来表示，也称为序列同一性。序列同一性经常以同一性百分比来衡量。百分比越高，两个序列越相似。当使用标准方法比对时，多肽的同源物、直向同源物或变体将具有相对高度的序列同一性。
用于比较的序列比对方法是本领域熟知的。各种程序和比对算法描述于：Smith和Waterman,Adv.Appl.Math.2:482,1981；Needleman和Wunsch,J.Mol.Biol.48:443,1970；Pearson和Lipman,Proc.Natl.Acad.Sci.USA 85:2444,1988；Higgins和Sharp,Gene,73:237-44,1988；Higgins和Sharp,CABIOS 5:151-3,1989；Corpet等人，Nuc.Acids Res.16:10881-90,1988；Huang等人Computer Appls.In the Biosciences 8,155-65,1992；和Pearson等人，Meth.Mol.Bio.24:307-31,1994.Altschul等人，J.Mol.Biol.215:403-10,1990，详细考虑了序列比对方法和同源性计算。
多肽的变体通常特征在于在与目的氨基酸序列的全长比对中计数具有至少约75％，例如，至少约80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％序列同一性。当通过该方法评估时，与参考序列具有甚至更大相似性的蛋白将显示出增加的同一性百分比，例如至少80％、至少85％、至少90％、至少95％、至少98％或至少99％序列同一性。当待比较少于整个序列的序列同一性时，同源物和变体通常在10-20个氨基酸的短窗口内具有至少80％序列同一性，并且可能具有至少85％或至少90％或95％序列同一性，这取决于它们与参考序列的相似性。在此类短窗口上确定序列同一性的方法可在互联网上的NCBI网站上获得。
如本文所用，提及“至少90％同一性”(或类似用语)是指与指定的参考序列具有“至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、至少99％或甚至100％同一性”。
信号肽：指导新合成的分泌或膜蛋白进入和穿过膜(例如，内质网膜)的短氨基酸序列(例如，长度为约18-25个氨基酸)。信号肽通常位于多肽的N-末端，并且在多肽穿过膜后被信号肽酶去除。信号肽序列通常包含三个常见的结构特征：N-末端极性碱性区域(n区域)、疏水性核心和亲水性c区域)。示例性信号肽序列以MKAFSVTCLSFAVFSSSIC(SEQ ID NO:2的残基1-19)示出。
特异性结合：当提及抗体：抗原蛋白复合物或蛋白：蛋白复合物的形成时，是指在蛋白和其他生物制剂的异质群存在下，确定靶蛋白、肽或多糖(例如，糖蛋白)的存在的结合反应。因此，在指定条件下，特定抗体或蛋白优先结合特定靶蛋白、肽或多糖(例如病原体表面上存在的抗原，例如，MuV F或MeV F胞外域三聚体的融合前构象的膜远端顶点处的抗原位点)并且未大量结合样本或受试者中存在的其他蛋白或多糖或相同蛋白的可选构象(例如，MuV或MeV F蛋白的融合后构象)。可以通过本领域已知的方法确定特异性结合。当相互作用的KD小于10-6摩尔，例如小于10-7摩尔、小于10-8摩尔、小于10-9或甚至小于10-10摩尔时，第一蛋白或抗体特异性结合靶蛋白。
可溶性蛋白：在室温下能够溶解在水性液体中并保持溶解状态的蛋白。蛋白的溶解度可能根据水基液体中蛋白的浓度、液体的缓冲条件、液体中其他溶质的浓度(例如盐和蛋白浓度)以及液体的热量而改变。在若干实施方案中，可溶性蛋白是在室温下在磷酸盐缓冲盐水(pH 7.4)中溶解至至少0.5mg/ml浓度并保持溶解至少48小时的蛋白。
受试者：活的多细胞脊椎动物生物，包括人和非人哺乳动物的类别。在实例中，受试者是人。在特定实例中，受试者是新生婴儿。在另一个实例中，选择需要抑制MuV或MeV感染的受试者。例如，受试者未感染并处于感染MuV或MeV的风险中，或者被感染需要治疗。
T4纤维蛋白三聚化结构域：也称为“折叠子”结构域，T4纤维蛋白三聚化结构域包含天然形成三聚体结构的氨基酸序列。在一些实例中，T4纤维蛋白三聚化结构域可以包括在公开的重组蛋白的氨基酸序列中，从而抗原将形成三聚体。在一个实例中，T4纤维蛋白三聚化结构域包含如GYIPEAPRDGQAYVRKDGEWVLLSTFL(SEQ ID NO:34)所示的氨基酸序列。若干实施方案包括可以从纯化的蛋白切割的T4纤维蛋白三聚化结构域，例如通过掺入与可用于切割目的的T4纤维蛋白三聚化结构域相邻的凝血酶切割位点。
跨膜结构域：插入脂质双层的氨基酸序列，例如细胞或病毒或病毒样颗粒的脂质双层。跨膜结构域可用于将抗原锚定至膜。在一些实例中，跨膜结构域是MuV F跨膜结构域。在其他实例中，跨膜结构域是MeV F跨膜结构域。
在足以满足以下的条件下：用于描述允许期望活性的任何环境的短语。
疫苗：能够刺激免疫应答的免疫原性材料制剂，用于预防、改善或治疗感染性或其他类型的疾病。免疫原性材料可以包括减毒或杀死的微生物(例如细菌或病毒)，或源自它们的抗原蛋白、肽或DNA。疫苗可以包括公开的免疫原(例如重组MuV或MeV F胞外域三聚体或编码它们的核酸分子)、病毒、细胞或一种或更多种细胞成分。疫苗可以引起预防性(预防性或保护性)和治疗性应答。施用方法根据疫苗而异，但可能包括接种、摄入、吸入或其他形式的施用。疫苗可以与佐剂一起施用以增强免疫应答。在一个具体的非限制性实例中，与对照相比，疫苗预防和/或降低与MuV感染相关的症状的严重程度和/或降低病毒载量。在另一个具体的非限制性实例中，与对照相比，疫苗预防和/或降低与MeV感染相关的症状的严重程度和/或降低病毒载量。
载体：含有携带启动子的DNA或RNA分子的实体，该启动子可操作地连接至目的抗原的编码序列并且可以表达编码序列。非限制性实例包括裸露的或包裹的(脂质和/或蛋白)DNA、裸露的或包裹的RNA、病毒或细菌或可能无法复制的其他微生物的亚组分，或病毒或细菌或可能具有复制能力的其他微生物。载体有时被称为构建体。重组DNA载体是具有重组DNA的载体。载体可以包括允许其在宿主细胞中复制的核酸序列，例如复制起点。载体还可以包括一种或更多种选择标志物基因和本领域已知的其他遗传元件。病毒载体是具有至少一些源自一种或更多种病毒的核酸序列的重组核酸载体。
病毒样颗粒(VLP)：非复制的病毒壳，源自多种病毒中的任何一种。VLP通常由一种或更多种病毒蛋白组成，例如但不限于，那些称为衣壳、外壳(coat)、壳(shell)、表面和/或包膜蛋白的蛋白，或源自这些蛋白的颗粒形成多肽。在适当的表达系统中重组表达蛋白后，VLP可以自发形成。产生特定VLP的方法是本领域已知的。可以使用本领域已知的常规技术，例如通过电子显微镜、生物物理表征等来检测病毒蛋白重组表达后VLP的存在。此外，可以通过已知技术分离VLP，例如密度梯度离心，并通过特征密度带进行鉴定。参见，例如，Baker等人(1991)Biophys.J.60:1445-1456；和Hagensee等人(1994)J.Virol.68:4503-4505；Vincente,J InvertebrPathol.,2011；Schneider-Ohrum和Ross,Curr.Top.Microbiol.Immunol.,354:53073,2012)。
II.免疫原
A.重组MuV F胞外域三聚体
本文公开了重组MuV F胞外域三聚体，其从天然形式修饰(例如，通过引入一个或更多个氨基酸取代)以融合前构象稳定。如实施例中所述，已通过多轮基于结构的设计选择公开的MuV F胞外域三聚体的实施方案以优化溶解度、稳定性、表达和免疫原性。重组MuV F胞外域三聚体可用于诱导脊椎动物(例如人)对MuV产生免疫应答。与以融合前构象不稳定的相应MuV F胞外域三聚体相比，示例性实施方案显示在动物模型中产生优异的免疫应答。
在一些实施方案中，免疫原包含重组MuV F胞外域三聚体，其包含包含使MuV F胞外域三聚体以融合前构象稳定的一个或更多个氨基酸取代或缺失的原体。
在一些实施方案中，免疫原包含重组MuV F胞外域三聚体，其通过三聚体原体中的一个或更多个氨基酸取代以融合前构象稳定，该氨基酸取代包括形成非天然二硫键以稳定以融合前构象的MuV胞外域三聚体的半胱氨酸取代。非天然二硫键是在天然MuV F蛋白中不存在的一种二硫键，并且通过蛋白工程引入(例如，通过包括形成非天然二硫键的一个或更多个取代的半胱氨酸残基)。例如，在一些实施方案中，任何公开的重组MuVF蛋白可以通过1、2、3、4、5、6、7、8、9或10个非天然二硫键中的任何一个以融合前构象稳定。
可以通过一个或更多个氨基酸取代将形成二硫键的半胱氨酸残基引入天然MuV F序列。例如，在一些实施方案中，单个氨基酸取代引入与天然MuV F序列中存在的半胱氨酸残基形成二硫键的半胱氨酸。可选地，可以将两个半胱氨酸残基引入天然MuV F序列以形成二硫键。非天然二硫键的一个半胱氨酸(或多个半胱氨酸)的位置可以由本领域普通技术人员使用公开的以融合前构象的MuV F胞外域三聚体的结构来确定。
半胱氨酸的氨基酸位置通常在足够近的距离内，以便在MuV F蛋白三聚体的融合前构象中形成二硫键。使用三维结构数据(例如，如表1中提供)来确定两个残基是否在足够近的距离内以形成二硫键的方法是已知的(参见，例如，Peterson等人，Proteinengineering,12:535-548,1999和Dombkowski,Bioinformatics,19:1852-1853,3002(公开了DISULFIDE BY DESIGNTM)，其各自通过引用并入本文)。基于本文提供的以融合前构象的MuVF三聚体的三维结构，可以手动选择残基，或可以使用软件，例如DISULFIDEBYDESIGNTM。不受理论束缚，形成二硫键的理想距离通常被认为对于Cα-Cα距离为约～
Figure GDA0003943442610000151
，对于Sγ-Sγ距离为约
Figure GDA0003943442610000152
，以及对于Cβ-Cβ距离为约
Figure GDA0003943442610000153
(使用最佳转子)。本领域普通技术人员将理解，当在三维结构中选择可取代半胱氨酸以引入二硫键的残基时，包括这些距离的变化。例如，在一些实施方案中，选择的残基具有小于
Figure GDA0003943442610000154
的Cα-Cα距离和/或小于
Figure GDA0003943442610000156
的Cβ-Cβ距离。在一些实施方案中，选择的残基具有
Figure GDA0003943442610000155
的Cα-Cα距离和/或
Figure GDA0003943442610000157
的Cβ-Cβ距离。
在一些实施方案中，重组MuV F胞外域三聚体的原体包含位于MuV F第86和215位的半胱氨酸取代(例如N86C和A215C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MuV F胞外域三聚体的原体包含位于MuV F第155和161位的半胱氨酸取代(例如K155C和L161C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MuV F胞外域三聚体的原体包含位于MuV F第165和231位的半胱氨酸取代(例如V165C和M231C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MuV F胞外域三聚体的原体包含位于MuV F第206和223位的半胱氨酸取代(例如V206C和A223C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MuV F胞外域三聚体的原体包含位于MuV F第209和214位的半胱氨酸取代(例如P209C和P214C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MuV F胞外域三聚体的原体包含位于MuV F第221和255位的半胱氨酸取代(例如I221C和M255C取代)，其形成非天然原体内二硫键以融合前构象稳定。
任何上述重组MuV F蛋白可以进一步包含修饰以消除F1和F2多肽之间的蛋白酶切割位点以产生“单链”重组F蛋白。例如，任何上述重组MuV蛋白可以包含缺失MuV F第101-103位，其中第100和104位通过肽接头融合。这种修饰去除F2/F1弗林蛋白酶切割位点，还去除融合肽的第一残基(其是疏水性)。可以使用融合F2和F1胞外域并使得胞外域折叠成融合前构象的任何合适的肽接头。在一些实施方案中，肽接头是甘氨酸、丝氨酸或甘氨酸-丝氨酸肽接头。在一些实施方案中，肽接头是Gly-Gly-Gly接头。
在非限制性实例中，提供了重组MuV F胞外域三聚体，其包括具有V206C和A223C取代以形成非天然二硫键和缺失MuV F第101-103位(其中第100和104位通过Gly-Gly-Gly肽接头融合)的原体。
在若干实施方案中，重组MuV F胞外域的原体可以包含一个或更多个另外的氨基酸取代，例如，以增加融合前构象的稳定性，或用于其他目的，例如增加溶解度或减少非期望的免疫应答。
以上列出的非天然二硫键将MuV F胞外域的膜远端部分以其融合前构象稳定。任何这些突变均可以与对MuV F胞外域的膜近端部分(例如茎部)的修饰组合，例如，以增加胞外域的三聚化。
在若干实施方案中，重组F2多肽在原体中的N-末端位置可以是MuV F第20-30位中的一个(例如第20位)，并且F1胞外域的C-末端位置可以来自胞外域的茎部区，例如MuV F第469-483位中的一个(例如第476位)。
在非限制性实例中，提供了包括原体的重组MuV F胞外域三聚体，该原体包括MuV第20-476位具有V206C和A223C取代以形成非天然二硫键和缺失MuV F第101-103位，其中第100和104位通过Gly-Gly-Gly肽接头融合。
本文提供了MuV F胞外域三聚体的原体的非限制性实例，该原体包括氨基酸取代用于以融合前构象稳定。在一些实施方案中，MuV F胞外域三聚体的原体包含与SEQ ID NO:3-8中任一项的残基20-483，SEQ ID NO:11-16、26或51中任一项的残基20-476，或SEQ IDNO:19-24中任一项的残基20-469具有至少90％同一性的氨基酸序列；其中该原体包含使MuV F胞外域三聚体以融合前构象稳定的一个或更多个氨基酸取代。在一些实施方案中，MuV F胞外域三聚体的原体包含SEQ ID NO:3-8中任一项的残基20-483，SEQ ID NO:11-16、26或51中任一项的残基20-476，或SEQ ID NO:19-4中任一项的残基20-469。
在若干实施方案中，重组MuV F胞外域三聚体是可溶性蛋白复合物，例如，用作重组亚基疫苗。在若干此类实施方案中，重组MuV F胞外域三聚体的原体可以各自包含与三聚化结构域(例如GCN4三聚化结构域或T4纤维蛋白三聚化结构域或两者)的C-末端连接。三聚化结构域促进重组MuV F胞外域三聚体的膜近端方面的三聚化和稳定化。例如，重组MuV F胞外域三聚体的原体的C-末端残基(例如三聚体的茎部区的残基)可以与三聚化结构域直接连接，或通过肽接头与三聚化结构域间接连接。示例性接头包括甘氨酸和甘氨酸-丝氨酸接头。促进可溶性重组蛋白的稳定三聚体的外源多聚化结构域的非限制性实例包括：GCN4亮氨酸拉链、T4纤维蛋白三聚化结构域、来自肺表面活性蛋白(Hoppe等人1994FEBS Lett344:191-195)或胶原蛋白(McAlinden等人2003J Biol Chem 278:42200-42207)的三聚化基序，只要重组MuV F胞外域三聚体保留融合前构象，其中任何一个均可以连接至重组MuVF胞外域的原体的C-末端以促进三聚化。在一些实例中，重组MuV F胞外域三聚体的原体可以连接至MuV三聚化结构域，例如，三聚体中的每个原体可以包括与GCN4三聚化结构域的C-末端连接，例如与以下中任何一个的连接：MuV F第469-483位，例如MuV F第469位、MuV F第476位或MuV F第483位。在具体实例中，GCN4三聚化结构域包含氨基酸序列IEDKIEEILSKIYHIENEIARIKKLIGEAP(SEQ ID NO:33)或由其组成。在具体实例中，T4纤维蛋白三聚化结构域包含氨基酸序列GYIPEAPRDGQAYVRKDGEWVLLSTFL(SEQ ID NO:34)或由其组成。在具体实例中，与纤维蛋白三聚化结构域融合的GCN4三聚化结构域包含氨基酸序列IEDKIEEILSKIYHIENEIIARIKKLIGEAPGSGYIPEAPRDGQAYVRKDGEWVLLSTFL(SEQ ID NO:35)或由其组成。
在非限制性实例中，提供了包括原体的重组MuV F胞外域三聚体，该原体包括MuV第20-476位具有V206C和A223C取代以形成非天然二硫键，缺失MuV F第101-103位(其中第100和104位通过Gly-Gly-Gly肽接头融合)和与胞外域中原体的C-末端连接的GCN4三聚化结构域。
本文提供了MuV F胞外域三聚体的原体的非限制性实例，该原体包括用于以融合前构象稳定的氨基酸取代以及与三聚化结构域的C-末端连接。在一些实施方案中，MuV F胞外域三聚体的原体包含与SEQ ID NO:3-8中任一项的残基20-513，SEQ IDNO:11-16、26或51中任一项的残基20-506，或SEQ ID NO:19-24中任一项的残基20-499具有至少90％同一性的氨基酸序列；并且其中该原体包含使MuV F胞外域三聚体以融合前构象稳定的一个或更多个氨基酸取代。在一些实施方案中，MuV F胞外域三聚体的原体包含SEQ ID NO:3-8中任一项的残基20-513，SEQ ID NO:11-16、26或51中任一项的残基20-506，或SEQ ID NO:19-24中任一项的残基20-499。
在一些实施方案中，重组MuV F胞外域三聚体可以是膜锚定蛋白复合物，例如，用于减毒病毒或病毒样颗粒疫苗。可以实现膜锚定，例如，通过重组MuV F胞外域三聚体的原体与跨膜结构域和任选的胞质尾(例如MuV F跨膜结构域和胞质尾)的C-末端连接。在一些实施方案中，可以使用一种或更多种肽接头(例如gly-ser接头，例如10个氨基酸的甘氨酸-丝氨酸肽接头)将重组MuV F胞外域三聚体的原体与跨膜结构域连接。与公开的实施方案一起使用的跨膜结构域的非限制性实例包括MuVF跨膜结构域，例如GAIIVAALVLSILSIIISLLFCCW(SEQ ID NO:44)。与公开的实施方案一起使用的跨膜结构域和胞质尾的非限制性实例包括MuVF跨膜结构域和胞质尾，例如GAIIVAALVLSILSIIISLLFCCWAYIATKEIRRINFKTNHINTISSSVDDLIRY(SEQ IDNO:99)。
来自不同MuV毒株的天然MuV F蛋白以及编码此类蛋白的核酸序列和方法是已知的并且可以使用本文提供的描述进行改变以产生重组MuV F胞外域三聚体。
重组MuV F胞外域三聚体可以衍生或连接至另一个分子(例如另一个肽或蛋白)。通常，重组MuV F胞外域被衍生化，使得中和抗体与重组MuV F蛋白的三聚体的结合不受衍生化或标记的不利影响。例如，重组MuV F胞外域可以功能性连接(通过化学偶联、基因融合、非共价缔合或其他方式)至一个或更多个其他分子实体，例如运载体蛋白、抗体、异源蛋白或检测标签。
在一些实施方案中，重组MuV F胞外域三聚体与一个或更多个MuV HN胞外域，例如如下所述的MuV HN序列的胞外域头部融合：
MuV HN胞外域头部(基因型G，Arkansas16，GenBank ARM65482.1(SEQ ID NO:30)
NIPLVNDLRFINGINKFIIEDYATHDFSIGHPLNMPSFIPTATSPNGCTRIPSFSLGKTHWCYTHNVINANCKDHTSSNQYVSMGILVQTASGYPMFKTLKIQYLSDGLNRKSCSIATVPDGCAMYCYVSTQLETDDYAGSSPPTQKLTLLFYNDTVTERTISPSGLEGNWATLVPGVGSGIYFENKLIFPAYGGVLPNSTLGVKSAREFFRPVNPYNPCSGPQQDLDQRALRSYFPSYFSNRRIQSAFLVCAWNQILVTNCELVVPSSNQTMMGAEGRVLLINNRLLYYQRSTSWWPYELLYEISFTFTNSGPSSVNMSWIPIYSFTRPGSGNCSGENVCPTACVSGVYLDPWPLTPYSHQSGINRNFYFTGALLNSSTTRVNPTLYVSALNNLKVLAPYGTQGLFASYTTTTCFQDTGDASVYCVYIMELASNIVGEFQILPVLTRLTIT
在一些实施方案中，重组MuV F胞外域三聚体与一个或更多个MuV HN胞外域融合，例如MuV HN第54-130位至MuV HN第582位(例如MuV HN第54-63位至MuV HN第582位，例如，第54-582、61-582、63-582或55-582位)中任一个的胞外域茎部和头部。在一些实施方案中，重组MuV F胞外域三聚体与一个或更多个MuV HN胞外域融合，例如如SEQ ID NO:90的残基22-550、SEQ ID NO:91的残基22-543、SEQ ID NO:92的残基22-541或SEQ ID NO:93的残基22-549所示的序列的胞外域茎部和头部。
例如，重组MuV F胞外域三聚体的原体各自与MuV HN胞外域融合。融合可以是直接的或通过肽接头。在一些实施方案中，MuV HN胞外域可以与MuV F胞外域三聚体的原体的C-末端直接融合或通过肽接头间接融合。在一些此类实施方案中，MuV HN胞外域可以与与MuVF胞外域三聚体的原体的C-末端融合的三聚化结构域(例如GCN4或T4纤维蛋白三聚化结构域)的C-末端直接融合或通过肽接头间接融合。在一些此类实施方案中，与三聚化结构域连接的MuV F胞外域三聚体和MuV HN胞外域的原体包含如SEQ ID NO:27的残基20-966所示的氨基酸序列，或与SEQ ID NO:27的残基20-966具有至少90％同一性的氨基酸序列。
在一些实施方案中，重组MuV F胞外域三聚体与一个或更多个MeV H胞外域，例如如下所示的H序列的胞外域头部的原体融合：
MeV H胞外域头部(SEQ ID NO:31)
FLAVSKGNCSGPTTIRGQFSNMSLSLLDLYLGRGYNVSSIVTMTSQGMYGGTYLVEKPNLSSKRSELSQLSMYRVFEVGVIRNPGLGAPVFHMTNYLEQPVSNDLSNCMVALGELKLAALCHGEDSITIPYQGSGKGVSFQLVKLGVWKSPADMQSWVPLSTDDPVIDRLYLSSHRGVIADNQAKWAVPTTRTDDKLRMETCFQQACKGKIQTLCENPEWAPLKDNRIPSYGVLSVDLSLTVELKIKIASGFGPLITHGSGMDLYKSNHNNVYWLTIPPMKNLALGVINTLEWIPRFKVSPYLFTVPIKEAGEDCHAPTYLPAEVDGDVKLSSNLVILPGQDLQYVLATYDTSRVEHAVVYYVYSPGRSFSYFYPFRLPIKGVPIELQVECFTWDQKLWCRHFCVLADSESGGHITHSGMVGMGVSCTVTREDGTNRR
MeV H胞外域头部(SEQ ID NO:32)
ADVAAEELMNALVNSTLLEARATNQFLAVSKGNCSGPTTIRGQFSNMSLSLLDLYLSRGYNVSSIVTMTSQGMYGGTYLVGKPNLSSKGSELSQLSMHRVFEVGVIRNPGLGAPVFHMTNYFEQPVSNDFSNCMVALGELKFAALCHREDSITIPYQGSGKGVSFQLVKLGVWKSPTDMRSWVPLSTDDPVIDRLYLSSHRGVIADNQAKWAVPTTRTDDKLRMETCFQQACKGKNQALCENPEWAPLKDNRIPSYGVLSVNLSLTVELKIKIASGFGPLITHGSGMDLYKTNHNNVYWLTIPPMKNLALGVINTLEWIPRFKVSPNLFTVPIKEAGEDCHAPTYLPAEVDGDVKLSSNLVILPGQDLQYVLATYDTSRVEHAVVYYVYSPSRSFSYFYPFRLPIKGVPIELQVECFTWDKKLWCRHFCVLADSESGGHITHSGMVGMGVSCTVTREDGTNRR
在一些实施方案中，重组MuV F胞外域三聚体与一个或更多个MeV H胞外域融合，例如MeV H第59-179位至MeV H第617位中任一位置(例如MeV H第59-67位至MeV H第617位，例如第59-617、62-617、60-617或67-617位中任一位置)的胞外域茎部和头部。在一些实施方案中，重组MuV F胞外域三聚体与一个或更多个MeV H胞外域融合，例如如SEQ ID NO:86的残基22-580、SEQ ID NO:87的残基22-577、SEQ ID NO:88的残基22-579或SEQ ID NO:89的残基22-572所示的序列的胞外域茎部和头部。
例如，以融合前构象稳定的重组MuV F胞外域三聚体的原体各自与MeV H胞外域融合。融合可以是直接的或通过肽接头。在一些实施方案中，MeV H胞外域可以与融合前MuV F胞外域三聚体的原体的C-末端直接融合或通过肽接头间接融合。在一些此类实施方案中，MeV H胞外域可以与与MuV F胞外域三聚体的原体的C-末端融合的三聚化结构域(例如GCN4或T4纤维蛋白三聚化结构域)的C-末端直接融合或通过肽接头间接融合。在一些此类实施方案中，连接至三聚化结构域的MuV F胞外域三聚体和MeV H胞外域的原体包含如SEQ IDNO:28的残基21-981或SEQ ID NO:29的残基20-1006所示的氨基酸序列，或与SEQ ID NO:28的残基21-981或SEQ ID NO:29的残基20-1006具有至少90％同一性的氨基酸序列。
如下提供了包含具有用于以融合前构象稳定的氨基酸取代的MuV F胞外域的序列的非限制性实例：
MuV F V206C/A223C-GGG-483-GCN4(SEQ ID NO:3)mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttCfqpqltnpalspisiqClrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsvsvnski IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F N86C/A215C-GGG-483-GCN4(SEQ ID NO:4)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeCinniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpClspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsvsvnski IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F P209C/P214C-GGG-483-GCN4(SEQ ID NO:5)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqCqltnCalspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiq
lpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsvsvnski IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F K155C/L161C-GGG-483-GCN4(SEQ ID NO:6)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevCegtqqCaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsvsvnski IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F V165C/M231C-GGG-483-GCN4(SEQ ID NO:7)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiaCqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsCtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsvsvnski IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F I221C/M255C-GGG-483-GCN4(SEQ ID NO:8)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisCqalrsllgsmtpavvqatlstsisaaeilsaglCegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsvsvnski IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F V206C/A223C-GGG-476-GCN4(SEQ ID NO:11)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttCfqpqltnpalspisiqClrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqs IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F N86C/A215C-GGG-476-GCN4(SEQ ID NO:12)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeCinniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpClspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqs IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F P209C/P214C-GGG-476-GCN4(SEQ ID NO:13)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqCqltnCalspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqs IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F K155C/L161C-GGG-476-GCN4(SEQ ID NO:14)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevCegtqqCaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqs IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F V165C/M231C-GGG-476-GCN4(SEQ ID NO:15)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiaCqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsCtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqs IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F I221C/M255C-GGG-476-GCN4(SEQ ID NO:16)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisCqalrsllgsmtpavvqatlstsisaaeilsaglCegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqs IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F V206C/A223C-GGG-469-GCN4(SEQ ID NO:19)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttCfqpqltnpalspisiqClrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyike IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F N86C/A215C-GGG-469-GCN4(SEQ ID NO:20)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeCinniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpClspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyike IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F P209C/P214C-GGG-469-GCN4(SEQ ID NO:21)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqCqltnCalspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyike IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F K155C/L161C-GGG-469-GCN4(SEQ ID NO:22)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevCegtqqCaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyike IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F V165C/M231C-GGG-469-GCN4(SEQ ID NO:23)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiaCqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsCtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyike IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F I221C/M255C-GGG-469-GCN4(SEQ ID NO:24)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisCqalrsllgsmtpavvqatlstsisaaeilsaglCegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyike IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV-JL F 206C/A223C-GGG-476-GCN4(SEQ ID NO:26)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyVvvkllpniqptdNscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttCfqpqlInpalspisiqClrsllgsmtpavvqatlstsisaaeilsaglmegqiVsvlldemqmivkinVptivtqsnalvidfysissfinnqesiiqlpdrileigneqwRypaknckStrhhifcqyneaerlsleTklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltScqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqs IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV-IL17 F 206C/A223C-GGG-476-GCN4(SEQ ID NO:51)
mkvslvtclgfavfsfsicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptdnscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqnhintimntqlnnmscqildnqlatslglylteLTTCfqpqlinpalspisiqCLRSllgsmtpavvqatlstsisaaeilsaglmegqivsvlldemqmivkiniptivtqsnalvidfysissfingqesiiqlpdrileigneqwsypaknckltrhnifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelikvnaslqnavkyikesnhqlqs IEDKIEEILSKIYHIENEIARIKKLIGEAP
以上序列包括N-末端信号肽、MuV F胞外域和GCN4三聚化结构域。应当理解，可以使用可选的三聚化结构域，例如T4纤维蛋白三聚化结构域。此外，许多以上序列包括GGG接头，以去除分离F1和F2亚基的天然弗林蛋白酶切割位点。还可以使用可选的甘氨酸接头，例如GSG、GGS或SGG。此外，在任何序列中，可以包括天然弗林蛋白酶切割位点，而不是GGG接头。应当理解，N-末端信号肽在细胞加工过程中被去除并且在纯化的蛋白中不存在。此外，在全长MuV F蛋白中可以包括任何以上序列的MuV F胞外域以提供融合前MuV F蛋白的膜锚定形式，例如用于mRNA免疫。
如下提供了含有MuV F胞外域的序列的非限制性实例，其中氨基酸取代用于以与MuV HN胞外域或MeV H胞外域连接的融合前构象稳定：
MuV F 206C/223C-GGG-476+GCN4+MuV HN_G(SEQ ID NO:27)
MkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttCfqpqltnpalspisiqClrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsIEDKIEEILSKIYHIENEIARIKKLIGEAP GSGGGGGGNiplvndlrfinginkfiiedyathdfsighplnmpsfiptatspngctripsfslgkthwcythnvinanckdhtssnqyvsmgilvqtasgypmfktlkiqylsdglnrkscsiatvpdgcamycyvstqletddyagsspptqkltllfyndtvtertispsglegnwatlvpgvgsgiyfenklifpayggvlpnstlgvksareffrpvnpynpcsgpqqdldqralrsyfpsyfsnrriqsaflvcawnqilvtncelvvpssnqtmmgaegrvllinnrllyyqrstswwpyellyeisftftnsgpssvnmswipiysftrpgsgncsgenvcptacvsgvyldpwpltpyshqsginrnfyftgallnssttrvnptlyvsalnnlkvlapygtqglfasyttttcfqdtgdasvycvyimelasnivgefqilpvltrltit
MuV F 206C/223C-GGG-476+GCN4/Fd+MeV_H(SEQ ID NO:28)
MkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttCfqpqltnpalspisiqClrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsIEDKIEEILSKIYHIENEIARIKKLIGEAP GSGYIPEAPRDGQAYVRKDGEWVLLSTFL GSGGGGGgFlavskgncsgpttirgqfsnmslslldlylgrgynvssivtmtsqgmyggtylvekpnlsskrselsqlsmyrvfevgvirnpglgapvfhmtnyleqpvsndlsncmvalgelklaalchgedsitipyqgsgkgvsfqlvklgvwkspadmqswvplstddpvidrlylsshrgviadnqakwavpttrtddklrmetcfqqackgkiqtlcenpewaplkdnripsygvlsvdlsltvelkikiasgfgplithgsgmdlyksnhnnvywltippmknlalgvintlewiprfkvspylftvpikeagedchaptylpaevdgdvklssnlvilpgqdlqyvlatydtsrvehavvyyvyspgrsfsyfypfrlpikgvpielqvecftwdqklwcrhfcvladsesgghithsgmvgmgvsctvtredgtnrr
MuV F 206C/223C-GGG-476+GCN4/Fd+MeV_H(SEQ ID NO:29)
MkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttCfqpqltnpalspisiqClrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsIEDKIEEILSKIYHIENEIARIKKLIGEAP GSGYIPEAPRDGQAYVRKDGEWVLLSTFL GSGGGGGgAdvaaeelmnalvnstllearatnqflavskgncsgpttirgqfsnmslslldlylsrgynvssivtmtsqgmyggtylvgkpnlsskgselsqlsmhrvfevgvirnpglgapvfhmtnyfeqpvsndfsncmvalgelkfaalchredsitipyqgsgkgvsfqlvklgvwksptdmrswvplstddpvidrlylsshrgviadnqakwavpttrtddklrmetcfqqackgknqalcenpewaplkdnripsygvlsvnlsltvelkikiasgfgplithgsgmdlyktnhnnvywltippmknlalgvintlewiprfkvspnlftvpikeagedchaptylpaevdgdvklssnlvilpgqdlqyvlatydtsrvehavvyyvyspsrsfsyfypfrlpikgvpielqvecftwdkklwcrhfcvladsesgghithsgmvgmgvsctvtredgtnrr
以上序列包括N-末端信号肽和MuV F胞外域与各种其他元件组合，包括GCN4三聚化结构域、T4-纤维蛋白三聚化结构域、肽切割位点(例如凝血酶)、HIS标签、Strep标签，以及区段之间的各种接头残基。这些蛋白的纯化形式通常缺乏通过肽切割去除的N-末端信号肽和C-末端残基。
B.重组MeV F胞外域三聚体
本文公开了重组MeV F胞外域三聚体，其从天然形式修饰(例如，通过引入一个或更多个氨基酸取代)以融合前构象稳定。如实施例中所述，已通过多轮基于结构的设计选择公开的MeV F胞外域三聚体的实施方案以优化溶解度、稳定性、表达和免疫原性。重组MeV F胞外域三聚体可用于诱导脊椎动物(例如人)对MeV产生免疫应答。与以融合前构象不稳定的相应MeV F胞外域三聚体相比，示例性实施方案显示在动物模型中产生优异的免疫应答。
在一些实施方案中，免疫原包括重组MeV F胞外域三聚体，其包含包含使MeV F胞外域三聚体以融合前构象稳定的一个或更多个氨基酸取代或缺失的原体。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含位于MeV F第48和284位的半胱氨酸取代(例如R48C和A284C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含处于MeV F第90和225位的半胱氨酸取代(例如A90C和I225C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含处于MeV F第141和270位的半胱氨酸取代(例如M141C和T270C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含位于MeV F第165和171位的半胱氨酸取代(例如R165C和M171C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含处于MeV F第173和245位的半胱氨酸取代(例如L173C和V245C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含位于MeV F第175和241位的半胱氨酸取代(例如V175C和D241C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含位于MeV F第212和236位的半胱氨酸取代(例如E212C和Y236C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含位于MeV F第216和233位的半胱氨酸取代(例如L216C和A233C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含位于MeV F第219和224位的半胱氨酸取代(例如P219C和P224C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含位于MeV F第99和117位的半胱氨酸取代(例如R99C和V117C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含位于MeV F第100和117位的半胱氨酸取代(例如P100C和V117C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含位于MeV F第101和117位的半胱氨酸取代(例如V101C和V117C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含位于MeV F第102和117位的半胱氨酸取代(例如Q102C和V117C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含位于MeV F第103和117位的半胱氨酸取代(例如S103C和V117C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含位于MeV F第165和171位(例如R165C和M171C取代)以及第141和270位的半胱氨酸取代(例如M141C和T270C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含位于MeV F第165和171位(例如R165C和M171C取代)以及第212和236位的半胱氨酸取代(例如E212C和Y236C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含位于MeV F第165和171位(例如R165C和M171C取代)和第48和284位的半胱氨酸取代(例如R48C和A284C取代)，其形成非天然原体内二硫键以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体在MeV F第175位包含苯丙氨酸(例如V175F取代)以定融合前构象稳。苯丙氨酸取代可以与任何公开的半胱氨酸取代或位于MeV F第194位的脯氨酸取代组合以使重组MeV F胞外域三聚体以融合前构象稳定。
在一些实施方案中，重组MeV F胞外域三聚体的原体包含位于MeV F第194位的脯氨酸取代(例如S194P取代)以融合前构象稳定。脯氨酸取代可以与任何公开的半胱氨酸取代组合以使重组MeV F胞外域三聚体以融合前构象稳定。
任何上述重组MeV F蛋白可进一步包含修饰以消除F1和F2多肽之间的蛋白酶切割位点以产生“单链”重组F蛋白。例如，任何上述重组MeV F蛋白可以包含缺失MeV F第111-113位，其中第110和114位通过肽接头融合。这种修饰去除F2/F1弗林蛋白酶切割位点，还去除融合肽的第一残基(其是疏水性)。可以使用融合F2和F1胞外域并使得将胞外域折叠成融合前构象的任何合适的肽接头。在一些实施方案中，肽接头是甘氨酸、丝氨酸或甘氨酸-丝氨酸肽接头。在一些实施方案中，肽接头是Gly-Gly-Gly接头。
在非限制性实例中，提供了重组MeV F胞外域三聚体，其包括具有R165C和M171C取代以形成非天然二硫键和缺失MeV F第111-113位(其中第110和114位通过Gl-Gly-Gly肽接头融合)的原体。
在若干实施方案中，重组MeV F胞外域的原体可以包含一个或更多个另外的氨基酸取代，例如，以增加融合前构象的稳定性，或用于其他目的，例如增加溶解度或减少和非期望的免疫应答。
以上列出的非天然二硫键将MeV F胞外域的膜远端部分以其融合前构象稳定。任何这些突变均可以与对MeV F胞外域的膜近端部分(例如茎部)的修饰相组合，例如，以增加胞外域的三聚化。
在若干实施方案中，重组F2多肽在原体中的N-末端位置可以是MeV F第24-34位中的一个(例如第24位)，并且F1胞外域的C-末端位置可以来自胞外域的茎部区，例如MeV F第472-486位中的一个(例如第486位)。
在非限制性实例中，提供了包括原体的重组MeV F胞外域三聚体，该原体包括MeV第24-486位具有R165C和M171C取代以形成非天然二硫键和缺失MeV F第111-113位，其中第110和114位通过Gly-Gly-Gly肽接头融合。
本文提供了MeV F胞外域三聚体的原体的非限制性实例，该原体包括氨基酸取代用于以融合前构象稳定。在一些实施方案中，MeV F胞外域三聚体的原体包含与SEQ ID NO:37-43或53-55中任一项的残基21-483具有至少90％同一性的氨基酸序列；其中该原体包含使MeV F胞外域三聚体以融合前构象稳定的一个或更多个氨基酸取代。在一些实施方案中，MeV F胞外域三聚体的原体包含SEQ ID NO:37-43或53-55中任一项的残基21-483。
在若干实施方案中，重组MeV F胞外域三聚体是可溶性蛋白复合物，例如，用作重组亚基疫苗。在若干此类实施方案中，重组MeV F胞外域三聚体的原体可以各自包含与三聚化结构域(例如GCN4三聚化结构域或T4纤维蛋白三聚化结构域或两者)的C-末端连接。三聚化结构域促进重组MeV F胞外域三聚体的膜近端方面的三聚化和稳定化。例如，重组MeV F胞外域三聚体的原体的C-末端残基(例如三聚体的茎部区的残基)可以与三聚化结构域直接连接，或通过肽接头与三聚化结构域间接连接。示例性接头包括甘氨酸和甘氨酸-丝氨酸接头。促进可溶性重组蛋白的稳定三聚体的外源多聚化结构域的非限制性实例包括：GCN4亮氨酸拉链、T4纤维蛋白三聚化结构域、来自肺表面活性蛋白(Hoppe等人1994FEBS Lett344:191-195)或胶原蛋白(McAlinden等人2003J Biol Chem 278:42200-42207)的三聚化基序，只要重组MeV F胞外域三聚体保留融合前构象，其中任何一个均可以连接至重组MeVF胞外域的原体的C-末端以促进三聚化。在一些实例中，重组MeV F胞外域三聚体的原体可以连接至MeV三聚化结构域，例如，三聚体中的每个原体可以包括与GCN4三聚化结构域的C-末端连接，例如与以下中任何一个的连接：MeV F第472-486位，例如MeV F第486位。在具体实例中，GCN4三聚化结构域包含氨基酸序列IEDKIEEILSKIYHIENEIARIKKLIGEAP(SEQ IDNO:33)或由其组成。在具体实例中，T4纤维蛋白三聚化结构域包含氨基酸序列GYIPEAPRDGQAYVRKDGEWVLLSTFL(SEQ ID NO:34)或由其组成。在具体实例中，与纤维蛋白三聚化结构域融合的GCN4三聚化结构域包含氨基酸序列IEDKIEEILSKIYHIENEIIARIKKLIGEAPGSGYIPEAPRDGQAYVRKDGEWVLLSTFL(SEQ ID NO:35)或由其组成。
在非限制性实例中，提供了包括原体的重组MeV F胞外域三聚体，该原体包括MeV第24-486位具有R165C和M171C取代以形成非天然二硫键和缺失MeV F第111-113位(其中第110和114位通过Gly-Gly-Gly肽接头融合)以及与胞外域中原体的C-末端连接的GCN4三聚化结构域。
本文提供了MeV F胞外域三聚体的原体的非限制性实例，该原体包括用于以融合前构象稳定的氨基酸取代以及与三聚化结构域的C-末端连接。在一些实施方案中，MeV F胞外域三聚体的原体包含与SEQ ID NO:37-43或53-55中任一项的残基21-513具有至少90％同一性的氨基酸序列；其中该原体包含使MeV F胞外域三聚体稳定以融合前构象稳定的一个或更多个氨基酸取代。在一些实施方案中，MeV F胞外域三聚体的原体包含SEQ ID NO:37-43或53-55中任一项的残基21-513。
在一些实施方案中，重组MeV F胞外域三聚体可以是膜锚定蛋白复合物，例如，用于减毒病毒或病毒样颗粒疫苗。可以实现膜锚定，例如，通过重组MeV F胞外域三聚体的原体与跨膜结构域和任选的胞质尾(例如MeV F跨膜结构域和胞质尾)的C-末端连接。在一些实施方案中，可以使用一种或更多种肽接头(例如gly-ser接头，例如10个氨基酸的甘氨酸-丝氨酸肽接头)将重组MeV F胞外域三聚体的原体与跨膜结构域连接。与公开的实施方案一起使用的跨膜结构域的非限制性实例包括MeV F跨膜结构域，例如CIVYILIAVCLGGLIGI(SEQID NO:100)。与公开的实施方案一起使用的跨膜结构域的非限制性实例包括MeV F跨膜结构域，例如CIVYILIAVCLGGLIGIPALICCCRGRCNKKGEQVGMSRPGLKPDLTGTSKSYVRSL(SEQ ID NO:101)。
来自不同MeV毒株的天然MeV F蛋白以及编码此类蛋白的核酸序列和方法是已知的并且可以使用本文提供的描述进行改变以产生重组MeV F胞外域三聚体。
重组MeV F胞外域三聚体可以衍生或连接至另一个分子(例如另一个肽或蛋白质)。通常，重组MeV F胞外域被衍生化，使得交叉中和抗体与重组MeV F蛋白的三聚体的的结合不受衍生化或标记的不利影响。例如，重组MeV F胞外域可以功能性连接(通过化学偶联、基因融合、非共价缔合或其他方式)至一个或更多个其他分子实体，例如运载体蛋白、抗体、异源蛋白或检测标签。
在一些实施方案中，重组MeV F胞外域三聚体与一个或更多个MuV HN胞外域，例如如下所示的MuV HN序列的胞外域融合：
MuV HN胞外域头部(SEQ ID NO:30)
NIPLVNDLRFINGINKFIIEDYATHDFSIGHPLNMPSFIPTATSPNGCTRIPSFSLGKTHWCYTHNVINANCKDHTSSNQYVSMGILVQTASGYPMFKTLKIQYLSDGLNRKSCSIATVPDGCAMYCYVSTQLETDDYAGSSPPTQKLTLLFYNDTVTERTISPSGLEGNWATLVPGVGSGIYFENKLIFPAYGGVLPNSTLGVKSAREFFRPVNPYNPCSGPQQDLDQRALRSYFPSYFSNRRIQSAFLVCAWNQILVTNCELVVPSSNQTMMGAEGRVLLINNRLLYYQRSTSWWPYELLYEISFTFTNSGPSSVNMSWIPIYSFTRPGSGNCSGENVCPTACVSGVYLDPWPLTPYSHQSGINRNFYFTGALLNSSTTRVNPTLYVSALNNLKVLAPYGTQGLFASYTTTTCFQDTGDASVYCVYIMELASNIVGEFQILPVLTRLTIT
例如，重组MeV F胞外域三聚体的原体各自与MuV HN胞外域融合。融合可以是直接的或通过肽接头。在一些实施方案中，MuV HN胞外域可以与MeV F胞外域三聚体的原体的C-末端直接融合或通过肽接头间接融合。在一些此类实施方案中，MuV HN胞外域可以与与MeVF胞外域三聚体的的原体的C-末端融合的三聚化结构域(例如GCN4或T4纤维蛋白三聚化结构域)的C-末端直接融合或通过肽接头间接融合。在一些此类实施方案中，与三聚化结构域连接的MeV F胞外域三聚体和MuV HN胞外域的原体包含如SEQ ID NO:27的残基20-966所示的氨基酸序列，或与SEQ ID NO:27的残基20-966具有至少90％同一性的氨基酸序列。
在一些实施方案中，重组MeV F胞外域三聚体与一个或更多个MeV H胞外域，例如如下所示的H序列的胞外域融合：
MeV胞外域(SEQ ID NO:31)
FLAVSKGNCSGPTTIRGQFSNMSLSLLDLYLGRGYNVSSIVTMTSQGMYGGTYLVEKPNLSSKRSELSQLSMYRVFEVGVIRNPGLGAPVFHMTNYLEQPVSNDLSNCMVALGELKLAALCHGEDSITIPYQGSGKGVSFQLVKLGVWKSPADMQSWVPLSTDDPVIDRLYLSSHRGVIADNQAKWAVPTTRTDDKLRMETCFQQACKGKIQTLCENPEWAPLKDNRIPSYGVLSVDLSLTVELKIKIASGFGPLITHGSGMDLYKSNHNNVYWLTIPPMKNLALGVINTLEWIPRFKVSPYLFTVPIKEAGEDCHAPTYLPAEVDGDVKLSSNLVILPGQDLQYVLATYDTSRVEHAVVYYVYSPGRSFSYFYPFRLPIKGVPIELQVECFTWDQKLWCRHFCVLADSESGGHITHSGMVGMGVSCTVTREDGTNRR
MeV胞外域(SEQ ID NO:32)
ADVAAEELMNALVNSTLLEARATNQFLAVSKGNCSGPTTIRGQFSNMSLSLLDLYLSRGYNVSSIVTMTSQGMYGGTYLVGKPNLSSKGSELSQLSMHRVFEVGVIRNPGLGAPVFHMTNYFEQPVSNDFSNCMVALGELKFAALCHREDSITIPYQGSGKGVSFQLVKLGVWKSPTDMRSWVPLSTDDPVIDRLYLSSHRGVIADNQAKWAVPTTRTDDKLRMETCFQQACKGKNQALCENPEWAPLKDNRIPSYGVLSVNLSLTVELKIKIASGFGPLITHGSGMDLYKTNHNNVYWLTIPPMKNLALGVINTLEWIPRFKVSPNLFTVPIKEAGEDCHAPTYLPAEVDGDVKLSSNLVILPGQDLQYVLATYDTSRVEHAVVYYVYSPSRSFSYFYPFRLPIKGVPIELQVECFTWDKKLWCRHFCVLADSESGGHITHSGMVGMGVSCTVTREDGTNRR
例如，重组MeV F胞外域三聚体的原体各自与MeV H胞外域融合。融合可以是直接的或通过肽接头。在一些实施方案中，MeV H胞外域可以与MeV F胞外域三聚体的原体的C-末端直接融合或通过肽接头间接融合。在一些此类实施方案中，MeV H胞外域可以与与MeVF胞外域三聚体的原体的C-末端融合的三聚化结构域(例如GCN4或T4纤维蛋白三聚化结构域)的C-末端直接融合或通过肽接头间接融合。在一些此类实施方案中，与三聚化结构域连接的MeV F胞外域三聚体和MeV H胞外域的原体包含如SEQ ID NO:56的残基21-959或SEQID NO 57的残基21-973所示的氨基酸序列，或与其具有至少90％同一性的氨基酸序列。
如下提供了含有具有用于以融合前构象稳定的氨基酸取代的MeV F胞外域的序列的非限制性实例：
MeV F R48C/A284C-GGG-486-GCN4(SEQ ID NO:53)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtCsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiCyptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F A90C/I225C-GGG-486-GCN4(SEQ ID NO:37)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdClnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpCsaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F M141C/T270C-GGG-486-GCN4(SEQ ID NO:54)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsClnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikariChvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R165C/M171C-GGG-486-GCN4(SEQ ID NO:38)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F L173C/V245C-GGG-486-GCN4(SEQ ID NO:39)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemiCavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkCleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F V175C/D241C-GGG-486-GCN4(SEQ ID NO:40)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilaCqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggCinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F E212C/Y236C-GGG-486-GCN4(SEQ ID NO:55)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryytCilslfgpslrdpisaeisiqalsCalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F L216C/A233C-GGG-486-GCN4(SEQ ID NO:41)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryyteilsCfgpslrdpisaeisiqClsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F P219C/P224C-GGG-486-GCN4(SEQ ID NO:42)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgCslrdCisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F S194P-GGG-486-GCN4(SEQ ID NO:43)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F S103C/V117C-GGG-486-GCN4(SEQ ID NO:62)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqCvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F V175F-GGG-486-GCN4(SEQ ID NO:63)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilaFqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F S103C/V117C,V175F,S194P-GGG-486-GCN4(SEQ ID NO:64)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqCvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F S103C/V117C,R165C/M171C,S194P-GGG-486-GCN4(SEQ ID NO:65)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqCvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C-GGG-486-GCN4(SEQ ID NO:66)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C,V175F,S194P-GGG-486-GCN4(SEQ ID NO:67)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C,R165C/M171C,S194P-GGG-486-GCN4(SEQ ID NO:68)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C-GGG-493-GCN4(SEQ ID NO:69)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqil rsmkglssa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C,V175F,S194P-GGG-493-GCN4(SEQ ID NO:70)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqil rsmkglssa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C,R165C/M171C,S194P-GGG-493-GCN4(SEQ ID NO:71)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqil rsmkglssa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F P100C/V117C,R165C/M171C,S194P-GGG-486-GCN4(SEQ ID NO:72)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirCvqsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F V101C/V117C,R165C/M171C,S194P-GGG-486-GCN4(SEQ ID NO:73)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpCqsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F Q102C/V117C,R165C/M171C,S194P-GGG-486-GCN4(SEQ ID NO:74)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvCsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R165C/M171C,M141C/T270C-GGG-486-GCN4(SEQ ID NO:75)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsClnsqaidnlraslettnqaieaiCqagqeCilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarichvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R165C/M171C,E212C/Y236C-GGG-486-GCN4(SEQ ID NO:76)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilavqgvqdyinnelipsmnqlscdligqklglkllryytCilslfgpslrdpisaeisiqalsCalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R165C/M171C,R48C/A284C-GGG-486-GCN4(SEQ ID NO:77)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtCsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiCyptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C,天然弗林-486-GCN4(SEQ ID NO:78)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhkrfagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C,V175F,S194P,天然弗林-486-GCN4(SEQ ID NO:79)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhkrfagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C,R165C/M171C,S194P,天然弗林-486-GCN4(SEQ ID NO:80)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhkrfagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
以上序列包括N-末端信号肽、MeV F胞外域和GCN4三聚化结构域。应当理解，可以使用可选的三聚化结构域，例如T4纤维蛋白三聚化结构域。此外，许多以上序列包括GGG接头，以去除分离F1和F2亚基的天然弗林蛋白酶切割位点。还可以使用可选的甘氨酸接头，例如GSG、GGS或SGG。此外，在任何序列中，可以包括天然弗林蛋白酶切割位点，而不是GGG接头。应当理解，N-末端信号肽在细胞加工过程中被去除并且在纯化的蛋白中不存在。此外，在全长MeV F蛋白中可以包括任何以上序列的MeV F胞外域以提供融合前MeV F蛋白的膜锚定形式，例如用于mRNA免疫。
如下提供了含有MeV F胞外域的序列的非限制性实例，其中氨基酸取代用于以与MuV HN胞外域或MeV H胞外域连接的融合前构象稳定：
MeV F R165C/M171C-GGG-486-GCN4+MeV_H(SEQ ID NO:56)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP GSGGGGGGflavskgncsgpttirgqfsnmslslldlylgrgynvssivtmtsqgmyggtylvekpnlsskrselsqlsmyrvfevgvirnpglgapvfhmtnyleqpvsndlsncmvalgelklaalchgedsitipyqgsgkgvsfqlvklgvwkspadmqswvplstddpvidrlylsshrgviadnqakwavpttrtddklrmetcfqqackgkiqtlcenpewaplkdnripsygvlsvdlsltvelkikiasgfgplithgsgmdlyksnhnnvywltippmknlalgvintlewiprfkvspylftvpikeagedchaptylpaevdgdvklssnlvilpgqdlqyvlatydtsrvehavvyyvyspgrsfsyfypfrlpikgvpielqvecftwdqklwcrhfcvladsesgghithsgmvgmgvsctvtredgtnrr
MeV F R165C/M171C-GGG-486-GCN4+MuV_HN(SEQ ID NO:57)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAPG sggggggniplvndlrfinginkfiiedyathdfsighplnmpsfiptatspngctripsfslgkthwcythnvinanckdhtssnqyvsmgilvqtasgypmfktlkiqylsdglnrkscsiatvpdgcamycyvstqletddyagsspptqkltllfyndtvtertispsglegnwatlvpgvgsgiyfenklifpayggvlpnstlgvksareffrpvnpynpcsgpqqdldqralrsyfpsyfsnrriqsaflvcawnqilvtncelvvpssnqtmmgaegrvllinnrllyyqrstswwpyellyeisftftnsgpssvnmswipiysftrpgsgncsgenvcptacvsgvyldpwpltpyshqsginrnfyftgallnssttrvnptlyvsalnnlkvlapygtqglfasyttttcfqdtgdasvycvyimelasnivgefqilpvltrltit
MeV F R165C/M171C-GGG-486-GCN4/Fd+MeV_H(SEQ ID NO:81)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP GSGYIPEAPRDGQAYVRKDGEWVLLSTFL GSGGGGGg flavskgncsgpttirgqfsnmslslldlylgrgynvssivtmtsqgmyggtylvekpnlsskrselsqlsmyrvfevgvirnpglgapvfhmtnyleqpvsndlsncmvalgelklaalchgedsitipyqgsgkgvsfqlvklgvwkspadmqswvplstddpvidrlylsshrgviadnqakwavpttrtddklrmetcfqqackgkiqtlcenpewaplkdnripsygvlsvdlsltvelkikiasgfgplithgsgmdlyksnhnnvywltippmknlalgvintlewiprfkvspylftvpikeagedchaptylpaevdgdvklssnlvilpgqdlqyvlatydtsrvehavvyyvyspgrsfsyfypfrlpikgvpielqvecftwdqklwcrhfcvladsesgghithsgmvgmgvsctvtredgtnrr
以上序列包括N-末端信号肽、MeV F胞外域、三聚化结构域(GCN4和/或T4纤维蛋白)和MeV H胞外域头部或MuV HN胞外域头部区。MeV F胞外域序列包括GGG接头，用于去除分离F1和F2亚基的天然弗林蛋白酶切割位点。还可以使用可选的甘氨酸接头，例如GSG、GGS或SGG。此外，在任何序列中，可以包括天然弗林蛋白酶切割位点，而不是GGG接头。应当理解，N-末端信号肽在细胞加工过程中被去除并且在纯化的蛋白中不存在。
C.MuV HN和MuV H多聚体
在一些实施方案中，提供了包含MuV HN胞外域和/或MeV H胞外域的多聚体的免疫原。H或HN胞外域可以包括胞外域头部或胞外域茎部和头部。
在一些实施方案中，免疫原包括融合蛋白的三聚体，每种融合蛋白包含一个或更多个MuV HN胞外域或MeV H胞外域和三聚化结构域(例如GCN4三聚化结构域、T4纤维蛋白三聚化结构域或与T4纤维蛋白三聚化结构域融合的GCN4三聚化结构域)。
在一些实施方案中，融合蛋白在N-末端至C-末端方向上包含三聚化结构域(例如GCN4三聚化结构域、T4纤维蛋白三聚化结构域或与T4纤维蛋白三聚化结构域融合的GCN4三聚化结构域)和一个或更多个(例如一个、两个或三个)Mev HN胞外域或Mev H胞外域。三聚化结构域相互作用形成三聚体。在一些实施方案中，包括胞外域的片段，例如MuV HN胞外域的头部区或MeV H胞外域的头部区可以任选地通过肽接头与三聚化结构域融合。在一些实施方案中，三聚体中的融合蛋白包含如以下所示的氨基酸序列或由其组成：SEQ ID NO:58的残基24-510或SEQ ID NO:59的残基24-496，或与SEQ ID NO:58的残基25-510或SEQ IDNO:59的残基25-496中的任一个具有至少90％同一性的序列。
在一些实施方案中，融合蛋白在N-末端至C-末端方向上包含一个或更多个(例如一个、两个或三个)MuV HN胞外域或Mev H胞外域、三聚化结构域(例如GCN4三聚化结构域、T4纤维蛋白三聚化结构域或融合到T4纤维蛋白三聚化结构域的GCN4三聚化结构域)和一个或更多个(例如一个、两个或三个)MuV HN胞外域或Mev H胞外域。三聚化结构域相互作用形成三聚体。在一些实施方案中，包括胞外域的片段，例如MuV HN胞外域的头部区或MeV H胞外域的头部区可以任选地通过肽接头与三聚化结构域融合。在一些实施方案中，三聚体中的融合蛋白包含如SEQ ID NO:82的残基22-950、SEQ ID NO:83的残基25-985、SEQ ID NO:84的残基22-948或SEQ ID NO:85的残基22-981所示的氨基酸序列，或与SEQ ID NO:82的残基22-950、SEQ ID NO:83的残基25-985、SEQ ID NO:84的残基22-948或SEQ ID NO:85的残基22-981中的任一个具有至少90％同一性的序列，或由其组成。
在一些实施方案中，多聚体是MeV H胞外域头部区的二聚体。MeV H胞外域头部区可以在哺乳动物细胞中表达，并且在生理溶液中自发形成二聚体。然后可以纯化该二聚体并用作免疫原。在一些实施方案中，二聚体的亚基包含如SEQ ID NO:60的残基22-459所示的氨基酸序列或与SEQ ID NO:60的残基22-459具有至少90％同一性的序列，或由其组成。
在一些实施方案中，多聚体是MeV H胞外域茎部和头部区的二聚体。MeV H胞外域茎部和头部区可以在哺乳动物细胞中表达，并且在生理溶液中自发形成二聚体。然后可以纯化该二聚体并用作免疫原。在一些实施方案中，二聚体的亚基包含MeV H第59-197位至MeV H第617位中任一位置的氨基酸序列(例如MeV H第59-67位至MeV H第617位中任一位置的氨基酸序列，例如第59-617位、第62-617位、第60-617位或第67-617位)，或由其组成。在一些实施方案中，二聚体的亚基包含如SEQ ID NO:86的残基22-580、SEQ ID NO:87的残基22-577、SEQ ID NO:88的残基22-579或SEQ ID NO:89的残基22-572所示氨基酸序列，或与SEQ ID NO:86的残基22-580、SEQ IDNO:87的残基22-577、SEQ ID NO:88的残基22-579或SEQ ID NO:89的残基22-572具有至少90％同一性的序列，或由其组成。
在一些实施方案中，多聚体是MuV HN胞外域头部区的二聚体。在一些实施方案中，二聚体的亚基包含如SEQ ID NO:30所示的氨基酸序列或与SEQ ID NO:30具有至少90％同一性的序列，或由其组成。
在一些实施方案中，多聚体是MuV H胞外域茎部和头部区的二聚体。在一些实施方案中，二聚体的亚基包含MuV H第54-130位至MuV HN第582位中任一位置(例如MuV H第54-63位至MuV HN第582中任一位置，例如第54-582位、第61-582位、第63-582位或第55-582位)的氨基酸序列，或由其组成。在一些实施方案中，二聚体的亚基包含如SEQ ID NO:90的残基22-550、SEQ ID NO:91的残基22-543、SEQ ID NO:92的残基22-541或SEQ ID NO:93的残基22-549所示的氨基酸序列，或与SEQ ID NO:90的残基22-550、SEQ ID NO:91的残基22-543、SEQ ID NO:92的残基22-541或SEQ ID NO:93的残基22-549具有至少90％同一性的序列，或由其组成。
D.另外的描述
除了上述那些之外，重组MuV或MeV F胞外域三聚体中的原体可以包含天然MuV F或MeV F序列的修饰，例如氨基酸取代、缺失或插入、糖基化和/或与不相关蛋白(例如蛋白标签)的共价连接，只要重组MuV或MeV F胞外域三聚体以融合前构象保持稳定并保持免疫原性。此外，在包括异源MuV HN胞外域或MeV H胞外域，或MuV HN胞外域或MeV H胞外域的多聚体的实施方案中，HN或H胞外域可以包括天然HN或H序列的修饰，例如氨基酸取代、缺失或插入、糖基化和/或与不相关蛋白(例如蛋白标签)的共价连接，只要HN或H胞外域保留免疫原性。这些序列变异可以是天然存在的变异，或它们可以通过使用本领域技术人员已知的基因工程技术进行工程改造。此类技术的实例参见，例如，Sambrook等人(分子克隆：实验室手册，第4版，Cold Spring Harbor,New York,2012)和Ausubel等人(见现代分子生物学实验指南，John Wiley&Sons，New York，至增刊104,2013)，两者均通过引用以其全文并入本文。
在一些实施方案中，与相应的天然MuV F序列相比，重组MuV F胞外域三聚体中的原体包含一个或更多个氨基酸取代。例如，在一些实施方案中，与天然MuV F序列相比，F2多肽、F1胞外域或两者可以包括至多20个(例如至多1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18或19个)氨基酸取代(例如保守氨基酸取代)。
在一些实施方案中，与相应的天然MeV F序列相比，重组MeV F胞外域三聚体中的原体包含一个或更多个氨基酸取代。例如，在一些实施方案中，与天然MeV F序列相比，F2多肽、F1胞外域或两者可以包括至多20个(例如至多1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18或19个)氨基酸取代(例如保守氨基酸取代)。
在一些实施方案中，与相应的天然MuV HN胞外域序列相比，MuV HN胞外域包含一个或更多个氨基酸取代。例如，在一些实施方案中，与天然MuV HN胞外域序列相比，MuV HN胞外域包括至多20个(例如至多1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18或19个)氨基酸取代(例如保守氨基酸取代)。
在一些实施方案中，与相应的天然MeV H胞外域序列相比，MeV H胞外域包含一个或更多个氨基酸取代。例如，在一些实施方案中，与天然MeV H胞外域序列相比，MeV H胞外域包括至多20个(例如至多1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18或19个)氨基酸取代(例如保守氨基酸取代)。
最简单的修饰涉及用一个或更多个氨基酸取代具有相似生化特性的氨基酸，例如保守氨基酸取代。此类取代可能对所得蛋白的活性影响极小。
在一些实施方案中，重组MeV F胞外域三聚体或MuV F胞外域三聚体中的原体可以在任一末端与其他不相关序列(例如非MuV F或MeV F蛋白序列、非病毒包膜或非病毒蛋白序列)
在若干实施方案中，重组MuV F胞外域三聚体或MeV F胞外域三聚体，或这些原体与异源蛋白(例如MuV HN胞外域或MeV H胞外域)的融合物可溶于水溶液中。在一些实施方案中，在室温(例如20-22摄氏度)下，重组MuV F胞外域三聚体MeV F胞外域三聚体，或与异源蛋白(例如MuV HN胞外域或MeV H胞外域)的相应融合物在水溶液(例如磷酸盐缓冲盐水(pH 7.4)或350mM NaCl(pH 7.0))中溶解至至少0.5mg/ml(例如至少1.0mg/ml、1.5mg/ml、2.0mg/ml、3.0mg/ml、4.0mg/ml或至少5.0mg/ml)的浓度并保持溶解至少12小时(例如至少24小时、至少48小时、至少一周、至少两周、至少一个月或更长时间)。在一个实施方案中，磷酸盐缓冲盐水包括NaCl(137mM)、KCl(2.7mM)、Na2HPO4(10mM)、KH2PO4(1.8mM)，pH 7.4。在一些实施方案中，磷酸盐缓冲盐水还包括CaCl2(1mM)和MgCl2(0.5mM。本领域技术人员熟悉确定蛋白是否随时间保留在溶液中的方法。例如，可以使用标准方法随时间测试溶解在水溶液中的蛋白浓度。
