CN115335390A - 基于SARS-CoV-2的S蛋白的疫苗和组合物 - Google Patents

基于SARS-CoV-2的S蛋白的疫苗和组合物 Download PDF

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CN115335390A
CN115335390A CN202280002200.3A CN202280002200A CN115335390A CN 115335390 A CN115335390 A CN 115335390A CN 202280002200 A CN202280002200 A CN 202280002200A CN 115335390 A CN115335390 A CN 115335390A
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张必良
马林
温剑
张虹
钟惠玲
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Guangzhou Ribobio Co ltd
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Abstract

本公开提供了基于SARS‑CoV‑2的S蛋白的疫苗和组合物，并具体涉及重组SARS‑CoV‑2刺突蛋白(S蛋白)及编码其的mRNA和DNA。本公开还涉及包含编码重组S蛋白的DNA序列的重组质粒。本公开进一步涉及包含前述重组S蛋白和/或mRNA的组合物、mRNA‑载体颗粒例如脂质纳米颗粒(LNP)和组合物例如疫苗组合物。

Description

基于SARS-CoV-2的S蛋白的疫苗和组合物
本国际专利申请要求2022年1月10日提交的中国专利申请号202210019169.6的权益，其全部内容通过引用而并入以用于所有目的。
技术领域
本公开属于生物医药和疫苗技术领域，尤其涉及针对SARS-CoV-2，例如SARS-CoV-2 Delta变异株(B.1.617.2)和Omicron变异株(B.1.1.529)的疫苗和组合物。
背景技术
SARS-CoV-2的基因组随着在不同宿主群体中的传播而不断发生变异，产生了多种亚型，其中，SARS-CoV-2 Delta变异株，即B.1.617.2，是最早在印度报道的一个新的变体；SARS-CoV-2 Omicron变异株(B.1.1.529)是最早在南非发现的具有高度传染性的另一变体。目前针对SARS-CoV-2 Delta变异株和Omicron变异株尚无成熟的特效药，迫切需要有效的疫苗来应对。
SARS-CoV-2 Delta变异株相对于早期报道的SARS-CoV-2，在基因组诸多位置发生了突变。这些突变触发了新冠病毒的免疫逃逸，导致该病毒相对于其他早期的新冠病毒亚型具有对人体适应性更强、传播速度更快、病毒载量更高、治疗所需时间更长、更容易发展成重症等特点。
SARS-CoV-2 Omicron变异株相对于早期报道的SARS-CoV-2也在基因组诸多位置发生了突变，包括在S蛋白、ORF1a、ORF1b、ORF9b、M蛋白、E蛋白和N蛋白中发生了突变。这些突变不仅导致Omicron变异株传播能力更强，而且导致该病毒亚型对抗体保护作用的抵制力增强，使其对目前的SARS-CoV-2疫苗更具有抵抗力，能够逃避疫苗诱导的免疫反应。因此，迫切需要开发相应的针对性疫苗。