在一些实施方案中，免疫原作为基本上以融合前构象的可溶性三聚体的同质群体提供，仅限于没有以融合后构象的MuV F胞外域三聚体和/或MeV F胞外域三聚体。MeV F胞外域三聚体或MuV F胞外域三聚体的构象可以例如通过负染色电子显微镜和/或通过适当的融合前或融合后特异性抗体的特异性结合来检测。在一些实施方案中，同质群体中至少约95％的重组MuV F胞外域三聚体或MeV F胞外域三聚体(例如至少约95％、96％、97％、98％、99％或99.9％的MuV或MeV F蛋白)以融合前构象稳定。
在一些实施方案中，重组MuV F胞外域三聚体或MeV F胞外域三聚体在磷酸盐缓冲盐水中于50℃下孵育一小时后保留对融合前特异性抗体的特异性结合。在一些实施方案中，重组MuV F胞外域三聚体或MeV F胞外域三聚体在磷酸盐缓冲盐水中于4℃下孵育六个月后保留对融合前特异性抗体的特异性结合。
在某些实施方案中，本文提供的免疫原可经进一步修饰以含有本领域已知且容易获得的另外的非蛋白质部分。适用于衍生免疫原的部分包括但不限于水溶性聚合物。水溶性聚合物的非限制性实例包括但不限于聚乙二醇(PEG)、乙二醇/丙二醇的共聚物、羧甲基纤维素、右旋糖、聚乙烯醇、聚乙烯吡咯烷酮、聚-1,3-二氧戊环、聚-1,3,6-三噁烷、乙烯/马来酸酐共聚物、聚氨基酸(均聚物或无规共聚物)和右旋糖或聚(n-乙烯基吡咯烷酮)聚乙二醇、丙二醇均聚物、丙二醇/环氧乙烷共聚物、聚氧乙烯化多元醇(例如甘油)、聚乙烯醇及其混合物。聚乙二醇丙醛由于其在水中的稳定性而在生产中可能具有优势。聚合物可以具有任何分子量，并且可以是支链或非支链的。附接至抗体的聚合物的数量可能不同，并且如果附接多于一种聚合物，它们可以是相同或不同的分子。通常，用于衍生化的聚合物的数量和/或类型可以基于以下考虑，包括但不限于待改进或改变的免疫原的特定性质或功能、免疫原衍生物是否将用于在规定条件下进行治疗等。
本文提供的包括重组MuV F胞外域三聚体、或MeV F胞外域三聚体、或MuV HN胞外域或MeV H胞外域的一些序列包括蛋白酶切割位点(例如凝血酶位点)、蛋白标签(例如His标签、Strep Tag II、Avi标签等)和信号肽的序列；此类序列可以从包括重组MuV F胞外域三聚体、或MeV F胞外域三聚体、或MuV HN胞外域或MeV H胞外域的分离的免疫原中去除以用于治疗用途。
E.蛋白纳米颗粒
在一些实施方案中，提供了蛋白纳米颗粒，其包括一种或更多种公开的重组MuV F胞外域三聚体或重组MeV F胞外域三聚体、或MuV HN、或MeV H多聚体，或它们的嵌合体。
在一些实施方案中，蛋白纳米颗粒包含在双组分自组装纳米颗粒平台上展示的MeV F胞外域三聚体或MuV F胞外域三聚体，如Marcandalli等人“Induction of potentneutralizing antibody responses by a designed protein nanoparticle vaccinefor respiratory syncytial virus,”Cell,176(6):1420-1431,2019中所述，其通过引用并入本文。
在纳米颗粒的另外的非限制性实例中，包括铁蛋白纳米颗粒、encapsulin纳米颗粒、硫加氧酶还原酶(SOR)纳米颗粒和lumazine合酶纳米颗粒，它们由一组单体亚基组成，分别包括铁蛋白、encapsulin、SOR蛋白和lumazine合酶。为了构建此类蛋白纳米颗粒，将重组MuV F胞外域三聚体或重组MeV F胞外域三聚体的原体，或MuV HN或MeV H多聚体的亚基连接至蛋白纳米颗粒的亚基(例如铁蛋白、encapsulin、SOR蛋白或lumazine合酶蛋白)并在适当条件下在细胞中表达。融合蛋白自组装成任何可以纯化的纳米颗粒。
在一些实施方案中，公开的重组MuV F胞外域三聚体或重组MeV F胞外域三聚体的原体，或MuV HN或MeV H多聚体的亚基，可以连接至铁蛋白亚基以构建铁蛋白纳米颗粒。铁蛋白纳米颗粒及其用于免疫目的(例如，用于针对流感抗原的免疫)的用途已在本领域中公开(参见，例如，Kanekiyo等人，Nature,499:102-106,2013，其通过引用以其全文并入本文)。铁蛋白纳米颗粒的球状形式由单体亚基组成，这些单体亚基是分子量为约17-20kDa的多肽。产生后，这些单体亚基蛋白自组装成球状铁蛋白。因此，铁蛋白的球状形式包含24个单体亚基蛋白，并且具有具有432个对称性的衣壳样结构。本文进一步描述了构建铁蛋白纳米颗粒的方法(参见，例如，Zhang,Int.J.Mol.Sci.,12:5406-5421,2011，其通过引用以其全文并入本文)。一个此类单体铁蛋白亚基的氨基酸序列的实例表示为：
DIIKLLNEQVNKEMQSSNLYMSMSSWCYTHSLDGAGLFLFDHAAEEYEHAKKLIIFLNENNVPVQLTSISAPEHKFEGLTQIFQKAYEHEQHISESINNIVDHAIKSKDHATFNFLQWYVAEQHEEEVLFKDILDKIELIGNENHGLYLADQYVKGIAKSRKS(SEQ ID NO:45)
在具体实例中，铁蛋白多肽是大肠杆菌铁蛋白、幽门螺杆菌铁蛋白、人轻链铁蛋白、牛蛙铁蛋白或其杂合体，例如大肠杆菌-人杂合铁蛋白、大肠杆菌-牛蛙杂合铁蛋白或人-牛蛙杂交铁蛋白。用于制备包括重组MuV或MeV F胞外域三聚体的铁蛋白纳米颗粒的铁蛋白多肽的示例性氨基酸序列和编码铁蛋白多肽的核酸序列可见于GENBANK
Figure GDA0003943442610000371
，例如登录号ZP_03085328、ZP_06990637、EJB64322.1、AAA35832、NP_000137AAA49532、AAA49525、AAA49524和AAA49523，它们于2015年4月10日通过引用以其全文并入本文。在一些实施方案中，重组MuV或MeV F胞外域三聚体的原体可以连接至包括与SEQ ID NO:45所示的氨基酸序列具有至少80％(例如至少85％、至少90％、至少95％或至少97％)同一性的氨基酸序列的铁蛋白亚基。
在一些实施方案中，公开的重组MuV F胞外域三聚体或重组MuV或MeV F胞外域三聚体的原体，或MuV HN或MeV H多聚体的亚基，可以连接至lumazine合酶亚基以构建lumazine合酶纳米颗粒。Lumazine合酶纳米颗粒的球状形式由单体亚基组成；一个此类lumazine合酶亚基的序列的实例作为如下所示的氨基酸序列提供：
MQIYEGKLTAEGLRFGIVASRFNHALVDRLVEGAIDCIVRHGGREEDITLVRVPGSWEIPVAAGELARKEDIDAVIAIGVLIRGATPHFDYIASEVSKGLADLSLELRKPITFGVITADTLEQAIERAGTKHGNKGWEAALSAIEMANLFKSLR(SEQ ID NO:46)。
在一些实施方案中，公开的重组MuV F胞外域三聚体或重组MeV F胞外域三聚体的原体，或MuV HN或MeV H多聚体的亚基，可以与包括与如SEQ ID NO:46所示的氨基酸序列具有至少80％(例如至少85％、至少90％、至少95％或至少97％)同一性的氨基酸序列的lumazine合酶亚基连接。
在一些实施方案中，公开的重组MuV F胞外域三聚体或重组MeV F胞外域三聚体的原体，或MuV HN或MeV H多聚体的亚基，可以连接至encapsulin纳米颗粒亚基以构建encapsulin纳米颗粒。Encapsulin纳米颗粒的球状形式由单体亚基组成；一个此类encapsulin亚基的序列的实例作为如下所示的氨基酸序列提供
MEFLKRSFAPLTEKQWQEIDNRAREIFKTQLYGRKFVDVEGPYGWEYAAHPLGEVEVLSDENEVVKWGLRKSLPLIELRATFTLDLWELDNLERGKPNVDLSSLEETVRKVAEFEDEVIFRGCEKSGVKGLLSFEERKIECGSTPKDLLEAIVRALSIFSKDGIEGPYTLVINTDRWINFLKEEAGHYPLEKRVEECLRGGKIITTPRIEDALVVSERGGDFKLILGQDLSIGYEDREKDAVRLFITETFTFQVVNPEALILLKF(SEQ ID NO:47)。
在一些实施方案中，公开的重组MuV F胞外域三聚体或重组MeV F胞外域三聚体的原体，或MuV HN或MeV H多聚体的亚基，可以与包括与如SEQ ID NO:47所示的氨基酸序列具有至少80％(例如至少85％、至少90％、至少95％或至少97％)同一性氨基酸序列的encapsulin亚基连接。
Encapsulin蛋白是保守的细菌蛋白家族，也称为linocin样蛋白，它们形成大蛋白组装体，充当包装酶的最小隔室。Encapsulin组装体由单体亚基组成，这些单体亚基是分子量为约30kDa的多肽。产生后，单体亚基自组装成球状encapsulin组装体，包括60个，或在一些情况下，180个单体亚基。进一步描述了构建encapsulin纳米颗粒的方法(参见，例如，Sutter等人，Nature Struct.and Mol.Biol.,15:939-947,2008，其通过引用以其全文并入本文)。在具体实例中，encapsulin多肽是细菌encapsulin，例如海栖热袍菌(Thermotogamaritime)或嗜热古细菌(Pyrococcusfuriosus)或红串红球菌(Rhodococcuserythropolis)或黄色粘球菌(Myxococcus xanthus)encapsulin。
在一些实施方案中，公开的重组MuV F胞外域三聚体或重组MeV F胞外域三聚体的原体，或MuV HN或MeV H多聚体的亚基，可以与硫加氧酶还原酶(SOR)亚基连接以构建重组SOR纳米颗粒。在一些实施方案中，SOR亚基可以包括如下所示的氨基酸序列
MEFLKRSFAPLTEKQWQEIDNRAREIFKTQLYGRKFVDVEGPYGWEYAAHPLGEVEVLSDENEVVKWGLRKSLPLIELRATFTLDLWELDNLERGKPNVDLSSLEETVRKVAEFEDEVIFRGCEKSGVKGLLSFEERKIECGSTPKDLLEAIVRALSIFSKDGIEGPYTLVINTDRWINFLKEEAGHYPLEKRVEECLRGGKIITTPRIEDALVVSERGGDFKLILGQDLSIGYEDREKDAVRLFITETFTFQVVNPEALILLKF(SEQ ID NO:48)。
在一些实施方案中，公开的重组MuV F胞外域三聚体或重组MeV F胞外域三聚体的原体，或MuV HN或MeV H多聚体的亚基，可以连接至包括与如SEQ ID NO:48所示的氨基酸序列具有至少80％(例如至少85％、至少90％、至少95％或至少97％)同一性的氨基酸序列的SOR亚基。
SOR蛋白是微生物蛋白(例如来自嗜热嗜酸古生菌Acidianusambivalens，其形成24个亚基蛋白组装体。构建SOR纳米颗粒的方法描述于Urich等人，Science,311:996-1000,2006，其通过引用以其全文并入本文。用于制备SOR纳米颗粒的SOR蛋白的氨基酸序列的实例阐述于Urich等人，Science,311:996-1000,2006，其通过引用以其全文并入本文。
出于产生目的，与纳米颗粒亚基连接的重组MuV F胞外域或重组MeV F胞外域，或MuV HN或MeV H多聚体的亚基，可以包括在细胞加工过程中被切割的N-末端信号肽。例如，与蛋白纳米颗粒亚基连接的重组MuV F胞外域原体或重组MeV F胞外域原体可以在其N-末端包括信号肽，包括例如天然MuV或MeV F信号肽。
蛋白纳米颗粒可以在适当的细胞(例如，HEK 293Freestyle细胞)中表达，并且融合蛋白从自组装成纳米颗粒的细胞中分泌。可以使用已知技术纯化纳米颗粒，例如通过一些不同色谱程序，例如Mono Q(阴离子交换)，然后进行尺寸排阻(SUPEROSE
Figure GDA0003943442610000391
6)色谱。
融合蛋白不需要包含铁蛋白、encapsulin、SOR或lumazine合酶蛋白的单体亚基多肽的全长序列。可以使用单体亚基多肽的部分或区域，只要该部分包含指导单体亚基自组装成蛋白的球状形式的氨基酸序列。
II.多核苷酸和表达
还提供了编码任何公开的免疫原的多核苷酸。例如，编码以融合前构象稳定的MuVF胞外域三聚体的原体的多核苷酸，以融合前构象稳定的MeV F胞外域三聚体的原体，与MuVHN或MeV H胞外域连接的这些原体中的一个的嵌合体，或含有重组MuV或MeV F胞外域的自组装蛋白纳米颗粒的亚基。这些多核苷酸包括DNA、cDNA和RNA序列，包括包括DNA、cDNA和RNA序列的载体，例如用于免疫的DNA或RNA载体。遗传密码可用于构建多种功能等同的核酸，例如序列不同但编码相同蛋白序列的核酸，或编码包括该核酸序列的缀合物或融合蛋白。
编码全长MeV F蛋白的示例性核酸序列提供为SEQ ID NO:94：
atgggtctcaaggtgaacgtctctgccatattcatggcagtactgttaactctccaaacacccaccggtcaaatccattggggcaatctctctaagataggggtggtaggaataggaagtgcaagctacaaagttatgactcgttccagccatcaatcattagtcataaaattaatgcccaatataactctcctcaataactgcacgagggtagagattgcagaatacaggagactactgagaacagttttggaaccaattagagatgcacttaatgcaatgacccagaatataagaccggttcagagtgtagcttcaagtaggagacacaagagatttgcgggagtagtcctggcaggtgcggccctaggcgttgccacagctgctcagataacagccggcattgcacttcaccagtccatgctgaactctcaagccatcgacaatctgagagcgagcctggaaactactaatcaggcaattgaggcaatcagacaagcagggcaggagatgatattggctgttcagggtgtccaagactacatcaataatgagctgataccgtctatgaaccaactatcttgtgatttaatcggccagaagctcgggctcaaattgctcagatactatacagaaatcctgtcattatttggccccagcttacgggaccccatatctgcggagatatctatccaggctttgagctatgcgcttggaggagacatcaataaggtgttagaaaagctcggatacagtggaggtgatttactgggcatcttagagagcagaggaataaaggcccggataactcacgtcgacacagagtcctacttaattgtcctcagtatagcctatccgacgctgtccgagattaagggggtgattgtccaccggctagagggggtctcgtacaacataggctctcaagagtggtataccactgtgcccaagtatgttgcaacccaagggtaccttatctcgaattttgatgagtcatcgtgtactttcatgccagaggggactgtgtgcagccaaaatgccttgtacccgatgagtcctctgctccaagaatgcctccgggggtccaccaagtcctgtgctcgtacactcgtatccgggtcttttgggaaccggttcattttatcacaagggaacctaatagccaattgtgcatcaatcctttgcaagtgttacacaacaggaacgatcattaatcaagaccctgacaagatcctaacatacattgctgccgatcactgcccggtagtcgaggtgaacggcgtgaccatccaagtcgggagcaggaggtatccagacgctgtgtacttgcacagaattgacctcggtcctcccatattattggagaggttggacgtagggacaaatctggggaatgcaattgctaagttggaggatgccaaggaattgttggagtcatcggaccagatattgaggagtatgaaaggtttatcgagcacttgcatagtctacatcctgattgcagtgtgtcttggagggttgatagggatccccgctttaatatgttgctgcagggggcgttgtaacaaaaagggagaacaagttggtatgtcaagaccaggcctaaagcctgatcttacgggaacatcaaaatcctatgtaaggtcgctctga
编码全长MuV F蛋白的示例性核酸序列提供为SEQ ID NO:95：
atgaaggccttttcagttacttgcttgggctttgcagtcttttcgtcttctatatgtgtgaatatcaacatcttgcagcaaattggatatatcaagcaacaagtcaggcaactaagctattactcacaaagttctagctcctacatagtggtcaagcttttaccgaatatccaacccactgataacagctgtgaatttaagagtgtaactcaatacaataagaccttgagtaacttgcttcttcccattgcagaaaacataaacaatattgcatcgccctcacctgggtcaagacgtcataaaaggtttgctggcattgccattggcattgctgcgctcggtgttgcgaccgcagcacaggtaactgccgctgtctcattagttcaagcacagacaaatgcacgtgcaatagcggcgatgaaaaattcaatacaggcaactaatcgggcaatcttcgaagtgaaggaaggcacccaacagttagctatagcggtacaagcaatacaagaccacatcaatactattatgaacacccaattgaacaatatgtcttgtcagatccttgataaccagcttgcaacctacctaggattatacctaacagaattaacaacagtgtttcagccacaattaattaatccggcattgtcaccgattagtatacaagccttgaggtctttgcttggaagtatgacgcctgcagtggttcaagcaacattatctacgtcaatttctgctgctgaaatactaagtgccggtctaatggagggtcagattgtttctgttctgctagatgagatgcagatgatagtcaagataaatattccaaccattgtcacacaatcaaatgcattggtgattgacttctactcaatttcgagctttattaataatcaagaatccataattcaattgccagacaggatcctagagatcgggaatgaacaatggagctatccagctaaaaattgtaagttgacaagacaccacatattctgccaatacaatgaggcagagaggctgagcctagaatcaaaactatgccttgcaggtaatataagtgcctgtgtgttctcacccatagcagggagttatatgaggcgatttgtagcactggatggaacaattgttgcaaactgtcgaagtctaacgtgtctatgcaagagtccatcttatcctatataccaacctgaccatcatgcagtcacgaccattgatctaaccacatgtcaaacattgtccctagacggattggacttcagcattgtctctctaagcaacatcacttacgctgagaaccttaccatttcattgtctcaaacaatcaatactcaacccattgacatatcaactgaactgagtaaagttaatgcatccctccaaaatgccgttaagtacataaaggagagcaaccatcaactccaatctgtgagtgtaaattccaaaattggagctataattgtagcagccttagttttgagcatcctgtcaattatcatttcgctattgttttgctgctgggcttacattgcaactaaagaaatcagaagaatcaacttcaaaacaaatcatatcaacacaatatcgagtagtgtcgatgatctcatcaggtactaa
编码全长MeV H蛋白的示例性核酸序列提供为SEQ ID NO:96：
atgtcaccacaacgagaccggataaatgccttctacaaagataacccccatcccaagggaagtaggatagtcattaacagagaacatcttatgattgatagaccttatgttttgctggctgttctgtttgtcatgtttctgagcttgatcgggttgctagccattgcaggcattagacttcatcgggcagccatctacaccgcagagatccataaaagcctcagcaccaatctagatgtaactaactcaatcgagcatcaggtcaaggacgtgctgacaccactcttcaaaatcatcggtgatgaagtgggcctgaggacacctcagagattcactgacctagtgaaattcatctctgacaagattaaattccttaatccggatagggagtacgacttcagagatctcacttggtgtatcaacccgccagagagaatcaaattggattatgatcaatactgtgcagatgtggctgctgaagagctcatgaatgcattggtgaactcaactctactggagaccagaacaaccaatcagttcctagctgtctcaaagggaaactgctcagggcccactacaatcagaggtcaattctcaaacatgtcgctgtccctgttagacttgtatttaggtcgaggttacaatgtgtcatctatagtcactatgacatcccagggaatgtatgggggaacttacctagtggaaaagcctaatctgagcagcaaaaggtcagagttgtcacaactgagcatgtaccgagtgtttgaagtaggtgttatcagaaatccgggtttgggggctccggtgttccatatgacaaactatcttgagcaaccagtcagtaatgatctcagcaactgtatggtggctttgggggagctcaaactcgcagccctttgtcacggggaagattctatcacaattccctatcagggatcagggaaaggtgtcagcttccagctcgtcaagctaggtgtctggaaatccccaaccgacatgcaatcctgggtccccttatcaacggatgatccagtgatagacaggctttacctctcatctcacagaggtgttatcgctgacaatcaagcaaaatgggctgtcccgacaacacgaacagatgacaagttgcgaatggagacatgcttccaacaggcgtgtaagggtaaaatccaagcactctgcgagaatcccgagtgggcaccattgaaggataacaggattccttcatacggggtcttgtctgttgatctgagtctgacagttgagcttaaaatcaaaattgcttcgggattcgggccattgatcacacacggttcagggatggacctatacaaatccaaccacaacaatgtgtattggctgactatcccgccaatgaagaacctagccttaggtgtaatcaacacattggagtggataccgagattcaaggttagtccctacctcttcactgtcccaattaaggaagcaggcgaagactgccatgccccaacatacctacctgcggaggtggatggtgatgtcaaactcagttccaatctggtgattctacctggtcaagatctccaatatgttttggcaacctacgatacttccagggttgaacatgctgtggtttattacgtttacagcccaagccgctcattttcttacttttatccttttaggttgcctataaagggggtccccatcgaattacaagtggaatgcttcacatgggaccaaaaactctggtgccgtcacttctgtgtgcttgcggactcagaatctggtggacatatcactcactctgggatggtgggcatgggagtcagctgcacagtcacccgggaagatggaaccaatcgcagatag
编码全长MuV HN蛋白的示例性核酸序列提供为SEQ ID NO:97：
atggagccctcgaaattcttcacaatatcggacagtgccacctttgcacctgggcctgtcagcaatgcggctgacaagaagacattccgaacctgcttccgaatactggtactatctgtacaagctgtcaccctcatattggttattgtcactttaggtgagcttgtaaggatgatcaatgatcaaggcttgagcaatcagttgtcttcaattacagacaagataagagagtcagctactatgattgcatctgctgtgggagtaatgaatcaagttattcatggagtaacggtatccttacccctacaaattgagggaaaccaaaatcaattgttagccacacttgccacaatctgcgccagccaaaaacaagtctcaaactgctctacaaacatccccttagtcaatgacctcaggtttataaatgggatcaataaatttattattgaagattacgcaactcatgatttctctatcggccatccactcaatatgcccagctttatcccaactgcaacttcacccaatggttgcacaagaattccatccttttctttaggtaagacacactggtgctacacacataatgtaattaatgccaactgcaaggaccatacttcgtctaaccaatatgtgtccatggggattctcgttcagaccgcgtcagggtatcctatgttcaaaaccttaaaaatccaatatctcagtgatggcctgaatcggaaaagctgctcaattgcaacagtccctgatgggtgcgcgatgtactgttatgtctctactcaacttgaaaccgacgactatgcggggtccagtccacccacccaaaaacttaccctgttattctataatgacaccgtcacagaaaggacaatatctccatctggtcttgaagggaattgggctactttggtgccaggagtggggagtgggatatattttgagaataagttgatcttccctgcatatgggggtgtcttgcccaatagtacactcggggttaaatcagcaagagaattttttcggcctgttaatccatataatccatgttcaggaccacaacaagatttagatcagcgtgctttgaggtcatacttcccaagttacttctctaatcgaagaatacagagtgcatttcttgtctgtgcctggaatcagatcctagttacaaattgtgagctagttgtcccctcaagcaatcagacaatgatgggtgcagaagggagagttttattgatcaataatcgactattatattatcagagaagtaccagctggtggccgtatgaactcctctacgagatatcattcacatttacaaactctggtccatcatctgtaaatatgtcctggatacctatatattcattcactcgtcctggttcaggcaattgcagtggtgaaaatgtgtgcccgactgcttgtgtgtcaggggtttatcttgatccctggccattaactccatatagccaccaatcaggtattaacagaaatttctatttcacaggtgcactattaaattcaagtacaactagagtaaatcctaccctttatgtctctgcccttaataatcttaaagtactagccccatatggtactcaaggactgtttgcctcgtacaccacaaccacctgctttcaagataccggtgatgctagtgtgtattgtgtttatattatggagctagcatcaaatattgttggagaattccaaattctacctgtgctaactagattgactatcacttga
这些示例性核酸序列(或相应的RNA序列)可以经修饰以编码本文提供的任何免疫原。
在若干实施方案中，核酸分子编码MuV或MeV F胞外域三聚体的原体的前体或MeVF胞外域三聚体的启动子或此类原体与MuV HN或MeV H胞外域的嵌合体，或MuV HN或MeV H多聚体或亚基，即，当在合适的细胞中表达时，其被加工成胞外域三聚体的原体，或MuV HN或MeV H多聚体的亚基，其可以自组装成相应的三聚体或多聚体。例如，核酸分子可以编码MuV或MeV F胞外域三聚体的原体或MeV F胞外域三聚体的启动子，包括用于进入细胞分泌系统的N-末端信号序列，其在细胞中重组F胞外域的加工过程中被蛋白水解切割。
在一些实施方案中，核酸分子编码F0多肽，当在合适的细胞中表达时，其被加工成MuV或MeV F胞外域三聚体的原体或MeV F胞外域三聚体的启动子，其包括与F1胞外域连接的F2多肽，其中重组F2-F1胞外域原体包括本文所述的任何融合前稳定化修饰，并且任选地可以与三聚化结构域连接，例如GCN4三聚化结构域和/或T4纤维蛋白三聚化结构域。
在一些实施方案中，核酸分子编码全长F0多肽，当在适当的细胞中表达时，其被加工成MuV或MeV F胞外域三聚体的原体或MuV或MeV F胞外域三聚体的启动子，其包括与F1多肽连接的F2多肽，包括F1跨膜尾和胞质尾，其中重组F2-F1胞外域原体包括本文所述的任何融合前稳定修饰。
示例性核酸可以通过克隆技术制备。适当的克隆和测序技术的实例，以及足以指导技术人员通过许多克隆练习的说明是已知的(参见，例如，Sambrook等人(分子克隆：实验室手册，第4版，Cold Spring Harbor,New York,2012)和Ausubel等人(见现代分子生物学实验指南，John Wiley&Sons，New York，至增刊104,2013))。
核酸还可以通过扩增方法制备。扩增方法包括聚合酶链反应(PCR)、连接酶链反应(LCR)、基于转录的扩增系统(TAS)、自持序列复制系统(3SR)。多种克隆方法、宿主细胞和体外扩增方法是本领域技术人员熟知的。
编码MuV或MeV F胞外域三聚体的原体，或MuV HN或MeV H多聚体的亚基的多核苷酸可以包括重组DNA，其被整合至载体(例如表达载体)，进入自主复制的质粒或病毒或进入原核生物或真核生物的基因组DNA，或作为独立于其他序列的单独分子(例如cDNA)存在。核苷酸可以是核糖核苷酸、脱氧核糖核苷酸或任一核苷酸的修饰形式。该术语包括单链和双链形式的DNA。
编码MuV或MeV F胞外域三聚体的原体或MeV F胞外域三聚体的启动子或此类原体与MuV HN或MeV H胞外域的嵌合体，或MuV HN或MeV H多聚体的亚基的多核苷酸序列，可以可操作地连接至表达控制序列。连接与编码序列可操作连接的表达控制序列，从而在与表达控制序列相容的条件下实现编码序列的表达。表达控制序列包括但不限于适当的启动子、增强子、转录终止子、蛋白编码基因前面的起始密码子(即ATG)、内含子的剪接信号、维持该基因的正确阅读框以允许正确翻译mRNA和终止密码子。
编码MuV或MeV F胞外域三聚体的原体或MeV F胞外域三聚体的启动子或此类原体与MuV HN或MeV H胞外域的嵌合体，或MuV HN或MeV H多聚体的亚基的DNA序列，可以通过将DNA转移至合适的宿主细胞中进行体外表达。该细胞可以是原核的或真核的。该术语还包括主题宿主细胞的任何后代。应当理解，所有后代可能与亲代细胞不同，因为在复制过程中可能发生突变。稳定转移的方法，意味着外源DNA在宿主中持续保持，是本领域已知的。
宿主可以包括微生物、酵母、昆虫和哺乳动物生物体。在原核生物中表达具有真核或病毒序列的DNA序列的方法是本领域熟知的。合适的宿主细胞的非限制性实例包括细菌、古生菌、昆虫、真菌(例如酵母)、植物和动物细胞(例如哺乳动物细胞，例如人)。使用的示例性细胞包括大肠杆菌、枯草芽孢杆菌、酿酒酵母、鼠伤寒沙门氏菌、SF9细胞、C129细胞、293细胞、脉孢菌和永生化哺乳动物骨髓和淋巴样细胞系。在培养物中繁殖哺乳动物细胞的技术是熟知的(参见，例如，Helgason和Miller(编辑)，2012，基本细胞培养方案(分子生物学方法)，第4版，Humana Press)。常用的哺乳动物宿主细胞系的实例是VERO和HeLa细胞、CHO细胞以及WI38、BHK和COS细胞系，尽管可以使用细胞系，例如旨在提供更高表达、期望的糖基化模式或其他特征的细胞。在一些实施方案中，宿主细胞包括HEK293细胞或其衍生物，例如GnTI-/-细胞(ATCC
Figure GDA0003943442610000421
No.CRL-3022)，或HEK-293F细胞。
可以通过常规技术用重组DNA转化宿主细胞。在宿主是原核生物(例如但不限于大肠杆菌)的一些实施方案中，能够从指数生长期后收获并随后通过CaCl2方法处理的细胞制备能够摄取DNA的感受态细胞。可选地，可以使用MgCl2或RbCl。如有需要，也可以在形成宿主细胞的原生质体之后或通过电穿孔进行转化。
当宿主是真核生物时，可以使用例如磷酸钙共沉淀物的DNA转染方法、常规的机械方法例如显微注射、电穿孔、插入包裹在脂质体中的质粒或病毒载体。真核细胞还可以与编码公开抗原的多核苷酸序列和编码可选择表型(例如单纯疱疹胸苷激酶基因)的第二外源DNA分子共转化。另一种方法是使用真核病毒载体，例如猿猴病毒40(SV40)或牛乳头瘤病毒，瞬时感染或转化真核细胞并表达蛋白(参见例如，病毒表达载体，Springer press，Muzyczka编辑，2011)。合适的表达系统(例如质粒和载体)用于在细胞(包括高等真核细胞，例如COS、CHO、HeLa和骨髓瘤细胞系)中产生蛋白。
在一个非限制性实例中，使用pVRC8400载体表达公开的免疫原(描述于Barouch等人，J.Virol.,79,8828-8834,2005，其通过引用并入本文)。
可以对编码公开的免疫原的核酸进行修饰而不降低其生物活性。可以进行一些修饰以促进靶向分子的克隆、表达或掺入融合蛋白中。示例性修饰包括终止密码子、在氨基末端添加的甲硫氨酸以提供起始位点、置于任一末端以产生方便定位的限制性位点的另外氨基酸，或有助于纯化步骤的另外氨基酸(例如聚组氨酸)。
在一些实施方案中，编码MuV或MeV F胞外域三聚体的原体或MeV F胞外域三聚体的启动子或此类原体与MuV HN或MeV H胞外域的嵌合体，或MuV HN或MeV H多聚体的亚基的核酸，可以在原体自组装成三聚体的条件下在细胞中表达，该三聚体从细胞分泌到细胞培养基中，例如关于RSV F蛋白所描述的(参见，例如，PCT公开WO2014160463，McLellan等人，Science,340:1113-1117,2013；McLellan等人，Science,342:592-598,2013，其各自通过引用以其全文并入本文)。在此类实施方案中，原体含有导致蛋白进入分泌系统的前导序列(信号肽)，并且信号肽在被分泌到细胞培养基中之前被切割并且原体形成三聚体。可以将培养基离心并从上清液纯化重组MuV或MeV F胞外域三聚体或重组MuV或MeV F胞外域三聚体或其与MuV HN或MeV H胞外域的嵌合体。
III.病毒载体
编码公开的免疫原的核酸分子可以包括在病毒载体中，例如，用于在宿主细胞中表达免疫原，或用于如本文公开的受试者的免疫。在一些实施方案中，将病毒载体作为初免-加强疫苗接种的一部分施用于受试者。通常此类病毒载体包括编码含有跨膜结构域的免疫原的核酸分子。在若干实施方案中，病毒载体包括在疫苗中，例如用于初免-加强疫苗接种的初免疫苗或加强疫苗。
在一些实例中，病毒载体可以是可复制的。例如，病毒载体可以在病毒基因组中具有突变(例如，插入编码原体的核酸)，该突变减弱但不完全阻断病毒在宿主细胞中的复制。
在若干实施方案中，病毒载体可以通过呼吸道递送。例如，hPIV载体，例如牛副流感病毒(BPIV)载体(例如，BPIV1、BPIV2或BPIV3载体)或人hPIV载体(例如，hPIV3载体)、偏肺病毒(MPV)载体、Sendia病毒载体、新城疫病毒(NCDV(载体))、腮腺炎病毒载体、麻疹病毒载体或另一种副粘病毒或肺病毒载体用于表达公开的抗原。
另外的病毒载体也可用于表达公开的抗原，包括多瘤病毒，即SV40(Madzak等人，1992,J.Gen.Virol.,73:15331536)、腺病毒(Berkner,1992,Cur.Top.Microbiol.Immunol.,158:39-6；Berliner等人，1988,Bio Techniques,6:616-629；Gorziglia等人，1992,J.Virol.,66:4407-4412；Quantin等人，1992,Proc.Natl.Acad.Sci.USA,89:2581-2584；Rosenfeld等人，1992,Cell,68:143-155；Wilkinson等人，1992,Nucl.Acids Res.,20:2233-2239；Stratford-Perricaudet等人，1990,Hum.Gene Ther.,1:241-256)、牛痘病毒(Mackett等人，1992,Biotechnology,24:495-499)、腺相关病毒(Muzyczka,1992,Curr.Top.Microbiol.Immunol.,158:91-123；On等人，1990,Gene,89:279-282)、疱疹病毒(包括HSV和EBV和CMV)(Margolskee,1992,Curr.Top.Microbiol.Immunol.,158:67-90；Johnson等人，1992,J.Virol.,66:29522965；Fink等人，1992,Hum.Gene Ther.3:11-19；Breakfield等人，1987,Mol.Neurobiol.,1:337-371；Fresse等人，1990,Biochem.Pharmacol.,40:2189-2199)、Sindbis病毒(H.Herweijer等人，1995,Human Gene Therapy 6:1161-1167；美国专利号5,091,309和5,2217,879)、甲病毒(S.Schlesinger,1993,Trends Biotechnol.11:18-22；I.Frolov等人，1996,Proc.Natl.Acad.Sci.USA 93:11371-11377)以及禽类逆转录病毒(Brandyopadhyay等人，1984,Mol.Cell Biol.,4:749-754；Petropouplos等人，1992,J.Virol.,66:3391-3397)、鼠类逆转录病毒(Miller,1992,Curr.Top.Microbiol.Immunol.,158:1-24；Miller等人，1985,Mol.Cell Biol.,5:431-437；Sorge等人，1984,Mol.Cell Biol.,4:1730-1737；Mann等人，1985,J.Virol.,54:401-407)和人源逆转录病毒(Page等人，1990,J.Virol.,64:5370-5276；Buchschalcher等人，1992,J.Virol.,66:2731-2739)。杆状病毒(苜蓿银纹夜蛾核型多角体病毒；AcMNPV)载体也是本领域已知的，并且可以从商业来源获得(例如PharMingen,San Diego,Calif.；Protein Sciences Corp.,Meriden,Conn.；Stratagene,La Jolla,Calif.)。
IV.病毒样颗粒
在一些实施方案中，提供了包括公开的免疫原的病毒样颗粒(VLP)。通常此类VLP包括含有跨膜结构域的免疫原，例如，具有含有MuV F跨膜结构域和胞质尾的原体的重组MuV F胞外域三聚体，或具有含有MeV F跨膜结构域和胞质尾的原体的重组MeV F胞外域三聚体。VLP缺乏病毒复制所需的病毒组分，因此代表高减毒、无法复制的病毒形式。然而，VLP可以展示类似于感染性病毒颗粒上表达的多肽(例如，重组MuV或MeV F胞外域三聚体)，并且当施用于受试者时可以引发对MuV或MeV的免疫应答。示例性病毒样颗粒及其产生方法，以及来自已知可形成VLP的几种病毒的病毒蛋白，包括人乳头瘤病毒、HIV(Kang等人，Biol.Chem.380:353-64(1999))、Semliki-Forest病毒(Notka等人，Biol.Chem.380:341-52(1999))、人多瘤病毒(Goldmann等人，J.Virol.73:4465-9(1999))、轮状病毒(Jiang等人，Vaccine 17:1005-13(1999))、细小病毒(Casal,Biotechnology and AppliedBiochemistry,Vol 29,Part 2,pp 141-150(1999))、犬细小病毒(Hurtado等人，J.Virol.70:5422-9(1996))、戊型肝炎病毒(Li等人，J.Virol.71:7207-13(1997))和新城疫病毒。此类VLP的形成可以通过任何合适的技术来检测。用于检测培养基中VLP的合适技术的实例包括，例如，电子显微镜技术、动态光散射(DLS)、选择性色谱分离(例如，离子交换、疏水相互作用和/或VLP的尺寸排阻色谱分离)和密度梯度离心。
V.免疫原性组合物
还提供了包含公开的免疫原(例如，重组MuV F胞外域三聚体、重组MeV F胞外域三聚体、或与MuV HN胞外域或MeV H胞外域的相应融合物、或MuV HN或MeV H多聚体)和药学上可接受的运载体的免疫原性组合物。此类组合物可以通过多种施用方式例如(肌内、皮下、静脉内、动脉内、关节内、腹腔或肠胃外途径)施用于受试者。在若干实施方案中，包括一种或更多种公开的免疫原的药物组合物是免疫原性组合物。制备可施用的组合物的实际方法在例如雷明顿制药科学，第19版，Mack Publishing Company,Easton,Pennsylvania,1995等出版物中更详细的描述。
因此，本文所述的免疫原可以与药学上可接受的运载体一起配制以助于保持生物活性，同时还促进在可接受的温度范围内在储存期间的稳定性增加。潜在的运载体包括但不限于生理平衡培养基、磷酸盐缓冲盐溶液、水、乳液(例如油/水或水/油乳液)、各种类型的润湿剂、防冻添加剂或稳定剂，例如蛋白、肽或水解产物(例如白蛋白、明胶)、糖(例如蔗糖、乳糖、山梨糖醇)、氨基酸(例如谷氨酸钠)或其他保护剂。得到的水溶液可以按原样包装使用或冻干。冻干制剂在用于单次或多次给药的施用前与无菌溶液组合。
配制的组合物，尤其是液体制剂，可以含有抑菌剂以防止或最小化储存期间的降解，包括但不限于有效浓度(通常≤1％w/v)的苯甲醇、苯酚、间甲酚、氯丁醇、对羟基苯甲酸甲酯和/或对羟基苯甲酸丙酯。一些患者可能禁用抑菌剂；因此，冻干制剂可以在含有或不含有此类组分的溶液中复溶。
本公开的免疫原性组合物可以含有作为近似生理条件所需的药学上可接受的溶媒物质，例如pH调节剂和缓冲剂、张力调节剂、润湿剂等，例如，乙酸钠、乳酸钠、氯化钠、氯化钾、氯化钙、脱水山梨糖醇单月桂酸酯和油酸三乙醇胺。
免疫原性组合物可以任选地包括佐剂以增强宿主的免疫应答。佐剂，例如氢氧化铝(例如，ALHYDROGEL
Figure GDA0003943442610000451
，可获自Brenntag Biosector,Copenhagen,Denmark，和Amphogel
Figure GDA0003943442610000452
，Wyeth Laboratories,Madison,NJ)、弗氏佐剂、MPLTM(3-O-脱酰基单磷酰脂质A；Corixa，Hamilton,IN)和IL-12(Genetics Institute,Cambridge,MA)、TLR激动剂(例如TLR-9激动剂，例如胞苷-磷酸-鸟苷寡脱氧核苷酸(CpG-ODN)1018)，以及本领域熟知的许多其他合适的佐剂，可以包括在组合物中。合适的佐剂是，例如，toll样受体激动剂、铝剂、AlPO4、水凝胶、脂质-A及其衍生物或变体、油乳剂、皂苷、中性脂质体、含有疫苗和细胞因子的脂质体、非离子嵌段共聚物和趋化因子。含有聚氧乙烯(POE)和聚二丙烯(POP)的非离子嵌段聚合物，例如POE-POP-POE嵌段共聚物、MPLTM(3-O-脱酰基单磷酰脂质A；Corixa，Hamilton，IN)和IL-12(Genetics Institute,Cambridge,MA)可用作佐剂(Newman等人，1998,CriticalReviews in Therapeutic Drug Carrier Systems 15:89-142)。这些佐剂的优势在于它们有助于以非特异性方式刺激免疫系统，从而增强对药物产品的免疫应答。
在一些实例中，佐剂制剂是无机盐，例如钙盐或铝(铝剂)盐，例如磷酸钙、磷酸铝或氢氧化铝。在一些实施方案中，公开的免疫原包含一种或更多种磷酸丝氨酸修饰并且与铝剂佐剂一起使用。在一些实施方案中，佐剂包括油水乳剂，例如水包油乳剂(例如MF59(Novartis)或AS03(GlaxoSmithKline))。水包油乳剂的一个实例包含在水性运载体中的可代谢油，例如角鲨烯、母育酚(例如生育酚，例如α-生育酚)和表面活性剂，例如山梨糖醇三油酸酯(Span 85)或聚氧乙烯山梨糖醇单油酸酯(Tween80)。
在一些实例中，可能期望将公开的免疫原与诱导对其他试剂的保护性应答的其他药物产品(例如疫苗)组合。例如，如本文所述的包括重组MuV F胞外域三聚体、重组MeV F胞外域三聚体或与MuV HN胞外域或MeV H胞外域的相应融合物的组合物可以与由免疫实践咨询委员会(ACIP；cdc.gov/vaccines/acip/index.html)建议用于目标年龄组(例如，约一个月龄至六个月龄的婴儿)的其他疫苗同时(通常单独地)或序贯施用。因此，本文所述的公开的免疫原可以与针对例如乙型肝炎(HepB)、白喉、破伤风和百日咳(DTaP)、肺炎球菌(PCV)、b型流感嗜血杆菌(Hib)、脊髓灰质炎、流感和轮状病毒的疫苗同时或序贯施用。
在一些实施方案中，组合物可以作为无菌组合物提供。免疫原性组合物通常含有有效量的公开的免疫原并且可以通过常规技术制备。通常，免疫原性组合物的每个剂量中的免疫原的量被选择为诱导免疫应答而没有显著的不利副作用的量。在一些实施方案中，组合物可以以单位剂型提供，用于诱导受试者产生免疫应答，例如，抑制受试者中的MuV和/或MeV感染。单位剂型包括用于施用于受试者的合适的单一预选剂量，或两个或更多个预选单位剂量的合适标记或测量的倍数，和/或用于施用单位剂量或其倍数的计量机制。
VI.诱导免疫应答的方法
可以将公开的免疫原(例如，重组MuV F胞外域三聚体、重组MeV F胞外域三聚体、或与MuV HN胞外域或MeV H胞外域的相应融合物、MuV HN或MeV H多聚体、编码公开的免疫原的核酸分子(例如RNA分子)、包含免疫原的蛋白纳米颗粒或病毒样颗粒)施用于受试者以诱导受试者对MuV和/或MeV产生免疫应答。在具体实例中，受试者是人。免疫应答可以是保护性免疫应答，例如抑制随后用MuV和/或MeV感染的应答。免疫应答的激发也可用于治疗或抑制MuV和/或MeV感染和与之相关的疾病。
可以选择例如因为暴露于或可能暴露于MuV或MeV而患有或有风险发生MuV或MeV感染的受试者进行治疗。在施用公开的免疫原后，可以监测受试者的MuV和/或MeV感染或与之相关的症状，或两者。
计划用本公开的疗法和方法进行治疗的典型受试者包括人。在一些实施方案中，受试者是对MuV和/或MeV特异性抗体呈血清阴性的人受试者。在一些实施方案中，受试者是对MuV和/或MeV特异性抗体呈血清阳性的人受试者，并且施用免疫原以增强受试者对MeV和/或MuV的免疫应答。为了根据本公开的方法识别用于治疗的受试者，采用公认的筛选方法来确定与目标或疑似疾病或病况相关的风险因素，或确定受试者中现有疾病或病况的状态。这些筛选方法包括，例如，确定可能与目标或疑似疾病或病况相关的环境、家庭、职业和其他此类风险因素的常规检查，以及诊断方法，例如各种ELISA和用于检测和/或表征MuV和/或MeV感染的其他免疫测定方法。这些和其他常规方法使得临床医生使用本公开的方法和免疫原性组合物来选择需要治疗的患者。根据这些方法和原理，可以根据本文的教导或其他常规方法施用组合物，作为独立的预防或治疗方案，或作为其他治疗的后续、辅助或协调治疗方案。
公开的免疫原的施用可以用于预防或治疗目的。当预防性提供时，可以在任何症状之前，例如在感染之前提供免疫原。预防性施用用于预防或改善任何随后的感染。在一些实施方案中，该方法可以涉及选择具有感染MuV和/或MeV感染风险的受试者，并且向该受试者施用治疗有效量的公开的免疫原。可以在预期暴露于MuV和/或MeV之前提供免疫原，以在暴露或疑似暴露于病毒之后，或在实际感染开始之后减轻感染和/或相关疾病症状的预期严重程度、持续时间或程度。从预防性使用公开的免疫原(例如，作为加强免疫)获益的人群包括进入学校的儿童(例如，5岁)和进入高中或大学或军队的青少年(例如，15-18岁)。移植受者可能还需要重新接种疫苗，或免疫功能低下的儿童(如HIV+)将从蛋白疫苗获益，与可能不安全的减毒活病毒相反，包括孕妇。
当在治疗上提供时，在MuV和/或MeV感染的症状发作时或之后，或在MuV和/或MeV感染的诊断之后提供公开的免疫原。通过抑制MuV复制或感染来治疗MuV可以包括延迟和/或减少受试者中MuV感染的体征或症状。通过抑制MeV复制或感染来治疗MeV可以包括延迟和/或减少受试者中MeV感染的体征或症状。在一些实例中，使用本文公开的方法的治疗延长受试者的生存时间。
在一些实施方案中，当受试者随后感染或再次感染野生型MuV和/或MeV时，向受试者施用公开的免疫原可以引发产生免疫应答，该免疫应答预防或减轻疾病症状。虽然自然循环的病毒可能仍然能够引起感染，但由于接种疫苗，可能降低出现严重或危及生命的症状的可能性，并且可能因随后感染野生型病毒而增强抵抗力。接种疫苗后，存在可检测水平的宿主产生的血清和分泌抗体，它们能够在体外和体内中和(同一亚组的)同源野生型病毒。在许多情况下，宿主抗体还将中和不同非疫苗亚组的野生型病毒。
本文所述的免疫原及其免疫原性组合物以有效诱导或增强受试者(优选人)对MuV和/或MeV的免疫应答的量提供给受试者。公开的免疫原的实际剂量将根据以下因素而变化，例如疾病适应症和受试者的特定状态(例如受试者的年龄、体型、健康、症状程度、易感因素等)、施用时间和途径、同时施用的其他药物或治疗，以及用于在受试者中引发期望活性或生物反应的组合物的特定药理学。可以调整剂量方案以提供最佳预防或治疗反应。
包括一种或更多种公开的免疫原的免疫原性组合物可用于协调(或初免-加强)疫苗接种方案或组合制剂。在某些实施方案中，新型组合免疫原性组合物和协同免疫方案采用单独的免疫原或制剂，其各自针对引发抗病毒免疫应答，例如针对MuV F蛋白和/或MeV F蛋白的免疫应答。引发抗病毒免疫应答的单独免疫原性组合物可以组合成多价免疫原性组合物，在单个免疫步骤中施用于受试者，或者它们可以在协同(或初免-加强)免疫方案中单独施用(在单价免疫原性组合物中)。
可以有几次加强，并且每次加强可以是不同的公开的免疫原。在一些实例中，加强可以是与另一加强或初免相同的免疫原。初免和加强可以作为单剂或多剂施用，例如可以在数天、数周或数月内向受试者施用两剂、三剂、四剂、五剂、六剂或更多剂。还可以给予多次加强，例如一次至五次(例如，1、2、3、4或5次加强)，或更多次。在一系列序贯免疫中可以使用不同的剂量。例如，在初次免疫中使用相对较大的剂量，然后以相对较小的剂量进行加强。
在一些实施方案中，可以在初免后约两周、约三至八周或约四周，或在初免后约数月施用加强。在一些实施方案中，可以在初免后约5、约6、约7、约8、约10、约12、约18、约24个月，或在初免后更多或更少时间施用加强。还可以在适当的时间点使用周期性的另外加强来增强受试者的“免疫记忆”。选择的疫苗接种参数的充分性，例如制剂、剂量、方案等，可以通过从受试者取出等分试样的血清并在免疫程序过程中测定抗体滴度来确定。此外，可以监测受试者的临床状况以获得期望的效果，例如抑制MuV和/或MeV感染或改善疾病状态(例如减少病毒载量)。如果这种监测表明疫苗接种是次优的，则可以用另外剂量的免疫原性组合物加强受试者，并且可以以预期增强免疫应答的方式修改疫苗接种参数。
在一些实施方案中，初免-加强方法可以包括用于受试者的DNA-初免和蛋白-加强疫苗接种方案。该方法可以包括两次或更多次施用核酸分子或蛋白。
对于蛋白疗法，通常，每人剂量将包含1-1000μg蛋白质，例如约1μg至约100μg，例如约1μg至约50μg，例如约1μg、约2μg、约5μg、约10μg、约15μg、约20μg、约25μg、约30μg、约40μg或约50μg。
免疫原性组合物中使用的量基于受试者群体(例如，婴儿或老年人)来选择。可以通过标准研究确定特定组合物的最佳量，这些研究涉及观察受试者的抗体滴度和其他应答。应当理解，免疫原性组合物中有效量的公开的免疫原，例如重组MuV或MeV F胞外域三聚体或重组MeV F胞外域三聚体或其与MuV HN或MeV H胞外域的嵌合体、病毒载体或核酸分子，可以包括在通过施用单剂量时对引发免疫应答无效但在施用多剂量时有效的量，例如在初免-加强施用方案中。
在施用公开的免疫原后，受试者的免疫系统通常通过产生对病毒蛋白特异的抗体来应答免疫原性组合物。此类应答表明免疫有效剂量被递送至受试者。
对于每名特定受试者，可以根据个体需要和施用或监督免疫原性组合物施用的人的专业判断来评价和调整特定的剂量方案。剂量和给药次数将取决于环境，例如，在成人或任何既往MuV和/或MeV感染或免疫接种的人中，单剂可以是足够的加强剂。在一些实例中，在空白受试者中，将给予至少两剂，例如，至少三剂。在一些实施方案中，例如与每年流感疫苗接种一起给予每年加强免疫。
在一些实施方案中，受试者的抗体应答将在评价有效剂量/免疫方案的背景下确定。在大多数情况下，评估从受试者获得的血清或血浆中的抗体滴度将足够。关于是否施用加强接种和/或改变向个体施用的治疗剂的量的决定可以至少部分地基于抗体滴度水平。抗体滴度水平可以基于例如免疫结合测定，该测定测量血清中与抗原(包括例如MuV F蛋白和/或MeV F蛋白)结合的抗体浓度。
有效剂量的确定通常基于动物模型研究，随后是人体临床试验，并且以显著降低受试者中目标疾病症状或病况的发生或严重程度，或诱导受试者产生期望的应答(例如中和免疫应答)的施用方案为指导。在这方面合适的模型包括，例如，鼠、大鼠、猪、猫、雪貂、非人灵长类动物和本领域已知的其他公认的动物模型受试者。可选地，可以使用体外模型(例如，免疫学和组织病理学分析)确定有效剂量。使用此类模型，只需要普通的计算和调整来确定合适的浓度和剂量以施用有效量的组合物(例如，有效引发期望的免疫应答或减轻目标疾病的一种或更多种症状的量)。在可选的实施方案中，有效量或有效剂量的组合物可以简单地抑制或增强一种或更多种选择的与疾病或病况相关的生物活性，如本文所述，用于治疗或诊断目的。
引发免疫应答以减少或预防感染的免疫原性组合物的施用可以但不一定完全消除这种感染，只要可测量地减少感染即可。例如，施用有效量的药剂可以减少MuV或MeV感染(例如，通过细胞感染进行测量，或通过期望量的MuV或MeV感染的受试者的数量或百分比进行测量，例如通过与合适的对照(也称为灭菌免疫)相比，至少10％、至少20％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％、至少98％或甚至至少100％消除或预防可检测的MuV或MeV感染)。
在一些实施方案中，向受试者施用有效量的一种或更多种公开的免疫原诱导受试者产生中和免疫应答。为了评估中和活性，在受试者免疫后，可以在适当的时间点从受试者收集血清，冷冻并储存用于中和检测。测定中和活性的方法包括但不限于空斑减少中和滴度(PRNT)测定、微量中和测定、基于流式细胞术的测定、单周期感染测定。在一些实施方案中，可以使用一组MuV和/或MeV假病毒来测定血清中和活性。
一种施用核酸的方法是用质粒DNA(例如用哺乳动物表达质粒)直接免疫。通过核酸构建体进行免疫是本领域熟知的，并且在例如美国专利号5,643,578(其描述了通过引入编码期望的抗原的DNA以引发细胞介导的或体液应答来免疫脊椎动物的方法)，以及美国专利号5,593,972和美国专利号5,817,637(描述了将编码抗原的核酸序列可操作地连接至能够表达的调节序列)中教导。美国专利号5,880,103描述了几种将编码免疫原性肽或其他抗原的核酸递送至生物体的方法。这些方法包括脂质体递送核酸(或合成肽本身的)，以及免疫刺激构建体或ISCOMSTM，在混合胆固醇和Quil ATM(皂苷)时自发形成的30-40nm大小的带负电荷的笼状结构。保护性免疫已在各种感染实验模型中产生，包括弓形虫病和Epstein-Barr病毒诱导的肿瘤，使用ISCOMSTM作为抗原的递送溶媒(Mowat和Donachie,Immunol.Today 12:383,1991)。已发现封装在ISCOMSTM中的低至1μg剂量的抗原可产生I类介导的CTL应答(Takahashi等人，Nature 344:873,1990)。
在一些实施方案中，质粒DNA疫苗用于在受试者中表达公开的免疫原。例如，编码公开的免疫原的核酸分子可以施用于受试者以引发对MuV或MeV的F蛋白的免疫应答。在一些实施方案中，核酸分子可以被包括在用于DNA免疫的质粒载体(例如pVRC8400载体)上(描述于Barouch等人，J.Virol,79,8828-8834,2005，其通过引用并入本文)。
在使用核酸进行免疫的另一种方法中，公开的免疫原可以由减毒病毒宿主(例如减毒MuV或MeV载体)或载体或细菌载体表达。重组牛痘病毒、腺相关病毒(AAV)、疱疹病毒、逆转录病毒、巨细胞病毒、副粘病毒、肺病毒或其他病毒载体可用于表达肽或蛋白，从而引发CTL应答。例如，用于免疫方案的牛痘载体和方法描述于美国专利号4,722,848。BCG(卡介苗芽孢杆菌(Bacillus Calmette Guerin))提供了另一种用于表达肽的载体(参见Stover,Nature 351:456-460,1991)。
在另一个实例中，可以使用RNA免疫，例如使用脂质包封的mRNA免疫平台，将公开的免疫原施用于受试者(参见，例如，Roth等人，“A Modified mRNA Vaccine TargetingImmunodominant NS Epitopes Protects Against Dengue Virus Infection in HLAClass I Transgenic Mice,”Frot Immunol.,June 21,2019,Vol.10,Article 1424；Jagger等人，J Infect Dis,“Protective Efficacy of Nucleic Acid Vaccines AgainstTransmission of Zika Virus During Pregnancy in Mice,”jiz338,Jul 1,2019；Feldman等人，“mRNA vaccines against H10N8 and H7N9 influenza viruses ofpandemic potential are immunogenic and well tolerated in healthy adults inphase 1randomized clinical trials,”Vaccine,37(25),3326-3334,2019；和Hasset等人，“Optimization of Lipid Nanoparticles for Intramuscular Administration ofmRNA Vaccines,”Mol Ther Nucleic Acids,15:1-11,2019)。
在一个实施方案中，将编码公开的MuV F或MeV F胞外域三聚体的原体的核酸直接引入细胞中。例如，可以通过标准方法将核酸负载至金微球上，然后通过装置(例如Bio-Rad的HeliosTM Gene Gun)引入皮肤。核酸可以是“裸露的”，由受强启动子控制的质粒组成。通常，DNA被注射到肌肉中，尽管它也可以直接注射到其他部位。注射剂量通常为约0.5μg/kg至约50mg/kg，以及通常为约0.005mg/kg至约5mg/kg(参见，例如，美国专利号5,589,466)。
在另一个实施方案中，基于mRNA的免疫方案可用于将编码公开的免疫原的核酸直接递送至细胞中。在一些实施方案中，基于mRNA的基于核酸的疫苗可以提供前述方法的有效可选方案。mRNA疫苗排除了DNA整合到宿主基因组中的安全性问题，并且可以直接在宿主细胞的细胞质中翻译。此外，RNA的简单无细胞体外合成避免了与病毒载体相关的生产并发症。可用于递送编码公开的免疫原的核酸的两种示例性形式的基于RNA的疫苗接种包括常规的非扩增性mRNA免疫(参见，例如，Petsch等人，“Protective efficacy of in vitrosynthesized,specific mRNA vaccines against influenza A virus infection,”Nature biotechnology,30(12):1210–6,2012)和自扩增mRNA免疫(参见，例如，Geall等人，“Nonviral delivery of self-amplifying RNA vaccines,”PNAS,109(36):14604-14609,2012；Magini等人，“Self-Amplifying mRNA Vaccines Expressing Multiple ConservedInfluenza Antigens Confer Protection against Homologous and HeterosubtypicViral Challenge,”PLoS One,11(8):e0161193,2016；和Brito等人，“Self-amplifyingmRNA vaccines,”Adv Genet.,89:179-233,2015)。
在一些实施方案中，包括编码公开的免疫原的mRNA的脂质纳米颗粒用于引发免疫应答的方法，例如，如WO2017070626、US2019/0192646中所述，以及关于mRNA-1273疫苗描述于Jackson等人“An mRNA vaccine against SARS-CoV2–preliminary report,”N.Engl.J.Med.,383(20):1920-1931,2020，其各自均通过引用并入本文。如WO2017070626中所述，编码免疫原的mRNA可以在具有50mol％可电离脂质、10mol％DSPC、38.5mol％胆固醇和1.5mol％(PEG2000 DMG)的脂质纳米颗粒中进行配制。此外，编码免疫原的mRNA可以是修饰的mRNA，用1-甲基假尿苷代替尿苷和7mG(5')ppp(5')N1mpNp帽(酶)，以及5'UTR、3'UTR和polyA尾。
实施例
提供以下实施例以说明某些实施方案的特定特征，但权利要求的范围不应限于示例的那些特征。
实施例1
以融合前构象稳定的MuV F蛋白及其与MuV HN胞外域的融合物
本实施例示出了通过一个或更多个氨基酸取代以融合前构象稳定的MuV F胞外域三聚体的实施方案。进一步提供了与MuV HN胞外域连接的MuV F胞外域三聚体。例如，融合前稳定的MuV F胞外域三聚体和与MuV HN胞外域的相应融合物可用于诱导受试者产生对MuV的中和免疫应答。
在20世纪60年代后期引入含有腮腺炎的疫苗之前，腮腺炎引起以发烧、腮腺炎为特征的广泛发病率，以及较少见的睾丸炎、脑膜炎、脑炎和耳聋。麻疹、腮腺炎和风疹联合疫苗(MMR)大大降低了全世界腮腺炎的发病率。两剂MMR疫苗对预防腮腺炎的有效率为约88％，但自2006年以来，世界上高度接种疫苗的人群中腮腺炎病例数量有所回升，在美国有＞30,000名受累个体。促成因素可能包括免疫力减弱、抗体效应效果不佳以及MMR疫苗中使用的Jeryl Lynn菌株与循环野生型菌株之间的抗原差异。近年来，在美国和欧洲爆发流行性腮腺炎的主要基因型是基因型G。
如本文所述，基于结构的设计用于工程化以融合前构象稳定的MuV F糖蛋白。
Figure GDA0003943442610000501
Figure GDA0003943442610000502
分辨率的腮腺炎融合糖蛋白晶体结构显示了融合前构象稳定的基础。有效的跨腮腺炎基因型斑块减少中和滴度(PRNT)在小鼠中从腮腺炎融合前稳定的F糖蛋白或与基因型G腮腺炎血凝素神经氨酸酶(HN)连接的融合前稳定的腮腺炎F三聚体的嵌合融合糖蛋白中引发。融合前的F-HN腮腺炎嵌合体可以引发对基因型A、G和H腮腺炎病毒的最高PRNT，超过所报道的人保护滴度的100倍。此外，从免疫小鼠中分离针对腮腺炎融合前F和HN的单克隆抗体，这些抗体能够中和具有一系列效力的基因型G腮腺炎病毒。这些融合前F特异性抗体的结构和结合分析显示了与四个离散的中和抗原位点的结合。工程化免疫原是流行性腮腺炎的候选疫苗，无论是新型疫苗还是加强疫苗。
结果
二硫键和膜-近端卷曲螺旋稳定强稳定并使得产生可溶性腮腺炎F三聚体。
当在细胞中产生时，与C-末端GCN4三聚化结构域连接的MuV F胞外域形成三聚体，其自发转变为融合前构象。不稳定的重组MuV F-GCN4非常不稳定，100％的分子在评价点(EM)已转变为融合后构象。在没有稳定化的情况下，蛋白表达也显著降低。因此，基于结构的疫苗设计用于识别突变以稳定融合前构象的MuV F胞外域，并消除F1/F2切割位点以产生表达增加的“单链”MuV F蛋白。
猿融合前副流感病毒5(PIV5)F糖蛋白(PDB IDs 4GIP,4WSG)(Welch,B.D.etal.Proc Natl Acad Sci U S A 109,16672-16677,2012)的晶体结构用于构建融合前腮腺炎F蛋白，其由三个相互缠绕的单体组成，形成DI、DII、DIII和HRB结构域的四元组装体。
采用多种稳定策略将MuV F胞外域“锁定”在融合前构象中，包括引入二硫键和脯氨酸取代。MuV F中突变为半胱氨酸的残基的选择基于来自PIV5融合前F结构(PDB 4WSG)的同源性设计，基于预测在从融合前构象到融合后构象的转变中经历构象变化的残基。残基对Cbeta原子被确定为在5埃以内，并且定向使得可能形成二硫键。总共对约60种不同突变体进行设计、表达、纯化、评估表达水平并通过负染色EM评估融合前构象。
将突变引入MuV F胞外域(基于MuV F第469、476或483位的C-末端截短)，并且连接至C-末端GCN4三聚化结构域，并且如上所述筛选所得突变体。胞外域还包括突变以去除F1/F2弗林蛋白酶切割位点。评估的融合前稳定突变包括：位于MuV F第86和215、155和161、163和235、165和231、206和223、209和214、221和255位中的一个或更多个位置的半胱氨酸取代，形成非天然二硫键；和位于MuV F第184位的脯氨酸取代。相关序列如下所示。在图1A中，报告了初步成功的融合前稳定突变的总结。
与未修饰的MuV F相比，单链和融合前稳定的MuV F蛋白的表达和纯化显示出表达水平显著增加。
如图1A和1B所示，负染色EM可用于区分处于融合前构象的MuV F胞外域三聚体与处于融合后构象的那些。
通过创建二硫键取代和C-末端卷曲螺旋-GCN4附接位置的矩阵，使用负染色EM评价蛋白表达水平和采用融合前或融合后三聚体构象的蛋白比例(图1A)。观察到虽然二硫键和GCN4附接位置的五种组合产生100％融合前三聚体，但其中一种组合从Expi293细胞产生约4.8mg/L的高产率蛋白表达(V206C-A223C与476-GCN4)。该设计的阴性染色EM 2D平均值(图1B，上部插图)与融合后腮腺炎F糖蛋白三聚体(图1B，下部插图)形成对比，并且与先前对副流感病毒F蛋白构象的观察结果一致。
融合前腮腺炎F糖蛋白三聚体的晶体结构的分辨率为
Figure GDA0003943442610000511
显示了稳定的二硫键设计和多态性残基的位置。融合前稳定的腮腺炎F蛋白三聚体(MuV F V206C-A223C-GGG-476-GCN4,SEQ ID NO:11)的尺寸排阻色谱显示了均质峰(图1C)，当在kifunensine存在下表达时，可以去糖基化和结晶。结构的3维坐标在表1中提供。分辨率为
Figure GDA0003943442610000512
的x射线结构(图1D和1E)显示了与融合前PIV5和其他副粘病毒融合前F三聚体相似的整体结构(Stewart-Jones等人，Proc Natl Acad Sci U S A 115,12265-12270,2018；Welch,B.D.等人Proc Natl Acad Sci U S A 109,16672-16677,2012；Xu K.等人PLoSPathog11(12):e1005322,2015)，然而在腮腺炎融合前F三聚体的三聚体顶点处采用由环N177-S184组成的“封闭帽”结构，其由侧链T178、Q179和N181之间的相互作用形成(图2A和2B)。来自位置N73、N182、N352、N427、N433和N457的六个聚糖的剩余N-乙酰氨基葡萄糖部分在电子密度和建立的糖基化模型中可见(图2C)。V206C-A223C二硫化物由Cα-Cα原子距离为
Figure GDA0003943442610000521
的电子密度明确定义，而在同源融合后PIV3 F三聚体结构[PDB ID 1ZTM]中，Cα-Cα原子相距
Figure GDA0003943442610000522
。DI-DIII结构域封闭了体积测量为～
Figure GDA0003943442610000523
的大水腔，并且原体掩埋
Figure GDA0003943442610000524
的界面，类似于在PIV5、PIV3和nipah融合前F结构中观察到的界面(Stewart-Jones等人，Proc NatlAcad Sci U S A 115,12265-12270,2018；Welch,B.D.等人Proc Natl Acad Sci U S A109,16672-16677,2012；Xu K.等人PLoSPathog11(12):e1005322,2015)。被认为在融合前和融合后构象之间经历构象变化的残基(基于PIV5 pre-F结构[PDB ID 4WSG]和PIV3post-F[PDB ID 1ZTM])在图1E中以黑色显示，对应于MuV F残基92-253。
虽然腮腺炎F和HN的序列同一性在基因型之间相对较高(图2A-2C)，但绘制融合前腮腺炎F和HN结构的多态性变异(图2D)。将所有基因型的变异映射到融合前F，特别是基因型G和Jeryl Lynn(基因型A)之间，暴露用于抗体识别的大部分蛋白表面是保守的，而许多可变氨基酸位于融合前三聚体的核心中的水腔内(图2D，左)。相反，腮腺炎HN二聚体结构显示大多数多态性氨基酸暴露于溶剂(图2D，右)，包括Jeryl Lynn和基因型G HN之间的位置N464聚糖变异。与融合前F相比，HN上溶剂暴露的多态性残基的优势表明，基因型G对JerylLynn疫苗引发的体液免疫的抗性可能更多地来自HN，而不是来自融合前F。
稳定突变的跨菌株有效性。为了表明融合前稳定突变对来自MuV毒株的F有效，这些突变在来自几种不同MuV毒株的F中进行检测。MuV F V206C-A223C-GGG-476-GCN4(SEQID NO:11)基于基因型C MuV F。将这些融合前稳定突变引入基因型A(Jeryl Lynn)MuV F蛋白(MuV-JL F 206C-A223C-GGG-476-GCN4(SEQ ID NO:26)和基因型G MuV F蛋白(MuV-IL17F 206C-A223C-GGG-476-GCN4(SEQ ID NO:51)也提供了融合前稳定性。此外，将这些来自以下MuV染色的融合前稳定突变引入F蛋白也导致融合前稳定，如通过负染色EM和/或融合前特异性抗体结合所测量：加拿大(URABE)、奥尔巴尼(基因型A)、Hoshino(基因型B)、印度(基因型C)、荷兰(基因型D)、中国(基因型F)、NethL11(基因型G)、NY14(基因型G)、IA14(基因型G)、MA16(基因型G)、LA17(基因型G)、IL17(基因型G)、弗吉尼亚(基因型H)、中国台湾(基因型J)、中国台湾(基因型K)、荷兰(基因型L)、MG15(基因型A)
F-HN嵌合体。为了增加MuV F胞外域三聚体的免疫原性足迹，将MuV HN胞外域基因融合至三聚体每个原体的三聚化结构域的C-末端。该形式如图4A所示。对应的序列如下所示。负染色EM显示F胞外域保持融合前构象，其中三个HN胞外域(一个与每个F原体连接)排列在三聚域的C-末端。这种设计从Expi293细胞产生约0.3mg/L，并且在尺寸排阻色谱上是单分散的，并且显示出来自负染色EM的预期组装(图1C和图4A)。
融合前稳定的腮腺炎F和融合前稳定的F-HN嵌合三聚体在小鼠中引发高滴度中和抗体。评估与其他腮腺炎免疫原相比，融合前稳定的腮腺炎F引发中和抗体的能力。10只CB6F1/J小鼠组在第0、3和10周用10μg剂量的腮腺炎糖蛋白联合10μg聚肌苷-聚胞苷酸(poly-I:C)佐剂进行免疫，并测量HEp-2细胞的血清预防腮腺炎病毒感染的能力(图3A)。
评估的免疫原是以融合后构象的MuV F胞外域三聚体(具有-476-GCN4的天然胞外域)、以融合前构象的MuV F胞外域三聚体(MuV F V206C-A223C-GGG-476-GCN4，SEQ ID NO:11)、MuV HN胞外域单体和以融合前构象的MuV F胞外域三聚体，其中三聚体的原体与MuVHN胞外域融合(MuV F 206C-223C-GGG-476+GCN4+MuV HN_G(SEQ ID NO:27))。
当用含融合前腮腺炎F的免疫原(preF或preF-HN)免疫小鼠时，在血清中检测到对preF的特异性反应，与融合后免疫小鼠的结合水平较低(图4B)。重组HN单体仅结合HN免疫的小鼠血清，然而单体HN免疫的小鼠显示几乎没有结合，而来自preF-HN免疫小鼠的血清显示大量的HN结合水平，表明HN的多价驱动强体液应答(图4C)。
为了分析来自用融合后F、融合前F或融合前F-HN的三种免疫的中和抗体滴度的引发，分析每次免疫后2周的PRNT(图3B)。每次免疫后观察到中和滴度的增量增加，但是第三次免疫显示比第二次免疫后更小的增加。在融合后免疫中观察到中和抗体，但在第二次和第三次免疫后，融合前F免疫原的中和性分别比融合后F高3.5和2.5倍(对基因型G腮腺炎的几何平均感染剂量(ID60值)分别为322和789病毒)。preF-HN嵌合变体在第三次免疫后引发中和滴度，比融合后F高12倍，比融合前F高5倍，对基因型G病毒的ID60值为3930。
接下来，对Jeryl Lynn和基因型H病毒进行PRNT评价，以表征从重组免疫原引发的交叉中和抗体。对于融合后和融合前F，观察到对Jeryl Lynn病毒的中和水平高于对基因型G病毒，而对于融合前F-HN嵌合体，观察到等同的PRNT。在第16周，来自preF和preF-HN组的血清对基因型H病毒显示出强PRNT。这表明这些重组免疫原可以引发可以交叉中和多种腮腺炎基因型的抗体。另监测PRNT对这三种腮腺炎病毒的持久性，持续6个月，发现虽然滴度降低，但ID60平台在3个月后形成，preF-HN具有基因型G、Jeryl Lynn和基因型H病毒的几何平均PRNT分别为约640、800和1700(图4D)。用针对基因型G、Jeryl Lynn和基因型H病毒的preF免疫原形成的ID60 PRNT平台为约100、660和3153(图4E)。总之，与HN相关的融合前稳定的F腮腺炎产生的中和滴度高于单独的F，并且代表可将两种病毒表面抗原以化学计量方式组合在单一免疫原中的设计策略。
讨论
MMR疫苗接种后人对腮腺炎的免疫力以PRNT为特征，Jeryl Lynn通常为约220，基因型G为约40(Rasheed等人，Proc Natl Acad Sci U S A 116(38):19071-19076,2019)。假设小鼠模型数据与人的应答相关，本文提供的关于公开的免疫原的结果似乎提供了通过PNRT测量的有效性增加。
由于两剂疫苗接种者之间爆发流行性腮腺炎，需要对目前流行性腮腺炎疫苗进行改进，以减少疾病发病率和公共卫生资源的负担。免疫实践咨询委员会(ACIP)建议在暴发地区有感染腮腺炎风险的个人接种第三剂含腮腺炎疫苗。第三剂减毒活MMR导致中和滴度暂时升高，持续约12个月(Fiebelkorn A.P.等人Open Forum Infect Dis1(3):ofu094,2014)。此外，与接受两剂MMR的个体相比，第三剂MMR后感染腮腺炎的风险降低了78％(Cardemil C.等人，Effectiveness of a Third Dose of MMR Vaccine for MumpsOutbreak Control.N Engl J Med.377(10):947–956,2017)。本实施例中描述的重组蛋白候选疫苗提供了可选的疫苗模式，而不是在流行性腮腺炎暴发环境中施用MMR，并提供更高的耐用性和有效性。
融合前F-HN嵌合体，包含腮腺炎病毒颗粒上的两个关键中和靶标，能够引发有效的跨基因型中和应答，包括对导致爆发的主要腮腺炎基因型G和其他两种基因型A和H代表全球流行性腮腺炎菌株的通用候选疫苗。
序列：
MuV F A163C/V235C-GGG-483-GCN4(SEQ ID NO:2)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaCavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsmtpaCvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsvsvnski IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F V206C/A223C-GGG-483-GCN4(SEQ ID NO:3)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttCfqpqltnpalspisiqClrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsvsvnski IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F N86C/A215C-GGG-483-GCN4(SEQ ID NO:4)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeCinniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpClspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsvsvnski IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F P209C/P214C-GGG-483-GCN4(SEQ ID NO:5)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqCqltnCalspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsvsvnski IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F K155C/L161C-GGG-483-GCN4(SEQ ID NO:6)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevCegtqqCaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsvsvnski IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F V165C/M231C-GGG-483-GCN4(SEQ ID NO:7)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiaCqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsCtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsvsvnski IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F I221C/M255C-GGG-483-GCN4(SEQ ID NO:8)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisCqalrsllgsmtpavvqatlstsisaaeilsaglCegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsvsvnski IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F S184P/GGG-483-GCN4(SEQ ID NO:9)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmPcqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsvsvnski IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F A163C/V235C-GGG-476-GCN4(SEQ ID NO:10)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaCavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsmtpaCvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqs IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F V206C/A223C-GGG-476-GCN4(SEQ ID NO:11)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttCfqpqltnpalspisiqClrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqs IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F N86C/A215C-GGG-476-GCN4(SEQ ID NO:12)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeCinniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpClspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqs IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F P209C/P214C-GGG-476-GCN4(SEQ ID NO:13)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqCqltnCalspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqs IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F K155C/L161C-GGG-476-GCN4(SEQ ID NO:14)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevCegtqqCaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqs IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F V165C/M231C-GGG-476-GCN4(SEQ ID NO:15)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiaCqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsCtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqs IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F I221C/M255C-GGG-476-GCN4(SEQ ID NO:16)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisCqalrsllgsmtpavvqatlstsisaaeilsaglCegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqs IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F S184P-GGG-476-GCN4(SEQ ID NO:17)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmPcqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqs IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F A163C/V235C-GGG-469-GCN4(SEQ ID NO:18)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaCavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsmtpaCvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyike IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F V206C/A223C-GGG-469-GCN4(SEQ ID NO:19)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttCfqpqltnpalspisiqClrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyike IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F N86C/A215C-GGG-469-GCN4(SEQ ID NO:20)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeCinniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpClspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyike IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F P209C/P214C-GGG-469-GCN4(SEQ ID NO:21)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqCqltnCalspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyike IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F K155C/L161C-GGG-469-GCN4(SEQ ID NO:22)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevCegtqqCaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyike IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F V165C/M231C-GGG-469-GCN4(SEQ ID NO:23)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiaCqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsCtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyike IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F I221C/M255C-GGG-469-GCN4(SEQ ID NO:24)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisCqalrsllgsmtpavvqatlstsisaaeilsaglCegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyike IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV F S184P-GGG-469-GCN4(SEQ ID NO:25)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmPcqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyike IEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV-JL F 206C/A223C-GGG-476-GCN4(SEQ ID NO:26)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyVvvkllpniqptdNscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttCfqpqlInpalspisiqClrsllgsmtpavvqatlstsisaaeilsaglmegqiVsvlldemqmivkinVptivtqsnalvidfysissfinnqesiiqlpdrileigneqwRypaknckStrhhifcqyneaerlsleTklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltScqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsIEDKIEEILSKIYHIENEIARIKKLIGEAP
MuV-IL17 F 206C/A223C-GGG-476-GCN4(SEQ ID NO:51
mkvslvtclgfavfsfsicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptdnscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqnhintimntqlnnmscqildnqlatslglylteLTTCfqpqlinpalspisiqCLRSllgsmtpavvqatlstsisaaeilsaglmegqivsvlldemqmivkiniptivtqsnalvidfysissfingqesiiqlpdrileigneqwsypaknckltrhnifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelikvnaslqnavkyikesnhqlqsIEDKIEEILSKIYHIENEIARIKKLIGEAP
融合后MuV胞外域(SEQ ID NO:52)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevCegtqqCaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttvfqpqltnpalspisiqalrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsvsvnskigaiIEDKIEEILSKIYHIENEIARIKKLIGEAP
MuVF 206C/223C-GGG-476+GCN4+MuVHN_G(SEQ ID NO:27)
mkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttCfqpqltnpalspisiqClrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsIEDKIEEILSKIYHIENEIARIKKLIGEAPGSGGGGGGniplvndlrfinginkfiiedyathdfsighplnmpsfiptatspngctripsfslgkthwcythnvinanckdhtssnqyvsmgilvqtasgypmfktlkiqylsdglnrkscsiatvpdgcamycyvstqletddyagsspptqkltllfyndtvtertispsglegnwatlvpgvgsgiyfenklifpayggvlpnstlgvksareffrpvnpynpcsgpqqdldqralrsyfpsyfsnrriqsaflvcawnqilvtncelvvpssnqtmmgaegrvllinnrllyyqrstswwpyellyeisftftnsgpssvnmswipiysftrpgsgncsgenvcptacvsgvyldpwpltpyshqsginrnfyftgallnssttrvnptlyvsalnnlkvlapygtqglfasyttttcfqdtgdasvycvyimelasnivgefqilpvltrltit
以上序列包括N-末端信号肽、MuVF胞外域、GCN4三聚化结构域、任选的MuV HN胞外域、凝血酶切割位点、HIS标签和Strep标签，以及区段之间的各种接头残基。
实施例2
以融合前构象稳定的MeVF蛋白及其与MeVH胞外域或MuVHN胞外域的融合物
本实施例示出了通过一个或更多个氨基酸取代以融合前构象稳定的MeVF胞外域三聚体的实施方案。进一步提供了与MeVH胞外域连接的MeVF胞外域三聚体。例如，融合前稳定的MeVF胞外域三聚体和与MeVH胞外域的相应融合物可用于诱导受试者对MeV的中和免疫应答。
当在细胞中产生时，连接至C-末端GCN4三聚化结构域的MeVF胞外域形成三聚体，这些三聚体自发地转变为融合前构象。不稳定的重组MeVF-GCN4非常不稳定，100％的分子在评价点(EM)已转变成融合后构象。在没有稳定化的情况下，蛋白表达也显著降低。
因此，基于结构的疫苗设计用于识别稳定以融合前构象的MeV F胞外域的突变(基于融合前PIV5 F结构PDB ID 4WSG和MeV F结构PDB ID 5YXW)，并消除F1/F2切割位点以产生表达增加的“单链”MeV F蛋白。采用多种稳定策略将MeV F胞外域“锁定”在融合前构象中，包括引入二硫键和脯氨酸取代。总共对约40种不同突变体进行设计、表达、纯化、评估表达水平并通过负染色EM评估融合前构象。
将突变引入MeV F胞外域(基于MeV F第486位的C-末端截短)，并且连接至C-末端GCN4三聚化结构域，并且如上所述筛选所得突变体。胞外域还包括突变以去除F1/F2弗林蛋白酶切割位点。评估的融合前稳定突变包括：位于MeV F第48和284、90和225、141和270、165和171、173和245、175和241、212和236、216和233以及219和224位中的一个或更多个位置的半胱氨酸取代，形成非天然二硫键；和位于MeV F第194位的脯氨酸取代。相关序列如下所示。
与未修饰的MeV F相比，单链和融合前稳定的MeV F蛋白的表达和纯化显示出表达水平显著增加。
如图5所示，负EM可用于区分处于融合前构象和融合后构象的MeV F胞外域三聚体。此外，MeV F R165C-M171C-486-GCN4(SEQ ID NO:38)显示出融合前稳定性和蛋白表达的出色组合，并通过S200凝胶过滤纯化为单分散蛋白质。通过电子显微镜分析该构建体和其他构建体以进一步确认融合前构象。
用以融合后构象的MeV F胞外域三聚体和以融合前构象的MeV F胞外域三聚体(MeV F R165C-M171C-486-GCN4(SEQ ID NO:38))进行免疫测定。免疫方案根据图3A所示。在第0周和第3周用在Poly IC佐剂中的10μg/剂量的蛋白免疫10只CB6F1/J小鼠的组，并评估来自免疫小鼠的第5周血清的中和滴度。在MeV中和试验中评估免疫血清(图5C)，这表明与来自用融合后MeV F免疫的动物的血清相比，来自用MeV FR165C-M171C-486-GCN4(SEQID NO:38)免疫的动物的免疫血清中和MeV高多于200倍，并且超过保护阈值。
序列：
MeV F A90C/I225C-GGG-486-GCN4(SEQ ID NO:37)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdClnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpCsaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R165C/M171C-GGG-486-GCN4(SEQ ID NO:38)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F L173C/V245C-GGG-486-GCN4(SEQ ID NO:39)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemiCavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkCleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F V175C/D241C-GGG-486-GCN4(SEQ ID NO:40)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilaCqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggCinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F L216C/A233C-GGG-486-GCN4(SEQ ID NO:41)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryyteilsCfgpslrdpisaeisiqClsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F P219C/P224C-GGG-486-GCN4(SEQ ID NO:42)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgCslrdCisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F S194P-GGG-486-GCN4(SEQ ID NO:43)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
融合后MeV F(SEQ ID NO:61)
MYSMQLASCVTLTLVLLVNSQIHWGNLSKIGVVGIGSASYKVMTRSSHQSLVIKLMPNITLLNNCTRVEIAEYRRLLRTVLEPIRDALNAVTQNIRPVQSVASSGSGGGSAQITAGIALHQSMLNSQAIDNLRASLETTNQAIEAIRQAGQEMILAVQGVQDYINNELIPSMNQLSCDLIGQKLGLKLLRYYTEILSLFGPSLRDPISAEISIQALSYALGGDINKVLEKLGYSGGDLLGILESRGIKARITHVDTESYFIVLSIAYPTLSEIKGVIVHRLEGVSYNIGSQEWYTTVPKYVATQGYLISNFDESSCTFMPEGTVCSQNALYPMSPLLQECLRGSTKSCARTLVSGSFGNRFILSQGNLIANCASILCKCYTTGTIINQDPDKILTYIAADHCPVVEVNGVTIQVGSRRYPDAVYLHRIDLGPPISLERLDVGTNLGNAIAKLEDAKELLESSDQILSAIGGYIPEAPRDGQAYVRKDGEWVLLSTFL
MeV F S103C/V117C-GGG-486-GCN4(SEQ ID NO:62)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqCvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F V175F-GGG-486-GCN4(SEQ ID NO:63)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilaFqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F S103C/V117C,V175F,S194P-GGG-486-GCN4(SEQ ID NO:64)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqCvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F S103C/V117C,R165C/M171C,S194P-GGG-486-GCN4(SEQ ID NO:65)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqCvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C-GGG-486-GCN4(SEQ ID NO:66)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C,V175F,S194P-GGG-486-GCN4(SEQ ID NO:67)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C,R165C/M171C,S194P-GGG-486-GCN4(SEQ ID NO:68)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C-GGG-493-GCN4(SEQ ID NO:69)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqil rsmkglssa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C,V175F,S194P-GGG-493-GCN4(SEQ ID NO:70)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqil rsmkglssa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C,R165C/M171C,S194P-GGG-493-GCN4(SEQ ID NO:71)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqil rsmkglssa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F P100C/V117C,R165C/M171C,S194P-GGG-486-GCN4(SEQ ID NO:72)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirCvqsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F V101C/V117C,R165C/M171C,S194P-GGG-486-GCN4(SEQ ID NO:73)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpCqsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F Q102C/V117C,R165C/M171C,S194P-GGG-486-GCN4(SEQ ID NO:74)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvCsvassrrhGGGagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R165C/M171C,M141C/T270C-GGG-486-GCN4(SEQ ID NO:75)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsClnsqaidnlraslettnqaieaiCqagqeCilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarichvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R165C/M171C,E212C/Y236C-GGG-486-GCN4(SEQ ID NO:76)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilavqgvqdyinnelipsmnqlscdligqklglkllryytCilslfgpslrdpisaeisiqalsCalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R165C/M171C,R48C/A284C-GGG-486-GCN4(SEQ ID NO:77)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtCsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiCyptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C,天然弗林-486-GCN4(SEQ ID NO:78)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhkrfagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C,V175F,S194P,天然弗林-486-GCN4(SEQ ID NO:79)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhkrfagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieairqagqemilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
MeV F R99C/V117C,R165C/M171C,S194P,天然弗林-486-GCN4(SEQ ID NO:80)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqniCpvqsvassrrhkrfagvClagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilaFqgvqdyinnelipsmnqlPcdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP
以上序列包括N-末端信号肽、MeV F胞外域、GCN4三聚化结构域，以及区段之间的各种接头残基。
此外，具有MeV F胞外域的嵌合构建体，其具有氨基酸取代以与MuV HN胞外域或MeV H胞外域连接的融合前构象稳定，设计如下：
MeV F R165C/M171C-GGG-486-GCN4+MeV_H(SEQ ID NO:56)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAP GSGGGGGGflavskgncsgpttirgqfsnmslslldlylgrgynvssivtmtsqgmyggtylvekpnlsskrselsqlsmyrvfevgvirnpglgapvfhmtnyleqpvsndlsncmvalgelklaalchgedsitipyqgsgkgvsfqlvklgvwkspadmqswvplstddpvidrlylsshrgviadnqakwavpttrtddklrmetcfqqackgkiqtlcenpewaplkdnripsygvlsvdlsltvelkikiasgfgplithgsgmdlyksnhnnvywltippmknlalgvintlewiprfkvspylftvpikeagedchaptylpaevdgdvklssnlvilpgqdlqyvlatydtsrvehavvyyvyspgrsfsyfypfrlpikgvpielqvecftwdqklwcrhfcvladsesgghithsgmvgmgvsctvtredgtnrr
MeV F R165C/M171C-GGG-486-GCN4+MuV_HN(SEQ ID NO:57)
mysmqlascvtltlvllvnsqihwgnlskigvvgigsasykvmtrsshqslviklmpnitllnnctrveiaeyrrllrtvlepirdalnavtqnirpvqsvassrrhGGGagvvlagaalgvataaqitagialhqsmlnsqaidnlraslettnqaieaiCqagqeCilavqgvqdyinnelipsmnqlscdligqklglkllryyteilslfgpslrdpisaeisiqalsyalggdinkvleklgysggdllgilesrgikarithvdtesyfivlsiayptlseikgvivhrlegvsynigsqewyttvpkyvatqgylisnfdessctfmpegtvcsqnalypmspllqeclrgstkscartlvsgsfgnrfilsqgnliancasilckcyttgtiinqdpdkiltyiaadhcpvvevngvtiqvgsrrypdavylhridlgppislerldvgtnlgnaiakledakellessdqilsa IEDKIEEILSKIYHIENEIARIKKLIGEAPG sggggggniplvndlrfinginkfiiedyathdfsighplnmpsfiptatspngctripsfslgkthwcythnvinanckdhtssnqyvsmgilvqtasgypmfktlkiqylsdglnrkscsiatvpdgcamycyvstqletddyagsspptqkltllfyndtvtertispsglegnwatlvpgvgsgiyfenklifpayggvlpnstlgvksareffrpvnpynpcsgpqqdldqralrsyfpsyfsnrriqsaflvcawnqilvtncelvvpssnqtmmgaegrvllinnrllyyqrstswwpyellyeisftftnsgpssvnmswipiysftrpgsgncsgenvcptacvsgvyldpwpltpyshqsginrnfyftgallnssttrvnptlyvsalnnlkvlapygtqglfasyttttcfqdtgdasvycvyimelasnivgefqilpvltrltit
MeV F R165C/M171C-GGG-486-GCN4/Fd+MeV_H(SEQ ID NO:81)
MYSMQLASCVTLTLVLLVNSQIHWGNLSKIGVVGIGSASYKVMTRSSHQSLVIKLMPNITLLNNCTRVEIAEYRRLLRTVLEPIRDALNAVTQNIRPVQSVASSRRHGGGAGVVLAGAALGVATAAQITAGIALHQSMLNSQAIDNLRASLETTNQAIEAICQAGQECILAVQGVQDYINNELIPSMNQLSCDLIGQKLGLKLLRYYTEILSLFGPSLRDPISAEISIQALSYALGGDINKVLEKLGYSGGDLLGILESRGIKARITHVDTESYFIVLSIAYPTLSEIKGVIVHRLEGVSYNIGSQEWYTTVPKYVATQGYLISNFDESSCTFMPEGTVCSQNALYPMSPLLQECLRGSTKSCARTLVSGSFGNRFILSQGNLIANCASILCKCYTTGTIINQDPDKILTYIAADHCPVVEVNGVTIQVGSRRYPDAVYLHRIDLGPPISLERLDVGTNLGNAIAKLEDAKELLESSDQILSA IEDKIEEILSKIYHIENEIARIKKLIGEAPGSGYIPEAPRDGQAYVRKDGEWVLLSTFL GSGGGGGgFLAVSKGNCSGPTTIRGQFSNMSLSLLDLYLGRGYNVSSIVTMTSQGMYGGTYLVEKPNLSSKRSELSQLSMYRVFEVGVIRNPGLGAPVFHMTNYLEQPVSNDLSNCMVALGELKLAALCHGEDSITIPYQGSGKGVSFQLVKLGVWKSPADMQSWVPLSTDDPVIDRLYLSSHRGVIADNQAKWAVPTTRTDDKLRMETCFQQACKGKIQTLCENPEWAPLKDNRIPSYGVLSVDLSLTVELKIKIASGFGPLITHGSGMDLYKSNHNNVYWLTIPPMKNLALGVINTLEWIPRFKVSPYLFTVPIKEAGEDCHAPTYLPAEVDGDVKLSSNLVILPGQDLQYVLATYDTSRVEHAVVYYVYSPGRSFSYFYPFRLPIKGVPIELQVECFTWDQKLWCRHFCVLADSESGGHITHSGMVGMGVSCTVTREDGTNRR
以上序列包括N-末端信号肽、MeV F胞外域、GCN4三聚化结构域、任选的T4纤维蛋白三聚化结构域、MeV H胞外域，以及区段之间的各种接头残基。
实施例3
多聚体MuV HN、多聚体MeV H和MuV pre-F-MeV H嵌合体
本实施例描述了与MeV H胞外域连接的重组MuV前F胞外域三聚体的实施方案，以提供引发对MeV和MuV的交叉中和免疫应答的嵌合免疫原。此外，还描述了多聚体MuV HN和多聚体MeV H。
为了增加MuV F胞外域三聚体的免疫原性足迹，将MeV H胞外域基因融合至三聚体每个原体的三聚化结构域的C-末端。评估的构建体包括含有V206C-A223C的MuV F胞外域、消除F1/F2弗林蛋白酶切割位点的突变、与胞外域第476位融合的三聚化结构域和与三聚化结构域的C-末端连接的MeV H胞外域。在本实施例中，三聚化结构域包括串联的GCN4三聚化结构域和T4纤维蛋白三聚化结构域；但是，任一这些结构域也可以单独使用。形式如图6A所示。相应的序列如下所示。
MuV F 206C/223C-476+GCN4/Fd+MeV-H(SEQ ID NO:28):
MkafsvtclsfavfsssicvninilqqigyikqqvrqlsyysqssssyivvkllpniqptddscefksvtqynktlsnlllpiaeninniaspspgsrrhGGGagiaigiaalgvataaqvtaavslvqaqtnaraiaamknsiqatnravfevkegtqqlaiavqaiqdhintimntqlnnmscqildnqlatslglyltelttCfqpqltnpalspisiqClrsllgsmtpavvqatlstsisaaeilsaglmegqiisvlldemqmivkiniptivtqsnalvidfysissfinnqesiiqlpdrileigneqwsypaknckltrhhifcqyneaerlslesklclagnisacvfspiagsymrrfvaldgtivancrsltclckspsypiyqpdhhavttidltacqtlsldgldfsivslsnityaenltislsqtintqpidistelskvnaslqnavkyikesnhqlqsIEDKIEEILSKIYHIENEIARIKKLIGEAP GSGYIPEAPRDGQAYVRKDGEWVLLSTFL GSGGGGGgFlavskgncsgpttirgqfsnmslslldlylgrgynvssivtmtsqgmyggtylvekpnlsskrselsqlsmyrvfevgvirnpglgapvfhmtnyleqpvsndlsncmvalgelklaalchgedsitipyqgsgkgvsfqlvklgvwkspadmqswvplstddpvidrlylsshrgviadnqakwavpttrtddklrmetcfqqackgkiqtlcenpewaplkdnripsygvlsvdlsltvelkikiasgfgplithgsgmdlyksnhnnvywltippmknlalgvintlewiprfkvspylftvpikeagedchaptylpaevdgdvklssnlvilpgqdlqyvlatydtsrvehavvyyvyspgrsfsyfypfrlpikgvpielqvecftwdqklwcrhfcvladsesgghithsgmvgmgvsctvtredgtnrr
纯化的MuV F 206C-223C-476+GCN4/Fd+MeV-H(SEQ ID NO:28)的负染色EM显示F胞外域保持融合前构象，其中具有三个H胞外域(一个与每个F原体相连)，它们排列在三聚化结构域的C-末端(图6A)。负染色EM显示该构建体组装有类似于实施例1中所述的MuV F-MuV HN的构象。
构建含有MeV H胞外域头部区或MuV HN胞外域头部区的多聚体的另外的免疫原。
三聚体MuV HN胞外域头部区和三聚体MeV H胞外域头部区通过将头部区的N-末端连接至T4纤维蛋白三聚化结构域进行构建。序列提供如下：
Fd-MuV HN(三聚体MuV HN,SEQ ID NO:58)
mysmqlascvtltlvllvnsq GS GYIPEAPRDGQAYVRKDGEWVLLSTFL GSGGGGGgNIPLVNDLRFINGINKFIIEDYATHDFSIGHPLNMPSFIPTATSPNGCTRIPSFSLGKTHWCYTHNVINANCKDHTSSNQYVSMGILVQTASGYPMFKTLKIQYLSDGLNRKSCSIATVPDGCAMYCYVSTQLETDDYAGSSPPTQKLTLLFYNDTVTERTISPSGLEGNWATLVPGVGSGIYFENKLIFPAYGGVLPNSTLGVKSAREFFRPVNPYNPCSGPQQDLDQRALRSYFPSYFSNRRIQSAFLVCAWNQILVTNCELVVPSSNQTMMGAEGRVLLINNRLLYYQRSTSWWPYELLYEISFTFTNSGPSSVNMSWIPIYSFTRPGSGNCSGENVCPTACVSGVYLDPWPLTPYSHQSGINRNFYFTGALLNSSTTRVNPTLYVSALNNLKVLAPYGTQGLFASYTTTTCFQDTGDASVYCVYIMELASNIVGEFQILPVLTRLTIT
Fd-MeV H(三聚体MeV H,SEQ ID NO:59)
mysmqlascvtltlvllvnsq GS GYIPEAPRDGQAYVRKDGEWVLLSTFL GSGGGGGgFLAVSKGNCSGPTTIRGQFSNMSLSLLDLYLGRGYNVSSIVTMTSQGMYGGTYLVEKPNLSSKRSELSQLSMYRVFEVGVIRNPGLGAPVFHMTNYLEQPVSNDLSNCMVALGELKLAALCHGEDSITIPYQGSGKGVSFQLVKLGVWKSPADMQSWVPLSTDDPVIDRLYLSSHRGVIADNQAKWAVPTTRTDDKLRMETCFQQACKGKIQTLCENPEWAPLKDNRIPSYGVLSVDLSLTVELKIKIASGFGPLITHGSGMDLYKSNHNNVYWLTIPPMKNLALGVINTLEWIPRFKVSPYLFTVPIKEAGEDCHAPTYLPAEVDGDVKLSSNLVILPGQDLQYVLATYDTSRVEHAVVYYVYSPGRSFSYFYPFRLPIKGVPIELQVECFTWDQKLWCRHFCVLADSESGGHITHSGMVGMGVSCTVTREDGTNRR
通过在哺乳动物细胞中表达MeV H胞外域头部区并纯化所得蛋白复合物来构建二聚体MeV H；MeV H头部在生理溶液中二聚化。MeV H头部区的序列提供如下：
二聚体MeV H头部区(SEQ ID NO:60)
Mysmqlascvtltlvllvnsq flavskgncsgpttirgqfsnmslslldlylgrgynvssivtmtsqgmyggtylvekpnlsskrselsqlsmyrvfevgvirnpglgapvfhmtnyleqpvsndlsncmvalgelklaalchgedsitipyqgsgkgvsfqlvklgvwkspadmqswvplstddpvidrlylsshrgviadnqakwavpttrtddklrmetcfqqackgkiqtlcenpewaplkdnripsygvlsvdlsltvelkikiasgfgplithgsgmdlyksnhnnvywltippmknlalgvintlewiprfkvspylftvpikeagedchaptylpaevdgdvklssnlvilpgqdlqyvlatydtsrvehavvyyvyspgrsfsyfypfrlpikgvpielqvecftwdqklwcrhfcvladsesgghithsgmvgmgvsctvtredgtnrr
通过在哺乳动物细胞中表达MeV H胞外域茎部和头部区并纯化所得蛋白复合物，可以构建包括茎部和头部区的二聚体MeV H；MeV H茎部和头部在生理溶液中二聚化。MeV H茎部和头部区的示例性序列提供如下：
MeV H茎部和头部(SEQ ID NO:86)
mysmqlascvtltlvllvnsq RLHraaiytaeihkslstnldvtnsiehqvkdvltplfkiigdevglrtpqrftdlvkfisdkikflnpdreydfrdltwcinpperikldydqycadvaaeelmnalvnstlletrttnqFLAVskgncsgpttirgqfsnmslslldlylgrgynvssivtmtsqgmyggtylvekpnlsskrselsqlsmyrvfevgvirnpglgapvfhmtnyleqpvsndlsncmvalgelklaalchgedsitipyqgsgkgvsfqlvklgvwksptdmqswvplstddpvidrlylsshrgviadnqakwavpttrtddklrmetcfqqackgkiqalcenpewaplkdnripsygvlsvdlsltvelkikiasgfgplithgsgmdlyksnhnnvywltippmknlalgvintlewiprfkvspylfnvpikeagedchaptylpaevdgdvklssnlvilpgqdlqyvlatydtsrvehavvyyvyspsrsfsyfypfrlpikgvpielqvecftwdqklwcrhfcvladsesgghithsgmegmgvsctvtredgtnrr
MeV H茎部和头部2(SEQ ID NO:87)
mysmqlascvtltlvllvnsq raaiytaeihkslstnldvtnsiehqvkdvltplfkiigdevglrtpqrftdlvkfisdkikflnpdreydfrdltwcinpperikldydqycadvaaeelmnalvnstlletrttnqFLAVskgncsgpttirgqfsnmslslldlylgrgynvssivtmtsqgmyggtylvekpnlsskrselsqlsmyrvfevgvirnpglgapvfhmtnyleqpvsndlsncmvalgelklaalchgedsitipyqgsgkgvsfqlvklgvwksptdmqswvplstddpvidrlylsshrgviadnqakwavpttrtddklrmetcfqqackgkiqalcenpewaplkdnripsygvlsvdlsltvelkikiasgfgplithgsgmdlyksnhnnvywltippmknlalgvintlewiprfkvspylfnvpikeagedchaptylpaevdgdvklssnlvilpgqdlqyvlatydtsrvehavvyyvyspsrsfsyfypfrlpikgvpielqvecftwdqklwcrhfcvladsesgghithsgmegmgvsctvtredgtnrr
MeV H茎部和头部3(SEQ ID NO:88)
mysmqlascvtltlvllvnsq LHraaiytaeihkslstnldvtnsiehqvkdvltplfkiigdevglrtpqrftdlvkfisdkikflnpdreydfrdltwcinpperikldydqycadvaaeelmnalvnstlletrttnqFLAVskgncsgpttirgqfsnmslslldlylgrgynvssivtmtsqgmyggtylvekpnlsskrselsqlsmyrvfevgvirnpglgapvfhmtnyleqpvsndlsncmvalgelklaalchgedsitipyqgsgkgvsfqlvklgvwksptdmqswvplstddpvidrlylsshrgviadnqakwavpttrtddklrmetcfqqackgkiqalcenpewaplkdnripsygvlsvdlsltvelkikiasgfgplithgsgmdlyksnhnnvywltippmknlalgvintlewiprfkvspylfnvpikeagedchaptylpaevdgdvklssnlvilpgqdlqyvlatydtsrvehavvyyvyspsrsfsyfypfrlpikgvpielqvecftwdqklwcrhfcvladsesgghithsgmegmgvsctvtredgtnrr
MeV H茎部和头部4(SEQ ID NO:89)
mysmqlascvtltlvllvnsq taeihkslstnldvtnsiehqvkdvltplfkiigdevglrtpqrftdlvkfisdkikflnpdreydfrdltwcinpperikldydqycadvaaeelmnalvnstlletrttnqFLAVskgncsgpttirgqfsnmslslldlylgrgynvssivtmtsqgmyggtylvekpnlsskrselsqlsmyrvfevgvirnpglgapvfhmtnyleqpvsndlsncmvalgelklaalchgedsitipyqgsgkgvsfqlvklgvwksptdmqswvplstddpvidrlylsshrgviadnqakwavpttrtddklrmetcfqqackgkiqalcenpewaplkdnripsygvlsvdlsltvelkikiasgfgplithgsgmdlyksnhnnvywltippmknlalgvintlewiprfkvspylfnvpikeagedchaptylpaevdgdvklssnlvilpgqdlqyvlatydtsrvehavvyyvyspsrsfsyfypfrlpikgvpielqvecftwdqklwcrhfcvladsesgghithsgmegmgvsctvtredgtnrr
可以通过在哺乳动物细胞中表达MuV HN胞外域茎部和头部区并纯化所得蛋白来构建包括茎部和头部区的MuV HN。MuV HN茎部和头部区的示例性序列提供如下：
MuV HN茎部和头部(SEQ ID NO:90)
mysmqlascvtltlvllvnsq ELVRmindqglsnqlssitdkiresatmiasavgvmnqvihgvtvslplqiegnqnqllatlatictsqkqvsncstNIPlvndlrfinginkfiiedyathdfsighplnmpsfiptatspngctripsfslgkthwcythnvinanckdhtssnqyvsmgilvqtasgypmfktlkiqylsdglnrkscsiatvpdgcamycyvstqletddyagsspptqkltllfyndtvtertispsglegnwatlvpgvgsgiyfenklifpayggvlpnstlgvksareffrpvnpynpcsgpqqdldqralrsyfpsyfsnrriqsaflvcawnqilvtncelvvpssnqtmmgaegrvllinnrllyyqrstswwpyellyeisftftnsgpssvnmswipiysftrpgsgncsgenvcptacvsgvyldpwpltpyshqsginrnfyftgallnssttrvnptlyvsalnnlkvlapygtqglfasyttttcfqdtgdasvycvyimelasnivgefqilpvltrltit
MuV HN茎部和头部2(SEQ ID NO:91)
mysmqlascvtltlvllvnsq dqglsnqlssitdkiresatmiasavgvmnqvihgvtvslplqiegnqnqllatlatictsqkqvsncstNIPlvndlrfinginkfiiedyathdfsighplnmpsfiptatspngctripsfslgkthwcythnvinanckdhtssnqyvsmgilvqtasgypmfktlkiqylsdglnrkscsiatvpdgcamycyvstqletddyagsspptqkltllfyndtvtertispsglegnwatlvpgvgsgiyfenklifpayggvlpnstlgvksareffrpvnpynpcsgpqqdldqralrsyfpsyfsnrriqsaflvcawnqilvtncelvvpssnqtmmgaegrvllinnrllyyqrstswwpyellyeisftftnsgpssvnmswipiysftrpgsgncsgenvcptacvsgvyldpwpltpyshqsginrnfyftgallnssttrvnptlyvsalnnlkvlapygtqglfasyttttcfqdtgdasvycvyimelasnivgefqilpvltrltit
MuV HN茎部和头部3(SEQ ID NO:92)
mysmqlascvtltlvllvnsq glsnqlssitdkiresatmiasavgvmnqvihgvtvslplqiegnqnqllatlatictsqkqvsncstNIPlvndlrfinginkfiiedyathdfsighplnmpsfiptatspngctripsfslgkthwcythnvinanckdhtssnqyvsmgilvqtasgypmfktlkiqylsdglnrkscsiatvpdgcamycyvstqletddyagsspptqkltllfyndtvtertispsglegnwatlvpgvgsgiyfenklifpayggvlpnstlgvksareffrpvnpynpcsgpqqdldqralrsyfpsyfsnrriqsaflvcawnqilvtncelvvpssnqtmmgaegrvllinnrllyyqrstswwpyellyeisftftnsgpssvnmswipiysftrpgsgncsgenvcptacvsgvyldpwpltpyshqsginrnfyftgallnssttrvnptlyvsalnnlkvlapygtqglfasyttttcfqdtgdasvycvyimelasnivgefqilpvltrltit
MuV HN茎部和头部4(SEQ ID NO:93)
mysmqlascvtltlvllvnsq LVRmindqglsnqlssitdkiresatmiasavgvmnqvihgvtvslplqiegnqnqllatlatictsqkqvsncstNIPlvndlrfinginkfiiedyathdfsighplnmpsfiptatspngctripsfslgkthwcythnvinanckdhtssnqyvsmgilvqtasgypmfktlkiqylsdglnrkscsiatvpdgcamycyvstqletddyagsspptqkltllfyndtvtertispsglegnwatlvpgvgsgiyfenklifpayggvlpnstlgvksareffrpvnpynpcsgpqqdldqralrsyfpsyfsnrriqsaflvcawnqilvtncelvvpssnqtmmgaegrvllinnrllyyqrstswwpyellyeisftftnsgpssvnmswipiysftrpgsgncsgenvcptacvsgvyldpwpltpyshqsginrnfyftgallnssttrvnptlyvsalnnlkvlapygtqglfasyttttcfqdtgdasvycvyimelasnivgefqilpvltrltit
此外，通过将这些分子连接至T4纤维蛋白三聚化结构域和/或GCN4三聚化结构域的N-末端和C-末端来构建三聚体MuV HN胞外域头部区和三聚体MeV H胞外域头部区的嵌合形式。序列提供如下：
MuV HN-Fd-MeV H(SEQ ID NO:82)
mysmqlascvtltlvllvnsq NIPlvndlrfinginkfiiedyathdfsighplnmpsfiptatspngctripsfslgkthwcythnvinanckdhtssnqyvsmgilvqtasgypmfktlkiqylsdglnrkscsiatvpdgcamycyvstqletddyagsspptqkltllfyndtvtertispsglegnwatlvpgvgsgiyfenklifpayggvlpnstlgvksareffrpvnpynpcsgpqqdldqralrsyfpsyfsnrriqsaflvcawnqilvtncelvvpssnqtmmgaegrvllinnrllyyqrstswwpyellyeisftftnsgpssvnmswipiysftrpgsgncsgenvcptacvsgvyldpwpltpyshqsginrnfyftgallnssttrvnptlyvsalnnlkvlapygtqglfasyttttcfqdtgdasvycvyimelasnivgefqilpvltrltitgg GSGYIPEAPRDGQAYVRKDGEWVLLSTFLGSGGGGGgflavskgncsgpttirgqfsnmslslldlylgrgynvssivtmtsqgmyggtylvekpnlsskrselsqlsmyrvfevgvirnpglgapvfhmtnyleqpvsndlsncmvalgelklaalchgedsitipyqgsgkgvsfqlvklgvwksptdmqswvplstddpvidrlylsshrgviadnqakwavpttrtddklrmetcfqqackgkiqalcenpewaplkdnripsygvlsvdlsltvelkikiasgfgplithgsgmdlyksnhnnvywltippmknlalgvintlewiprfkvspylfnvpikeagedchaptylpaevdgdvklssnlvilpgqdlqyvlatydtsrvehavvyyvyspsrsfsyfypfrlpikgvpielqvecftwdqklwcrhfcvladsesgghithsgmegmgvsctvtredgtnrr
MuV HN-GCN4/Fd-MeV H(SEQ ID NO:83)
mysmqlascvtltlvllvnsq NIPlvndlrfinginkfiiedyathdfsighplnmpsfiptatspngctripsfslgkthwcythnvinanckdhtssnqyvsmgilvqtasgypmfktlkiqylsdglnrkscsiatvpdgcamycyvstqletddyagsspptqkltllfyndtvtertispsglegnwatlvpgvgsgiyfenklifpayggvlpnstlgvksareffrpvnpynpcsgpqqdldqralrsyfpsyfsnrriqsaflvcawnqilvtncelvvpssnqtmmgaegrvllinnrllyyqrstswwpyellyeisftftnsgpssvnmswipiysftrpgsgncsgenvcptacvsgvyldpwpltpyshqsginrnfyftgallnssttrvnptlyvsalnnlkvlapygtqglfasyttttcfqdtgdasvycvyimelasnivgefqilpvltrltit gg KLMKQIEDKIEEILSKIYHIENEIARIKKLIGEAPGSGYIPEAPRDGQAYVRKDGEWVLLSTFL GSGGGGGgflavskgncsgpttirgqfsnmslslldlylgrgynvssivtmtsqgmyggtylvekpnlsskrselsqlsmyrvfevgvirnpglgapvfhmtnyleqpvsndlsncmvalgelklaalchgedsitipyqgsgkgvsfqlvklgvwksptdmqswvplstddpvidrlylsshrgviadnqakwavpttrtddklrmetcfqqackgkiqalcenpewaplkdnripsygvlsvdlsltvelkikiasgfgplithgsgmdlyksnhnnvywltippmknlalgvintlewiprfkvspylfnvpikeagedchaptylpaevdgdvklssnlvilpgqdlqyvlatydtsrvehavvyyvyspsrsfsyfypfrlpikgvpielqvecftwdqklwcrhfcvladsesgghithsgmegmgvsctvtredgtnrr
MeV H-Fd-MuV HN(SEQ ID NO:84)
mysmqlascvtltlvllvnsq FLAvskgncsgpttirgqfsnmslslldlylgrgynvssivtmtsqgmyggtylvekpnlsskrselsqlsmyrvfevgvirnpglgapvfhmtnyleqpvsndlsncmvalgelklaalchgedsitipyqgsgkgvsfqlvklgvwksptdmqswvplstddpvidrlylsshrgviadnqakwavpttrtddklrmetcfqqackgkiqalcenpewaplkdnripsygvlsvdlsltvelkikiasgfgplithgsgmdlyksnhnnvywltippmknlalgvintlewiprfkvspylfnvpikeagedchaptylpaevdgdvklssnlvilpgqdlqyvlatydtsrvehavvyyvyspsrsfsyfypfrlpikgvpielqvecftwdqklwcrhfcvladsesgghithsgmegmgvsctvtredgtnrr GSGYIPEAPRDGQAYVRKDGEWVLLSTFL GSGGGGGgNIPlvndlrfinginkfiiedyathdfsighplnmpsfiptatspngctripsfslgkthwcythnvinanckdhtssnqyvsmgilvqtasgypmfktlkiqylsdglnrkscsiatvpdgcamycyvstqletddyagsspptqkltllfyndtvtertispsglegnwatlvpgvgsgiyfenklifpayggvlpnstlgvksareffrpvnpynpcsgpqqdldqralrsyfpsyfsnrriqsaflvcawnqilvtncelvvpssnqtmmgaegrvllinnrllyyqrstswwpyellyeisftftnsgpssvnmswipiysftrpgsgncsgenvcptacvsgvyldpwpltpyshqsginrnfyftgallnssttrvnptlyvsalnnlkvlapygtqglfasyttttcfqdtgdasvycvyimelasnivgefqilpvltrltit
MeV H-GCN4/Fd-MuV HN(SEQ ID NO:85)
mysmqlascvtltlvllvnsq FLAvskgncsgpttirgqfsnmslslldlylgrgynvssivtmtsqgmyggtylvekpnlsskrselsqlsmyrvfevgvirnpglgapvfhmtnyleqpvsndlsncmvalgelklaalchgedsitipyqgsgkgvsfqlvklgvwksptdmqswvplstddpvidrlylsshrgviadnqakwavpttrtddklrmetcfqqackgkiqalcenpewaplkdnripsygvlsvdlsltvelkikiasgfgplithgsgmdlyksnhnnvywltippmknlalgvintlewiprfkvspylfnvpikeagedchaptylpaevdgdvklssnlvilpgqdlqyvlatydtsrvehavvyyvyspsrsfsyfypfrlpikgvpielqvecftwdqklwcrhfcvladsesgghithsgmegmgvsctvtredgtnrr MKQIEDKIEEILSKIYHIENEIARIKKLIGEAP GSGYIPEAPRDGQAYVRKDGEWVLLSTFLGSGGGGGgNIPlvndlrfinginkfiiedyathdfsighplnmpsfiptatspngctripsfslgkthwcythnvinanckdhtssnqyvsmgilvqtasgypmfktlkiqylsdglnrkscsiatvpdgcamycyvstqletddyagsspptqkltllfyndtvtertispsglegnwatlvpgvgsgiyfenklifpayggvlpnstlgvksareffrpvnpynpcsgpqqdldqralrsyfpsyfsnrriqsaflvcawnqilvtncelvvpssnqtmmgaegrvllinnrllyyqrstswwpyellyeisftftnsgpssvnmswipiysftrpgsgncsgenvcptacvsgvyldpwpltpyshqsginrnfyftgallnssttrvnptlyvsalnnlkvlapygtqglfasyttttcfqdtgdasvycvyimelasnivgefqilpvltrltit
对MeV H胞外域头部二聚体(SEQ ID NO:60)、MeV H胞外域头部三聚体(SEQ IDNO:59)和MuV HN胞外域头部三聚体(SEQ ID NO:58)进行设计、表达、纯化和用负染色EM表征(参见图6A)。
用纯化的构建体免疫小鼠，并通过PRNT评估血清的MeV和MuV中和。评估的免疫原是MeV H二聚体或三聚体(SEQ ID NO:59或60)、融合前MuV F胞外域三聚体(MuV F V206C-A223C-GGG-476-GCN4,SEQ ID NO:11)、MuV pre-F-MeV H嵌合体(MuV F 206C-223C-476+GCN4/Fd+MeV_H_3INB-tHS(SEQ ID NO:28)和MuV HN三聚体(SEQ ID NO:58)。
对于MeV中和(图6B)，MeV H二聚体引发的PRNT ID60平均为66,000，MeV H三聚体为30,000，并且融合前MuV F的C-末端的三聚化H为35,000。考虑到用融合前MeV F三聚体免疫观察到的中和滴度低得多，这些结果非常出乎意料(PRNT为873；参见图5C)。2×MMR疫苗接种后的人PRNT滴度平均为约650。MeV H设计的MeV PRNT滴度不是预期值，因为最初认为融合前F胞外域三聚体提供更佳免疫应答(类似于其他副粘病毒，例如RSV)。出乎意料的是，多聚体MeV H免疫原显示出极佳免疫原性，比融合前稳定的MeV F胞外域三聚体强75倍，比MMR疫苗接种后的人应答高约100倍。
对于MuV中和(图6C)，每种评估的免疫原引发高于保护阈值的免疫应答，并且可溶性三聚体MuV HN引发非常有效的免疫应答，考虑到可溶性MuV HN单体引发的极小应答，这是非常出人意料的(图4C)。出人意料的是，MuV HN胞外域单体和三聚体MuV HN胞外域之间的中和效力增加了13.4倍。
如上所述进行蛋白产生、分析和免疫。
显然，所描述的方法或组合物的准确详情可以在不背离所描述实施方案的精神的情况下改变或修改。我们要求所有落入以下权利要求的范围和精神内的修改和变化。
序列表
<110> 美国政府（由卫生和人类服务部的部长所代表）
<120> 腮腺炎和麻疹病毒免疫原及其用途
<130> 4239-102936-02
<150> 62/946,902
<151> 2019-12-11
<160> 106
<170> PatentIn 3.5版
<210> 1
<211> 538
<212> PRT
<213> 腮腺炎病毒
<400> 1
Met Lys Ala Phe Ser Val Thr Cys Leu Gly Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asn Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Lys Arg Phe Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Ile Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Tyr Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Ile Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Val Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Thr Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Val Ser Val Asn
465 470 475 480
Ser Lys Ile Gly Ala Ile Ile Val Ala Ala Leu Val Leu Ser Ile Leu
485 490 495
Ser Ile Ile Ile Ser Leu Leu Phe Cys Cys Trp Ala Tyr Ile Ala Thr
500 505 510
Lys Glu Ile Arg Arg Ile Asn Phe Lys Thr Asn His Ile Asn Thr Ile
515 520 525
Ser Ser Ser Val Asp Asp Leu Ile Arg Tyr
530 535
<210> 2
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 2
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Cys Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Cys Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Val Ser Val Asn
465 470 475 480
Ser Lys Ile Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 3
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 3
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Cys Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Cys Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Val Ser Val Asn
465 470 475 480
Ser Lys Ile Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 4
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 4
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Cys Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Cys Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Val Ser Val Asn
465 470 475 480
Ser Lys Ile Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 5
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 5
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Cys Gln Leu Thr Asn Cys Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Val Ser Val Asn
465 470 475 480
Ser Lys Ile Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 6
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 6
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Cys Glu Gly Thr Gln Gln
145 150 155 160
Cys Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Val Ser Val Asn
465 470 475 480
Ser Lys Ile Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 7
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 7
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Cys Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Cys Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Val Ser Val Asn
465 470 475 480
Ser Lys Ile Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 8
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 8
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Cys Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Cys Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Val Ser Val Asn
465 470 475 480
Ser Lys Ile Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 9
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 9
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Pro Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Val Ser Val Asn
465 470 475 480
Ser Lys Ile Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 10
<211> 506
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 10
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Cys Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Cys Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Ile Glu Asp Lys
465 470 475 480
Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu Ile Ala
485 490 495
Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro
500 505
<210> 11
<211> 506
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 11
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Cys Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Cys Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Ile Glu Asp Lys
465 470 475 480
Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu Ile Ala
485 490 495
Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro
500 505
<210> 12
<211> 506
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 12
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Cys Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Cys Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Ile Glu Asp Lys
465 470 475 480
Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu Ile Ala
485 490 495
Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro
500 505
<210> 13
<211> 506
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 13
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Cys Gln Leu Thr Asn Cys Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Ile Glu Asp Lys
465 470 475 480
Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu Ile Ala
485 490 495
Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro
500 505
<210> 14
<211> 506
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 14
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Cys Glu Gly Thr Gln Gln
145 150 155 160
Cys Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Ile Glu Asp Lys
465 470 475 480
Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu Ile Ala
485 490 495
Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro
500 505
<210> 15
<211> 506
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 15
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Cys Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Cys Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Ile Glu Asp Lys
465 470 475 480
Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu Ile Ala
485 490 495
Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro
500 505
<210> 16
<211> 506
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 16
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Cys Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Cys Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Ile Glu Asp Lys
465 470 475 480
Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu Ile Ala
485 490 495
Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro
500 505
<210> 17
<211> 506
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 17
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Pro Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Ile Glu Asp Lys
465 470 475 480
Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu Ile Ala
485 490 495
Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro
500 505
<210> 18
<211> 499
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 18
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Cys Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Cys Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys
465 470 475 480
Ile Tyr His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly
485 490 495
Glu Ala Pro
<210> 19
<211> 499
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 19
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Cys Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Cys Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys
465 470 475 480
Ile Tyr His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly
485 490 495
Glu Ala Pro
<210> 20
<211> 499
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 20
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Cys Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Cys Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys
465 470 475 480
Ile Tyr His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly
485 490 495
Glu Ala Pro
<210> 21
<211> 499
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 21
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Cys Gln Leu Thr Asn Cys Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys
465 470 475 480
Ile Tyr His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly
485 490 495
Glu Ala Pro
<210> 22
<211> 499
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 22
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Cys Glu Gly Thr Gln Gln
145 150 155 160
Cys Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys
465 470 475 480
Ile Tyr His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly
485 490 495
Glu Ala Pro
<210> 23
<211> 499
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 23
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Cys Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Cys Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys
465 470 475 480
Ile Tyr His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly
485 490 495
Glu Ala Pro
<210> 24
<211> 499
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 24
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Cys Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Cys Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys
465 470 475 480
Ile Tyr His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly
485 490 495
Glu Ala Pro
<210> 25
<211> 499
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 25
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Pro Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys
465 470 475 480
Ile Tyr His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly
485 490 495
Glu Ala Pro
<210> 26
<211> 506
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 26
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Val Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asn Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Cys Phe Gln
195 200 205
Pro Gln Leu Ile Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Cys Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Val Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Val Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Arg Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Ser Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Thr Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ser Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Ile Glu Asp Lys
465 470 475 480
Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu Ile Ala
485 490 495
Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro
500 505
<210> 27
<211> 966
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 27
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Cys Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Cys Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Ile Glu Asp Lys
465 470 475 480
Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu Ile Ala
485 490 495
Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro Gly Ser Gly Gly Gly Gly
500 505 510
Gly Gly Asn Ile Pro Leu Val Asn Asp Leu Arg Phe Ile Asn Gly Ile
515 520 525
Asn Lys Phe Ile Ile Glu Asp Tyr Ala Thr His Asp Phe Ser Ile Gly
530 535 540
His Pro Leu Asn Met Pro Ser Phe Ile Pro Thr Ala Thr Ser Pro Asn
545 550 555 560
Gly Cys Thr Arg Ile Pro Ser Phe Ser Leu Gly Lys Thr His Trp Cys
565 570 575
Tyr Thr His Asn Val Ile Asn Ala Asn Cys Lys Asp His Thr Ser Ser
580 585 590
Asn Gln Tyr Val Ser Met Gly Ile Leu Val Gln Thr Ala Ser Gly Tyr
595 600 605