mRNA疫苗作为第三代疫苗，可以诱导机体同时产生体液免疫和细胞免疫，通过多种机制来保护机体，并且由于其自身的特性，免疫后在转染的细胞的细胞质中能够很快被降解，从而降低安全性风险。在变异型新冠病毒疫情应对中，mRNA疫苗相对于其他类型的疫苗已彰显出了独特的优势。临床试验数据表明，针对变异株设计的增强型mRNA疫苗对变异病毒的中和能力更高。此外，mRNA疫苗的研发和生产周期相对于传统疫苗更短，容易实现量产，疫苗的产能较高。
联合疫苗由两种以上疫苗原液按特定比例配合制成，可以预防多种疾病或由同一病原微生物的不同亚型感染引起的疾病，前者成为多联疫苗，后者成为多价疫苗。联合疫苗并不等于任意单种疫苗的简单叠加，其不仅不会加重注射后的副作用，还能有效降低因多次接种而可能出现的不良反应的风险。
基于当前不同SARS-CoV-2变异株肆虐的形势之下，联合型免疫策略为预防不同变异株的感染、减少疫苗注射次数以及降低免疫不良反应提供了新思路。因此，迫切需要能够同时有效针对不同SARS-CoV-2变异株的疫苗。
发明内容
本发明提供了一种针对SARS-CoV-2，特别是针对SARS-CoV-2 Delta变异株(B.1.617.2)的mRNA疫苗，其递送至小鼠体内后能够在体内表达预融合稳定形式的重组S蛋白，触发机体的细胞免疫和体液免疫反应，从而诱导体内产生特异性抗体。与第1代针对SARS-CoV-2的mRNA疫苗相比，经本发明的疫苗免疫的血清对SARS-CoV-2 Delta变异株S蛋白的效价更高，而且对SARS-CoV-2 Delta变异株的中和能力更强。
本发明还提供了一种针对SARS-CoV-2，特别是针对SARS-CoV-2 Omicron变异株的mRNA疫苗，其递送至小鼠体内后能够在体内表达预融合稳定形式的重组S蛋白，触发机体的细胞免疫和体液免疫反应，从而诱导体内产生特异性抗体。与第1代针对SARS-CoV-2的mRNA疫苗、以及第2代针对SARS-CoV-2 Delta变异株的mRNA疫苗相比，经本发明的疫苗免疫的血清对SARS-CoV-2 Omicron变异株S蛋白的效价更高，而且对SARS-CoV-2 Omicron变异株的中和能力更强，同时对野生型和Delta变异株均有一定的抑制效果。
本发明还提供了一种针对SARS-CoV-2及其变异株(例如Delta和Omicron变异株)的mRNA疫苗组合物，经本发明的mRNA疫苗组合物免疫的血清能够对多种SARS-CoV-2变异株起到抑制效果，其中对抗野生型、Delta型和Omicron型SARS-CoV-2均具有较强的中和能力。
SARS-CoV-2与SARS-CoV和MERS-CoV同属于冠状病毒科的β-冠状病毒属。SARS-CoV-2基因组序列全长29903bp，与SARS-CoV基因序列同一性为79.5％，与MERS-CoV序列同一性为40％。SARS-CoV-2病毒颗粒的主要结构包括单股正链核酸(ssRNA)、刺突蛋白(spikeprotein，S)、膜蛋白(membrane protein，M)、包膜蛋白(envelop protein，E)和核衣壳蛋白(nucelocapsid protein，N)。与其他β-冠状病毒类似，SARS-CoV-2病毒的对细胞的吸附入侵过程主要依赖于S蛋白；在该过程中，S蛋白以同源三聚体的形式装配，其具有短的胞质尾和疏水跨膜结构域，将蛋白质锚定到膜中。
S蛋白可分为受体结合亚基S1和膜融合亚基S2，S1亚基可进一步分成信号肽区域(signal peptide，SP)、N-端区域(N-terminal domain，NTD)和受体结合域(receptorbinding domain，RBD)。S2亚基通过跨膜区锚定在膜上，它含有膜融合过程所需要的基本元件，包括：内在的膜融合肽(fusion peptide，FP)、两个7肽重复序列(heptad repeat，HR)、跨膜结构域(transmembrane domain，TM)，以及C末端的胞质结构域(cytoplasmic domain，CP)。