Pro Met Phe Lys Thr Leu Lys Ile Gln Tyr Leu Ser Asp Gly Leu Asn
610 615 620
Arg Lys Ser Cys Ser Ile Ala Thr Val Pro Asp Gly Cys Ala Met Tyr
625 630 635 640
Cys Tyr Val Ser Thr Gln Leu Glu Thr Asp Asp Tyr Ala Gly Ser Ser
645 650 655
Pro Pro Thr Gln Lys Leu Thr Leu Leu Phe Tyr Asn Asp Thr Val Thr
660 665 670
Glu Arg Thr Ile Ser Pro Ser Gly Leu Glu Gly Asn Trp Ala Thr Leu
675 680 685
Val Pro Gly Val Gly Ser Gly Ile Tyr Phe Glu Asn Lys Leu Ile Phe
690 695 700
Pro Ala Tyr Gly Gly Val Leu Pro Asn Ser Thr Leu Gly Val Lys Ser
705 710 715 720
Ala Arg Glu Phe Phe Arg Pro Val Asn Pro Tyr Asn Pro Cys Ser Gly
725 730 735
Pro Gln Gln Asp Leu Asp Gln Arg Ala Leu Arg Ser Tyr Phe Pro Ser
740 745 750
Tyr Phe Ser Asn Arg Arg Ile Gln Ser Ala Phe Leu Val Cys Ala Trp
755 760 765
Asn Gln Ile Leu Val Thr Asn Cys Glu Leu Val Val Pro Ser Ser Asn
770 775 780
Gln Thr Met Met Gly Ala Glu Gly Arg Val Leu Leu Ile Asn Asn Arg
785 790 795 800
Leu Leu Tyr Tyr Gln Arg Ser Thr Ser Trp Trp Pro Tyr Glu Leu Leu
805 810 815
Tyr Glu Ile Ser Phe Thr Phe Thr Asn Ser Gly Pro Ser Ser Val Asn
820 825 830
Met Ser Trp Ile Pro Ile Tyr Ser Phe Thr Arg Pro Gly Ser Gly Asn
835 840 845
Cys Ser Gly Glu Asn Val Cys Pro Thr Ala Cys Val Ser Gly Val Tyr
850 855 860
Leu Asp Pro Trp Pro Leu Thr Pro Tyr Ser His Gln Ser Gly Ile Asn
865 870 875 880
Arg Asn Phe Tyr Phe Thr Gly Ala Leu Leu Asn Ser Ser Thr Thr Arg
885 890 895
Val Asn Pro Thr Leu Tyr Val Ser Ala Leu Asn Asn Leu Lys Val Leu
900 905 910
Ala Pro Tyr Gly Thr Gln Gly Leu Phe Ala Ser Tyr Thr Thr Thr Thr
915 920 925
Cys Phe Gln Asp Thr Gly Asp Ala Ser Val Tyr Cys Val Tyr Ile Met
930 935 940
Glu Leu Ala Ser Asn Ile Val Gly Glu Phe Gln Ile Leu Pro Val Leu
945 950 955 960
Thr Arg Leu Thr Ile Thr
965
<210> 28
<211> 981
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 28
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Cys Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Cys Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Ile Glu Asp Lys
465 470 475 480
Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu Ile Ala
485 490 495
Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro Gly Ser Gly Tyr Ile Pro
500 505 510
Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg Lys Asp Gly Glu Trp
515 520 525
Val Leu Leu Ser Thr Phe Leu Gly Ser Gly Gly Gly Gly Gly Gly Phe
530 535 540
Leu Ala Val Ser Lys Gly Asn Cys Ser Gly Pro Thr Thr Ile Arg Gly
545 550 555 560
Gln Phe Ser Asn Met Ser Leu Ser Leu Leu Asp Leu Tyr Leu Gly Arg
565 570 575
Gly Tyr Asn Val Ser Ser Ile Val Thr Met Thr Ser Gln Gly Met Tyr
580 585 590
Gly Gly Thr Tyr Leu Val Glu Lys Pro Asn Leu Ser Ser Lys Arg Ser
595 600 605
Glu Leu Ser Gln Leu Ser Met Tyr Arg Val Phe Glu Val Gly Val Ile
610 615 620
Arg Asn Pro Gly Leu Gly Ala Pro Val Phe His Met Thr Asn Tyr Leu
625 630 635 640
Glu Gln Pro Val Ser Asn Asp Leu Ser Asn Cys Met Val Ala Leu Gly
645 650 655
Glu Leu Lys Leu Ala Ala Leu Cys His Gly Glu Asp Ser Ile Thr Ile
660 665 670
Pro Tyr Gln Gly Ser Gly Lys Gly Val Ser Phe Gln Leu Val Lys Leu
675 680 685
Gly Val Trp Lys Ser Pro Ala Asp Met Gln Ser Trp Val Pro Leu Ser
690 695 700
Thr Asp Asp Pro Val Ile Asp Arg Leu Tyr Leu Ser Ser His Arg Gly
705 710 715 720
Val Ile Ala Asp Asn Gln Ala Lys Trp Ala Val Pro Thr Thr Arg Thr
725 730 735
Asp Asp Lys Leu Arg Met Glu Thr Cys Phe Gln Gln Ala Cys Lys Gly
740 745 750
Lys Ile Gln Thr Leu Cys Glu Asn Pro Glu Trp Ala Pro Leu Lys Asp
755 760 765
Asn Arg Ile Pro Ser Tyr Gly Val Leu Ser Val Asp Leu Ser Leu Thr
770 775 780
Val Glu Leu Lys Ile Lys Ile Ala Ser Gly Phe Gly Pro Leu Ile Thr
785 790 795 800
His Gly Ser Gly Met Asp Leu Tyr Lys Ser Asn His Asn Asn Val Tyr
805 810 815
Trp Leu Thr Ile Pro Pro Met Lys Asn Leu Ala Leu Gly Val Ile Asn
820 825 830
Thr Leu Glu Trp Ile Pro Arg Phe Lys Val Ser Pro Tyr Leu Phe Thr
835 840 845
Val Pro Ile Lys Glu Ala Gly Glu Asp Cys His Ala Pro Thr Tyr Leu
850 855 860
Pro Ala Glu Val Asp Gly Asp Val Lys Leu Ser Ser Asn Leu Val Ile
865 870 875 880
Leu Pro Gly Gln Asp Leu Gln Tyr Val Leu Ala Thr Tyr Asp Thr Ser
885 890 895
Arg Val Glu His Ala Val Val Tyr Tyr Val Tyr Ser Pro Gly Arg Ser
900 905 910
Phe Ser Tyr Phe Tyr Pro Phe Arg Leu Pro Ile Lys Gly Val Pro Ile
915 920 925
Glu Leu Gln Val Glu Cys Phe Thr Trp Asp Gln Lys Leu Trp Cys Arg
930 935 940
His Phe Cys Val Leu Ala Asp Ser Glu Ser Gly Gly His Ile Thr His
945 950 955 960
Ser Gly Met Val Gly Met Gly Val Ser Cys Thr Val Thr Arg Glu Asp
965 970 975
Gly Thr Asn Arg Arg
980
<210> 29
<211> 1006
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 29
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Cys Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Cys Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Ile Glu Asp Lys
465 470 475 480
Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu Ile Ala
485 490 495
Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro Gly Ser Gly Tyr Ile Pro
500 505 510
Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg Lys Asp Gly Glu Trp
515 520 525
Val Leu Leu Ser Thr Phe Leu Gly Ser Gly Gly Gly Gly Gly Gly Ala
530 535 540
Asp Val Ala Ala Glu Glu Leu Met Asn Ala Leu Val Asn Ser Thr Leu
545 550 555 560
Leu Glu Ala Arg Ala Thr Asn Gln Phe Leu Ala Val Ser Lys Gly Asn
565 570 575
Cys Ser Gly Pro Thr Thr Ile Arg Gly Gln Phe Ser Asn Met Ser Leu
580 585 590
Ser Leu Leu Asp Leu Tyr Leu Ser Arg Gly Tyr Asn Val Ser Ser Ile
595 600 605
Val Thr Met Thr Ser Gln Gly Met Tyr Gly Gly Thr Tyr Leu Val Gly
610 615 620
Lys Pro Asn Leu Ser Ser Lys Gly Ser Glu Leu Ser Gln Leu Ser Met
625 630 635 640
His Arg Val Phe Glu Val Gly Val Ile Arg Asn Pro Gly Leu Gly Ala
645 650 655
Pro Val Phe His Met Thr Asn Tyr Phe Glu Gln Pro Val Ser Asn Asp
660 665 670
Phe Ser Asn Cys Met Val Ala Leu Gly Glu Leu Lys Phe Ala Ala Leu
675 680 685
Cys His Arg Glu Asp Ser Ile Thr Ile Pro Tyr Gln Gly Ser Gly Lys
690 695 700
Gly Val Ser Phe Gln Leu Val Lys Leu Gly Val Trp Lys Ser Pro Thr
705 710 715 720
Asp Met Arg Ser Trp Val Pro Leu Ser Thr Asp Asp Pro Val Ile Asp
725 730 735
Arg Leu Tyr Leu Ser Ser His Arg Gly Val Ile Ala Asp Asn Gln Ala
740 745 750
Lys Trp Ala Val Pro Thr Thr Arg Thr Asp Asp Lys Leu Arg Met Glu
755 760 765
Thr Cys Phe Gln Gln Ala Cys Lys Gly Lys Asn Gln Ala Leu Cys Glu
770 775 780
Asn Pro Glu Trp Ala Pro Leu Lys Asp Asn Arg Ile Pro Ser Tyr Gly
785 790 795 800
Val Leu Ser Val Asn Leu Ser Leu Thr Val Glu Leu Lys Ile Lys Ile
805 810 815
Ala Ser Gly Phe Gly Pro Leu Ile Thr His Gly Ser Gly Met Asp Leu
820 825 830
Tyr Lys Thr Asn His Asn Asn Val Tyr Trp Leu Thr Ile Pro Pro Met
835 840 845
Lys Asn Leu Ala Leu Gly Val Ile Asn Thr Leu Glu Trp Ile Pro Arg
850 855 860
Phe Lys Val Ser Pro Asn Leu Phe Thr Val Pro Ile Lys Glu Ala Gly
865 870 875 880
Glu Asp Cys His Ala Pro Thr Tyr Leu Pro Ala Glu Val Asp Gly Asp
885 890 895
Val Lys Leu Ser Ser Asn Leu Val Ile Leu Pro Gly Gln Asp Leu Gln
900 905 910
Tyr Val Leu Ala Thr Tyr Asp Thr Ser Arg Val Glu His Ala Val Val
915 920 925
Tyr Tyr Val Tyr Ser Pro Ser Arg Ser Phe Ser Tyr Phe Tyr Pro Phe
930 935 940
Arg Leu Pro Ile Lys Gly Val Pro Ile Glu Leu Gln Val Glu Cys Phe
945 950 955 960
Thr Trp Asp Lys Lys Leu Trp Cys Arg His Phe Cys Val Leu Ala Asp
965 970 975
Ser Glu Ser Gly Gly His Ile Thr His Ser Gly Met Val Gly Met Gly
980 985 990
Val Ser Cys Thr Val Thr Arg Glu Asp Gly Thr Asn Arg Arg
995 1000 1005
<210> 30
<211> 452
<212> PRT
<213> 腮腺炎病毒
<400> 30
Asn Ile Pro Leu Val Asn Asp Leu Arg Phe Ile Asn Gly Ile Asn Lys
1 5 10 15
Phe Ile Ile Glu Asp Tyr Ala Thr His Asp Phe Ser Ile Gly His Pro
20 25 30
Leu Asn Met Pro Ser Phe Ile Pro Thr Ala Thr Ser Pro Asn Gly Cys
35 40 45
Thr Arg Ile Pro Ser Phe Ser Leu Gly Lys Thr His Trp Cys Tyr Thr
50 55 60
His Asn Val Ile Asn Ala Asn Cys Lys Asp His Thr Ser Ser Asn Gln
65 70 75 80
Tyr Val Ser Met Gly Ile Leu Val Gln Thr Ala Ser Gly Tyr Pro Met
85 90 95
Phe Lys Thr Leu Lys Ile Gln Tyr Leu Ser Asp Gly Leu Asn Arg Lys
100 105 110
Ser Cys Ser Ile Ala Thr Val Pro Asp Gly Cys Ala Met Tyr Cys Tyr
115 120 125
Val Ser Thr Gln Leu Glu Thr Asp Asp Tyr Ala Gly Ser Ser Pro Pro
130 135 140
Thr Gln Lys Leu Thr Leu Leu Phe Tyr Asn Asp Thr Val Thr Glu Arg
145 150 155 160
Thr Ile Ser Pro Ser Gly Leu Glu Gly Asn Trp Ala Thr Leu Val Pro
165 170 175
Gly Val Gly Ser Gly Ile Tyr Phe Glu Asn Lys Leu Ile Phe Pro Ala
180 185 190
Tyr Gly Gly Val Leu Pro Asn Ser Thr Leu Gly Val Lys Ser Ala Arg
195 200 205
Glu Phe Phe Arg Pro Val Asn Pro Tyr Asn Pro Cys Ser Gly Pro Gln
210 215 220
Gln Asp Leu Asp Gln Arg Ala Leu Arg Ser Tyr Phe Pro Ser Tyr Phe
225 230 235 240
Ser Asn Arg Arg Ile Gln Ser Ala Phe Leu Val Cys Ala Trp Asn Gln
245 250 255
Ile Leu Val Thr Asn Cys Glu Leu Val Val Pro Ser Ser Asn Gln Thr
260 265 270
Met Met Gly Ala Glu Gly Arg Val Leu Leu Ile Asn Asn Arg Leu Leu
275 280 285
Tyr Tyr Gln Arg Ser Thr Ser Trp Trp Pro Tyr Glu Leu Leu Tyr Glu
290 295 300
Ile Ser Phe Thr Phe Thr Asn Ser Gly Pro Ser Ser Val Asn Met Ser
305 310 315 320
Trp Ile Pro Ile Tyr Ser Phe Thr Arg Pro Gly Ser Gly Asn Cys Ser
325 330 335
Gly Glu Asn Val Cys Pro Thr Ala Cys Val Ser Gly Val Tyr Leu Asp
340 345 350
Pro Trp Pro Leu Thr Pro Tyr Ser His Gln Ser Gly Ile Asn Arg Asn
355 360 365
Phe Tyr Phe Thr Gly Ala Leu Leu Asn Ser Ser Thr Thr Arg Val Asn
370 375 380
Pro Thr Leu Tyr Val Ser Ala Leu Asn Asn Leu Lys Val Leu Ala Pro
385 390 395 400
Tyr Gly Thr Gln Gly Leu Phe Ala Ser Tyr Thr Thr Thr Thr Cys Phe
405 410 415
Gln Asp Thr Gly Asp Ala Ser Val Tyr Cys Val Tyr Ile Met Glu Leu
420 425 430
Ala Ser Asn Ile Val Gly Glu Phe Gln Ile Leu Pro Val Leu Thr Arg
435 440 445
Leu Thr Ile Thr
450
<210> 31
<211> 438
<212> PRT
<213> 麻疹病毒
<400> 31
Phe Leu Ala Val Ser Lys Gly Asn Cys Ser Gly Pro Thr Thr Ile Arg
1 5 10 15
Gly Gln Phe Ser Asn Met Ser Leu Ser Leu Leu Asp Leu Tyr Leu Gly
20 25 30
Arg Gly Tyr Asn Val Ser Ser Ile Val Thr Met Thr Ser Gln Gly Met
35 40 45
Tyr Gly Gly Thr Tyr Leu Val Glu Lys Pro Asn Leu Ser Ser Lys Arg
50 55 60
Ser Glu Leu Ser Gln Leu Ser Met Tyr Arg Val Phe Glu Val Gly Val
65 70 75 80
Ile Arg Asn Pro Gly Leu Gly Ala Pro Val Phe His Met Thr Asn Tyr
85 90 95
Leu Glu Gln Pro Val Ser Asn Asp Leu Ser Asn Cys Met Val Ala Leu
100 105 110
Gly Glu Leu Lys Leu Ala Ala Leu Cys His Gly Glu Asp Ser Ile Thr
115 120 125
Ile Pro Tyr Gln Gly Ser Gly Lys Gly Val Ser Phe Gln Leu Val Lys
130 135 140
Leu Gly Val Trp Lys Ser Pro Ala Asp Met Gln Ser Trp Val Pro Leu
145 150 155 160
Ser Thr Asp Asp Pro Val Ile Asp Arg Leu Tyr Leu Ser Ser His Arg
165 170 175
Gly Val Ile Ala Asp Asn Gln Ala Lys Trp Ala Val Pro Thr Thr Arg
180 185 190
Thr Asp Asp Lys Leu Arg Met Glu Thr Cys Phe Gln Gln Ala Cys Lys
195 200 205
Gly Lys Ile Gln Thr Leu Cys Glu Asn Pro Glu Trp Ala Pro Leu Lys
210 215 220
Asp Asn Arg Ile Pro Ser Tyr Gly Val Leu Ser Val Asp Leu Ser Leu
225 230 235 240
Thr Val Glu Leu Lys Ile Lys Ile Ala Ser Gly Phe Gly Pro Leu Ile
245 250 255
Thr His Gly Ser Gly Met Asp Leu Tyr Lys Ser Asn His Asn Asn Val
260 265 270
Tyr Trp Leu Thr Ile Pro Pro Met Lys Asn Leu Ala Leu Gly Val Ile
275 280 285
Asn Thr Leu Glu Trp Ile Pro Arg Phe Lys Val Ser Pro Tyr Leu Phe
290 295 300
Thr Val Pro Ile Lys Glu Ala Gly Glu Asp Cys His Ala Pro Thr Tyr
305 310 315 320
Leu Pro Ala Glu Val Asp Gly Asp Val Lys Leu Ser Ser Asn Leu Val
325 330 335
Ile Leu Pro Gly Gln Asp Leu Gln Tyr Val Leu Ala Thr Tyr Asp Thr
340 345 350
Ser Arg Val Glu His Ala Val Val Tyr Tyr Val Tyr Ser Pro Gly Arg
355 360 365
Ser Phe Ser Tyr Phe Tyr Pro Phe Arg Leu Pro Ile Lys Gly Val Pro
370 375 380
Ile Glu Leu Gln Val Glu Cys Phe Thr Trp Asp Gln Lys Leu Trp Cys
385 390 395 400
Arg His Phe Cys Val Leu Ala Asp Ser Glu Ser Gly Gly His Ile Thr
405 410 415
His Ser Gly Met Val Gly Met Gly Val Ser Cys Thr Val Thr Arg Glu
420 425 430
Asp Gly Thr Asn Arg Arg
435
<210> 32
<211> 463
<212> PRT
<213> 麻疹病毒
<400> 32
Ala Asp Val Ala Ala Glu Glu Leu Met Asn Ala Leu Val Asn Ser Thr
1 5 10 15
Leu Leu Glu Ala Arg Ala Thr Asn Gln Phe Leu Ala Val Ser Lys Gly
20 25 30
Asn Cys Ser Gly Pro Thr Thr Ile Arg Gly Gln Phe Ser Asn Met Ser
35 40 45
Leu Ser Leu Leu Asp Leu Tyr Leu Ser Arg Gly Tyr Asn Val Ser Ser
50 55 60
Ile Val Thr Met Thr Ser Gln Gly Met Tyr Gly Gly Thr Tyr Leu Val
65 70 75 80
Gly Lys Pro Asn Leu Ser Ser Lys Gly Ser Glu Leu Ser Gln Leu Ser
85 90 95
Met His Arg Val Phe Glu Val Gly Val Ile Arg Asn Pro Gly Leu Gly
100 105 110
Ala Pro Val Phe His Met Thr Asn Tyr Phe Glu Gln Pro Val Ser Asn
115 120 125
Asp Phe Ser Asn Cys Met Val Ala Leu Gly Glu Leu Lys Phe Ala Ala
130 135 140
Leu Cys His Arg Glu Asp Ser Ile Thr Ile Pro Tyr Gln Gly Ser Gly
145 150 155 160
Lys Gly Val Ser Phe Gln Leu Val Lys Leu Gly Val Trp Lys Ser Pro
165 170 175
Thr Asp Met Arg Ser Trp Val Pro Leu Ser Thr Asp Asp Pro Val Ile
180 185 190
Asp Arg Leu Tyr Leu Ser Ser His Arg Gly Val Ile Ala Asp Asn Gln
195 200 205
Ala Lys Trp Ala Val Pro Thr Thr Arg Thr Asp Asp Lys Leu Arg Met
210 215 220
Glu Thr Cys Phe Gln Gln Ala Cys Lys Gly Lys Asn Gln Ala Leu Cys
225 230 235 240
Glu Asn Pro Glu Trp Ala Pro Leu Lys Asp Asn Arg Ile Pro Ser Tyr
245 250 255
Gly Val Leu Ser Val Asn Leu Ser Leu Thr Val Glu Leu Lys Ile Lys
260 265 270
Ile Ala Ser Gly Phe Gly Pro Leu Ile Thr His Gly Ser Gly Met Asp
275 280 285
Leu Tyr Lys Thr Asn His Asn Asn Val Tyr Trp Leu Thr Ile Pro Pro
290 295 300
Met Lys Asn Leu Ala Leu Gly Val Ile Asn Thr Leu Glu Trp Ile Pro
305 310 315 320
Arg Phe Lys Val Ser Pro Asn Leu Phe Thr Val Pro Ile Lys Glu Ala
325 330 335
Gly Glu Asp Cys His Ala Pro Thr Tyr Leu Pro Ala Glu Val Asp Gly
340 345 350
Asp Val Lys Leu Ser Ser Asn Leu Val Ile Leu Pro Gly Gln Asp Leu
355 360 365
Gln Tyr Val Leu Ala Thr Tyr Asp Thr Ser Arg Val Glu His Ala Val
370 375 380
Val Tyr Tyr Val Tyr Ser Pro Ser Arg Ser Phe Ser Tyr Phe Tyr Pro
385 390 395 400
Phe Arg Leu Pro Ile Lys Gly Val Pro Ile Glu Leu Gln Val Glu Cys
405 410 415
Phe Thr Trp Asp Lys Lys Leu Trp Cys Arg His Phe Cys Val Leu Ala
420 425 430
Asp Ser Glu Ser Gly Gly His Ile Thr His Ser Gly Met Val Gly Met
435 440 445
Gly Val Ser Cys Thr Val Thr Arg Glu Asp Gly Thr Asn Arg Arg
450 455 460
<210> 33
<211> 30
<212> PRT
<213> 人工序列
<220>
<223> GCN4三聚化结构域
<400> 33
Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu
1 5 10 15
Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro
20 25 30
<210> 34
<211> 27
<212> PRT
<213> 人工序列
<220>
<223> T4纤维蛋白三聚化结构域
<400> 34
Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg Lys
1 5 10 15
Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu
20 25
<210> 35
<211> 59
<212> PRT
<213> 人工序列
<220>
<223> GCN4/T4纤维蛋白三聚化结构域
<400> 35
Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu
1 5 10 15
Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro Gly Ser
20 25 30
Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg Lys
35 40 45
Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu
50 55
<210> 36
<211> 550
<212> PRT
<213> 麻疹病毒
<400> 36
Met Gly Leu Lys Val Asn Val Ser Ala Ile Phe Met Ala Val Leu Leu
1 5 10 15
Thr Leu Gln Thr Pro Thr Gly Gln Ile His Trp Gly Asn Leu Ser Lys
20 25 30
Ile Gly Val Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg
35 40 45
Ser Ser His Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu
50 55 60
Leu Asn Asn Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu
65 70 75 80
Arg Thr Val Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Met Thr Gln
85 90 95
Asn Ile Arg Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Lys Arg
100 105 110
Phe Ala Gly Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala
115 120 125
Ala Gln Ile Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser
130 135 140
Gln Ala Ile Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala
145 150 155 160
Ile Glu Ala Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Val Gln
165 170 175
Gly Val Gln Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln
180 185 190
Leu Ser Cys Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg
195 200 205
Tyr Tyr Thr Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro
210 215 220
Ile Ser Ala Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly
225 230 235 240
Asp Ile Asn Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu
245 250 255
Leu Gly Ile Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val
260 265 270
Asp Thr Glu Ser Tyr Leu Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu
275 280 285
Ser Glu Ile Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr
290 295 300
Asn Ile Gly Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala
305 310 315 320
Thr Gln Gly Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe
325 330 335
Met Pro Glu Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser
340 345 350
Pro Leu Leu Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg
355 360 365
Thr Leu Val Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly
370 375 380
Asn Leu Ile Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr
385 390 395 400
Gly Thr Ile Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala
405 410 415
Ala Asp His Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val
420 425 430
Gly Ser Arg Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu
435 440 445
Gly Pro Pro Ile Leu Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly
450 455 460
Asn Ala Ile Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser
465 470 475 480
Asp Gln Ile Leu Arg Ser Met Lys Gly Leu Ser Ser Thr Cys Ile Val
485 490 495
Tyr Ile Leu Ile Ala Val Cys Leu Gly Gly Leu Ile Gly Ile Pro Ala
500 505 510
Leu Ile Cys Cys Cys Arg Gly Arg Cys Asn Lys Lys Gly Glu Gln Val
515 520 525
Gly Met Ser Arg Pro Gly Leu Lys Pro Asp Leu Thr Gly Thr Ser Lys
530 535 540
Ser Tyr Val Arg Ser Leu
545 550
<210> 37
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 37
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Cys Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Cys Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 38
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 38
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Cys Gln Ala Gly Gln Glu Cys Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 39
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 39
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Cys Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Cys Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 40
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 40
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Cys Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Cys Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 41
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 41
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Cys Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Cys Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 42
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 42
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Cys Ser Leu Arg Asp Cys Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 43
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 43
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Pro Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 44
<211> 24
<212> PRT
<213> 人工序列
<220>
<223> 跨膜结构域
<400> 44
Gly Ala Ile Ile Val Ala Ala Leu Val Leu Ser Ile Leu Ser Ile Ile
1 5 10 15
Ile Ser Leu Leu Phe Cys Cys Trp
20
<210> 45
<211> 163
<212> PRT
<213> 人工序列
<220>
<223> 铁蛋白亚基
<400> 45
Asp Ile Ile Lys Leu Leu Asn Glu Gln Val Asn Lys Glu Met Gln Ser
1 5 10 15
Ser Asn Leu Tyr Met Ser Met Ser Ser Trp Cys Tyr Thr His Ser Leu
20 25 30
Asp Gly Ala Gly Leu Phe Leu Phe Asp His Ala Ala Glu Glu Tyr Glu
35 40 45
His Ala Lys Lys Leu Ile Ile Phe Leu Asn Glu Asn Asn Val Pro Val
50 55 60
Gln Leu Thr Ser Ile Ser Ala Pro Glu His Lys Phe Glu Gly Leu Thr
65 70 75 80
Gln Ile Phe Gln Lys Ala Tyr Glu His Glu Gln His Ile Ser Glu Ser
85 90 95
Ile Asn Asn Ile Val Asp His Ala Ile Lys Ser Lys Asp His Ala Thr
100 105 110
Phe Asn Phe Leu Gln Trp Tyr Val Ala Glu Gln His Glu Glu Glu Val
115 120 125
Leu Phe Lys Asp Ile Leu Asp Lys Ile Glu Leu Ile Gly Asn Glu Asn
130 135 140
His Gly Leu Tyr Leu Ala Asp Gln Tyr Val Lys Gly Ile Ala Lys Ser
145 150 155 160
Arg Lys Ser
<210> 46
<211> 154
<212> PRT
<213> 人工序列
<220>
<223> lumazine合酶亚基
<400> 46
Met Gln Ile Tyr Glu Gly Lys Leu Thr Ala Glu Gly Leu Arg Phe Gly
1 5 10 15
Ile Val Ala Ser Arg Phe Asn His Ala Leu Val Asp Arg Leu Val Glu
20 25 30
Gly Ala Ile Asp Cys Ile Val Arg His Gly Gly Arg Glu Glu Asp Ile
35 40 45
Thr Leu Val Arg Val Pro Gly Ser Trp Glu Ile Pro Val Ala Ala Gly
50 55 60
Glu Leu Ala Arg Lys Glu Asp Ile Asp Ala Val Ile Ala Ile Gly Val
65 70 75 80
Leu Ile Arg Gly Ala Thr Pro His Phe Asp Tyr Ile Ala Ser Glu Val
85 90 95
Ser Lys Gly Leu Ala Asp Leu Ser Leu Glu Leu Arg Lys Pro Ile Thr
100 105 110
Phe Gly Val Ile Thr Ala Asp Thr Leu Glu Gln Ala Ile Glu Arg Ala
115 120 125
Gly Thr Lys His Gly Asn Lys Gly Trp Glu Ala Ala Leu Ser Ala Ile
130 135 140
Glu Met Ala Asn Leu Phe Lys Ser Leu Arg
145 150
<210> 47
<211> 265
<212> PRT
<213> 人工序列
<220>
<223> encapsulin亚基
<400> 47
Met Glu Phe Leu Lys Arg Ser Phe Ala Pro Leu Thr Glu Lys Gln Trp
1 5 10 15
Gln Glu Ile Asp Asn Arg Ala Arg Glu Ile Phe Lys Thr Gln Leu Tyr
20 25 30
Gly Arg Lys Phe Val Asp Val Glu Gly Pro Tyr Gly Trp Glu Tyr Ala
35 40 45
Ala His Pro Leu Gly Glu Val Glu Val Leu Ser Asp Glu Asn Glu Val
50 55 60
Val Lys Trp Gly Leu Arg Lys Ser Leu Pro Leu Ile Glu Leu Arg Ala
65 70 75 80
Thr Phe Thr Leu Asp Leu Trp Glu Leu Asp Asn Leu Glu Arg Gly Lys
85 90 95
Pro Asn Val Asp Leu Ser Ser Leu Glu Glu Thr Val Arg Lys Val Ala
100 105 110
Glu Phe Glu Asp Glu Val Ile Phe Arg Gly Cys Glu Lys Ser Gly Val
115 120 125
Lys Gly Leu Leu Ser Phe Glu Glu Arg Lys Ile Glu Cys Gly Ser Thr
130 135 140
Pro Lys Asp Leu Leu Glu Ala Ile Val Arg Ala Leu Ser Ile Phe Ser
145 150 155 160
Lys Asp Gly Ile Glu Gly Pro Tyr Thr Leu Val Ile Asn Thr Asp Arg
165 170 175
Trp Ile Asn Phe Leu Lys Glu Glu Ala Gly His Tyr Pro Leu Glu Lys
180 185 190
Arg Val Glu Glu Cys Leu Arg Gly Gly Lys Ile Ile Thr Thr Pro Arg
195 200 205
Ile Glu Asp Ala Leu Val Val Ser Glu Arg Gly Gly Asp Phe Lys Leu
210 215 220
Ile Leu Gly Gln Asp Leu Ser Ile Gly Tyr Glu Asp Arg Glu Lys Asp
225 230 235 240
Ala Val Arg Leu Phe Ile Thr Glu Thr Phe Thr Phe Gln Val Val Asn
245 250 255
Pro Glu Ala Leu Ile Leu Leu Lys Phe
260 265
<210> 48
<211> 265
<212> PRT
<213> 人工序列
<220>
<223> 硫加氧酶还原酶（SOR）亚基
<400> 48
Met Glu Phe Leu Lys Arg Ser Phe Ala Pro Leu Thr Glu Lys Gln Trp
1 5 10 15
Gln Glu Ile Asp Asn Arg Ala Arg Glu Ile Phe Lys Thr Gln Leu Tyr
20 25 30
Gly Arg Lys Phe Val Asp Val Glu Gly Pro Tyr Gly Trp Glu Tyr Ala
35 40 45
Ala His Pro Leu Gly Glu Val Glu Val Leu Ser Asp Glu Asn Glu Val
50 55 60
Val Lys Trp Gly Leu Arg Lys Ser Leu Pro Leu Ile Glu Leu Arg Ala
65 70 75 80
Thr Phe Thr Leu Asp Leu Trp Glu Leu Asp Asn Leu Glu Arg Gly Lys
85 90 95
Pro Asn Val Asp Leu Ser Ser Leu Glu Glu Thr Val Arg Lys Val Ala
100 105 110
Glu Phe Glu Asp Glu Val Ile Phe Arg Gly Cys Glu Lys Ser Gly Val
115 120 125
Lys Gly Leu Leu Ser Phe Glu Glu Arg Lys Ile Glu Cys Gly Ser Thr
130 135 140
Pro Lys Asp Leu Leu Glu Ala Ile Val Arg Ala Leu Ser Ile Phe Ser
145 150 155 160
Lys Asp Gly Ile Glu Gly Pro Tyr Thr Leu Val Ile Asn Thr Asp Arg
165 170 175
Trp Ile Asn Phe Leu Lys Glu Glu Ala Gly His Tyr Pro Leu Glu Lys
180 185 190
Arg Val Glu Glu Cys Leu Arg Gly Gly Lys Ile Ile Thr Thr Pro Arg
195 200 205
Ile Glu Asp Ala Leu Val Val Ser Glu Arg Gly Gly Asp Phe Lys Leu
210 215 220
Ile Leu Gly Gln Asp Leu Ser Ile Gly Tyr Glu Asp Arg Glu Lys Asp
225 230 235 240
Ala Val Arg Leu Phe Ile Thr Glu Thr Phe Thr Phe Gln Val Val Asn
245 250 255
Pro Glu Ala Leu Ile Leu Leu Lys Phe
260 265
<210> 49
<211> 617
<212> PRT
<213> 麻疹病毒
<400> 49
Met Ser Pro Gln Arg Asp Arg Ile Asn Ala Phe Tyr Lys Asp Asn Pro
1 5 10 15
His Pro Lys Gly Ser Arg Ile Val Ile Asn Arg Glu His Leu Met Ile
20 25 30
Asp Arg Pro Tyr Val Leu Leu Ala Val Leu Phe Val Met Phe Leu Ser
35 40 45
Leu Ile Gly Leu Leu Ala Ile Ala Gly Ile Arg Leu His Arg Ala Ala
50 55 60
Ile Tyr Thr Ala Glu Ile His Lys Ser Leu Ser Thr Asn Leu Asp Val
65 70 75 80
Thr Asn Ser Ile Glu His Gln Val Lys Asp Val Leu Thr Pro Leu Phe
85 90 95
Lys Ile Ile Gly Asp Glu Val Gly Leu Arg Thr Pro Gln Arg Phe Thr
100 105 110
Asp Leu Val Lys Phe Ile Ser Asp Lys Ile Lys Phe Leu Asn Pro Asp
115 120 125
Arg Glu Tyr Asp Phe Arg Asp Leu Thr Trp Cys Ile Asn Pro Pro Glu
130 135 140
Arg Ile Lys Leu Asp Tyr Asp Gln Tyr Cys Ala Asp Val Ala Ala Glu
145 150 155 160
Glu Leu Met Asn Ala Leu Val Asn Ser Thr Leu Leu Glu Thr Arg Thr
165 170 175
Thr Asn Gln Phe Leu Ala Val Ser Lys Gly Asn Cys Ser Gly Pro Thr
180 185 190
Thr Ile Arg Gly Gln Phe Ser Asn Met Ser Leu Ser Leu Leu Asp Leu
195 200 205
Tyr Leu Gly Arg Gly Tyr Asn Val Ser Ser Ile Val Thr Met Thr Ser
210 215 220
Gln Gly Met Tyr Gly Gly Thr Tyr Leu Val Glu Lys Pro Asn Leu Ser
225 230 235 240
Ser Lys Arg Ser Glu Leu Ser Gln Leu Ser Met Tyr Arg Val Phe Glu
245 250 255
Val Gly Val Ile Arg Asn Pro Gly Leu Gly Ala Pro Val Phe His Met
260 265 270
Thr Asn Tyr Leu Glu Gln Pro Val Ser Asn Asp Leu Ser Asn Cys Met
275 280 285
Val Ala Leu Gly Glu Leu Lys Leu Ala Ala Leu Cys His Gly Glu Asp
290 295 300
Ser Ile Thr Ile Pro Tyr Gln Gly Ser Gly Lys Gly Val Ser Phe Gln
305 310 315 320
Leu Val Lys Leu Gly Val Trp Lys Ser Pro Thr Asp Met Gln Ser Trp
325 330 335
Val Pro Leu Ser Thr Asp Asp Pro Val Ile Asp Arg Leu Tyr Leu Ser
340 345 350
Ser His Arg Gly Val Ile Ala Asp Asn Gln Ala Lys Trp Ala Val Pro
355 360 365
Thr Thr Arg Thr Asp Asp Lys Leu Arg Met Glu Thr Cys Phe Gln Gln
370 375 380
Ala Cys Lys Gly Lys Ile Gln Ala Leu Cys Glu Asn Pro Glu Trp Ala
385 390 395 400
Pro Leu Lys Asp Asn Arg Ile Pro Ser Tyr Gly Val Leu Ser Val Asp
405 410 415
Leu Ser Leu Thr Val Glu Leu Lys Ile Lys Ile Ala Ser Gly Phe Gly
420 425 430
Pro Leu Ile Thr His Gly Ser Gly Met Asp Leu Tyr Lys Ser Asn His
435 440 445
Asn Asn Val Tyr Trp Leu Thr Ile Pro Pro Met Lys Asn Leu Ala Leu
450 455 460
Gly Val Ile Asn Thr Leu Glu Trp Ile Pro Arg Phe Lys Val Ser Pro
465 470 475 480
Tyr Leu Phe Thr Val Pro Ile Lys Glu Ala Gly Glu Asp Cys His Ala
485 490 495
Pro Thr Tyr Leu Pro Ala Glu Val Asp Gly Asp Val Lys Leu Ser Ser
500 505 510
Asn Leu Val Ile Leu Pro Gly Gln Asp Leu Gln Tyr Val Leu Ala Thr
515 520 525
Tyr Asp Thr Ser Arg Val Glu His Ala Val Val Tyr Tyr Val Tyr Ser
530 535 540
Pro Ser Arg Ser Phe Ser Tyr Phe Tyr Pro Phe Arg Leu Pro Ile Lys
545 550 555 560
Gly Val Pro Ile Glu Leu Gln Val Glu Cys Phe Thr Trp Asp Gln Lys
565 570 575
Leu Trp Cys Arg His Phe Cys Val Leu Ala Asp Ser Glu Ser Gly Gly
580 585 590
His Ile Thr His Ser Gly Met Val Gly Met Gly Val Ser Cys Thr Val
595 600 605
Thr Arg Glu Asp Gly Thr Asn Arg Arg
610 615
<210> 50
<211> 582
<212> PRT
<213> 腮腺炎病毒
<400> 50
Met Glu Pro Ser Lys Phe Phe Thr Ile Ser Asp Ser Ala Thr Phe Ala
1 5 10 15
Pro Gly Pro Val Ser Asn Ala Ala Asp Lys Lys Thr Phe Arg Thr Cys
20 25 30
Phe Arg Ile Leu Val Leu Ser Val Gln Ala Val Thr Leu Ile Leu Val
35 40 45
Ile Val Thr Leu Gly Glu Leu Val Arg Met Ile Asn Asp Gln Gly Leu
50 55 60
Ser Asn Gln Leu Ser Ser Ile Thr Asp Lys Ile Arg Glu Ser Ala Thr
65 70 75 80
Met Ile Ala Ser Ala Val Gly Val Met Asn Gln Val Ile His Gly Val
85 90 95
Thr Val Ser Leu Pro Leu Gln Ile Glu Gly Asn Gln Asn Gln Leu Leu
100 105 110
Ala Thr Leu Ala Thr Ile Cys Ala Ser Gln Lys Gln Val Ser Asn Cys
115 120 125
Ser Thr Asn Ile Pro Leu Val Asn Asp Leu Arg Phe Ile Asn Gly Ile
130 135 140
Asn Lys Phe Ile Ile Glu Asp Tyr Ala Thr His Asp Phe Ser Ile Gly
145 150 155 160
His Pro Leu Asn Met Pro Ser Phe Ile Pro Thr Ala Thr Ser Pro Asn
165 170 175
Gly Cys Thr Arg Ile Pro Ser Phe Ser Leu Gly Lys Thr His Trp Cys
180 185 190
Tyr Thr His Asn Val Ile Asn Ala Asn Cys Lys Asp His Thr Ser Ser
195 200 205
Asn Gln Tyr Val Ser Met Gly Ile Leu Val Gln Thr Ala Ser Gly Tyr
210 215 220
Pro Met Phe Lys Thr Leu Lys Ile Gln Tyr Leu Ser Asp Gly Leu Asn
225 230 235 240
Arg Lys Ser Cys Ser Ile Ala Thr Val Pro Asp Gly Cys Ala Met Tyr
245 250 255
Cys Tyr Val Ser Thr Gln Leu Glu Thr Asp Asp Tyr Ala Gly Ser Ser
260 265 270
Pro Pro Thr Gln Lys Leu Thr Leu Leu Phe Tyr Asn Asp Thr Val Thr
275 280 285
Glu Arg Thr Ile Ser Pro Ser Gly Leu Glu Gly Asn Trp Ala Thr Leu
290 295 300
Val Pro Gly Val Gly Ser Gly Ile Tyr Phe Glu Asn Lys Leu Ile Phe
305 310 315 320
Pro Ala Tyr Gly Gly Val Leu Pro Asn Ser Thr Leu Gly Val Lys Ser
325 330 335
Ala Arg Glu Phe Phe Arg Pro Val Asn Pro Tyr Asn Pro Cys Ser Gly
340 345 350
Pro Gln Gln Asp Leu Asp Gln Arg Ala Leu Arg Ser Tyr Phe Pro Ser
355 360 365
Tyr Phe Ser Asn Arg Arg Ile Gln Ser Ala Phe Leu Val Cys Ala Trp
370 375 380
Asn Gln Ile Leu Val Thr Asn Cys Glu Leu Val Val Pro Ser Ser Asn
385 390 395 400
Gln Thr Met Met Gly Ala Glu Gly Arg Val Leu Leu Ile Asn Asn Arg
405 410 415
Leu Leu Tyr Tyr Gln Arg Ser Thr Ser Trp Trp Pro Tyr Glu Leu Leu
420 425 430
Tyr Glu Ile Ser Phe Thr Phe Thr Asn Ser Gly Pro Ser Ser Val Asn
435 440 445
Met Ser Trp Ile Pro Ile Tyr Ser Phe Thr Arg Pro Gly Ser Gly Asn
450 455 460
Cys Ser Gly Glu Asn Val Cys Pro Thr Ala Cys Val Ser Gly Val Tyr
465 470 475 480
Leu Asp Pro Trp Pro Leu Thr Pro Tyr Ser His Gln Ser Gly Ile Asn
485 490 495
Arg Asn Phe Tyr Phe Thr Gly Ala Leu Leu Asn Ser Ser Thr Thr Arg
500 505 510
Val Asn Pro Thr Leu Tyr Val Ser Ala Leu Asn Asn Leu Lys Val Leu
515 520 525
Ala Pro Tyr Gly Thr Gln Gly Leu Phe Ala Ser Tyr Thr Thr Thr Thr
530 535 540
Cys Phe Gln Asp Thr Gly Asp Ala Ser Val Tyr Cys Val Tyr Ile Met
545 550 555 560
Glu Leu Ala Ser Asn Ile Val Gly Glu Phe Gln Ile Leu Pro Val Leu
565 570 575
Thr Arg Leu Thr Ile Thr
580
<210> 51
<211> 506
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 51
Met Lys Val Ser Leu Val Thr Cys Leu Gly Phe Ala Val Phe Ser Phe
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asn Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asn His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Cys Phe Gln
195 200 205
Pro Gln Leu Ile Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Cys Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Val Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Gly Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His Asn Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ile Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Ile Glu Asp Lys
465 470 475 480
Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu Ile Ala
485 490 495
Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro
500 505
<210> 52
<211> 516
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 52
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Gly Gly Gly Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Val Phe Glu Val Cys Glu Gly Thr Gln Gln
145 150 155 160
Cys Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Ser Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Thr Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Ile Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ala Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser Val Ser Val Asn
465 470 475 480
Ser Lys Ile Gly Ala Ile Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser
485 490 495
Lys Ile Tyr His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile
500 505 510
Gly Glu Ala Pro
515
<210> 53
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 53
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Cys Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Cys Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 54
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 54
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Cys Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Cys His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 55
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 55
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Cys Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Cys Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 56
<211> 959
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 56
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Cys Gln Ala Gly Gln Glu Cys Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro Gly Ser Gly Gly Gly Gly Gly Gly Phe Leu Ala Val Ser Lys Gly
515 520 525
Asn Cys Ser Gly Pro Thr Thr Ile Arg Gly Gln Phe Ser Asn Met Ser
530 535 540
Leu Ser Leu Leu Asp Leu Tyr Leu Gly Arg Gly Tyr Asn Val Ser Ser
545 550 555 560
Ile Val Thr Met Thr Ser Gln Gly Met Tyr Gly Gly Thr Tyr Leu Val
565 570 575
Glu Lys Pro Asn Leu Ser Ser Lys Arg Ser Glu Leu Ser Gln Leu Ser
580 585 590
Met Tyr Arg Val Phe Glu Val Gly Val Ile Arg Asn Pro Gly Leu Gly
595 600 605
Ala Pro Val Phe His Met Thr Asn Tyr Leu Glu Gln Pro Val Ser Asn
610 615 620
Asp Leu Ser Asn Cys Met Val Ala Leu Gly Glu Leu Lys Leu Ala Ala
625 630 635 640
Leu Cys His Gly Glu Asp Ser Ile Thr Ile Pro Tyr Gln Gly Ser Gly
645 650 655
Lys Gly Val Ser Phe Gln Leu Val Lys Leu Gly Val Trp Lys Ser Pro
660 665 670
Ala Asp Met Gln Ser Trp Val Pro Leu Ser Thr Asp Asp Pro Val Ile
675 680 685
Asp Arg Leu Tyr Leu Ser Ser His Arg Gly Val Ile Ala Asp Asn Gln
690 695 700
Ala Lys Trp Ala Val Pro Thr Thr Arg Thr Asp Asp Lys Leu Arg Met
705 710 715 720
Glu Thr Cys Phe Gln Gln Ala Cys Lys Gly Lys Ile Gln Thr Leu Cys
725 730 735
Glu Asn Pro Glu Trp Ala Pro Leu Lys Asp Asn Arg Ile Pro Ser Tyr
740 745 750
Gly Val Leu Ser Val Asp Leu Ser Leu Thr Val Glu Leu Lys Ile Lys
755 760 765
Ile Ala Ser Gly Phe Gly Pro Leu Ile Thr His Gly Ser Gly Met Asp
770 775 780
Leu Tyr Lys Ser Asn His Asn Asn Val Tyr Trp Leu Thr Ile Pro Pro
785 790 795 800
Met Lys Asn Leu Ala Leu Gly Val Ile Asn Thr Leu Glu Trp Ile Pro
805 810 815
Arg Phe Lys Val Ser Pro Tyr Leu Phe Thr Val Pro Ile Lys Glu Ala
820 825 830
Gly Glu Asp Cys His Ala Pro Thr Tyr Leu Pro Ala Glu Val Asp Gly
835 840 845
Asp Val Lys Leu Ser Ser Asn Leu Val Ile Leu Pro Gly Gln Asp Leu
850 855 860
Gln Tyr Val Leu Ala Thr Tyr Asp Thr Ser Arg Val Glu His Ala Val
865 870 875 880
Val Tyr Tyr Val Tyr Ser Pro Gly Arg Ser Phe Ser Tyr Phe Tyr Pro
885 890 895
Phe Arg Leu Pro Ile Lys Gly Val Pro Ile Glu Leu Gln Val Glu Cys
900 905 910
Phe Thr Trp Asp Gln Lys Leu Trp Cys Arg His Phe Cys Val Leu Ala
915 920 925
Asp Ser Glu Ser Gly Gly His Ile Thr His Ser Gly Met Val Gly Met
930 935 940
Gly Val Ser Cys Thr Val Thr Arg Glu Asp Gly Thr Asn Arg Arg
945 950 955
<210> 57
<211> 973
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 57
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Cys Gln Ala Gly Gln Glu Cys Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro Gly Ser Gly Gly Gly Gly Gly Gly Asn Ile Pro Leu Val Asn Asp
515 520 525
Leu Arg Phe Ile Asn Gly Ile Asn Lys Phe Ile Ile Glu Asp Tyr Ala
530 535 540
Thr His Asp Phe Ser Ile Gly His Pro Leu Asn Met Pro Ser Phe Ile
545 550 555 560
Pro Thr Ala Thr Ser Pro Asn Gly Cys Thr Arg Ile Pro Ser Phe Ser
565 570 575
Leu Gly Lys Thr His Trp Cys Tyr Thr His Asn Val Ile Asn Ala Asn
580 585 590
Cys Lys Asp His Thr Ser Ser Asn Gln Tyr Val Ser Met Gly Ile Leu
595 600 605
Val Gln Thr Ala Ser Gly Tyr Pro Met Phe Lys Thr Leu Lys Ile Gln
610 615 620
Tyr Leu Ser Asp Gly Leu Asn Arg Lys Ser Cys Ser Ile Ala Thr Val
625 630 635 640
Pro Asp Gly Cys Ala Met Tyr Cys Tyr Val Ser Thr Gln Leu Glu Thr
645 650 655
Asp Asp Tyr Ala Gly Ser Ser Pro Pro Thr Gln Lys Leu Thr Leu Leu
660 665 670
Phe Tyr Asn Asp Thr Val Thr Glu Arg Thr Ile Ser Pro Ser Gly Leu
675 680 685
Glu Gly Asn Trp Ala Thr Leu Val Pro Gly Val Gly Ser Gly Ile Tyr
690 695 700
Phe Glu Asn Lys Leu Ile Phe Pro Ala Tyr Gly Gly Val Leu Pro Asn
705 710 715 720
Ser Thr Leu Gly Val Lys Ser Ala Arg Glu Phe Phe Arg Pro Val Asn
725 730 735
Pro Tyr Asn Pro Cys Ser Gly Pro Gln Gln Asp Leu Asp Gln Arg Ala
740 745 750
Leu Arg Ser Tyr Phe Pro Ser Tyr Phe Ser Asn Arg Arg Ile Gln Ser
755 760 765
Ala Phe Leu Val Cys Ala Trp Asn Gln Ile Leu Val Thr Asn Cys Glu
770 775 780
Leu Val Val Pro Ser Ser Asn Gln Thr Met Met Gly Ala Glu Gly Arg
785 790 795 800
Val Leu Leu Ile Asn Asn Arg Leu Leu Tyr Tyr Gln Arg Ser Thr Ser
805 810 815
Trp Trp Pro Tyr Glu Leu Leu Tyr Glu Ile Ser Phe Thr Phe Thr Asn
820 825 830
Ser Gly Pro Ser Ser Val Asn Met Ser Trp Ile Pro Ile Tyr Ser Phe
835 840 845
Thr Arg Pro Gly Ser Gly Asn Cys Ser Gly Glu Asn Val Cys Pro Thr
850 855 860
Ala Cys Val Ser Gly Val Tyr Leu Asp Pro Trp Pro Leu Thr Pro Tyr
865 870 875 880
Ser His Gln Ser Gly Ile Asn Arg Asn Phe Tyr Phe Thr Gly Ala Leu
885 890 895
Leu Asn Ser Ser Thr Thr Arg Val Asn Pro Thr Leu Tyr Val Ser Ala
900 905 910
Leu Asn Asn Leu Lys Val Leu Ala Pro Tyr Gly Thr Gln Gly Leu Phe
915 920 925
Ala Ser Tyr Thr Thr Thr Thr Cys Phe Gln Asp Thr Gly Asp Ala Ser
930 935 940
Val Tyr Cys Val Tyr Ile Met Glu Leu Ala Ser Asn Ile Val Gly Glu
945 950 955 960
Phe Gln Ile Leu Pro Val Leu Thr Arg Leu Thr Ile Thr
965 970
<210> 58
<211> 510
<212> PRT
<213> 人工序列
<220>
<223> 重组HN蛋白
<400> 58
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Gly Ser Gly Tyr Ile Pro Glu Ala Pro Arg Asp
20 25 30
Gly Gln Ala Tyr Val Arg Lys Asp Gly Glu Trp Val Leu Leu Ser Thr
35 40 45
Phe Leu Gly Ser Gly Gly Gly Gly Gly Gly Asn Ile Pro Leu Val Asn
50 55 60
Asp Leu Arg Phe Ile Asn Gly Ile Asn Lys Phe Ile Ile Glu Asp Tyr
65 70 75 80
Ala Thr His Asp Phe Ser Ile Gly His Pro Leu Asn Met Pro Ser Phe
85 90 95
Ile Pro Thr Ala Thr Ser Pro Asn Gly Cys Thr Arg Ile Pro Ser Phe
100 105 110
Ser Leu Gly Lys Thr His Trp Cys Tyr Thr His Asn Val Ile Asn Ala
115 120 125
Asn Cys Lys Asp His Thr Ser Ser Asn Gln Tyr Val Ser Met Gly Ile
130 135 140
Leu Val Gln Thr Ala Ser Gly Tyr Pro Met Phe Lys Thr Leu Lys Ile
145 150 155 160
Gln Tyr Leu Ser Asp Gly Leu Asn Arg Lys Ser Cys Ser Ile Ala Thr
165 170 175
Val Pro Asp Gly Cys Ala Met Tyr Cys Tyr Val Ser Thr Gln Leu Glu
180 185 190
Thr Asp Asp Tyr Ala Gly Ser Ser Pro Pro Thr Gln Lys Leu Thr Leu
195 200 205
Leu Phe Tyr Asn Asp Thr Val Thr Glu Arg Thr Ile Ser Pro Ser Gly
210 215 220
Leu Glu Gly Asn Trp Ala Thr Leu Val Pro Gly Val Gly Ser Gly Ile
225 230 235 240
Tyr Phe Glu Asn Lys Leu Ile Phe Pro Ala Tyr Gly Gly Val Leu Pro
245 250 255
Asn Ser Thr Leu Gly Val Lys Ser Ala Arg Glu Phe Phe Arg Pro Val
260 265 270
Asn Pro Tyr Asn Pro Cys Ser Gly Pro Gln Gln Asp Leu Asp Gln Arg
275 280 285
Ala Leu Arg Ser Tyr Phe Pro Ser Tyr Phe Ser Asn Arg Arg Ile Gln
290 295 300
Ser Ala Phe Leu Val Cys Ala Trp Asn Gln Ile Leu Val Thr Asn Cys
305 310 315 320
Glu Leu Val Val Pro Ser Ser Asn Gln Thr Met Met Gly Ala Glu Gly
325 330 335
Arg Val Leu Leu Ile Asn Asn Arg Leu Leu Tyr Tyr Gln Arg Ser Thr
340 345 350
Ser Trp Trp Pro Tyr Glu Leu Leu Tyr Glu Ile Ser Phe Thr Phe Thr
355 360 365
Asn Ser Gly Pro Ser Ser Val Asn Met Ser Trp Ile Pro Ile Tyr Ser
370 375 380
Phe Thr Arg Pro Gly Ser Gly Asn Cys Ser Gly Glu Asn Val Cys Pro
385 390 395 400
Thr Ala Cys Val Ser Gly Val Tyr Leu Asp Pro Trp Pro Leu Thr Pro
405 410 415
Tyr Ser His Gln Ser Gly Ile Asn Arg Asn Phe Tyr Phe Thr Gly Ala
420 425 430
Leu Leu Asn Ser Ser Thr Thr Arg Val Asn Pro Thr Leu Tyr Val Ser
435 440 445
Ala Leu Asn Asn Leu Lys Val Leu Ala Pro Tyr Gly Thr Gln Gly Leu
450 455 460
Phe Ala Ser Tyr Thr Thr Thr Thr Cys Phe Gln Asp Thr Gly Asp Ala
465 470 475 480
Ser Val Tyr Cys Val Tyr Ile Met Glu Leu Ala Ser Asn Ile Val Gly
485 490 495
Glu Phe Gln Ile Leu Pro Val Leu Thr Arg Leu Thr Ile Thr
500 505 510
<210> 59
<211> 496
<212> PRT
<213> 人工序列
<220>
<223> 重组H蛋白
<400> 59
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Gly Ser Gly Tyr Ile Pro Glu Ala Pro Arg Asp
20 25 30
Gly Gln Ala Tyr Val Arg Lys Asp Gly Glu Trp Val Leu Leu Ser Thr
35 40 45
Phe Leu Gly Ser Gly Gly Gly Gly Gly Gly Phe Leu Ala Val Ser Lys
50 55 60
Gly Asn Cys Ser Gly Pro Thr Thr Ile Arg Gly Gln Phe Ser Asn Met
65 70 75 80
Ser Leu Ser Leu Leu Asp Leu Tyr Leu Gly Arg Gly Tyr Asn Val Ser
85 90 95
Ser Ile Val Thr Met Thr Ser Gln Gly Met Tyr Gly Gly Thr Tyr Leu
100 105 110
Val Glu Lys Pro Asn Leu Ser Ser Lys Arg Ser Glu Leu Ser Gln Leu
115 120 125
Ser Met Tyr Arg Val Phe Glu Val Gly Val Ile Arg Asn Pro Gly Leu
130 135 140
Gly Ala Pro Val Phe His Met Thr Asn Tyr Leu Glu Gln Pro Val Ser
145 150 155 160
Asn Asp Leu Ser Asn Cys Met Val Ala Leu Gly Glu Leu Lys Leu Ala
165 170 175
Ala Leu Cys His Gly Glu Asp Ser Ile Thr Ile Pro Tyr Gln Gly Ser
180 185 190
Gly Lys Gly Val Ser Phe Gln Leu Val Lys Leu Gly Val Trp Lys Ser
195 200 205
Pro Ala Asp Met Gln Ser Trp Val Pro Leu Ser Thr Asp Asp Pro Val
210 215 220
Ile Asp Arg Leu Tyr Leu Ser Ser His Arg Gly Val Ile Ala Asp Asn
225 230 235 240
Gln Ala Lys Trp Ala Val Pro Thr Thr Arg Thr Asp Asp Lys Leu Arg
245 250 255
Met Glu Thr Cys Phe Gln Gln Ala Cys Lys Gly Lys Ile Gln Thr Leu
260 265 270
Cys Glu Asn Pro Glu Trp Ala Pro Leu Lys Asp Asn Arg Ile Pro Ser
275 280 285
Tyr Gly Val Leu Ser Val Asp Leu Ser Leu Thr Val Glu Leu Lys Ile
290 295 300
Lys Ile Ala Ser Gly Phe Gly Pro Leu Ile Thr His Gly Ser Gly Met
305 310 315 320
Asp Leu Tyr Lys Ser Asn His Asn Asn Val Tyr Trp Leu Thr Ile Pro
325 330 335
Pro Met Lys Asn Leu Ala Leu Gly Val Ile Asn Thr Leu Glu Trp Ile
340 345 350
Pro Arg Phe Lys Val Ser Pro Tyr Leu Phe Thr Val Pro Ile Lys Glu
355 360 365
Ala Gly Glu Asp Cys His Ala Pro Thr Tyr Leu Pro Ala Glu Val Asp
370 375 380
Gly Asp Val Lys Leu Ser Ser Asn Leu Val Ile Leu Pro Gly Gln Asp
385 390 395 400
Leu Gln Tyr Val Leu Ala Thr Tyr Asp Thr Ser Arg Val Glu His Ala
405 410 415
Val Val Tyr Tyr Val Tyr Ser Pro Gly Arg Ser Phe Ser Tyr Phe Tyr
420 425 430
Pro Phe Arg Leu Pro Ile Lys Gly Val Pro Ile Glu Leu Gln Val Glu
435 440 445
Cys Phe Thr Trp Asp Gln Lys Leu Trp Cys Arg His Phe Cys Val Leu
450 455 460
Ala Asp Ser Glu Ser Gly Gly His Ile Thr His Ser Gly Met Val Gly
465 470 475 480
Met Gly Val Ser Cys Thr Val Thr Arg Glu Asp Gly Thr Asn Arg Arg
485 490 495
<210> 60
<211> 459
<212> PRT
<213> 人工序列
<220>
<223> 重组H蛋白
<400> 60
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Phe Leu Ala Val Ser Lys Gly Asn Cys Ser Gly
20 25 30
Pro Thr Thr Ile Arg Gly Gln Phe Ser Asn Met Ser Leu Ser Leu Leu
35 40 45
Asp Leu Tyr Leu Gly Arg Gly Tyr Asn Val Ser Ser Ile Val Thr Met
50 55 60
Thr Ser Gln Gly Met Tyr Gly Gly Thr Tyr Leu Val Glu Lys Pro Asn
65 70 75 80
Leu Ser Ser Lys Arg Ser Glu Leu Ser Gln Leu Ser Met Tyr Arg Val
85 90 95
Phe Glu Val Gly Val Ile Arg Asn Pro Gly Leu Gly Ala Pro Val Phe
100 105 110
His Met Thr Asn Tyr Leu Glu Gln Pro Val Ser Asn Asp Leu Ser Asn
115 120 125
Cys Met Val Ala Leu Gly Glu Leu Lys Leu Ala Ala Leu Cys His Gly
130 135 140
Glu Asp Ser Ile Thr Ile Pro Tyr Gln Gly Ser Gly Lys Gly Val Ser
145 150 155 160
Phe Gln Leu Val Lys Leu Gly Val Trp Lys Ser Pro Ala Asp Met Gln
165 170 175
Ser Trp Val Pro Leu Ser Thr Asp Asp Pro Val Ile Asp Arg Leu Tyr
180 185 190
Leu Ser Ser His Arg Gly Val Ile Ala Asp Asn Gln Ala Lys Trp Ala
195 200 205
Val Pro Thr Thr Arg Thr Asp Asp Lys Leu Arg Met Glu Thr Cys Phe
210 215 220
Gln Gln Ala Cys Lys Gly Lys Ile Gln Thr Leu Cys Glu Asn Pro Glu
225 230 235 240
Trp Ala Pro Leu Lys Asp Asn Arg Ile Pro Ser Tyr Gly Val Leu Ser
245 250 255
Val Asp Leu Ser Leu Thr Val Glu Leu Lys Ile Lys Ile Ala Ser Gly
260 265 270
Phe Gly Pro Leu Ile Thr His Gly Ser Gly Met Asp Leu Tyr Lys Ser
275 280 285
Asn His Asn Asn Val Tyr Trp Leu Thr Ile Pro Pro Met Lys Asn Leu
290 295 300
Ala Leu Gly Val Ile Asn Thr Leu Glu Trp Ile Pro Arg Phe Lys Val
305 310 315 320
Ser Pro Tyr Leu Phe Thr Val Pro Ile Lys Glu Ala Gly Glu Asp Cys
325 330 335
His Ala Pro Thr Tyr Leu Pro Ala Glu Val Asp Gly Asp Val Lys Leu
340 345 350
Ser Ser Asn Leu Val Ile Leu Pro Gly Gln Asp Leu Gln Tyr Val Leu
355 360 365
Ala Thr Tyr Asp Thr Ser Arg Val Glu His Ala Val Val Tyr Tyr Val
370 375 380
Tyr Ser Pro Gly Arg Ser Phe Ser Tyr Phe Tyr Pro Phe Arg Leu Pro
385 390 395 400
Ile Lys Gly Val Pro Ile Glu Leu Gln Val Glu Cys Phe Thr Trp Asp
405 410 415
Gln Lys Leu Trp Cys Arg His Phe Cys Val Leu Ala Asp Ser Glu Ser
420 425 430
Gly Gly His Ile Thr His Ser Gly Met Val Gly Met Gly Val Ser Cys
435 440 445
Thr Val Thr Arg Glu Asp Gly Thr Asn Arg Arg
450 455
<210> 61
<211> 497
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 61
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Gly Ser Gly Gly Gly Ser Ala Gln
100 105 110
Ile Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala
115 120 125
Ile Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu
130 135 140
Ala Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Val Gln Gly Val
145 150 155 160
Gln Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser
165 170 175
Cys Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr
180 185 190
Thr Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser
195 200 205
Ala Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile
210 215 220
Asn Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly
225 230 235 240
Ile Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr
245 250 255
Glu Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu
260 265 270
Ile Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile
275 280 285
Gly Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln
290 295 300
Gly Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro
305 310 315 320
Glu Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu
325 330 335
Leu Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu
340 345 350
Val Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu
355 360 365
Ile Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr
370 375 380
Ile Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp
385 390 395 400
His Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser
405 410 415
Arg Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro
420 425 430
Pro Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala
435 440 445
Ile Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln
450 455 460
Ile Leu Ser Ala Ile Gly Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly
465 470 475 480
Gln Ala Tyr Val Arg Lys Asp Gly Glu Trp Val Leu Leu Ser Thr Phe
485 490 495
Leu
<210> 62
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 62
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Cys Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Cys Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 63
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 63
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Phe Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 64
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 64
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Cys Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Cys Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Phe Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Pro Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 65
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 65
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Cys Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Cys Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Cys Gln Ala Gly Gln Glu Cys Ile Leu Ala Phe Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Pro Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 66
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 66
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Cys
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Cys Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 67
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 67
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Cys
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Cys Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Phe Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Pro Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 68
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 68
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Cys
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Cys Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Cys Gln Ala Gly Gln Glu Cys Ile Leu Ala Phe Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Pro Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 69
<211> 520
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 69
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Cys
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Cys Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Arg Ser Met Lys Gly Leu Ser Ser Ala Ile Glu Asp Lys Ile Glu
485 490 495
Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu Ile Ala Arg Ile
500 505 510
Lys Lys Leu Ile Gly Glu Ala Pro
515 520
<210> 70
<211> 520
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 70
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Cys
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Cys Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Phe Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Pro Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Arg Ser Met Lys Gly Leu Ser Ser Ala Ile Glu Asp Lys Ile Glu
485 490 495
Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu Ile Ala Arg Ile
500 505 510
Lys Lys Leu Ile Gly Glu Ala Pro
515 520
<210> 71
<211> 520
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 71
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Cys
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Cys Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Cys Gln Ala Gly Gln Glu Cys Ile Leu Ala Phe Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Pro Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Arg Ser Met Lys Gly Leu Ser Ser Ala Ile Glu Asp Lys Ile Glu
485 490 495
Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu Ile Ala Arg Ile
500 505 510
Lys Lys Leu Ile Gly Glu Ala Pro
515 520
<210> 72
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 72
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Cys Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Cys Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Cys Gln Ala Gly Gln Glu Cys Ile Leu Ala Phe Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Pro Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 73
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 73
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Cys Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Cys Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Cys Gln Ala Gly Gln Glu Cys Ile Leu Ala Phe Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Pro Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 74
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 74
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Cys Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Cys Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Cys Gln Ala Gly Gln Glu Cys Ile Leu Ala Phe Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Pro Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 75
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 75
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Cys Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Cys Gln Ala Gly Gln Glu Cys Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Cys His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 76
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 76
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Cys Gln Ala Gly Gln Glu Cys Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Cys Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Cys Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 77
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 77
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Cys Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Cys Gln Ala Gly Gln Glu Cys Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Cys Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 78
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 78
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Cys
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Lys Arg Phe Ala Gly
100 105 110
Val Cys Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 79
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 79
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Cys
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Lys Arg Phe Ala Gly
100 105 110
Val Cys Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Arg Gln Ala Gly Gln Glu Met Ile Leu Ala Phe Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Pro Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 80
<211> 513
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 80
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Cys
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Lys Arg Phe Ala Gly
100 105 110
Val Cys Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Cys Gln Ala Gly Gln Glu Cys Ile Leu Ala Phe Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Pro Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro
<210> 81
<211> 988
<212> PRT
<213> 人工序列
<220>
<223> 重组F蛋白
<400> 81
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Ile His Trp Gly Asn Leu Ser Lys Ile Gly Val
20 25 30
Val Gly Ile Gly Ser Ala Ser Tyr Lys Val Met Thr Arg Ser Ser His
35 40 45
Gln Ser Leu Val Ile Lys Leu Met Pro Asn Ile Thr Leu Leu Asn Asn
50 55 60
Cys Thr Arg Val Glu Ile Ala Glu Tyr Arg Arg Leu Leu Arg Thr Val
65 70 75 80
Leu Glu Pro Ile Arg Asp Ala Leu Asn Ala Val Thr Gln Asn Ile Arg
85 90 95
Pro Val Gln Ser Val Ala Ser Ser Arg Arg His Gly Gly Gly Ala Gly
100 105 110
Val Val Leu Ala Gly Ala Ala Leu Gly Val Ala Thr Ala Ala Gln Ile
115 120 125
Thr Ala Gly Ile Ala Leu His Gln Ser Met Leu Asn Ser Gln Ala Ile
130 135 140
Asp Asn Leu Arg Ala Ser Leu Glu Thr Thr Asn Gln Ala Ile Glu Ala
145 150 155 160
Ile Cys Gln Ala Gly Gln Glu Cys Ile Leu Ala Val Gln Gly Val Gln
165 170 175
Asp Tyr Ile Asn Asn Glu Leu Ile Pro Ser Met Asn Gln Leu Ser Cys
180 185 190
Asp Leu Ile Gly Gln Lys Leu Gly Leu Lys Leu Leu Arg Tyr Tyr Thr
195 200 205
Glu Ile Leu Ser Leu Phe Gly Pro Ser Leu Arg Asp Pro Ile Ser Ala
210 215 220
Glu Ile Ser Ile Gln Ala Leu Ser Tyr Ala Leu Gly Gly Asp Ile Asn
225 230 235 240
Lys Val Leu Glu Lys Leu Gly Tyr Ser Gly Gly Asp Leu Leu Gly Ile
245 250 255
Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp Thr Glu
260 265 270
Ser Tyr Phe Ile Val Leu Ser Ile Ala Tyr Pro Thr Leu Ser Glu Ile
275 280 285
Lys Gly Val Ile Val His Arg Leu Glu Gly Val Ser Tyr Asn Ile Gly
290 295 300
Ser Gln Glu Trp Tyr Thr Thr Val Pro Lys Tyr Val Ala Thr Gln Gly
305 310 315 320
Tyr Leu Ile Ser Asn Phe Asp Glu Ser Ser Cys Thr Phe Met Pro Glu
325 330 335
Gly Thr Val Cys Ser Gln Asn Ala Leu Tyr Pro Met Ser Pro Leu Leu
340 345 350
Gln Glu Cys Leu Arg Gly Ser Thr Lys Ser Cys Ala Arg Thr Leu Val
355 360 365
Ser Gly Ser Phe Gly Asn Arg Phe Ile Leu Ser Gln Gly Asn Leu Ile
370 375 380
Ala Asn Cys Ala Ser Ile Leu Cys Lys Cys Tyr Thr Thr Gly Thr Ile
385 390 395 400
Ile Asn Gln Asp Pro Asp Lys Ile Leu Thr Tyr Ile Ala Ala Asp His
405 410 415
Cys Pro Val Val Glu Val Asn Gly Val Thr Ile Gln Val Gly Ser Arg
420 425 430
Arg Tyr Pro Asp Ala Val Tyr Leu His Arg Ile Asp Leu Gly Pro Pro
435 440 445
Ile Ser Leu Glu Arg Leu Asp Val Gly Thr Asn Leu Gly Asn Ala Ile
450 455 460
Ala Lys Leu Glu Asp Ala Lys Glu Leu Leu Glu Ser Ser Asp Gln Ile
465 470 475 480
Leu Ser Ala Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr
485 490 495
His Ile Glu Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala
500 505 510
Pro Gly Ser Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr
515 520 525
Val Arg Lys Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu Gly Ser
530 535 540
Gly Gly Gly Gly Gly Gly Phe Leu Ala Val Ser Lys Gly Asn Cys Ser
545 550 555 560
Gly Pro Thr Thr Ile Arg Gly Gln Phe Ser Asn Met Ser Leu Ser Leu
565 570 575
Leu Asp Leu Tyr Leu Gly Arg Gly Tyr Asn Val Ser Ser Ile Val Thr
580 585 590
Met Thr Ser Gln Gly Met Tyr Gly Gly Thr Tyr Leu Val Glu Lys Pro
595 600 605
Asn Leu Ser Ser Lys Arg Ser Glu Leu Ser Gln Leu Ser Met Tyr Arg
610 615 620
Val Phe Glu Val Gly Val Ile Arg Asn Pro Gly Leu Gly Ala Pro Val
625 630 635 640
Phe His Met Thr Asn Tyr Leu Glu Gln Pro Val Ser Asn Asp Leu Ser
645 650 655
Asn Cys Met Val Ala Leu Gly Glu Leu Lys Leu Ala Ala Leu Cys His
660 665 670
Gly Glu Asp Ser Ile Thr Ile Pro Tyr Gln Gly Ser Gly Lys Gly Val
675 680 685
Ser Phe Gln Leu Val Lys Leu Gly Val Trp Lys Ser Pro Ala Asp Met
690 695 700
Gln Ser Trp Val Pro Leu Ser Thr Asp Asp Pro Val Ile Asp Arg Leu
705 710 715 720
Tyr Leu Ser Ser His Arg Gly Val Ile Ala Asp Asn Gln Ala Lys Trp
725 730 735
Ala Val Pro Thr Thr Arg Thr Asp Asp Lys Leu Arg Met Glu Thr Cys
740 745 750
Phe Gln Gln Ala Cys Lys Gly Lys Ile Gln Thr Leu Cys Glu Asn Pro
755 760 765
Glu Trp Ala Pro Leu Lys Asp Asn Arg Ile Pro Ser Tyr Gly Val Leu
770 775 780
Ser Val Asp Leu Ser Leu Thr Val Glu Leu Lys Ile Lys Ile Ala Ser
785 790 795 800
Gly Phe Gly Pro Leu Ile Thr His Gly Ser Gly Met Asp Leu Tyr Lys
805 810 815
Ser Asn His Asn Asn Val Tyr Trp Leu Thr Ile Pro Pro Met Lys Asn
820 825 830
Leu Ala Leu Gly Val Ile Asn Thr Leu Glu Trp Ile Pro Arg Phe Lys
835 840 845
Val Ser Pro Tyr Leu Phe Thr Val Pro Ile Lys Glu Ala Gly Glu Asp
850 855 860
Cys His Ala Pro Thr Tyr Leu Pro Ala Glu Val Asp Gly Asp Val Lys
865 870 875 880
Leu Ser Ser Asn Leu Val Ile Leu Pro Gly Gln Asp Leu Gln Tyr Val
885 890 895
Leu Ala Thr Tyr Asp Thr Ser Arg Val Glu His Ala Val Val Tyr Tyr
900 905 910
Val Tyr Ser Pro Gly Arg Ser Phe Ser Tyr Phe Tyr Pro Phe Arg Leu
915 920 925
Pro Ile Lys Gly Val Pro Ile Glu Leu Gln Val Glu Cys Phe Thr Trp
930 935 940
Asp Gln Lys Leu Trp Cys Arg His Phe Cys Val Leu Ala Asp Ser Glu
945 950 955 960
Ser Gly Gly His Ile Thr His Ser Gly Met Val Gly Met Gly Val Ser
965 970 975
Cys Thr Val Thr Arg Glu Asp Gly Thr Asn Arg Arg
980 985
<210> 82
<211> 950
<212> PRT
<213> 人工序列
<220>
<223> 重组HN/H蛋白
<400> 82
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Asn Ile Pro Leu Val Asn Asp Leu Arg Phe Ile
20 25 30
Asn Gly Ile Asn Lys Phe Ile Ile Glu Asp Tyr Ala Thr His Asp Phe
35 40 45
Ser Ile Gly His Pro Leu Asn Met Pro Ser Phe Ile Pro Thr Ala Thr
50 55 60
Ser Pro Asn Gly Cys Thr Arg Ile Pro Ser Phe Ser Leu Gly Lys Thr
65 70 75 80
His Trp Cys Tyr Thr His Asn Val Ile Asn Ala Asn Cys Lys Asp His
85 90 95
Thr Ser Ser Asn Gln Tyr Val Ser Met Gly Ile Leu Val Gln Thr Ala
100 105 110
Ser Gly Tyr Pro Met Phe Lys Thr Leu Lys Ile Gln Tyr Leu Ser Asp
115 120 125
Gly Leu Asn Arg Lys Ser Cys Ser Ile Ala Thr Val Pro Asp Gly Cys
130 135 140
Ala Met Tyr Cys Tyr Val Ser Thr Gln Leu Glu Thr Asp Asp Tyr Ala
145 150 155 160
Gly Ser Ser Pro Pro Thr Gln Lys Leu Thr Leu Leu Phe Tyr Asn Asp
165 170 175
Thr Val Thr Glu Arg Thr Ile Ser Pro Ser Gly Leu Glu Gly Asn Trp
180 185 190
Ala Thr Leu Val Pro Gly Val Gly Ser Gly Ile Tyr Phe Glu Asn Lys
195 200 205
Leu Ile Phe Pro Ala Tyr Gly Gly Val Leu Pro Asn Ser Thr Leu Gly
210 215 220
Val Lys Ser Ala Arg Glu Phe Phe Arg Pro Val Asn Pro Tyr Asn Pro
225 230 235 240
Cys Ser Gly Pro Gln Gln Asp Leu Asp Gln Arg Ala Leu Arg Ser Tyr
245 250 255
Phe Pro Ser Tyr Phe Ser Asn Arg Arg Ile Gln Ser Ala Phe Leu Val
260 265 270
Cys Ala Trp Asn Gln Ile Leu Val Thr Asn Cys Glu Leu Val Val Pro
275 280 285
Ser Ser Asn Gln Thr Met Met Gly Ala Glu Gly Arg Val Leu Leu Ile
290 295 300
Asn Asn Arg Leu Leu Tyr Tyr Gln Arg Ser Thr Ser Trp Trp Pro Tyr
305 310 315 320
Glu Leu Leu Tyr Glu Ile Ser Phe Thr Phe Thr Asn Ser Gly Pro Ser
325 330 335
Ser Val Asn Met Ser Trp Ile Pro Ile Tyr Ser Phe Thr Arg Pro Gly
340 345 350
Ser Gly Asn Cys Ser Gly Glu Asn Val Cys Pro Thr Ala Cys Val Ser
355 360 365
Gly Val Tyr Leu Asp Pro Trp Pro Leu Thr Pro Tyr Ser His Gln Ser
370 375 380
Gly Ile Asn Arg Asn Phe Tyr Phe Thr Gly Ala Leu Leu Asn Ser Ser
385 390 395 400
Thr Thr Arg Val Asn Pro Thr Leu Tyr Val Ser Ala Leu Asn Asn Leu
405 410 415
Lys Val Leu Ala Pro Tyr Gly Thr Gln Gly Leu Phe Ala Ser Tyr Thr
420 425 430
Thr Thr Thr Cys Phe Gln Asp Thr Gly Asp Ala Ser Val Tyr Cys Val
435 440 445
Tyr Ile Met Glu Leu Ala Ser Asn Ile Val Gly Glu Phe Gln Ile Leu
450 455 460
Pro Val Leu Thr Arg Leu Thr Ile Thr Gly Gly Gly Ser Gly Tyr Ile
465 470 475 480
Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg Lys Asp Gly Glu
485 490 495
Trp Val Leu Leu Ser Thr Phe Leu Gly Ser Gly Gly Gly Gly Gly Gly
500 505 510
Phe Leu Ala Val Ser Lys Gly Asn Cys Ser Gly Pro Thr Thr Ile Arg
515 520 525
Gly Gln Phe Ser Asn Met Ser Leu Ser Leu Leu Asp Leu Tyr Leu Gly
530 535 540
Arg Gly Tyr Asn Val Ser Ser Ile Val Thr Met Thr Ser Gln Gly Met
545 550 555 560
Tyr Gly Gly Thr Tyr Leu Val Glu Lys Pro Asn Leu Ser Ser Lys Arg
565 570 575
Ser Glu Leu Ser Gln Leu Ser Met Tyr Arg Val Phe Glu Val Gly Val
580 585 590
Ile Arg Asn Pro Gly Leu Gly Ala Pro Val Phe His Met Thr Asn Tyr
595 600 605
Leu Glu Gln Pro Val Ser Asn Asp Leu Ser Asn Cys Met Val Ala Leu
610 615 620
Gly Glu Leu Lys Leu Ala Ala Leu Cys His Gly Glu Asp Ser Ile Thr
625 630 635 640
Ile Pro Tyr Gln Gly Ser Gly Lys Gly Val Ser Phe Gln Leu Val Lys
645 650 655
Leu Gly Val Trp Lys Ser Pro Thr Asp Met Gln Ser Trp Val Pro Leu
660 665 670
Ser Thr Asp Asp Pro Val Ile Asp Arg Leu Tyr Leu Ser Ser His Arg
675 680 685
Gly Val Ile Ala Asp Asn Gln Ala Lys Trp Ala Val Pro Thr Thr Arg
690 695 700
Thr Asp Asp Lys Leu Arg Met Glu Thr Cys Phe Gln Gln Ala Cys Lys
705 710 715 720
Gly Lys Ile Gln Ala Leu Cys Glu Asn Pro Glu Trp Ala Pro Leu Lys
725 730 735
Asp Asn Arg Ile Pro Ser Tyr Gly Val Leu Ser Val Asp Leu Ser Leu
740 745 750
Thr Val Glu Leu Lys Ile Lys Ile Ala Ser Gly Phe Gly Pro Leu Ile
755 760 765
Thr His Gly Ser Gly Met Asp Leu Tyr Lys Ser Asn His Asn Asn Val
770 775 780
Tyr Trp Leu Thr Ile Pro Pro Met Lys Asn Leu Ala Leu Gly Val Ile
785 790 795 800
Asn Thr Leu Glu Trp Ile Pro Arg Phe Lys Val Ser Pro Tyr Leu Phe
805 810 815
Asn Val Pro Ile Lys Glu Ala Gly Glu Asp Cys His Ala Pro Thr Tyr
820 825 830
Leu Pro Ala Glu Val Asp Gly Asp Val Lys Leu Ser Ser Asn Leu Val
835 840 845
Ile Leu Pro Gly Gln Asp Leu Gln Tyr Val Leu Ala Thr Tyr Asp Thr
850 855 860
Ser Arg Val Glu His Ala Val Val Tyr Tyr Val Tyr Ser Pro Ser Arg
865 870 875 880
Ser Phe Ser Tyr Phe Tyr Pro Phe Arg Leu Pro Ile Lys Gly Val Pro
885 890 895
Ile Glu Leu Gln Val Glu Cys Phe Thr Trp Asp Gln Lys Leu Trp Cys
900 905 910
Arg His Phe Cys Val Leu Ala Asp Ser Glu Ser Gly Gly His Ile Thr
915 920 925
His Ser Gly Met Glu Gly Met Gly Val Ser Cys Thr Val Thr Arg Glu
930 935 940
Asp Gly Thr Asn Arg Arg
945 950
<210> 83
<211> 985
<212> PRT
<213> 人工序列
<220>
<223> 重组HN/H蛋白
<400> 83
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Asn Ile Pro Leu Val Asn Asp Leu Arg Phe Ile
20 25 30
Asn Gly Ile Asn Lys Phe Ile Ile Glu Asp Tyr Ala Thr His Asp Phe
35 40 45
Ser Ile Gly His Pro Leu Asn Met Pro Ser Phe Ile Pro Thr Ala Thr
50 55 60
Ser Pro Asn Gly Cys Thr Arg Ile Pro Ser Phe Ser Leu Gly Lys Thr
65 70 75 80
His Trp Cys Tyr Thr His Asn Val Ile Asn Ala Asn Cys Lys Asp His
85 90 95
Thr Ser Ser Asn Gln Tyr Val Ser Met Gly Ile Leu Val Gln Thr Ala
100 105 110
Ser Gly Tyr Pro Met Phe Lys Thr Leu Lys Ile Gln Tyr Leu Ser Asp
115 120 125
Gly Leu Asn Arg Lys Ser Cys Ser Ile Ala Thr Val Pro Asp Gly Cys
130 135 140
Ala Met Tyr Cys Tyr Val Ser Thr Gln Leu Glu Thr Asp Asp Tyr Ala
145 150 155 160
Gly Ser Ser Pro Pro Thr Gln Lys Leu Thr Leu Leu Phe Tyr Asn Asp
165 170 175
Thr Val Thr Glu Arg Thr Ile Ser Pro Ser Gly Leu Glu Gly Asn Trp
180 185 190
Ala Thr Leu Val Pro Gly Val Gly Ser Gly Ile Tyr Phe Glu Asn Lys
195 200 205
Leu Ile Phe Pro Ala Tyr Gly Gly Val Leu Pro Asn Ser Thr Leu Gly
210 215 220
Val Lys Ser Ala Arg Glu Phe Phe Arg Pro Val Asn Pro Tyr Asn Pro
225 230 235 240
Cys Ser Gly Pro Gln Gln Asp Leu Asp Gln Arg Ala Leu Arg Ser Tyr
245 250 255
Phe Pro Ser Tyr Phe Ser Asn Arg Arg Ile Gln Ser Ala Phe Leu Val
260 265 270
Cys Ala Trp Asn Gln Ile Leu Val Thr Asn Cys Glu Leu Val Val Pro
275 280 285
Ser Ser Asn Gln Thr Met Met Gly Ala Glu Gly Arg Val Leu Leu Ile
290 295 300
Asn Asn Arg Leu Leu Tyr Tyr Gln Arg Ser Thr Ser Trp Trp Pro Tyr
305 310 315 320
Glu Leu Leu Tyr Glu Ile Ser Phe Thr Phe Thr Asn Ser Gly Pro Ser
325 330 335
Ser Val Asn Met Ser Trp Ile Pro Ile Tyr Ser Phe Thr Arg Pro Gly
340 345 350
Ser Gly Asn Cys Ser Gly Glu Asn Val Cys Pro Thr Ala Cys Val Ser
355 360 365
Gly Val Tyr Leu Asp Pro Trp Pro Leu Thr Pro Tyr Ser His Gln Ser
370 375 380
Gly Ile Asn Arg Asn Phe Tyr Phe Thr Gly Ala Leu Leu Asn Ser Ser
385 390 395 400
Thr Thr Arg Val Asn Pro Thr Leu Tyr Val Ser Ala Leu Asn Asn Leu
405 410 415
Lys Val Leu Ala Pro Tyr Gly Thr Gln Gly Leu Phe Ala Ser Tyr Thr
420 425 430
Thr Thr Thr Cys Phe Gln Asp Thr Gly Asp Ala Ser Val Tyr Cys Val
435 440 445
Tyr Ile Met Glu Leu Ala Ser Asn Ile Val Gly Glu Phe Gln Ile Leu
450 455 460
Pro Val Leu Thr Arg Leu Thr Ile Thr Gly Gly Lys Leu Met Lys Gln
465 470 475 480
Ile Glu Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu
485 490 495
Asn Glu Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro Gly Ser
500 505 510
Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg Lys
515 520 525
Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu Gly Ser Gly Gly Gly
530 535 540
Gly Gly Gly Phe Leu Ala Val Ser Lys Gly Asn Cys Ser Gly Pro Thr
545 550 555 560
Thr Ile Arg Gly Gln Phe Ser Asn Met Ser Leu Ser Leu Leu Asp Leu
565 570 575
Tyr Leu Gly Arg Gly Tyr Asn Val Ser Ser Ile Val Thr Met Thr Ser
580 585 590
Gln Gly Met Tyr Gly Gly Thr Tyr Leu Val Glu Lys Pro Asn Leu Ser
595 600 605
Ser Lys Arg Ser Glu Leu Ser Gln Leu Ser Met Tyr Arg Val Phe Glu
610 615 620
Val Gly Val Ile Arg Asn Pro Gly Leu Gly Ala Pro Val Phe His Met
625 630 635 640
Thr Asn Tyr Leu Glu Gln Pro Val Ser Asn Asp Leu Ser Asn Cys Met
645 650 655
Val Ala Leu Gly Glu Leu Lys Leu Ala Ala Leu Cys His Gly Glu Asp
660 665 670
Ser Ile Thr Ile Pro Tyr Gln Gly Ser Gly Lys Gly Val Ser Phe Gln
675 680 685
Leu Val Lys Leu Gly Val Trp Lys Ser Pro Thr Asp Met Gln Ser Trp
690 695 700
Val Pro Leu Ser Thr Asp Asp Pro Val Ile Asp Arg Leu Tyr Leu Ser
705 710 715 720
Ser His Arg Gly Val Ile Ala Asp Asn Gln Ala Lys Trp Ala Val Pro
725 730 735
Thr Thr Arg Thr Asp Asp Lys Leu Arg Met Glu Thr Cys Phe Gln Gln
740 745 750
Ala Cys Lys Gly Lys Ile Gln Ala Leu Cys Glu Asn Pro Glu Trp Ala
755 760 765
Pro Leu Lys Asp Asn Arg Ile Pro Ser Tyr Gly Val Leu Ser Val Asp
770 775 780
Leu Ser Leu Thr Val Glu Leu Lys Ile Lys Ile Ala Ser Gly Phe Gly
785 790 795 800
Pro Leu Ile Thr His Gly Ser Gly Met Asp Leu Tyr Lys Ser Asn His
805 810 815
Asn Asn Val Tyr Trp Leu Thr Ile Pro Pro Met Lys Asn Leu Ala Leu
820 825 830
Gly Val Ile Asn Thr Leu Glu Trp Ile Pro Arg Phe Lys Val Ser Pro
835 840 845
Tyr Leu Phe Asn Val Pro Ile Lys Glu Ala Gly Glu Asp Cys His Ala
850 855 860
Pro Thr Tyr Leu Pro Ala Glu Val Asp Gly Asp Val Lys Leu Ser Ser
865 870 875 880
Asn Leu Val Ile Leu Pro Gly Gln Asp Leu Gln Tyr Val Leu Ala Thr
885 890 895
Tyr Asp Thr Ser Arg Val Glu His Ala Val Val Tyr Tyr Val Tyr Ser
900 905 910
Pro Ser Arg Ser Phe Ser Tyr Phe Tyr Pro Phe Arg Leu Pro Ile Lys
915 920 925
Gly Val Pro Ile Glu Leu Gln Val Glu Cys Phe Thr Trp Asp Gln Lys
930 935 940
Leu Trp Cys Arg His Phe Cys Val Leu Ala Asp Ser Glu Ser Gly Gly
945 950 955 960
His Ile Thr His Ser Gly Met Glu Gly Met Gly Val Ser Cys Thr Val
965 970 975
Thr Arg Glu Asp Gly Thr Asn Arg Arg
980 985
<210> 84
<211> 948
<212> PRT
<213> 人工序列
<220>
<223> 重组HN/H蛋白
<400> 84
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Phe Leu Ala Val Ser Lys Gly Asn Cys Ser Gly
20 25 30
Pro Thr Thr Ile Arg Gly Gln Phe Ser Asn Met Ser Leu Ser Leu Leu
35 40 45
Asp Leu Tyr Leu Gly Arg Gly Tyr Asn Val Ser Ser Ile Val Thr Met
50 55 60
Thr Ser Gln Gly Met Tyr Gly Gly Thr Tyr Leu Val Glu Lys Pro Asn
65 70 75 80
Leu Ser Ser Lys Arg Ser Glu Leu Ser Gln Leu Ser Met Tyr Arg Val
85 90 95
Phe Glu Val Gly Val Ile Arg Asn Pro Gly Leu Gly Ala Pro Val Phe
100 105 110
His Met Thr Asn Tyr Leu Glu Gln Pro Val Ser Asn Asp Leu Ser Asn
115 120 125
Cys Met Val Ala Leu Gly Glu Leu Lys Leu Ala Ala Leu Cys His Gly
130 135 140
Glu Asp Ser Ile Thr Ile Pro Tyr Gln Gly Ser Gly Lys Gly Val Ser
145 150 155 160
Phe Gln Leu Val Lys Leu Gly Val Trp Lys Ser Pro Thr Asp Met Gln
165 170 175
Ser Trp Val Pro Leu Ser Thr Asp Asp Pro Val Ile Asp Arg Leu Tyr
180 185 190
Leu Ser Ser His Arg Gly Val Ile Ala Asp Asn Gln Ala Lys Trp Ala
195 200 205
Val Pro Thr Thr Arg Thr Asp Asp Lys Leu Arg Met Glu Thr Cys Phe
210 215 220
Gln Gln Ala Cys Lys Gly Lys Ile Gln Ala Leu Cys Glu Asn Pro Glu
225 230 235 240
Trp Ala Pro Leu Lys Asp Asn Arg Ile Pro Ser Tyr Gly Val Leu Ser
245 250 255
Val Asp Leu Ser Leu Thr Val Glu Leu Lys Ile Lys Ile Ala Ser Gly
260 265 270
Phe Gly Pro Leu Ile Thr His Gly Ser Gly Met Asp Leu Tyr Lys Ser
275 280 285
Asn His Asn Asn Val Tyr Trp Leu Thr Ile Pro Pro Met Lys Asn Leu
290 295 300
Ala Leu Gly Val Ile Asn Thr Leu Glu Trp Ile Pro Arg Phe Lys Val
305 310 315 320
Ser Pro Tyr Leu Phe Asn Val Pro Ile Lys Glu Ala Gly Glu Asp Cys
325 330 335
His Ala Pro Thr Tyr Leu Pro Ala Glu Val Asp Gly Asp Val Lys Leu
340 345 350
Ser Ser Asn Leu Val Ile Leu Pro Gly Gln Asp Leu Gln Tyr Val Leu
355 360 365
Ala Thr Tyr Asp Thr Ser Arg Val Glu His Ala Val Val Tyr Tyr Val
370 375 380
Tyr Ser Pro Ser Arg Ser Phe Ser Tyr Phe Tyr Pro Phe Arg Leu Pro
385 390 395 400
Ile Lys Gly Val Pro Ile Glu Leu Gln Val Glu Cys Phe Thr Trp Asp
405 410 415
Gln Lys Leu Trp Cys Arg His Phe Cys Val Leu Ala Asp Ser Glu Ser
420 425 430
Gly Gly His Ile Thr His Ser Gly Met Glu Gly Met Gly Val Ser Cys
435 440 445
Thr Val Thr Arg Glu Asp Gly Thr Asn Arg Arg Gly Ser Gly Tyr Ile
450 455 460
Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg Lys Asp Gly Glu
465 470 475 480
Trp Val Leu Leu Ser Thr Phe Leu Gly Ser Gly Gly Gly Gly Gly Gly
485 490 495
Asn Ile Pro Leu Val Asn Asp Leu Arg Phe Ile Asn Gly Ile Asn Lys
500 505 510
Phe Ile Ile Glu Asp Tyr Ala Thr His Asp Phe Ser Ile Gly His Pro
515 520 525
Leu Asn Met Pro Ser Phe Ile Pro Thr Ala Thr Ser Pro Asn Gly Cys
530 535 540
Thr Arg Ile Pro Ser Phe Ser Leu Gly Lys Thr His Trp Cys Tyr Thr
545 550 555 560
His Asn Val Ile Asn Ala Asn Cys Lys Asp His Thr Ser Ser Asn Gln
565 570 575
Tyr Val Ser Met Gly Ile Leu Val Gln Thr Ala Ser Gly Tyr Pro Met
580 585 590
Phe Lys Thr Leu Lys Ile Gln Tyr Leu Ser Asp Gly Leu Asn Arg Lys
595 600 605
Ser Cys Ser Ile Ala Thr Val Pro Asp Gly Cys Ala Met Tyr Cys Tyr
610 615 620
Val Ser Thr Gln Leu Glu Thr Asp Asp Tyr Ala Gly Ser Ser Pro Pro
625 630 635 640
Thr Gln Lys Leu Thr Leu Leu Phe Tyr Asn Asp Thr Val Thr Glu Arg
645 650 655
Thr Ile Ser Pro Ser Gly Leu Glu Gly Asn Trp Ala Thr Leu Val Pro
660 665 670
Gly Val Gly Ser Gly Ile Tyr Phe Glu Asn Lys Leu Ile Phe Pro Ala
675 680 685
Tyr Gly Gly Val Leu Pro Asn Ser Thr Leu Gly Val Lys Ser Ala Arg
690 695 700
Glu Phe Phe Arg Pro Val Asn Pro Tyr Asn Pro Cys Ser Gly Pro Gln
705 710 715 720
Gln Asp Leu Asp Gln Arg Ala Leu Arg Ser Tyr Phe Pro Ser Tyr Phe
725 730 735
Ser Asn Arg Arg Ile Gln Ser Ala Phe Leu Val Cys Ala Trp Asn Gln
740 745 750
Ile Leu Val Thr Asn Cys Glu Leu Val Val Pro Ser Ser Asn Gln Thr
755 760 765
Met Met Gly Ala Glu Gly Arg Val Leu Leu Ile Asn Asn Arg Leu Leu
770 775 780
Tyr Tyr Gln Arg Ser Thr Ser Trp Trp Pro Tyr Glu Leu Leu Tyr Glu
785 790 795 800
Ile Ser Phe Thr Phe Thr Asn Ser Gly Pro Ser Ser Val Asn Met Ser
805 810 815
Trp Ile Pro Ile Tyr Ser Phe Thr Arg Pro Gly Ser Gly Asn Cys Ser
820 825 830
Gly Glu Asn Val Cys Pro Thr Ala Cys Val Ser Gly Val Tyr Leu Asp
835 840 845
Pro Trp Pro Leu Thr Pro Tyr Ser His Gln Ser Gly Ile Asn Arg Asn
850 855 860
Phe Tyr Phe Thr Gly Ala Leu Leu Asn Ser Ser Thr Thr Arg Val Asn
865 870 875 880
Pro Thr Leu Tyr Val Ser Ala Leu Asn Asn Leu Lys Val Leu Ala Pro
885 890 895
Tyr Gly Thr Gln Gly Leu Phe Ala Ser Tyr Thr Thr Thr Thr Cys Phe
900 905 910
Gln Asp Thr Gly Asp Ala Ser Val Tyr Cys Val Tyr Ile Met Glu Leu
915 920 925
Ala Ser Asn Ile Val Gly Glu Phe Gln Ile Leu Pro Val Leu Thr Arg
930 935 940
Leu Thr Ile Thr
945
<210> 85
<211> 981
<212> PRT
<213> 人工序列
<220>
<223> 重组HN/H蛋白
<400> 85
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Phe Leu Ala Val Ser Lys Gly Asn Cys Ser Gly
20 25 30
Pro Thr Thr Ile Arg Gly Gln Phe Ser Asn Met Ser Leu Ser Leu Leu
35 40 45
Asp Leu Tyr Leu Gly Arg Gly Tyr Asn Val Ser Ser Ile Val Thr Met
50 55 60
Thr Ser Gln Gly Met Tyr Gly Gly Thr Tyr Leu Val Glu Lys Pro Asn
65 70 75 80
Leu Ser Ser Lys Arg Ser Glu Leu Ser Gln Leu Ser Met Tyr Arg Val
85 90 95
Phe Glu Val Gly Val Ile Arg Asn Pro Gly Leu Gly Ala Pro Val Phe
100 105 110
His Met Thr Asn Tyr Leu Glu Gln Pro Val Ser Asn Asp Leu Ser Asn
115 120 125
Cys Met Val Ala Leu Gly Glu Leu Lys Leu Ala Ala Leu Cys His Gly
130 135 140
Glu Asp Ser Ile Thr Ile Pro Tyr Gln Gly Ser Gly Lys Gly Val Ser
145 150 155 160
Phe Gln Leu Val Lys Leu Gly Val Trp Lys Ser Pro Thr Asp Met Gln
165 170 175
Ser Trp Val Pro Leu Ser Thr Asp Asp Pro Val Ile Asp Arg Leu Tyr
180 185 190
Leu Ser Ser His Arg Gly Val Ile Ala Asp Asn Gln Ala Lys Trp Ala
195 200 205
Val Pro Thr Thr Arg Thr Asp Asp Lys Leu Arg Met Glu Thr Cys Phe
210 215 220
Gln Gln Ala Cys Lys Gly Lys Ile Gln Ala Leu Cys Glu Asn Pro Glu
225 230 235 240
Trp Ala Pro Leu Lys Asp Asn Arg Ile Pro Ser Tyr Gly Val Leu Ser
245 250 255
Val Asp Leu Ser Leu Thr Val Glu Leu Lys Ile Lys Ile Ala Ser Gly
260 265 270
Phe Gly Pro Leu Ile Thr His Gly Ser Gly Met Asp Leu Tyr Lys Ser
275 280 285
Asn His Asn Asn Val Tyr Trp Leu Thr Ile Pro Pro Met Lys Asn Leu
290 295 300
Ala Leu Gly Val Ile Asn Thr Leu Glu Trp Ile Pro Arg Phe Lys Val
305 310 315 320
Ser Pro Tyr Leu Phe Asn Val Pro Ile Lys Glu Ala Gly Glu Asp Cys
325 330 335
His Ala Pro Thr Tyr Leu Pro Ala Glu Val Asp Gly Asp Val Lys Leu
340 345 350
Ser Ser Asn Leu Val Ile Leu Pro Gly Gln Asp Leu Gln Tyr Val Leu
355 360 365
Ala Thr Tyr Asp Thr Ser Arg Val Glu His Ala Val Val Tyr Tyr Val
370 375 380
Tyr Ser Pro Ser Arg Ser Phe Ser Tyr Phe Tyr Pro Phe Arg Leu Pro
385 390 395 400
Ile Lys Gly Val Pro Ile Glu Leu Gln Val Glu Cys Phe Thr Trp Asp
405 410 415
Gln Lys Leu Trp Cys Arg His Phe Cys Val Leu Ala Asp Ser Glu Ser
420 425 430
Gly Gly His Ile Thr His Ser Gly Met Glu Gly Met Gly Val Ser Cys
435 440 445
Thr Val Thr Arg Glu Asp Gly Thr Asn Arg Arg Met Lys Gln Ile Glu
450 455 460
Asp Lys Ile Glu Glu Ile Leu Ser Lys Ile Tyr His Ile Glu Asn Glu
465 470 475 480
Ile Ala Arg Ile Lys Lys Leu Ile Gly Glu Ala Pro Gly Ser Gly Tyr
485 490 495
Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg Lys Asp Gly
500 505 510
Glu Trp Val Leu Leu Ser Thr Phe Leu Gly Ser Gly Gly Gly Gly Gly
515 520 525
Gly Asn Ile Pro Leu Val Asn Asp Leu Arg Phe Ile Asn Gly Ile Asn
530 535 540
Lys Phe Ile Ile Glu Asp Tyr Ala Thr His Asp Phe Ser Ile Gly His
545 550 555 560
Pro Leu Asn Met Pro Ser Phe Ile Pro Thr Ala Thr Ser Pro Asn Gly
565 570 575
Cys Thr Arg Ile Pro Ser Phe Ser Leu Gly Lys Thr His Trp Cys Tyr
580 585 590
Thr His Asn Val Ile Asn Ala Asn Cys Lys Asp His Thr Ser Ser Asn
595 600 605
Gln Tyr Val Ser Met Gly Ile Leu Val Gln Thr Ala Ser Gly Tyr Pro
610 615 620
Met Phe Lys Thr Leu Lys Ile Gln Tyr Leu Ser Asp Gly Leu Asn Arg
625 630 635 640
Lys Ser Cys Ser Ile Ala Thr Val Pro Asp Gly Cys Ala Met Tyr Cys
645 650 655
Tyr Val Ser Thr Gln Leu Glu Thr Asp Asp Tyr Ala Gly Ser Ser Pro
660 665 670
Pro Thr Gln Lys Leu Thr Leu Leu Phe Tyr Asn Asp Thr Val Thr Glu
675 680 685
Arg Thr Ile Ser Pro Ser Gly Leu Glu Gly Asn Trp Ala Thr Leu Val
690 695 700
Pro Gly Val Gly Ser Gly Ile Tyr Phe Glu Asn Lys Leu Ile Phe Pro
705 710 715 720
Ala Tyr Gly Gly Val Leu Pro Asn Ser Thr Leu Gly Val Lys Ser Ala
725 730 735
Arg Glu Phe Phe Arg Pro Val Asn Pro Tyr Asn Pro Cys Ser Gly Pro
740 745 750
Gln Gln Asp Leu Asp Gln Arg Ala Leu Arg Ser Tyr Phe Pro Ser Tyr
755 760 765
Phe Ser Asn Arg Arg Ile Gln Ser Ala Phe Leu Val Cys Ala Trp Asn
770 775 780
Gln Ile Leu Val Thr Asn Cys Glu Leu Val Val Pro Ser Ser Asn Gln
785 790 795 800
Thr Met Met Gly Ala Glu Gly Arg Val Leu Leu Ile Asn Asn Arg Leu
805 810 815
Leu Tyr Tyr Gln Arg Ser Thr Ser Trp Trp Pro Tyr Glu Leu Leu Tyr
820 825 830
Glu Ile Ser Phe Thr Phe Thr Asn Ser Gly Pro Ser Ser Val Asn Met
835 840 845