S蛋白从N端到C端由信号肽区域(SP)、胞外结构域(ECD)、跨膜结构域(TM)和胞质结构域(CP)组成。其中，胞外结构域可进一步分成N-端区域(NTD)、受体结合域(RBD)、一个内在的膜融合肽(FP)和两个7肽重复序列(HR1和HR2)，属于第一类病毒膜融合蛋白。S蛋白的信号肽区域对应于S蛋白的氨基酸位置1-13的区域；胞外结构域对应于S蛋白的氨基酸位置14-1213的区域；跨膜结构域对应于S蛋白的氨基酸位置1214-1237的区域；胞质结构域对应于S蛋白的氨基酸位置1238-1273的区域。S蛋白的氨基酸序列如SEQ ID NO.29所示。在本文中，除非另有说明，否则重组S蛋白的氨基酸位置按照如SEQ ID NO.29所示的野生型S蛋白的氨基酸序列进行编号。
通过对S蛋白预融合结构进行解析，发现S1亚基的RBD结构域经历铰链类似的构象移动来隐藏或暴露受体结合的关键位点，“向下”为受体不可结合状态，“向上”为受体可结合状态，处于较为不稳定的状态。这种构象使得S蛋白能够容易地与宿主受体血管紧张素转换酶2(ACE2)结合。当RBD与受体结合后，S2亚基通过将FP插入宿主细胞膜而改变为融合后构象，HR1和HR2形成一个反平行的六螺旋束(6HB)，共同组成一个融合核心，最终导致病毒膜与细胞膜融合。使用冷冻电镜实验，在预融合构象中确定了大量的三聚体S糖蛋白结构域，预融合S蛋白上存在大量中和抗体敏感表位，而融合后构象使仅存在于预融合构象上的中和敏感表位的暴露量降至最低。因此，表达预融合稳定形式的SARS-CoV-2 S三聚体蛋白，是开发安全有效的SARS-CoV-2疫苗的关键。优化的疫苗抗原将保留存在于S蛋白的预融合构象形式中的表位，并诱导能够抑制病毒融合的抗体。
如本文所用的术语“SARS-CoV-2 Delta变异株”、“B.1.617.2”、“Delta型新冠病毒”、“德尔塔型新冠病毒”可互换使用，并且是指2020年10月在印度首次出现的SARS-CoV-2亚型，其相对于SARS-CoV-2，在基因组诸多位置发生了突变。这些突变触发了新冠病毒的免疫逃逸，导致该病毒相对于其他早期的新冠病毒亚型具有对人体适应性更强、传播速度更快、病毒载量更高、治疗所需时间更长、更容易发展成重症等特点。
英国公共卫生局的报告显示，Delta变异株的传播能力比Alpha变异株高60％(HAllen et.al.,Increased household transmission of COVID-19 cases associatedwith SARS-CoV-2 Variant of Concern B.1.617.2:a national case-control study2021)。研究表明，Delta变异株对疫苗诱导的中和抗体产生抗性(A Saito et.al.,SARS-CoV-2 spike P681R mutation enhances and accelerates viral fusion)。检测了250个辉瑞的BNT162b2疫苗接种者对各种VOC的中和抗体滴度，结果显示，与野生型相比，针对Delta变异株的中和抗体滴度下降了5.8倍，而Alpha变异株只下降了2.6倍(EC Wallet.al.,Neutralising antibody activity against SARS-CoV-2 VOCs B.1.617.2 andB.1.351 by BNT162b2 vaccination.Lancet.2021；397(10292):2331-3)。