Ser Trp Ile Pro Ile Tyr Ser Phe Thr Arg Pro Gly Ser Gly Asn Cys
850 855 860
Ser Gly Glu Asn Val Cys Pro Thr Ala Cys Val Ser Gly Val Tyr Leu
865 870 875 880
Asp Pro Trp Pro Leu Thr Pro Tyr Ser His Gln Ser Gly Ile Asn Arg
885 890 895
Asn Phe Tyr Phe Thr Gly Ala Leu Leu Asn Ser Ser Thr Thr Arg Val
900 905 910
Asn Pro Thr Leu Tyr Val Ser Ala Leu Asn Asn Leu Lys Val Leu Ala
915 920 925
Pro Tyr Gly Thr Gln Gly Leu Phe Ala Ser Tyr Thr Thr Thr Thr Cys
930 935 940
Phe Gln Asp Thr Gly Asp Ala Ser Val Tyr Cys Val Tyr Ile Met Glu
945 950 955 960
Leu Ala Ser Asn Ile Val Gly Glu Phe Gln Ile Leu Pro Val Leu Thr
965 970 975
Arg Leu Thr Ile Thr
980
<210> 86
<211> 580
<212> PRT
<213> 人工序列
<220>
<223> 重组H蛋白
<400> 86
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Arg Leu His Arg Ala Ala Ile Tyr Thr Ala Glu
20 25 30
Ile His Lys Ser Leu Ser Thr Asn Leu Asp Val Thr Asn Ser Ile Glu
35 40 45
His Gln Val Lys Asp Val Leu Thr Pro Leu Phe Lys Ile Ile Gly Asp
50 55 60
Glu Val Gly Leu Arg Thr Pro Gln Arg Phe Thr Asp Leu Val Lys Phe
65 70 75 80
Ile Ser Asp Lys Ile Lys Phe Leu Asn Pro Asp Arg Glu Tyr Asp Phe
85 90 95
Arg Asp Leu Thr Trp Cys Ile Asn Pro Pro Glu Arg Ile Lys Leu Asp
100 105 110
Tyr Asp Gln Tyr Cys Ala Asp Val Ala Ala Glu Glu Leu Met Asn Ala
115 120 125
Leu Val Asn Ser Thr Leu Leu Glu Thr Arg Thr Thr Asn Gln Phe Leu
130 135 140
Ala Val Ser Lys Gly Asn Cys Ser Gly Pro Thr Thr Ile Arg Gly Gln
145 150 155 160
Phe Ser Asn Met Ser Leu Ser Leu Leu Asp Leu Tyr Leu Gly Arg Gly
165 170 175
Tyr Asn Val Ser Ser Ile Val Thr Met Thr Ser Gln Gly Met Tyr Gly
180 185 190
Gly Thr Tyr Leu Val Glu Lys Pro Asn Leu Ser Ser Lys Arg Ser Glu
195 200 205
Leu Ser Gln Leu Ser Met Tyr Arg Val Phe Glu Val Gly Val Ile Arg
210 215 220
Asn Pro Gly Leu Gly Ala Pro Val Phe His Met Thr Asn Tyr Leu Glu
225 230 235 240
Gln Pro Val Ser Asn Asp Leu Ser Asn Cys Met Val Ala Leu Gly Glu
245 250 255
Leu Lys Leu Ala Ala Leu Cys His Gly Glu Asp Ser Ile Thr Ile Pro
260 265 270
Tyr Gln Gly Ser Gly Lys Gly Val Ser Phe Gln Leu Val Lys Leu Gly
275 280 285
Val Trp Lys Ser Pro Thr Asp Met Gln Ser Trp Val Pro Leu Ser Thr
290 295 300
Asp Asp Pro Val Ile Asp Arg Leu Tyr Leu Ser Ser His Arg Gly Val
305 310 315 320
Ile Ala Asp Asn Gln Ala Lys Trp Ala Val Pro Thr Thr Arg Thr Asp
325 330 335
Asp Lys Leu Arg Met Glu Thr Cys Phe Gln Gln Ala Cys Lys Gly Lys
340 345 350
Ile Gln Ala Leu Cys Glu Asn Pro Glu Trp Ala Pro Leu Lys Asp Asn
355 360 365
Arg Ile Pro Ser Tyr Gly Val Leu Ser Val Asp Leu Ser Leu Thr Val
370 375 380
Glu Leu Lys Ile Lys Ile Ala Ser Gly Phe Gly Pro Leu Ile Thr His
385 390 395 400
Gly Ser Gly Met Asp Leu Tyr Lys Ser Asn His Asn Asn Val Tyr Trp
405 410 415
Leu Thr Ile Pro Pro Met Lys Asn Leu Ala Leu Gly Val Ile Asn Thr
420 425 430
Leu Glu Trp Ile Pro Arg Phe Lys Val Ser Pro Tyr Leu Phe Asn Val
435 440 445
Pro Ile Lys Glu Ala Gly Glu Asp Cys His Ala Pro Thr Tyr Leu Pro
450 455 460
Ala Glu Val Asp Gly Asp Val Lys Leu Ser Ser Asn Leu Val Ile Leu
465 470 475 480
Pro Gly Gln Asp Leu Gln Tyr Val Leu Ala Thr Tyr Asp Thr Ser Arg
485 490 495
Val Glu His Ala Val Val Tyr Tyr Val Tyr Ser Pro Ser Arg Ser Phe
500 505 510
Ser Tyr Phe Tyr Pro Phe Arg Leu Pro Ile Lys Gly Val Pro Ile Glu
515 520 525
Leu Gln Val Glu Cys Phe Thr Trp Asp Gln Lys Leu Trp Cys Arg His
530 535 540
Phe Cys Val Leu Ala Asp Ser Glu Ser Gly Gly His Ile Thr His Ser
545 550 555 560
Gly Met Glu Gly Met Gly Val Ser Cys Thr Val Thr Arg Glu Asp Gly
565 570 575
Thr Asn Arg Arg
580
<210> 87
<211> 577
<212> PRT
<213> 人工序列
<220>
<223> 重组H蛋白
<400> 87
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Arg Ala Ala Ile Tyr Thr Ala Glu Ile His Lys
20 25 30
Ser Leu Ser Thr Asn Leu Asp Val Thr Asn Ser Ile Glu His Gln Val
35 40 45
Lys Asp Val Leu Thr Pro Leu Phe Lys Ile Ile Gly Asp Glu Val Gly
50 55 60
Leu Arg Thr Pro Gln Arg Phe Thr Asp Leu Val Lys Phe Ile Ser Asp
65 70 75 80
Lys Ile Lys Phe Leu Asn Pro Asp Arg Glu Tyr Asp Phe Arg Asp Leu
85 90 95
Thr Trp Cys Ile Asn Pro Pro Glu Arg Ile Lys Leu Asp Tyr Asp Gln
100 105 110
Tyr Cys Ala Asp Val Ala Ala Glu Glu Leu Met Asn Ala Leu Val Asn
115 120 125
Ser Thr Leu Leu Glu Thr Arg Thr Thr Asn Gln Phe Leu Ala Val Ser
130 135 140
Lys Gly Asn Cys Ser Gly Pro Thr Thr Ile Arg Gly Gln Phe Ser Asn
145 150 155 160
Met Ser Leu Ser Leu Leu Asp Leu Tyr Leu Gly Arg Gly Tyr Asn Val
165 170 175
Ser Ser Ile Val Thr Met Thr Ser Gln Gly Met Tyr Gly Gly Thr Tyr
180 185 190
Leu Val Glu Lys Pro Asn Leu Ser Ser Lys Arg Ser Glu Leu Ser Gln
195 200 205
Leu Ser Met Tyr Arg Val Phe Glu Val Gly Val Ile Arg Asn Pro Gly
210 215 220
Leu Gly Ala Pro Val Phe His Met Thr Asn Tyr Leu Glu Gln Pro Val
225 230 235 240
Ser Asn Asp Leu Ser Asn Cys Met Val Ala Leu Gly Glu Leu Lys Leu
245 250 255
Ala Ala Leu Cys His Gly Glu Asp Ser Ile Thr Ile Pro Tyr Gln Gly
260 265 270
Ser Gly Lys Gly Val Ser Phe Gln Leu Val Lys Leu Gly Val Trp Lys
275 280 285
Ser Pro Thr Asp Met Gln Ser Trp Val Pro Leu Ser Thr Asp Asp Pro
290 295 300
Val Ile Asp Arg Leu Tyr Leu Ser Ser His Arg Gly Val Ile Ala Asp
305 310 315 320
Asn Gln Ala Lys Trp Ala Val Pro Thr Thr Arg Thr Asp Asp Lys Leu
325 330 335
Arg Met Glu Thr Cys Phe Gln Gln Ala Cys Lys Gly Lys Ile Gln Ala
340 345 350
Leu Cys Glu Asn Pro Glu Trp Ala Pro Leu Lys Asp Asn Arg Ile Pro
355 360 365
Ser Tyr Gly Val Leu Ser Val Asp Leu Ser Leu Thr Val Glu Leu Lys
370 375 380
Ile Lys Ile Ala Ser Gly Phe Gly Pro Leu Ile Thr His Gly Ser Gly
385 390 395 400
Met Asp Leu Tyr Lys Ser Asn His Asn Asn Val Tyr Trp Leu Thr Ile
405 410 415
Pro Pro Met Lys Asn Leu Ala Leu Gly Val Ile Asn Thr Leu Glu Trp
420 425 430
Ile Pro Arg Phe Lys Val Ser Pro Tyr Leu Phe Asn Val Pro Ile Lys
435 440 445
Glu Ala Gly Glu Asp Cys His Ala Pro Thr Tyr Leu Pro Ala Glu Val
450 455 460
Asp Gly Asp Val Lys Leu Ser Ser Asn Leu Val Ile Leu Pro Gly Gln
465 470 475 480
Asp Leu Gln Tyr Val Leu Ala Thr Tyr Asp Thr Ser Arg Val Glu His
485 490 495
Ala Val Val Tyr Tyr Val Tyr Ser Pro Ser Arg Ser Phe Ser Tyr Phe
500 505 510
Tyr Pro Phe Arg Leu Pro Ile Lys Gly Val Pro Ile Glu Leu Gln Val
515 520 525
Glu Cys Phe Thr Trp Asp Gln Lys Leu Trp Cys Arg His Phe Cys Val
530 535 540
Leu Ala Asp Ser Glu Ser Gly Gly His Ile Thr His Ser Gly Met Glu
545 550 555 560
Gly Met Gly Val Ser Cys Thr Val Thr Arg Glu Asp Gly Thr Asn Arg
565 570 575
Arg
<210> 88
<211> 579
<212> PRT
<213> 人工序列
<220>
<223> 重组H蛋白
<400> 88
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Leu His Arg Ala Ala Ile Tyr Thr Ala Glu Ile
20 25 30
His Lys Ser Leu Ser Thr Asn Leu Asp Val Thr Asn Ser Ile Glu His
35 40 45
Gln Val Lys Asp Val Leu Thr Pro Leu Phe Lys Ile Ile Gly Asp Glu
50 55 60
Val Gly Leu Arg Thr Pro Gln Arg Phe Thr Asp Leu Val Lys Phe Ile
65 70 75 80
Ser Asp Lys Ile Lys Phe Leu Asn Pro Asp Arg Glu Tyr Asp Phe Arg
85 90 95
Asp Leu Thr Trp Cys Ile Asn Pro Pro Glu Arg Ile Lys Leu Asp Tyr
100 105 110
Asp Gln Tyr Cys Ala Asp Val Ala Ala Glu Glu Leu Met Asn Ala Leu
115 120 125
Val Asn Ser Thr Leu Leu Glu Thr Arg Thr Thr Asn Gln Phe Leu Ala
130 135 140
Val Ser Lys Gly Asn Cys Ser Gly Pro Thr Thr Ile Arg Gly Gln Phe
145 150 155 160
Ser Asn Met Ser Leu Ser Leu Leu Asp Leu Tyr Leu Gly Arg Gly Tyr
165 170 175
Asn Val Ser Ser Ile Val Thr Met Thr Ser Gln Gly Met Tyr Gly Gly
180 185 190
Thr Tyr Leu Val Glu Lys Pro Asn Leu Ser Ser Lys Arg Ser Glu Leu
195 200 205
Ser Gln Leu Ser Met Tyr Arg Val Phe Glu Val Gly Val Ile Arg Asn
210 215 220
Pro Gly Leu Gly Ala Pro Val Phe His Met Thr Asn Tyr Leu Glu Gln
225 230 235 240
Pro Val Ser Asn Asp Leu Ser Asn Cys Met Val Ala Leu Gly Glu Leu
245 250 255
Lys Leu Ala Ala Leu Cys His Gly Glu Asp Ser Ile Thr Ile Pro Tyr
260 265 270
Gln Gly Ser Gly Lys Gly Val Ser Phe Gln Leu Val Lys Leu Gly Val
275 280 285
Trp Lys Ser Pro Thr Asp Met Gln Ser Trp Val Pro Leu Ser Thr Asp
290 295 300
Asp Pro Val Ile Asp Arg Leu Tyr Leu Ser Ser His Arg Gly Val Ile
305 310 315 320
Ala Asp Asn Gln Ala Lys Trp Ala Val Pro Thr Thr Arg Thr Asp Asp
325 330 335
Lys Leu Arg Met Glu Thr Cys Phe Gln Gln Ala Cys Lys Gly Lys Ile
340 345 350
Gln Ala Leu Cys Glu Asn Pro Glu Trp Ala Pro Leu Lys Asp Asn Arg
355 360 365
Ile Pro Ser Tyr Gly Val Leu Ser Val Asp Leu Ser Leu Thr Val Glu
370 375 380
Leu Lys Ile Lys Ile Ala Ser Gly Phe Gly Pro Leu Ile Thr His Gly
385 390 395 400
Ser Gly Met Asp Leu Tyr Lys Ser Asn His Asn Asn Val Tyr Trp Leu
405 410 415
Thr Ile Pro Pro Met Lys Asn Leu Ala Leu Gly Val Ile Asn Thr Leu
420 425 430
Glu Trp Ile Pro Arg Phe Lys Val Ser Pro Tyr Leu Phe Asn Val Pro
435 440 445
Ile Lys Glu Ala Gly Glu Asp Cys His Ala Pro Thr Tyr Leu Pro Ala
450 455 460
Glu Val Asp Gly Asp Val Lys Leu Ser Ser Asn Leu Val Ile Leu Pro
465 470 475 480
Gly Gln Asp Leu Gln Tyr Val Leu Ala Thr Tyr Asp Thr Ser Arg Val
485 490 495
Glu His Ala Val Val Tyr Tyr Val Tyr Ser Pro Ser Arg Ser Phe Ser
500 505 510
Tyr Phe Tyr Pro Phe Arg Leu Pro Ile Lys Gly Val Pro Ile Glu Leu
515 520 525
Gln Val Glu Cys Phe Thr Trp Asp Gln Lys Leu Trp Cys Arg His Phe
530 535 540
Cys Val Leu Ala Asp Ser Glu Ser Gly Gly His Ile Thr His Ser Gly
545 550 555 560
Met Glu Gly Met Gly Val Ser Cys Thr Val Thr Arg Glu Asp Gly Thr
565 570 575
Asn Arg Arg
<210> 89
<211> 572
<212> PRT
<213> 人工序列
<220>
<223> 重组H蛋白
<400> 89
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Thr Ala Glu Ile His Lys Ser Leu Ser Thr Asn
20 25 30
Leu Asp Val Thr Asn Ser Ile Glu His Gln Val Lys Asp Val Leu Thr
35 40 45
Pro Leu Phe Lys Ile Ile Gly Asp Glu Val Gly Leu Arg Thr Pro Gln
50 55 60
Arg Phe Thr Asp Leu Val Lys Phe Ile Ser Asp Lys Ile Lys Phe Leu
65 70 75 80
Asn Pro Asp Arg Glu Tyr Asp Phe Arg Asp Leu Thr Trp Cys Ile Asn
85 90 95
Pro Pro Glu Arg Ile Lys Leu Asp Tyr Asp Gln Tyr Cys Ala Asp Val
100 105 110
Ala Ala Glu Glu Leu Met Asn Ala Leu Val Asn Ser Thr Leu Leu Glu
115 120 125
Thr Arg Thr Thr Asn Gln Phe Leu Ala Val Ser Lys Gly Asn Cys Ser
130 135 140
Gly Pro Thr Thr Ile Arg Gly Gln Phe Ser Asn Met Ser Leu Ser Leu
145 150 155 160
Leu Asp Leu Tyr Leu Gly Arg Gly Tyr Asn Val Ser Ser Ile Val Thr
165 170 175
Met Thr Ser Gln Gly Met Tyr Gly Gly Thr Tyr Leu Val Glu Lys Pro
180 185 190
Asn Leu Ser Ser Lys Arg Ser Glu Leu Ser Gln Leu Ser Met Tyr Arg
195 200 205
Val Phe Glu Val Gly Val Ile Arg Asn Pro Gly Leu Gly Ala Pro Val
210 215 220
Phe His Met Thr Asn Tyr Leu Glu Gln Pro Val Ser Asn Asp Leu Ser
225 230 235 240
Asn Cys Met Val Ala Leu Gly Glu Leu Lys Leu Ala Ala Leu Cys His
245 250 255
Gly Glu Asp Ser Ile Thr Ile Pro Tyr Gln Gly Ser Gly Lys Gly Val
260 265 270
Ser Phe Gln Leu Val Lys Leu Gly Val Trp Lys Ser Pro Thr Asp Met
275 280 285
Gln Ser Trp Val Pro Leu Ser Thr Asp Asp Pro Val Ile Asp Arg Leu
290 295 300
Tyr Leu Ser Ser His Arg Gly Val Ile Ala Asp Asn Gln Ala Lys Trp
305 310 315 320
Ala Val Pro Thr Thr Arg Thr Asp Asp Lys Leu Arg Met Glu Thr Cys
325 330 335
Phe Gln Gln Ala Cys Lys Gly Lys Ile Gln Ala Leu Cys Glu Asn Pro
340 345 350
Glu Trp Ala Pro Leu Lys Asp Asn Arg Ile Pro Ser Tyr Gly Val Leu
355 360 365
Ser Val Asp Leu Ser Leu Thr Val Glu Leu Lys Ile Lys Ile Ala Ser
370 375 380
Gly Phe Gly Pro Leu Ile Thr His Gly Ser Gly Met Asp Leu Tyr Lys
385 390 395 400
Ser Asn His Asn Asn Val Tyr Trp Leu Thr Ile Pro Pro Met Lys Asn
405 410 415
Leu Ala Leu Gly Val Ile Asn Thr Leu Glu Trp Ile Pro Arg Phe Lys
420 425 430
Val Ser Pro Tyr Leu Phe Asn Val Pro Ile Lys Glu Ala Gly Glu Asp
435 440 445
Cys His Ala Pro Thr Tyr Leu Pro Ala Glu Val Asp Gly Asp Val Lys
450 455 460
Leu Ser Ser Asn Leu Val Ile Leu Pro Gly Gln Asp Leu Gln Tyr Val
465 470 475 480
Leu Ala Thr Tyr Asp Thr Ser Arg Val Glu His Ala Val Val Tyr Tyr
485 490 495
Val Tyr Ser Pro Ser Arg Ser Phe Ser Tyr Phe Tyr Pro Phe Arg Leu
500 505 510
Pro Ile Lys Gly Val Pro Ile Glu Leu Gln Val Glu Cys Phe Thr Trp
515 520 525
Asp Gln Lys Leu Trp Cys Arg His Phe Cys Val Leu Ala Asp Ser Glu
530 535 540
Ser Gly Gly His Ile Thr His Ser Gly Met Glu Gly Met Gly Val Ser
545 550 555 560
Cys Thr Val Thr Arg Glu Asp Gly Thr Asn Arg Arg
565 570
<210> 90
<211> 550
<212> PRT
<213> 人工序列
<220>
<223> 重组HN蛋白
<400> 90
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Glu Leu Val Arg Met Ile Asn Asp Gln Gly Leu
20 25 30
Ser Asn Gln Leu Ser Ser Ile Thr Asp Lys Ile Arg Glu Ser Ala Thr
35 40 45
Met Ile Ala Ser Ala Val Gly Val Met Asn Gln Val Ile His Gly Val
50 55 60
Thr Val Ser Leu Pro Leu Gln Ile Glu Gly Asn Gln Asn Gln Leu Leu
65 70 75 80
Ala Thr Leu Ala Thr Ile Cys Thr Ser Gln Lys Gln Val Ser Asn Cys
85 90 95
Ser Thr Asn Ile Pro Leu Val Asn Asp Leu Arg Phe Ile Asn Gly Ile
100 105 110
Asn Lys Phe Ile Ile Glu Asp Tyr Ala Thr His Asp Phe Ser Ile Gly
115 120 125
His Pro Leu Asn Met Pro Ser Phe Ile Pro Thr Ala Thr Ser Pro Asn
130 135 140
Gly Cys Thr Arg Ile Pro Ser Phe Ser Leu Gly Lys Thr His Trp Cys
145 150 155 160
Tyr Thr His Asn Val Ile Asn Ala Asn Cys Lys Asp His Thr Ser Ser
165 170 175
Asn Gln Tyr Val Ser Met Gly Ile Leu Val Gln Thr Ala Ser Gly Tyr
180 185 190
Pro Met Phe Lys Thr Leu Lys Ile Gln Tyr Leu Ser Asp Gly Leu Asn
195 200 205
Arg Lys Ser Cys Ser Ile Ala Thr Val Pro Asp Gly Cys Ala Met Tyr
210 215 220
Cys Tyr Val Ser Thr Gln Leu Glu Thr Asp Asp Tyr Ala Gly Ser Ser
225 230 235 240
Pro Pro Thr Gln Lys Leu Thr Leu Leu Phe Tyr Asn Asp Thr Val Thr
245 250 255
Glu Arg Thr Ile Ser Pro Ser Gly Leu Glu Gly Asn Trp Ala Thr Leu
260 265 270
Val Pro Gly Val Gly Ser Gly Ile Tyr Phe Glu Asn Lys Leu Ile Phe
275 280 285
Pro Ala Tyr Gly Gly Val Leu Pro Asn Ser Thr Leu Gly Val Lys Ser
290 295 300
Ala Arg Glu Phe Phe Arg Pro Val Asn Pro Tyr Asn Pro Cys Ser Gly
305 310 315 320
Pro Gln Gln Asp Leu Asp Gln Arg Ala Leu Arg Ser Tyr Phe Pro Ser
325 330 335
Tyr Phe Ser Asn Arg Arg Ile Gln Ser Ala Phe Leu Val Cys Ala Trp
340 345 350
Asn Gln Ile Leu Val Thr Asn Cys Glu Leu Val Val Pro Ser Ser Asn
355 360 365
Gln Thr Met Met Gly Ala Glu Gly Arg Val Leu Leu Ile Asn Asn Arg
370 375 380
Leu Leu Tyr Tyr Gln Arg Ser Thr Ser Trp Trp Pro Tyr Glu Leu Leu
385 390 395 400
Tyr Glu Ile Ser Phe Thr Phe Thr Asn Ser Gly Pro Ser Ser Val Asn
405 410 415
Met Ser Trp Ile Pro Ile Tyr Ser Phe Thr Arg Pro Gly Ser Gly Asn
420 425 430
Cys Ser Gly Glu Asn Val Cys Pro Thr Ala Cys Val Ser Gly Val Tyr
435 440 445
Leu Asp Pro Trp Pro Leu Thr Pro Tyr Ser His Gln Ser Gly Ile Asn
450 455 460
Arg Asn Phe Tyr Phe Thr Gly Ala Leu Leu Asn Ser Ser Thr Thr Arg
465 470 475 480
Val Asn Pro Thr Leu Tyr Val Ser Ala Leu Asn Asn Leu Lys Val Leu
485 490 495
Ala Pro Tyr Gly Thr Gln Gly Leu Phe Ala Ser Tyr Thr Thr Thr Thr
500 505 510
Cys Phe Gln Asp Thr Gly Asp Ala Ser Val Tyr Cys Val Tyr Ile Met
515 520 525
Glu Leu Ala Ser Asn Ile Val Gly Glu Phe Gln Ile Leu Pro Val Leu
530 535 540
Thr Arg Leu Thr Ile Thr
545 550
<210> 91
<211> 543
<212> PRT
<213> 人工序列
<220>
<223> 重组HN蛋白
<400> 91
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Asp Gln Gly Leu Ser Asn Gln Leu Ser Ser Ile
20 25 30
Thr Asp Lys Ile Arg Glu Ser Ala Thr Met Ile Ala Ser Ala Val Gly
35 40 45
Val Met Asn Gln Val Ile His Gly Val Thr Val Ser Leu Pro Leu Gln
50 55 60
Ile Glu Gly Asn Gln Asn Gln Leu Leu Ala Thr Leu Ala Thr Ile Cys
65 70 75 80
Thr Ser Gln Lys Gln Val Ser Asn Cys Ser Thr Asn Ile Pro Leu Val
85 90 95
Asn Asp Leu Arg Phe Ile Asn Gly Ile Asn Lys Phe Ile Ile Glu Asp
100 105 110
Tyr Ala Thr His Asp Phe Ser Ile Gly His Pro Leu Asn Met Pro Ser
115 120 125
Phe Ile Pro Thr Ala Thr Ser Pro Asn Gly Cys Thr Arg Ile Pro Ser
130 135 140
Phe Ser Leu Gly Lys Thr His Trp Cys Tyr Thr His Asn Val Ile Asn
145 150 155 160
Ala Asn Cys Lys Asp His Thr Ser Ser Asn Gln Tyr Val Ser Met Gly
165 170 175
Ile Leu Val Gln Thr Ala Ser Gly Tyr Pro Met Phe Lys Thr Leu Lys
180 185 190
Ile Gln Tyr Leu Ser Asp Gly Leu Asn Arg Lys Ser Cys Ser Ile Ala
195 200 205
Thr Val Pro Asp Gly Cys Ala Met Tyr Cys Tyr Val Ser Thr Gln Leu
210 215 220
Glu Thr Asp Asp Tyr Ala Gly Ser Ser Pro Pro Thr Gln Lys Leu Thr
225 230 235 240
Leu Leu Phe Tyr Asn Asp Thr Val Thr Glu Arg Thr Ile Ser Pro Ser
245 250 255
Gly Leu Glu Gly Asn Trp Ala Thr Leu Val Pro Gly Val Gly Ser Gly
260 265 270
Ile Tyr Phe Glu Asn Lys Leu Ile Phe Pro Ala Tyr Gly Gly Val Leu
275 280 285
Pro Asn Ser Thr Leu Gly Val Lys Ser Ala Arg Glu Phe Phe Arg Pro
290 295 300
Val Asn Pro Tyr Asn Pro Cys Ser Gly Pro Gln Gln Asp Leu Asp Gln
305 310 315 320
Arg Ala Leu Arg Ser Tyr Phe Pro Ser Tyr Phe Ser Asn Arg Arg Ile
325 330 335
Gln Ser Ala Phe Leu Val Cys Ala Trp Asn Gln Ile Leu Val Thr Asn
340 345 350
Cys Glu Leu Val Val Pro Ser Ser Asn Gln Thr Met Met Gly Ala Glu
355 360 365
Gly Arg Val Leu Leu Ile Asn Asn Arg Leu Leu Tyr Tyr Gln Arg Ser
370 375 380
Thr Ser Trp Trp Pro Tyr Glu Leu Leu Tyr Glu Ile Ser Phe Thr Phe
385 390 395 400
Thr Asn Ser Gly Pro Ser Ser Val Asn Met Ser Trp Ile Pro Ile Tyr
405 410 415
Ser Phe Thr Arg Pro Gly Ser Gly Asn Cys Ser Gly Glu Asn Val Cys
420 425 430
Pro Thr Ala Cys Val Ser Gly Val Tyr Leu Asp Pro Trp Pro Leu Thr
435 440 445
Pro Tyr Ser His Gln Ser Gly Ile Asn Arg Asn Phe Tyr Phe Thr Gly
450 455 460
Ala Leu Leu Asn Ser Ser Thr Thr Arg Val Asn Pro Thr Leu Tyr Val
465 470 475 480
Ser Ala Leu Asn Asn Leu Lys Val Leu Ala Pro Tyr Gly Thr Gln Gly
485 490 495
Leu Phe Ala Ser Tyr Thr Thr Thr Thr Cys Phe Gln Asp Thr Gly Asp
500 505 510
Ala Ser Val Tyr Cys Val Tyr Ile Met Glu Leu Ala Ser Asn Ile Val
515 520 525
Gly Glu Phe Gln Ile Leu Pro Val Leu Thr Arg Leu Thr Ile Thr
530 535 540
<210> 92
<211> 541
<212> PRT
<213> 人工序列
<220>
<223> 重组HN蛋白
<400> 92
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Gly Leu Ser Asn Gln Leu Ser Ser Ile Thr Asp
20 25 30
Lys Ile Arg Glu Ser Ala Thr Met Ile Ala Ser Ala Val Gly Val Met
35 40 45
Asn Gln Val Ile His Gly Val Thr Val Ser Leu Pro Leu Gln Ile Glu
50 55 60
Gly Asn Gln Asn Gln Leu Leu Ala Thr Leu Ala Thr Ile Cys Thr Ser
65 70 75 80
Gln Lys Gln Val Ser Asn Cys Ser Thr Asn Ile Pro Leu Val Asn Asp
85 90 95
Leu Arg Phe Ile Asn Gly Ile Asn Lys Phe Ile Ile Glu Asp Tyr Ala
100 105 110
Thr His Asp Phe Ser Ile Gly His Pro Leu Asn Met Pro Ser Phe Ile
115 120 125
Pro Thr Ala Thr Ser Pro Asn Gly Cys Thr Arg Ile Pro Ser Phe Ser
130 135 140
Leu Gly Lys Thr His Trp Cys Tyr Thr His Asn Val Ile Asn Ala Asn
145 150 155 160
Cys Lys Asp His Thr Ser Ser Asn Gln Tyr Val Ser Met Gly Ile Leu
165 170 175
Val Gln Thr Ala Ser Gly Tyr Pro Met Phe Lys Thr Leu Lys Ile Gln
180 185 190
Tyr Leu Ser Asp Gly Leu Asn Arg Lys Ser Cys Ser Ile Ala Thr Val
195 200 205
Pro Asp Gly Cys Ala Met Tyr Cys Tyr Val Ser Thr Gln Leu Glu Thr
210 215 220
Asp Asp Tyr Ala Gly Ser Ser Pro Pro Thr Gln Lys Leu Thr Leu Leu
225 230 235 240
Phe Tyr Asn Asp Thr Val Thr Glu Arg Thr Ile Ser Pro Ser Gly Leu
245 250 255
Glu Gly Asn Trp Ala Thr Leu Val Pro Gly Val Gly Ser Gly Ile Tyr
260 265 270
Phe Glu Asn Lys Leu Ile Phe Pro Ala Tyr Gly Gly Val Leu Pro Asn
275 280 285
Ser Thr Leu Gly Val Lys Ser Ala Arg Glu Phe Phe Arg Pro Val Asn
290 295 300
Pro Tyr Asn Pro Cys Ser Gly Pro Gln Gln Asp Leu Asp Gln Arg Ala
305 310 315 320
Leu Arg Ser Tyr Phe Pro Ser Tyr Phe Ser Asn Arg Arg Ile Gln Ser
325 330 335
Ala Phe Leu Val Cys Ala Trp Asn Gln Ile Leu Val Thr Asn Cys Glu
340 345 350
Leu Val Val Pro Ser Ser Asn Gln Thr Met Met Gly Ala Glu Gly Arg
355 360 365
Val Leu Leu Ile Asn Asn Arg Leu Leu Tyr Tyr Gln Arg Ser Thr Ser
370 375 380
Trp Trp Pro Tyr Glu Leu Leu Tyr Glu Ile Ser Phe Thr Phe Thr Asn
385 390 395 400
Ser Gly Pro Ser Ser Val Asn Met Ser Trp Ile Pro Ile Tyr Ser Phe
405 410 415
Thr Arg Pro Gly Ser Gly Asn Cys Ser Gly Glu Asn Val Cys Pro Thr
420 425 430
Ala Cys Val Ser Gly Val Tyr Leu Asp Pro Trp Pro Leu Thr Pro Tyr
435 440 445
Ser His Gln Ser Gly Ile Asn Arg Asn Phe Tyr Phe Thr Gly Ala Leu
450 455 460
Leu Asn Ser Ser Thr Thr Arg Val Asn Pro Thr Leu Tyr Val Ser Ala
465 470 475 480
Leu Asn Asn Leu Lys Val Leu Ala Pro Tyr Gly Thr Gln Gly Leu Phe
485 490 495
Ala Ser Tyr Thr Thr Thr Thr Cys Phe Gln Asp Thr Gly Asp Ala Ser
500 505 510
Val Tyr Cys Val Tyr Ile Met Glu Leu Ala Ser Asn Ile Val Gly Glu
515 520 525
Phe Gln Ile Leu Pro Val Leu Thr Arg Leu Thr Ile Thr
530 535 540
<210> 93
<211> 549
<212> PRT
<213> 人工序列
<220>
<223> 重组HN蛋白
<400> 93
Met Tyr Ser Met Gln Leu Ala Ser Cys Val Thr Leu Thr Leu Val Leu
1 5 10 15
Leu Val Asn Ser Gln Leu Val Arg Met Ile Asn Asp Gln Gly Leu Ser
20 25 30
Asn Gln Leu Ser Ser Ile Thr Asp Lys Ile Arg Glu Ser Ala Thr Met
35 40 45
Ile Ala Ser Ala Val Gly Val Met Asn Gln Val Ile His Gly Val Thr
50 55 60
Val Ser Leu Pro Leu Gln Ile Glu Gly Asn Gln Asn Gln Leu Leu Ala
65 70 75 80
Thr Leu Ala Thr Ile Cys Thr Ser Gln Lys Gln Val Ser Asn Cys Ser
85 90 95
Thr Asn Ile Pro Leu Val Asn Asp Leu Arg Phe Ile Asn Gly Ile Asn
100 105 110
Lys Phe Ile Ile Glu Asp Tyr Ala Thr His Asp Phe Ser Ile Gly His
115 120 125
Pro Leu Asn Met Pro Ser Phe Ile Pro Thr Ala Thr Ser Pro Asn Gly
130 135 140
Cys Thr Arg Ile Pro Ser Phe Ser Leu Gly Lys Thr His Trp Cys Tyr
145 150 155 160
Thr His Asn Val Ile Asn Ala Asn Cys Lys Asp His Thr Ser Ser Asn
165 170 175
Gln Tyr Val Ser Met Gly Ile Leu Val Gln Thr Ala Ser Gly Tyr Pro
180 185 190
Met Phe Lys Thr Leu Lys Ile Gln Tyr Leu Ser Asp Gly Leu Asn Arg
195 200 205
Lys Ser Cys Ser Ile Ala Thr Val Pro Asp Gly Cys Ala Met Tyr Cys
210 215 220
Tyr Val Ser Thr Gln Leu Glu Thr Asp Asp Tyr Ala Gly Ser Ser Pro
225 230 235 240
Pro Thr Gln Lys Leu Thr Leu Leu Phe Tyr Asn Asp Thr Val Thr Glu
245 250 255
Arg Thr Ile Ser Pro Ser Gly Leu Glu Gly Asn Trp Ala Thr Leu Val
260 265 270
Pro Gly Val Gly Ser Gly Ile Tyr Phe Glu Asn Lys Leu Ile Phe Pro
275 280 285
Ala Tyr Gly Gly Val Leu Pro Asn Ser Thr Leu Gly Val Lys Ser Ala
290 295 300
Arg Glu Phe Phe Arg Pro Val Asn Pro Tyr Asn Pro Cys Ser Gly Pro
305 310 315 320
Gln Gln Asp Leu Asp Gln Arg Ala Leu Arg Ser Tyr Phe Pro Ser Tyr
325 330 335
Phe Ser Asn Arg Arg Ile Gln Ser Ala Phe Leu Val Cys Ala Trp Asn
340 345 350
Gln Ile Leu Val Thr Asn Cys Glu Leu Val Val Pro Ser Ser Asn Gln
355 360 365
Thr Met Met Gly Ala Glu Gly Arg Val Leu Leu Ile Asn Asn Arg Leu
370 375 380
Leu Tyr Tyr Gln Arg Ser Thr Ser Trp Trp Pro Tyr Glu Leu Leu Tyr
385 390 395 400
Glu Ile Ser Phe Thr Phe Thr Asn Ser Gly Pro Ser Ser Val Asn Met
405 410 415
Ser Trp Ile Pro Ile Tyr Ser Phe Thr Arg Pro Gly Ser Gly Asn Cys
420 425 430
Ser Gly Glu Asn Val Cys Pro Thr Ala Cys Val Ser Gly Val Tyr Leu
435 440 445
Asp Pro Trp Pro Leu Thr Pro Tyr Ser His Gln Ser Gly Ile Asn Arg
450 455 460
Asn Phe Tyr Phe Thr Gly Ala Leu Leu Asn Ser Ser Thr Thr Arg Val
465 470 475 480
Asn Pro Thr Leu Tyr Val Ser Ala Leu Asn Asn Leu Lys Val Leu Ala
485 490 495
Pro Tyr Gly Thr Gln Gly Leu Phe Ala Ser Tyr Thr Thr Thr Thr Cys
500 505 510
Phe Gln Asp Thr Gly Asp Ala Ser Val Tyr Cys Val Tyr Ile Met Glu
515 520 525
Leu Ala Ser Asn Ile Val Gly Glu Phe Gln Ile Leu Pro Val Leu Thr
530 535 540
Arg Leu Thr Ile Thr
545
<210> 94
<211> 1653
<212> DNA
<213> 麻疹病毒
<400> 94
atgggtctca aggtgaacgt ctctgccata ttcatggcag tactgttaac tctccaaaca 60
cccaccggtc aaatccattg gggcaatctc tctaagatag gggtggtagg aataggaagt 120
gcaagctaca aagttatgac tcgttccagc catcaatcat tagtcataaa attaatgccc 180
aatataactc tcctcaataa ctgcacgagg gtagagattg cagaatacag gagactactg 240
agaacagttt tggaaccaat tagagatgca cttaatgcaa tgacccagaa tataagaccg 300
gttcagagtg tagcttcaag taggagacac aagagatttg cgggagtagt cctggcaggt 360
gcggccctag gcgttgccac agctgctcag ataacagccg gcattgcact tcaccagtcc 420
atgctgaact ctcaagccat cgacaatctg agagcgagcc tggaaactac taatcaggca 480
attgaggcaa tcagacaagc agggcaggag atgatattgg ctgttcaggg tgtccaagac 540
tacatcaata atgagctgat accgtctatg aaccaactat cttgtgattt aatcggccag 600
aagctcgggc tcaaattgct cagatactat acagaaatcc tgtcattatt tggccccagc 660
ttacgggacc ccatatctgc ggagatatct atccaggctt tgagctatgc gcttggagga 720
gacatcaata aggtgttaga aaagctcgga tacagtggag gtgatttact gggcatctta 780
gagagcagag gaataaaggc ccggataact cacgtcgaca cagagtccta cttaattgtc 840
ctcagtatag cctatccgac gctgtccgag attaaggggg tgattgtcca ccggctagag 900
ggggtctcgt acaacatagg ctctcaagag tggtatacca ctgtgcccaa gtatgttgca 960
acccaagggt accttatctc gaattttgat gagtcatcgt gtactttcat gccagagggg 1020
actgtgtgca gccaaaatgc cttgtacccg atgagtcctc tgctccaaga atgcctccgg 1080
gggtccacca agtcctgtgc tcgtacactc gtatccgggt cttttgggaa ccggttcatt 1140
ttatcacaag ggaacctaat agccaattgt gcatcaatcc tttgcaagtg ttacacaaca 1200
ggaacgatca ttaatcaaga ccctgacaag atcctaacat acattgctgc cgatcactgc 1260
ccggtagtcg aggtgaacgg cgtgaccatc caagtcggga gcaggaggta tccagacgct 1320
gtgtacttgc acagaattga cctcggtcct cccatattat tggagaggtt ggacgtaggg 1380
acaaatctgg ggaatgcaat tgctaagttg gaggatgcca aggaattgtt ggagtcatcg 1440
gaccagatat tgaggagtat gaaaggttta tcgagcactt gcatagtcta catcctgatt 1500
gcagtgtgtc ttggagggtt gatagggatc cccgctttaa tatgttgctg cagggggcgt 1560
tgtaacaaaa agggagaaca agttggtatg tcaagaccag gcctaaagcc tgatcttacg 1620
ggaacatcaa aatcctatgt aaggtcgctc tga 1653
<210> 95
<211> 1617
<212> DNA
<213> 腮腺炎病毒
<400> 95
atgaaggcct tttcagttac ttgcttgggc tttgcagtct tttcgtcttc tatatgtgtg 60
aatatcaaca tcttgcagca aattggatat atcaagcaac aagtcaggca actaagctat 120
tactcacaaa gttctagctc ctacatagtg gtcaagcttt taccgaatat ccaacccact 180
gataacagct gtgaatttaa gagtgtaact caatacaata agaccttgag taacttgctt 240
cttcccattg cagaaaacat aaacaatatt gcatcgccct cacctgggtc aagacgtcat 300
aaaaggtttg ctggcattgc cattggcatt gctgcgctcg gtgttgcgac cgcagcacag 360
gtaactgccg ctgtctcatt agttcaagca cagacaaatg cacgtgcaat agcggcgatg 420
aaaaattcaa tacaggcaac taatcgggca atcttcgaag tgaaggaagg cacccaacag 480
ttagctatag cggtacaagc aatacaagac cacatcaata ctattatgaa cacccaattg 540
aacaatatgt cttgtcagat ccttgataac cagcttgcaa cctacctagg attataccta 600
acagaattaa caacagtgtt tcagccacaa ttaattaatc cggcattgtc accgattagt 660
atacaagcct tgaggtcttt gcttggaagt atgacgcctg cagtggttca agcaacatta 720
tctacgtcaa tttctgctgc tgaaatacta agtgccggtc taatggaggg tcagattgtt 780
tctgttctgc tagatgagat gcagatgata gtcaagataa atattccaac cattgtcaca 840
caatcaaatg cattggtgat tgacttctac tcaatttcga gctttattaa taatcaagaa 900
tccataattc aattgccaga caggatccta gagatcggga atgaacaatg gagctatcca 960
gctaaaaatt gtaagttgac aagacaccac atattctgcc aatacaatga ggcagagagg 1020
ctgagcctag aatcaaaact atgccttgca ggtaatataa gtgcctgtgt gttctcaccc 1080
atagcaggga gttatatgag gcgatttgta gcactggatg gaacaattgt tgcaaactgt 1140
cgaagtctaa cgtgtctatg caagagtcca tcttatccta tataccaacc tgaccatcat 1200
gcagtcacga ccattgatct aaccacatgt caaacattgt ccctagacgg attggacttc 1260
agcattgtct ctctaagcaa catcacttac gctgagaacc ttaccatttc attgtctcaa 1320
acaatcaata ctcaacccat tgacatatca actgaactga gtaaagttaa tgcatccctc 1380
caaaatgccg ttaagtacat aaaggagagc aaccatcaac tccaatctgt gagtgtaaat 1440
tccaaaattg gagctataat tgtagcagcc ttagttttga gcatcctgtc aattatcatt 1500
tcgctattgt tttgctgctg ggcttacatt gcaactaaag aaatcagaag aatcaacttc 1560
aaaacaaatc atatcaacac aatatcgagt agtgtcgatg atctcatcag gtactaa 1617
<210> 96
<211> 1854
<212> DNA
<213> 麻疹病毒
<400> 96
atgtcaccac aacgagaccg gataaatgcc ttctacaaag ataaccccca tcccaaggga 60
agtaggatag tcattaacag agaacatctt atgattgata gaccttatgt tttgctggct 120
gttctgtttg tcatgtttct gagcttgatc gggttgctag ccattgcagg cattagactt 180
catcgggcag ccatctacac cgcagagatc cataaaagcc tcagcaccaa tctagatgta 240
actaactcaa tcgagcatca ggtcaaggac gtgctgacac cactcttcaa aatcatcggt 300
gatgaagtgg gcctgaggac acctcagaga ttcactgacc tagtgaaatt catctctgac 360
aagattaaat tccttaatcc ggatagggag tacgacttca gagatctcac ttggtgtatc 420
aacccgccag agagaatcaa attggattat gatcaatact gtgcagatgt ggctgctgaa 480
gagctcatga atgcattggt gaactcaact ctactggaga ccagaacaac caatcagttc 540
ctagctgtct caaagggaaa ctgctcaggg cccactacaa tcagaggtca attctcaaac 600
atgtcgctgt ccctgttaga cttgtattta ggtcgaggtt acaatgtgtc atctatagtc 660
actatgacat cccagggaat gtatggggga acttacctag tggaaaagcc taatctgagc 720
agcaaaaggt cagagttgtc acaactgagc atgtaccgag tgtttgaagt aggtgttatc 780
agaaatccgg gtttgggggc tccggtgttc catatgacaa actatcttga gcaaccagtc 840
agtaatgatc tcagcaactg tatggtggct ttgggggagc tcaaactcgc agccctttgt 900
cacggggaag attctatcac aattccctat cagggatcag ggaaaggtgt cagcttccag 960
ctcgtcaagc taggtgtctg gaaatcccca accgacatgc aatcctgggt ccccttatca 1020
acggatgatc cagtgataga caggctttac ctctcatctc acagaggtgt tatcgctgac 1080
aatcaagcaa aatgggctgt cccgacaaca cgaacagatg acaagttgcg aatggagaca 1140
tgcttccaac aggcgtgtaa gggtaaaatc caagcactct gcgagaatcc cgagtgggca 1200
ccattgaagg ataacaggat tccttcatac ggggtcttgt ctgttgatct gagtctgaca 1260
gttgagctta aaatcaaaat tgcttcggga ttcgggccat tgatcacaca cggttcaggg 1320
atggacctat acaaatccaa ccacaacaat gtgtattggc tgactatccc gccaatgaag 1380
aacctagcct taggtgtaat caacacattg gagtggatac cgagattcaa ggttagtccc 1440
tacctcttca ctgtcccaat taaggaagca ggcgaagact gccatgcccc aacataccta 1500
cctgcggagg tggatggtga tgtcaaactc agttccaatc tggtgattct acctggtcaa 1560
gatctccaat atgttttggc aacctacgat acttccaggg ttgaacatgc tgtggtttat 1620
tacgtttaca gcccaagccg ctcattttct tacttttatc cttttaggtt gcctataaag 1680
ggggtcccca tcgaattaca agtggaatgc ttcacatggg accaaaaact ctggtgccgt 1740
cacttctgtg tgcttgcgga ctcagaatct ggtggacata tcactcactc tgggatggtg 1800
ggcatgggag tcagctgcac agtcacccgg gaagatggaa ccaatcgcag atag 1854
<210> 97
<211> 1749
<212> DNA
<213> 腮腺炎病毒
<400> 97
atggagccct cgaaattctt cacaatatcg gacagtgcca cctttgcacc tgggcctgtc 60
agcaatgcgg ctgacaagaa gacattccga acctgcttcc gaatactggt actatctgta 120
caagctgtca ccctcatatt ggttattgtc actttaggtg agcttgtaag gatgatcaat 180
gatcaaggct tgagcaatca gttgtcttca attacagaca agataagaga gtcagctact 240
atgattgcat ctgctgtggg agtaatgaat caagttattc atggagtaac ggtatcctta 300
cccctacaaa ttgagggaaa ccaaaatcaa ttgttagcca cacttgccac aatctgcgcc 360
agccaaaaac aagtctcaaa ctgctctaca aacatcccct tagtcaatga cctcaggttt 420
ataaatggga tcaataaatt tattattgaa gattacgcaa ctcatgattt ctctatcggc 480
catccactca atatgcccag ctttatccca actgcaactt cacccaatgg ttgcacaaga 540
attccatcct tttctttagg taagacacac tggtgctaca cacataatgt aattaatgcc 600
aactgcaagg accatacttc gtctaaccaa tatgtgtcca tggggattct cgttcagacc 660
gcgtcagggt atcctatgtt caaaacctta aaaatccaat atctcagtga tggcctgaat 720
cggaaaagct gctcaattgc aacagtccct gatgggtgcg cgatgtactg ttatgtctct 780
actcaacttg aaaccgacga ctatgcgggg tccagtccac ccacccaaaa acttaccctg 840
ttattctata atgacaccgt cacagaaagg acaatatctc catctggtct tgaagggaat 900
tgggctactt tggtgccagg agtggggagt gggatatatt ttgagaataa gttgatcttc 960
cctgcatatg ggggtgtctt gcccaatagt acactcgggg ttaaatcagc aagagaattt 1020
tttcggcctg ttaatccata taatccatgt tcaggaccac aacaagattt agatcagcgt 1080
gctttgaggt catacttccc aagttacttc tctaatcgaa gaatacagag tgcatttctt 1140
gtctgtgcct ggaatcagat cctagttaca aattgtgagc tagttgtccc ctcaagcaat 1200
cagacaatga tgggtgcaga agggagagtt ttattgatca ataatcgact attatattat 1260
cagagaagta ccagctggtg gccgtatgaa ctcctctacg agatatcatt cacatttaca 1320
aactctggtc catcatctgt aaatatgtcc tggataccta tatattcatt cactcgtcct 1380
ggttcaggca attgcagtgg tgaaaatgtg tgcccgactg cttgtgtgtc aggggtttat 1440
cttgatccct ggccattaac tccatatagc caccaatcag gtattaacag aaatttctat 1500
ttcacaggtg cactattaaa ttcaagtaca actagagtaa atcctaccct ttatgtctct 1560
gcccttaata atcttaaagt actagcccca tatggtactc aaggactgtt tgcctcgtac 1620
accacaacca cctgctttca agataccggt gatgctagtg tgtattgtgt ttatattatg 1680
gagctagcat caaatattgt tggagaattc caaattctac ctgtgctaac tagattgact 1740
atcacttga 1749
<210> 98
<211> 476
<212> PRT
<213> 腮腺炎病毒
<400> 98
Met Lys Ala Tyr Pro Val Ile Cys Leu Gly Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Val Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asn Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Lys Arg Phe Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Ile Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Tyr Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Ile Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Val Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Ser Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Thr Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Ser Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser
465 470 475
<210> 99
<211> 55
<212> PRT
<213> 人工序列
<220>
<223> 跨膜结构域和胞质尾
<400> 99
Gly Ala Ile Ile Val Ala Ala Leu Val Leu Ser Ile Leu Ser Ile Ile
1 5 10 15
Ile Ser Leu Leu Phe Cys Cys Trp Ala Tyr Ile Ala Thr Lys Glu Ile
20 25 30
Arg Arg Ile Asn Phe Lys Thr Asn His Ile Asn Thr Ile Ser Ser Ser
35 40 45
Val Asp Asp Leu Ile Arg Tyr
50 55
<210> 100
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> 跨膜结构域
<400> 100
Cys Ile Val Tyr Ile Leu Ile Ala Val Cys Leu Gly Gly Leu Ile Gly
1 5 10 15
Ile
<210> 101
<211> 57
<212> PRT
<213> 人工序列
<220>
<223> 跨膜结构域和胞质尾
<400> 101
Cys Ile Val Tyr Ile Leu Ile Ala Val Cys Leu Gly Gly Leu Ile Gly
1 5 10 15
Ile Pro Ala Leu Ile Cys Cys Cys Arg Gly Arg Cys Asn Lys Lys Gly
20 25 30
Glu Gln Val Gly Met Ser Arg Pro Gly Leu Lys Pro Asp Leu Thr Gly
35 40 45
Thr Ser Lys Ser Tyr Val Arg Ser Leu
50 55
<210> 102
<211> 476
<212> PRT
<213> 腮腺炎病毒
<400> 102
Met Lys Ala Phe Leu Val Thr Cys Leu Gly Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asn Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Lys Arg Phe Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Ile Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Tyr Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Ile Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Val Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Thr Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser
465 470 475
<210> 103
<211> 476
<212> PRT
<213> 腮腺炎病毒
<400> 103
Met Lys Ala Phe Ser Val Thr Cys Leu Ser Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Glu Asn Ser Cys
50 55 60
Glu Phe Lys Ser Ile Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Lys Arg Phe Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Ile Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Leu Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Tyr Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Ile Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Val Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Thr Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser
465 470 475
<210> 104
<211> 476
<212> PRT
<213> 腮腺炎病毒
<400> 104
Met Lys Ala Phe Leu Val Ile Cys Leu Gly Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Val Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asn Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Gly Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Lys Arg Phe Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Ile Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Ser Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Tyr Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Ile Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Val Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His Asn Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Thr Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn Gln Gln Leu Gln Ser
465 470 475
<210> 105
<211> 476
<212> PRT
<213> 腮腺炎病毒
<400> 105
Met Lys Ala Ser Leu Val Thr Cys Leu Gly Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Thr Asp Asn Ser Cys
50 55 60
Glu Phe Lys Ser Val Thr Gln Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Lys Arg Phe Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Ile Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asn His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Tyr Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Ile Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Thr Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Val Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His Asn Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Ile Asp Leu Thr Thr Cys Gln Met Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ile Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser
465 470 475
<210> 106
<211> 476
<212> PRT
<213> 腮腺炎病毒
<400> 106
Met Lys Ala Phe Ser Val Thr Cys Leu Gly Phe Ala Val Phe Ser Ser
1 5 10 15
Ser Ile Cys Val Asn Ile Asn Ile Leu Gln Gln Ile Gly Tyr Ile Lys
20 25 30
Gln Gln Val Arg Gln Leu Ser Tyr Tyr Ser Gln Ser Ser Ser Ser Tyr
35 40 45
Ile Val Val Lys Leu Leu Pro Asn Ile Gln Pro Pro Asp Asp Ser Cys
50 55 60
Glu Phe Lys Ser Val Ile Ser Tyr Asn Lys Thr Leu Ser Asn Leu Leu
65 70 75 80
Leu Pro Ile Ala Glu Asn Ile Asn Asn Ile Ala Ser Pro Ser Pro Gly
85 90 95
Ser Arg Arg His Lys Arg Phe Ala Gly Ile Ala Ile Gly Ile Ala Ala
100 105 110
Leu Gly Val Ala Thr Ala Ala Gln Val Thr Ala Ala Val Ser Leu Val
115 120 125
Gln Ala Gln Thr Asn Ala Arg Ala Ile Ala Ala Met Lys Asn Ser Ile
130 135 140
Gln Ala Thr Asn Arg Ala Ile Phe Glu Val Lys Glu Gly Thr Gln Gln
145 150 155 160
Leu Ala Ile Ala Val Gln Ala Ile Gln Asp His Ile Asn Thr Ile Met
165 170 175
Asn Thr Gln Leu Asn Asn Met Ser Cys Gln Ile Leu Asp Asn Gln Leu
180 185 190
Ala Thr Tyr Leu Gly Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe Gln
195 200 205
Pro Gln Leu Ile Asn Pro Ala Leu Ser Pro Ile Ser Ile Gln Ala Leu
210 215 220
Arg Ser Leu Leu Gly Ser Met Thr Pro Ala Val Val Gln Ala Ala Leu
225 230 235 240
Ser Thr Ser Ile Ser Ala Ala Glu Ile Leu Ser Ala Gly Leu Met Glu
245 250 255
Gly Gln Ile Val Ser Val Leu Leu Asp Glu Met Gln Met Ile Val Lys
260 265 270
Ile Asn Ile Pro Thr Ile Val Thr Gln Ser Asn Ala Leu Val Ile Asp
275 280 285
Phe Tyr Ser Ile Ser Ser Phe Ile Asn Asn Gln Glu Ser Ile Ile Gln
290 295 300
Leu Pro Asp Arg Ile Leu Glu Ile Gly Asn Glu Gln Trp Ser Tyr Pro
305 310 315 320
Ala Lys Asn Cys Lys Leu Thr Arg His His Ile Phe Cys Gln Tyr Asn
325 330 335
Glu Ala Glu Arg Leu Ser Leu Glu Ser Lys Leu Cys Leu Ala Gly Asn
340 345 350
Ile Ser Ala Cys Val Phe Ser Pro Ile Ala Gly Ser Tyr Met Arg Arg
355 360 365
Phe Val Ala Leu Asp Gly Thr Ile Val Ala Asn Cys Arg Ser Leu Thr
370 375 380
Cys Leu Cys Lys Ser Pro Ser Tyr Pro Ile Tyr Gln Pro Asp His His
385 390 395 400
Ala Val Thr Thr Val Asp Leu Thr Thr Cys Gln Thr Leu Ser Leu Asp
405 410 415
Gly Leu Asp Phe Ser Ile Val Ser Leu Ser Asn Ile Thr Tyr Ala Glu
420 425 430
Asn Leu Thr Ile Ser Leu Ser Gln Thr Ile Asn Thr Gln Pro Ile Asp
435 440 445
Ile Ser Thr Glu Leu Ser Lys Val Asn Ala Ser Leu Gln Asn Ala Val
450 455 460
Lys Tyr Ile Lys Glu Ser Asn His Gln Leu Gln Ser
465 470 475