如本文所用的术语“SARS-CoV-2 Omicron变异株”、“B.1.1.529”、“Omicron型新冠病毒”、“奥密克戎型新冠病毒”可互换使用，并且是指2021年11月在南非首次出现的SARS-CoV-2亚型，其相对于野生型SARS-CoV-2，在基因组诸多位置发生了突变，包括在S蛋白、ORF1a、ORF1b、ORF9b、M蛋白、E蛋白、N蛋白中发生了突变。这些突变不仅导致Omicron变异株传播能力更强，而且导致该病毒亚型对抗体保护作用的抵制力增强，使其对目前的SARS-CoV-2疫苗更具有抵抗力，能够逃避疫苗诱导的免疫反应。
在第一方面，本发明提供了重组SARS-CoV-2刺突蛋白(S蛋白)，所述重组S蛋白相对于野生型S蛋白在胞外结构域中包括以下突变：T19R、G142D、E156G、F157和R158缺失突变、A222V、L452R、T478K、D614G、P681R和D950N；其中，氨基酸位置按照如SEQ ID NO.29所示的野生型S蛋白的氨基酸序列进行编号。
在一些实施方案中，重组S蛋白的胞外结构域中的S1/S2切割位点RRAR(对应于S蛋白的氨基酸位置682-685)可以经突变以失去被蛋白酶例如弗林样(Furin-like)蛋白酶和溶酶体蛋白酶切割的能力。在一些实施方案中，可以将重组S蛋白中的S1/S2切割位点RRAR突变为GGSG。
在一些实施方案中，重组S蛋白的胞外结构域中的S2切割位点KR(对应于S蛋白的氨基酸位置814-815)可以经突变以失去被蛋白酶例如弗林样蛋白酶和溶酶体蛋白酶切割的能力。在一些实施方案中，可以将重组S蛋白或其抗原性片段中的S2切割位点KR突变为AN。
SARS-CoV-2病毒在细胞内包装过程中，S蛋白可能会被蛋白酶例如弗林样蛋白酶或溶酶体蛋白切割，分泌出S1亚基和S2亚基呈非融合状态的S蛋白。通过对重组S蛋白中的切割位点例如S1/S2切割位点和/或S2切割位点进行突变，可以避免重组S蛋白被蛋白酶切割，从而进一步提高其稳定性。
在一些实施方案中，重组S蛋白还包括K986P和V987P突变。在重组S蛋白的胞外结构域中引入2个脯氨酸突变K986P和V987P可以提高预融合构象的稳定性。
在一些实施方案中，重组S蛋白可以不包含功能性融合肽结构域(FP结构域：对应于S蛋白的氨基酸位置788-806)。例如，重组S蛋白可以包含经突变的融合肽结构域，例如通过一个或多个氨基酸残基的取代、缺失、插入和/或添加，导致融合肽结构域失去其天然功能，例如介导病毒与宿主细胞膜进行融合的功能。或者，在一些实施方案中，重组S蛋白可以不包含融合肽结构域。
通过在重组S蛋白中去除功能性融合肽结构域，能够提高预融合构象的稳定性，使得保留和暴露S蛋白的预融合构象上存在的大量中和抗体敏感表位。
在一些实施方案中，重组S蛋白可以不包含跨膜结构域(对应于S蛋白的氨基酸位置1214-1237的区域)和/或胞质结构域(对应于S蛋白的氨基酸位置1238-1273的区域)。在一些实施方案中，重组S蛋白可以不包含胞质结构域。在一些实施方案中，重组S蛋白可以不包含跨膜结构域和胞质结构域。在一些实施方案中，重组S蛋白还包含三聚体结构域，所述三聚体结构域当表达时促进所述重组S蛋白形成三聚体。
如本文所用，“三聚体结构域”是指当表达时，能够自发或诱导形成三聚体的蛋白质或多肽结构域。本领域已知多种此类三聚体结构域。通过在重组S蛋白中包括三聚体结构域(例如通过构建融合蛋白)，能够促进重组S蛋白形成三聚体构象，和/或稳定化重组S蛋白的三聚体构象。
在一些实施方案中，重组S蛋白中的三聚体结构域可以包含T4噬菌体fibritin三聚体基序。在一些实施方案中，所述T4噬菌体fibritin三聚体基序可以具有SEQ ID NO.18(GYIPEAPRDGQAYVRKDGEWVLLSTFL)所示的氨基酸序列。
在一些实施方案中，三聚体结构域可以与重组S蛋白直接融合。在另一些实施方案中，三聚体结构域可以通过接头与重组S蛋白融合。在一些实施方案中，三聚体结构域可以与重组S蛋白的N端融合。在另一些实施方案中，三聚体结构域可以与重组S蛋白的C端融合。例如，三聚体结构域可以通过接头与重组S蛋白的C端融合。在一些实施方案中，所述接头序列可以包含SEQ ID NO.19(SAIG)所示的序列。
在一些实施方案中，重组S蛋白还包含信号序列；优选地，所述信号序列包含免疫球蛋白重链可变区(IGHV)信号序列。例如，所述信号序列可以具有SEQ ID NO.17(MDWIWRILFLVGAATGAHS)所示的氨基酸序列。
在一些实施方案中，重组S蛋白从N端到C端由以下中的任一项组成：
i)胞外结构域；
ii)胞外结构域、跨膜结构域和任选的胞质结构域；
iii)胞外结构域和三聚体结构域；
iv)胞外结构域、跨膜结构域、任选的胞质结构域、和三聚体结构域；
v)信号序列、胞外结构域；
vi)信号序列、胞外结构域、跨膜结构域和任选的胞质结构域；
vii)信号序列、胞外结构域和三聚体结构域；和
viii)信号序列、胞外结构域、跨膜结构域、任选的胞质结构域、和三聚体结构域。
在一些实施方案中，所述胞外结构域相对于野生型序列具有以下突变中的一种或多种：
1)T19R、G142D、E156G、F157和R158缺失突变、A222V、L452R、T478K、D614G、P681R和D950N；
2)S1/S2切割位点RRAR经突变以失去被弗林样蛋白酶和溶酶体蛋白酶切割的能力，优选地，S1/S2切割位点突变为GGSG；
3)S2切割位点KR经突变以失去被弗林样蛋白酶和溶酶体蛋白酶切割的能力，优选地，S2切割位点突变为AN；
4)K986P和/或V987P突变；
5)融合肽结构域经突变以失去介导病毒与宿主细胞膜进行融合的功能；优选融合肽结构域缺失突变。
在一些实施方案中，所述信号序列包含免疫球蛋白重链可变区(IGHV)信号序列。例如，所述信号序列具有SEQ ID NO.17所示的氨基酸序列。
在一些实施方案中，所述三聚体结构域为T4噬菌体fibritin三聚体基序。在一些实施方案中，所述T4噬菌体fibritin三聚体基序具有SEQ ID NO.18所示的氨基酸序列。
在优选的实施方案中，重组S蛋白从N端到C端由信号序列、胞外结构域、跨膜结构域和胞质结构域组成，所述胞外结构域相对于野生型序列具有以下突变：T19R、G142D、E156G、F157和R158缺失突变、A222V、L452R、T478K、D614G、P681R和D950N。例如，重组S蛋白具有如SEQ ID NO.1所示的氨基酸序列。
在另一优选的实施方案中，重组S蛋白从N端到C端由信号序列、胞外结构域、跨膜结构域和胞质结构域组成，所述胞外结构域相对于野生型序列具有以下突变：T19R、G142D、E156G、F157和R158缺失突变、A222V、L452R、T478K、D614G、P681R、D950N、S1/S2切割位点RRAR突变为GGSG和S2切割位点KR突变为AN。例如，重组S蛋白具有如SEQ ID NO.2所示的氨基酸序列。
在另一优选的实施方案中，重组S蛋白从N端到C端由信号序列、胞外结构域、跨膜结构域和胞质结构域组成，所述胞外结构域相对于野生型序列具有以下突变：T19R、G142D、E156G、F157和R158缺失突变、A222V、L452R、T478K、D614G、P681R、D950N、K986P、V987P、S1/S2切割位点RRAR突变为GGSG和S2切割位点KR突变为AN。例如，重组S蛋白具有如SEQ IDNO.3所示的氨基酸序列。
在另一优选的实施方案中，重组S蛋白从N端到C端由信号序列、胞外结构域和三聚体结构域组成，所述信号序列为免疫球蛋白重链可变区(IGHV)信号序列，优选如SEQ IDNO.17所示的序列(MDWIWRILFLVGAATGAHS)；所述胞外结构域相对于野生型序列具有以下突变：T19R、G142D、E156G、F157和R158缺失突变、A222V、L452R、T478K、D614G、P681R、D950N、S1/S2切割位点RRAR突变为GGSG、S2切割位点KR突变为AN、和融合肽结构域缺失突变；并且所述三聚体结构域为T4噬菌体fibritin三聚体基序，优选如SEQ ID NO.18所示的序列(GYIPEAPRDGQAYVRKDGEWVLLSTFL)。优选地，三聚体结构域通过接头与胞外结构域的C端融合。所述接头序列可以为SEQ ID NO.19所示的序列(SAIG)。例如，重组S蛋白具有如SEQ IDNO.4所示的氨基酸序列。
在另一优选的实施方案中，重组S蛋白从N端到C端由信号序列、胞外结构域和三聚体结构域组成，所述信号序列为免疫球蛋白重链可变区(IGHV)信号序列，优选如SEQ IDNO.17所示的序列(MDWIWRILFLVGAATGAHS)；所述胞外结构域相对于野生型序列具有以下突变：T19R、G142D、E156G、F157和R158缺失突变、A222V、L452R、T478K、D614G、P681R、D950N、K986P、V987P、S1/S2切割位点RRAR突变为GGSG、S2切割位点KR突变为AN、和融合肽结构域缺失突变；并且所述三聚体结构域为T4噬菌体fibritin三聚体基序，优选如SEQ ID NO.18所示的序列(GYIPEAPRDGQAYVRKDGEWVLLSTFL)。优选地，三聚体结构域通过接头与胞外结构域的C端融合。所述接头序列可以为SEQ ID NO.19所示的序列(SAIG)。例如，重组S蛋白具有如SEQ ID NO.5所示的氨基酸序列。
在一些实施方案中，重组S蛋白具有选自SEQ ID NO.1-5中任一项所示的氨基酸序列。在优选的实施方案中，重组S蛋白具有选自SEQ ID NO.3-5中任一项所示的氨基酸序列。
在第二方面，本发明提供了mRNA，其编码本发明第一方面的重组S蛋白。
在一些实施方案中，mRNA从5’到3’方向包含帽结构、5’-UTR、编码本发明的重组S蛋白的开放阅读框(ORF)、3’-UTR和polyA尾。
在一些实施方案中，帽结构可以具有m7G5’ppp5’(2’-OMe)NpG，其中m7G为N7-甲基鸟嘌呤核苷；p为磷酸；ppp为三磷酸；2’-OMe为2’-甲氧基修饰；N为任意核苷，例如腺嘌呤核苷(A)、鸟嘌呤核苷(G)、胞嘧啶核苷(C)和尿嘧啶核苷(U)，或天然存在的其它核苷或经修饰的核苷。
在一些实施方案中，5’-UTR可以包含选自下组的基因的5’-UTR或其同源物、片段或变体：β-珠蛋白(HBB)基因、热休克蛋白70(Hsp70)基因、轴丝动力蛋白重链2(DNAH2)基因、17β-羟基类固醇脱氢酶4(HSD17B4)基因。例如，变体序列可以与相应基因的野生型5’-UTR序列具有至少80％，至少85％，至少90％，至少95％，至少98％，或至少99％的序列同一性。
在一些实施方案中，5’-UTR包含衍生自17β-羟基类固醇脱氢酶4(HSD17B4)基因的5’-UTR或其同源物、片段或变体。在一些实施方案中，5’-UTR包含KOZAK序列。在一些实施方案中，5’-UTR包含衍生自17β-羟基类固醇脱氢酶4(HSD17B4)基因的5’-UTR或其同源物、片段或变体，和KOZAK序列。在一些实施方案中，5’-UTR包含如SEQ ID NO.8(GTCCCGCAGTCGGCGTCCAGCGGCTCTGCTTGTTCGTGTGTGTGTCGTTGCAGGCCTTATTC)和/或SEQ ID NO.9(AGATCTACCGGTGGTACCGCCACC)所示的序列。