Claims (62)

1.一种免疫原，所述免疫原包含：
重组腮腺炎病毒(MuV)F胞外域三聚体，其通过所述三聚体的原体中的一个或更多个氨基酸取代以融合前构象稳定，所述氨基酸取代包括形成非天然二硫键以使所述MuV F胞外域三聚体以融合前构象稳定的半胱氨酸取代。
2.根据权利要求1所述的免疫原，其中所述半胱氨酸取代位于MuV F第86和215位、第155和161位、第165和231位、第206和223位、第209和214位以及第221和255位中的一个或更多个位置。
3.根据权利要求1或2所述的免疫原，其中所述重组MuV F胞外域三聚体通过位于MuV F第206和223位的所述三聚体的原体中的所述半胱氨酸取代之间的非天然二硫键以融合前构象稳定。
4.根据前述权利要求中任一项所述的免疫原，其中位于：
MuV F第86和215位的所述半胱氨酸取代是N86C和A215C取代；
MuV F第155和161位的所述半胱氨酸取代是K155C和L161C取代；
MuV F第165和231位的所述半胱氨酸取代是V165C和M231C取代；
MuV F第206和223位的所述半胱氨酸取代是V206C和A223C取代；
MuV F第209和214位的所述半胱氨酸取代是P209C和P214C取代；以及
MuV F第221和255位的所述半胱氨酸取代是I221C和M255C取代。
5.根据前述权利要求中任一项所述的免疫原，其中所述重组MuV F胞外域三聚体的原体包含F2蛋白和F1胞外域，所述F2蛋白包含MuV F第20-100位或由其组成，所述F1胞外域包含MuV F第104-469、104-476或104-483位或由其组成。
6.根据前述权利要求中任一项所述的免疫原，其中所述三聚体的原体进一步包含去除所述MuV F胞外域的F1/F2弗林蛋白酶切割位点和任选地F1胞外域的融合肽的第一个残基的突变。
7.根据权利要求6所述的免疫原，其中去除所述F1/F2弗林蛋白酶切割位点和所述融合肽的第一个残基的所述突变包含MuV F第101-103位的缺失，其中第100和104位通过肽接头融合。
8.根据权利要求7所述的免疫原，其中所述肽接头是Gly-Gly-Gly接头。
9.根据前述权利要求中任一项所述的免疫原，其中所述氨基酸取代的定位是根据如SEQ ID NO:1所示的参考MuV F蛋白序列。
10.根据前述权利要求中任一项所述的免疫原，其中所述MuV F胞外域三聚体的原体包含与以下项具有至少90％同一性的氨基酸序列
SEQ ID NO:3-8中任一项的残基20-483，
SEQ ID NO:11-16、26或51中任一项的残基20-476，或
SEQ ID NO:19-24中任一项的残基20-469；以及
其中所述原体包含使所述MuV F胞外域三聚体以融合前构象稳定的一个或更多个氨基酸取代。
11.根据权利要求10所述的免疫原，其中所述MuV F胞外域三聚体的原体包含如下所示的氨基酸序列或由其组成
SEQ ID NO:3-8中任一项的残基20-483，
SEQ ID NO:11-16、26或51中任一项的残基20-476，或
SEQ ID NO:19-24中任一项的残基20-469。
12.根据前述权利要求中任一项所述的免疫原，其中所述重组MuV F胞外域三聚体的原体在C-末端与三聚化结构域融合。
13.根据权利要求12所述的免疫原，其中所述三聚化结构域包含GCN4三聚化结构域、T4纤维蛋白三聚化结构域或两者。
14.根据权利要求13所述的免疫原，其中
所述GCN4三聚化结构域包含如IEDKIEEILSKIYHIENEIARIKKLIGEAP(SEQ ID NO:33)所示的氨基酸序列，
所述T4纤维蛋白三聚化结构域包含如GYIPEAPRDGQAYVRKDGEWVLLSTFL SEQ ID NO:34)所示的氨基酸序列，
包含所述GCN4和T4纤维蛋白三聚化结构域的三聚化结构域包含如IEDKIEEILSKIYHIENEIARIKKLIGEAPGSGYIPEAPRDGQAYVRKDGEWVLLSTFL(SEQ ID NO:35)所示的氨基酸序列。
15.根据权利要求12-14中任一项所述的免疫原，其中与所述三聚化结构域融合的所述MuV F胞外域三聚体的原体包含与以下项具有至少90％同一性的氨基酸序列
SEQ ID NO:3-8中任一项的残基20-513，
SEQ ID NO:11-16、26或51中任一项的残基20-506，或
SEQ ID NO:19-24中任一项的残基20-499；以及
其中所述原体包含使所述MuV F胞外域三聚体以融合前构象稳定的一个或更多个氨基酸取代。
16.根据权利要求15所述的免疫原，其中与所述三聚化结构域融合的所述MuV F胞外域三聚体的原体包含如下所示的氨基酸序列或由其组成
SEQ ID NO:3-8中任一项的残基20-513，
SEQ ID NO:11-16、26或51中任一项的残基20-506，或
SEQ ID NO:19-24中任一项的残基20-499。
17.根据前述权利要求中任一项所述的免疫原，其中所述重组MuV F胞外域三聚体的原体与异源蛋白连接。
18.根据权利要求17所述的免疫原，其中所述异源蛋白是MeV H蛋白或MuV HN蛋白的胞外域头部或胞外域茎部和头部。
19.根据权利要求18所述的免疫原，其中所述MeV H蛋白或所述MuV HN蛋白的胞外域头部或胞外域茎部和头部在C-末端与三聚化结构域融合，所述三聚化结构域在C-末端与所述重组MuV F胞外域三聚体的原体融合。
20.根据权利要求19所述的免疫原，其中与所述MeV H蛋白的三聚化结构域和胞外域或MuV HN蛋白的胞外域连接的所述MuV F胞外域三聚体的原体包含如SEQ ID NO:27的残基20-966、SEQ ID NO:28的残基21-981或SEQ ID NO:29的残基20-1006所示的氨基酸序列。
21.一种免疫原，所述免疫原包含：
重组麻疹病毒(MeV)F胞外域三聚体，其通过所述三聚体的原体中的一个或更多个氨基酸取代以融合前构象稳定，所述氨基酸取代包括以下项中的一项或多项：
位于MeV F第48和284位、第90和225位、第141和270位、第165和171位、第173和245位、第175和241位、第212和236位、第216和233位、第219和224位、第99和117位、第100和117位、第101和117位、第102和117位以及第103和117位的形成非天然二硫键的半胱氨酸取代；
位于MeV F第175位的苯丙氨酸取代；以及
位于MeV F第194位的脯氨酸取代。
22.根据权利要求21所述的免疫原，其中所述重组MeV F胞外域三聚体通过位于MeV F第165和171位的所述三聚体的原体中的所述半胱氨酸取代之间的非天然二硫键以融合前构象稳定。
23.根据权利要求21或22所述的免疫原，其中：
位于：
MeV F第48和284位的所述半胱氨酸取代是R48C和A284C取代；
MeV F第90和225位的所述半胱氨酸取代是A90C和I225C取代；
MeV F第141和270位的所述半胱氨酸取代是M141C和T270C取代；
MeV F第165和171位的所述半胱氨酸取代是R165C和M171C取代；
MeV F第173和245位的所述半胱氨酸取代是L173C和V245C取代；
MeV F第175和241位的所述半胱氨酸取代是V175C和D241C取代；
MeV F第212和236位的所述半胱氨酸取代是E212C和Y236C取代；
MeV F第216和233位的所述半胱氨酸取代是L216C和A233C取代；
MeV F第219和224位的所述半胱氨酸取代是P219C和P224C取代；
MeV F第99和117位的所述半胱氨酸取代是R99C-V117C取代；
MeV F第100和117位的所述半胱氨酸取代是P100C-V117C取代；
MeV F第101和117位的所述半胱氨酸取代是V101C-V117C取代；
MeV F第102和117位的所述半胱氨酸取代是Q102C-V117C取代；
MeV F第103和117位的所述半胱氨酸取代是S103C-V117C取代；
位于MeV F第175位的所述苯丙氨酸取代是V175F取代；以及
位于MeV F第194位的所述脯氨酸取代是S194P取代。
24.根据权利要求21-23中任一项所述的免疫原，其中所述重组MeV F胞外域三聚体的原体包含F2蛋白和F1胞外域，所述F2蛋白包含MeV F第24-110位或由其组成，所述F1胞外域包含MeV F第114-486位或由其组成。
25.根据权利要求21-24中任一项所述的免疫原，其中所述三聚体的原体进一步包含去除所述MeV F胞外域的F1/F2弗林蛋白酶切割位点和任选地F1胞外域的融合肽的第一个残基的突变。
26.根据权利要求25所述的免疫原，其中去除所述F1/F2弗林蛋白酶切割位点和所述融合肽的第一个残基的所述突变包含MeV F第111-113位的缺失，其中第110和114位通过肽接头融合。
27.根据权利要求26所述的免疫原，其中所述肽接头是Gly-Gly-Gly接头。
28.根据权利要求21-27中任一项所述的免疫原，其中所述氨基酸取代的定位是根据SEQ ID NO:36所示的参考MeV F蛋白序列。
29.根据权利要求21-28中任一项所述的免疫原，其中所述MeV F胞外域三聚体的原体包含与SEQ ID NO:37-43、53-55、62-68或72-80中任一项的残基21-483或SEQ ID NO:70-72中任一项的残基21-490具有至少90％同一性的氨基酸序列；以及
其中所述原体包含使所述MeV F胞外域三聚体以融合前构象稳定的一个或更多个氨基酸取代。
30.根据权利要求29所述的免疫原，其中所述MeV F胞外域三聚体的原体包含如SEQ IDNO:37-43、53-55、62-68或72-80中任一项的21-483，或SEQ ID NO:70-72中任一项的残基21-490所示的氨基酸序列或由其组成。
31.根据权利要求21-30中任一项所述的免疫原，其中所述重组MeV F胞外域三聚体的原体在C-末端与三聚化结构域融合。
32.根据权利要求31所述的免疫原，其中所述三聚化结构域包含GCN4三聚化结构域、T4纤维蛋白三聚化结构域或两者。
33.根据权利要求32所述的免疫原，其中
所述GCN4三聚化结构域包含如IEDKIEEILSKIYHIENEIARIKKLIGEAP(SEQ ID NO:33)所示的氨基酸序列，
所述T4纤维蛋白三聚化结构域包含如GYIPEAPRDGQAYVRKDGEWVLLSTFL SEQ ID NO:34)所示的氨基酸序列，
包含所述GCN4和T4纤维蛋白三聚化结构域的三聚化结构域包含如IEDKIEEILSKIYHIENEIARIKKLIGEAPGSGYIPEAPRDGQAYVRKDGEWVLLSTFL(SEQ ID NO:34)所示的氨基酸序列。
34.根据权利要求31-33中任一项所述的免疫原，其中与所述三聚化结构域融合的所述MeV F胞外域三聚体的原体包含与SEQ ID NO:37-43、53-55、62-68或72-80中任一项的残基21-513或SEQ ID NO:70-72中任一项的残基21-520具有至少90％同一性的氨基酸序列；以及
其中所述原体包含使所述MeV F胞外域三聚体以融合前构象稳定的一个或更多个氨基酸取代。
35.根据权利要求34所述的免疫原，其中与所述三聚化结构域融合的所述MeV F胞外域三聚体的原体包含如SEQ ID NO:37-43、53-55、62-68或72-80中任一项的残基21-513或SEQID NO:70-72中任一项的残基21-520所示的氨基酸序列或由其组成。
36.根据权利要求21-35中任一项所述的免疫原，其中所述重组MeV F胞外域三聚体的原体与异源蛋白连接。
37.根据权利要求36所述的免疫原，其中所述异源蛋白是MeV H蛋白或MuV HN蛋白的胞外域头部或胞外域茎部和头部。
38.根据权利要求37所述的免疫原，其中所述MeV H蛋白或所述MuV HN蛋白的胞外域头部或胞外域茎部和头部在C-末端与三聚化结构域融合，所述三聚化结构域在C-末端与所述重组MuV F的原体胞外域三聚体融合。
39.根据权利要求38所述的免疫原，其中与所述MeV H蛋白或所述MuV HN蛋白的三聚化结构域和胞外域连接的所述MeV F胞外域三聚体的原体包含如SEQ ID NO:56的残基21-959或SEQ ID NO 57的残基21-973或SEQ ID NO:81的残基21-988所示的氨基酸序列。
40.根据前述权利要求中任一项所述的免疫原，其中所述重组MeV F胞外域三聚体或所述重组MuV F胞外域三聚体的原体进一步包含一个或更多个另外的氨基酸取代。
41.一种分离的免疫原，所述分离的免疫原包含：
融合蛋白的三聚体，每个融合蛋白在N-末端至C-末端方向上包含以下或由以下组成：三聚化结构域、任选的肽接头和一个或更多个拷贝的MuV HN胞外域头部或MeV H胞外域头部；或
融合蛋白的三聚体，每个融合蛋白在N-末端至C-末端方向上包含以下或由以下组成：一个或更多个拷贝的MuV HN胞外域头部或MeV H胞外域头部、任选的肽接头、三聚化结构域、任选的肽接头，以及一个或更多个拷贝的MuV HN胞外域头部或MeV H胞外域头部；或
融合蛋白的三聚体，每个融合蛋白在N-末端至C-末端方向上包含以下或由以下组成：一个或更多个拷贝的MuV HN胞外域头部或茎部和头部或MeV H胞外域头部或茎部和头部、任选的肽接头、三聚化结构域、任选的肽接头，以及一个或更多个拷贝的MuV HN胞外域头部或茎部和头部或MeV H胞外域头部或茎部和头部。
42.根据权利要求41所述的免疫原，其中
所述MuV HN胞外域头部包含如SEQ ID NO:58的残基59-510所示的氨基酸序列，或与SEQ ID NO:58的残基59-510具有至少90％同一性的序列，或由其组成；
所述MuV HN胞外域茎部和头部包含SEQ ID NO:90的残基22-550、SEQ ID NO:91的残基22-543、SEQ ID NO:92的残基22-541、或SEQ ID NO:93的残基22-549，或与SEQ ID NO:90的残基22-550、SEQ ID NO:91的残基22-543、SEQ ID NO:92的残基22-541、或SEQ ID NO:93的残基22-549具有至少90％同一性的序列；
所述MeV H胞外域头部包含如SEQ ID NO:59的残基59-496所示的氨基酸序列，或与SEQID NO:59的残基59-496具有至少90％同一性的序列，或由其组成；或
所述MeV H胞外域茎部和头部包含SEQ ID NO:86的残基22-580、SEQ ID NO:87的残基22-577、SEQ ID NO:88的残基22-579、或SEQ ID NO:89的残基22-572，或与SEQ ID NO:86的残基22-580、SEQ ID NO:87的残基22-577、SEQ ID NO:88的残基22-579、或SEQ ID NO:89的残基22-572具有至少90％同一性的序列。
43.根据权利要求41或42所述的免疫原，其中
所述GCN4三聚化结构域包含如IEDKIEEILSKIYHIENEIARIKKLIGEAP(SEQ ID NO:33)所示的氨基酸序列，
所述T4纤维蛋白三聚化结构域包含如GYIPEAPRDGQAYVRKDGEWVLLSTFL SEQ ID NO:34)所示的氨基酸序列，
包含所述GCN4和T4纤维蛋白三聚化结构域的三聚化结构域包含如IEDKIEEILSKIYHIENEIARIKKLIGEAPGSGYIPEAPRDGQAYVRKDGEWVLLSTFL(SEQ ID NO:35)所示的氨基酸序列。
44.根据权利要求41-43中任一项所述的免疫原，其中所述三聚体中的融合蛋白包含以下项或由以下项组成：SEQ ID NO:58的残基24-510、SEQ ID NO:59的残基24-496、SEQ IDNO:82的残基22-950、SEQ ID NO:83的残基25-985、SEQ ID NO:84的残基22-948、或SEQ IDNO:85的残基22-981，或与SEQ ID NO:58的残基24-510、SEQ ID NO:59的残基24-496、SEQID NO:82的残基22-950、SEQ ID NO:83的残基25-985、SEQ ID NO:84的残基22-948、或SEQID NO:85的残基22-981中的任一项具有至少90％同一性的序列。
45.一种分离的免疫原，分离的免疫原包含：
MeV H胞外域头部的二聚体；
MeV H胞外域茎部和头部的二聚体；
MuV HN胞外域头部的二聚体；或
MuV HN胞外域茎部和头部的二聚体。
46.根据权利要求45所述的免疫原，其中
所述MeV H胞外域头部包含如SEQ ID NO:60的残基22-459所示的氨基酸序列，或与SEQID NO:60的残基22-459具有至少90％同一性的序列，或由其组成；
所述MeV H胞外域茎部和头部包含SEQ ID NO:86的残基22-580、SEQ ID NO:87的残基22-577、SEQ ID NO:88的残基22-579、或SEQ ID NO:89的残基22-572，或与SEQ ID NO:86的残基22-580、SEQ ID NO:87的残基22-577、SEQ ID NO:88的残基22-579、或SEQ ID NO:89残基22-572具有至少90％同一性的序列，
所述MuV H胞外域头部包含如SEQ ID NO:30所示的氨基酸序列，或与SEQ ID NO:30具有至少90％同一性的序列，或由其组成；或
所述MuV HN胞外域茎部和头部包含以下项或由以下项组成：SEQ ID NO:90的残基22-550、SEQ ID NO:91的残基22-543、SEQ ID NO:92的残基22-541、或SEQ ID NO:93的残基22-549，或与SEQ ID NO:90的残基22-550、SEQ ID NO:91的残基22-543、SEQ ID NO:92的残基22-541、或SEQ ID NO:93的残基22-549具有至少90％同一性的序列。
47.根据前述权利要求中任一项所述的免疫原，其与异源运载体缀合。
48.根据前述权利要求中任一项所述的免疫原，其中所述免疫原是可溶的。
49.根据前述权利要求中任一项所述的免疫原，其中所述重组MeV F胞外域三聚体的原体、所述重组MuV F胞外域三聚体的原体、融合蛋白的所述三聚体或所述MeV H胞外域头部的二聚体通过肽接头与跨膜结构域融合，或直接与所述跨膜结构域融合。
50.根据权利要求49所述的免疫原，其中所述重组MeV F胞外域三聚体或所述重组MuVF胞外域三聚体的原体包含全长F1蛋白。
51.一种病毒样颗粒，所述病毒样颗粒包含前述权利要求中任一项或多项的免疫原。
52.一种自组装蛋白纳米颗粒，所述自组装蛋白纳米颗粒包含前述权利要求中任一项或多项的免疫原。
53.一种编码前述权利要求中任一项的免疫原的核酸分子。
54.根据权利要求53所述的核酸分子，其可操作地连接至启动子。
55.一种包含权利要求54的核酸分子的载体。
56.根据权利要求55所述的载体，其中所述载体是RNA载体。
57.一种产生免疫原的方法，所述方法包括：
在宿主细胞中表达权利要求55-56中任一项的核酸分子；和
纯化所述免疫原。
58.通过权利要求57的方法产生的免疫原。
59.一种免疫原性组合物，所述免疫原性组合物包含权利要求1-56和58中任一项的免疫原、核酸分子、载体或病毒样颗粒，以及药学上可接受的运载体。
60.一种引发受试者对MuV F、MeV F、MuV NH和/或MeV H产生免疫应答的方法，所述方法包括向所述受试者施用有效量的权利要求59的免疫原性组合物以引发所述免疫应答。
61.根据权利要求60所述的方法，其中所述免疫应答抑制所述受试者中的MuV和/或MeV感染。
62.根据权利要求60或61所述的方法，其中所述受试者是先前用针对MeV和/或MuV的减毒活疫苗免疫的成年人，并且所述施用增强对所述MuV和/或MeV的免疫应答。
CN202080096383.0A 2019-12-11 2020-12-11 腮腺炎和麻疹病毒免疫原及其用途 Pending CN115515628A (zh)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US201962946902P 2019-12-11 2019-12-11
US62/946,902 2019-12-11
PCT/US2020/064619 WO2021119497A1 (en) 2019-12-11 2020-12-11 Mumps and measles virus immunogens and their use