在一些实施方案中，3’-UTR可以包含选自下组的基因的3’-UTR或其同源物、片段或变体：白蛋白(ALB)基因、α-珠蛋白基因、β-珠蛋白(HBB)基因、酪氨酸羟化酶基因、热休克蛋白70(Hsp70)基因，脂氧合酶基因和胶原蛋白α基因。例如，变体序列可以与相应基因的野生型3’-UTR序列具有至少80％，至少85％，至少90％，至少95％，至少98％，或至少99％的序列同一性。在一些实施方案中，3’-UTR包含衍生自白蛋白(ALB)基因的3’-UTR或其同源物、片段或变体。优选地，3’-UTR包含如SEQ ID NO.10(AGCCAACACCCTGTCTAAAAAACATAAATTTCTTTAATCATTTTGCCTCTTTTCTCTGTGCTTCAATTAATAAAAAATGGAAAGAACCT)所示的序列。
在一些实施方案中，poly A尾的长度可以为100-200个核苷酸，例如约100个核苷酸，约110个核苷酸，约120个核苷酸，约130个核苷酸，约140个核苷酸，约150个核苷酸，约160个核苷酸，约170个核苷酸，约180个核苷酸，约190个核苷酸，或约200个核苷酸。在一些实施方案中，polyA尾的长度可以为100-150个核苷酸。在一些实施方案中，所述poly A尾的长度可以为约120个核苷酸。
在一些实施方案中，本发明的mRNA包含如SEQ ID NO.14-16中任一项所示的序列，或由SEQ ID NO.14-16中任一项所示的序列组成。
在一些实施方案中，mRNA中的一个或多个核苷酸可以是经修饰的。例如，mRNA中的一个或多个核苷酸(例如所有核苷酸)可以各自独立替换为天然存在的核苷酸类似物或人工合成的核苷酸类似物。
在一些实施方案中，天然存在的核苷酸类似物选自假尿苷、2-硫尿苷、5-甲基尿苷、5-甲基胞苷和N6-甲基腺苷。在一些实施方案中，人工合成的核苷酸类似物选自N1-甲基假尿苷和5-乙炔基尿苷。
在一些实施方案中，mRNA中的一个或多个尿苷三磷酸可以各自独立地替换为假尿苷三磷酸、2-硫尿苷三磷酸、5-甲基尿苷三磷酸、N1-甲基假尿苷三磷酸或5-乙炔基尿苷三磷酸，和/或一个或多个胞苷三磷酸可以各自独立地替换为5-甲基胞苷三磷酸，和/或一个或多个腺苷三磷酸可以各自独立地替换为N6-甲基腺苷三磷酸。
在一些实施方案中，mRNA中的一个或多个尿苷三磷酸可以各自独立替换为假尿苷三磷酸、1-甲基-假尿苷三磷酸或5-乙炔基尿苷三磷酸。在一些实施方案中，mRNA中的一个或多个胞苷三磷酸可以各自独立替换为5-甲基胞苷三磷酸。
在第三方面，本发明提供了重组SARS-CoV-2刺突蛋白(S蛋白)，所述重组S蛋白相对于野生型S蛋白在胞外结构域中包括以下突变：NTD区域的A67V、H69缺失突变、V70缺失突变、T95I、G142缺失突变、V143缺失突变、Y144缺失突变、Y145D、N211缺失突变、L212I、以及在R214和D215之间三个氨基酸E、P、E的插入突变；RBD区域的G339D、S371L、S373P、S375F、K417N、N440K、G446S、S477N、T478K、E484A、Q493R、G496S、Q498R、N501Y、Y505H；SD1区域的T547K突变；SD2区域的D614G、H655Y、N679K、P681H；SD2和HR1间隔区域的N764K、D796Y、N856K；HR1区域的Q954H、N969K和L981F；其中，氨基酸位置按照如SEQ ID NO.29所示的野生型S蛋白的氨基酸序列进行编号。
在一些实施方案中，重组S蛋白的胞外结构域中的S1/S2切割位点RRAR(对应于S蛋白的氨基酸位置682-685)可以经突变以失去被蛋白酶例如弗林样(Furin-like)蛋白酶和溶酶体蛋白酶切割的能力。在一些实施方案中，可以将重组S蛋白中的S1/S2切割位点RRAR突变为GGSG。
在一些实施方案中，重组S蛋白的胞外结构域中的S2切割位点KR(对应于S蛋白的氨基酸位置814-815)可以经突变以失去被蛋白酶例如弗林样蛋白酶和溶酶体蛋白酶切割的能力。在一些实施方案中，可以将重组S蛋白或其抗原性片段中的S2切割位点KR突变为AN。
SARS-CoV-2病毒在细胞内包装过程中，S蛋白可能会被蛋白酶例如弗林样蛋白酶或溶酶体蛋白切割，分泌出S1亚基和S2亚基呈非融合状态的S蛋白。通过对重组S蛋白中的切割位点例如S1/S2切割位点和/或S2切割位点进行突变，可以避免重组S蛋白被蛋白酶切割，从而进一步提高其稳定性。
在一些实施方案中，重组S蛋白还包括K986P和V987P突变。在重组S蛋白的胞外结构域中引入2个脯氨酸突变K986P和V987P可以提高预融合构象的稳定性。
在一些实施方案中，重组S蛋白可以不包含功能性融合肽结构域(FP结构域：对应于S蛋白的氨基酸位置788-806)。例如，重组S蛋白可以包含经突变的融合肽结构域，例如通过一个或多个氨基酸残基的取代、缺失、插入和/或添加，导致融合肽结构域失去其天然功能，例如介导病毒与宿主细胞膜进行融合的功能。或者，在一些实施方案中，重组S蛋白可以不包含融合肽结构域。
在一些实施方案中，重组S蛋白可以不包含跨膜结构域(对应于S蛋白的氨基酸位置1214-1237的区域)和/或胞质结构域(对应于S蛋白的氨基酸位置1238-1273的区域)。在一些实施方案中，重组S蛋白可以不包含胞质结构域。在一些实施方案中，重组S蛋白可以不包含跨膜结构域和胞质结构域。在一些实施方案中，重组S蛋白还包含三聚体结构域，所述三聚体结构域当表达时促进所述重组S蛋白形成三聚体。
在一些实施方案中，重组S蛋白中的三聚体结构域可以包含T4噬菌体fibritin三聚体基序。在一些实施方案中，所述T4噬菌体fibritin三聚体基序可以具有SEQ ID NO.18所示的氨基酸序列。
在一些实施方案中，三聚体结构域可以与重组S蛋白直接融合。在另一些实施方案中，三聚体结构域可以通过接头与重组S蛋白融合。在一些实施方案中，三聚体结构域可以与重组S蛋白的N端融合。在另一些实施方案中，三聚体结构域可以与重组S蛋白的C端融合。例如，三聚体结构域可以通过接头与重组S蛋白的C端融合。在一些实施方案中，所述接头序列可以包含SEQ ID NO.19所示的序列。在一些实施方案中，重组S蛋白还包含信号序列；优选地，所述信号序列包含免疫球蛋白重链可变区(IGHV)信号序列。例如，所述信号序列可以具有SEQ ID NO.17所示的氨基酸序列。
在一些实施方案中，重组S蛋白从N端到C端由以下组成：任选的信号序列、胞外结构域、任选的跨膜结构域、任选的胞质结构域和任选的三聚体结构域。
在一些实施方案中，重组S蛋白从N端到C端由以下组成：胞外结构域、任选的跨膜结构域和任选的胞质结构域。
在优选的实施方案中，重组S蛋白从N端到C端由以下组成：信号序列、胞外结构域、跨膜结构域和胞质结构域。
在一些实施方案中，所述胞外结构域相对于野生型序列具有以下突变中的一种或多种：
1)A67V、H69缺失突变、V70缺失突变、T95I、G142缺失突变、V143缺失突变、Y144缺失突变、Y145D、N211缺失突变、L212I、在R214和D