Publications (1)

Publication Number Publication Date
CN115515628A true CN115515628A (zh) 2022-12-23

Family

ID=74183509

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202080096383.0A Pending CN115515628A (zh) 2019-12-11 2020-12-11 腮腺炎和麻疹病毒免疫原及其用途

Country Status (10)

Country Link
US (1) US20230053555A1 (zh)
EP (1) EP4072581A1 (zh)
JP (1) JP2023506170A (zh)
KR (1) KR20220141787A (zh)
CN (1) CN115515628A (zh)
AU (1) AU2020402106A1 (zh)
BR (1) BR112022011422A2 (zh)
CA (1) CA3164343A1 (zh)
IL (1) IL293666A (zh)
WO (1) WO2021119497A1 (zh)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116406375A (zh) * 2021-11-05 2023-07-07 上海科技大学 抗SARS-CoV-2刺突蛋白的三聚体抗体

Family Cites Families (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4722848A (en) 1982-12-08 1988-02-02 Health Research, Incorporated Method for immunizing animals with synthetically modified vaccinia virus
US5091309A (en) 1986-01-16 1992-02-25 Washington University Sindbis virus vectors
US5217879A (en) 1989-01-12 1993-06-08 Washington University Infectious Sindbis virus vectors
US5703055A (en) 1989-03-21 1997-12-30 Wisconsin Alumni Research Foundation Generation of antibodies through lipid mediated DNA delivery
US5643578A (en) 1992-03-23 1997-07-01 University Of Massachusetts Medical Center Immunization by inoculation of DNA transcription unit
EP0671926B1 (en) 1992-08-11 2002-11-13 President And Fellows Of Harvard College Immunomodulatory peptides
US5593972A (en) 1993-01-26 1997-01-14 The Wistar Institute Genetic immunization
GB9308963D0 (en) * 1993-04-30 1993-06-16 Smithkline Beecham Biolog Novel compounds
WO2000018929A2 (en) * 1998-09-25 2000-04-06 Smithkline Beecham Biologicals S.A. Paramyxovirus vaccines
MX2015013065A (es) 2013-03-13 2016-06-06 Us Health Proteinas f de rsv de prefusion y su uso.
US10420834B2 (en) * 2014-12-24 2019-09-24 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Recombinant metapneumovirus F proteins and their use
SI3718565T1 (sl) 2015-10-22 2022-08-31 Modernatx, Inc. Cepiva za respiratorni virus
US11174292B2 (en) * 2016-03-29 2021-11-16 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Substitutions-modified prefusion RSV F proteins and their use
WO2018081289A2 (en) * 2016-10-25 2018-05-03 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Prefusion piv f immunogens and their use
US20190192646A1 (en) 2017-11-03 2019-06-27 Modernatx, Inc. Salmonella vaccines

Also Published As

Publication number Publication date
BR112022011422A2 (pt) 2022-08-30
AU2020402106A1 (en) 2022-07-07
KR20220141787A (ko) 2022-10-20
IL293666A (en) 2022-08-01
WO2021119497A1 (en) 2021-06-17
JP2023506170A (ja) 2023-02-15
CA3164343A1 (en) 2021-06-17
EP4072581A1 (en) 2022-10-19
US20230053555A1 (en) 2023-02-23

Similar Documents

Publication Publication Date Title
US11964010B2 (en) Prefusion coronavirus spike proteins and their use
US20230113170A1 (en) Sars-cov-2 vaccine
US11890339B2 (en) Nipah virus immunogens and their use
US11845778B2 (en) Prefusion PIV F immunogens and their use
CN115960263A (zh) 取代修饰的融合前rsv f蛋白及其用途
CN112851766A (zh) 融合前rsv f蛋白和其用途
US20230174587A1 (en) Recombinant human metapneumovirus f proteins and their use
WO2016103238A1 (en) Recombinant metapneumovirus f proteins and their use
CA3207878A1 (en) Coronavirus spike protein designs, compositions and methods for their use
CN115515628A (zh) 腮腺炎和麻疹病毒免疫原及其用途
WO2022232648A1 (en) Prefusion-stabilized lassa virus glycoprotein complex and its use

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination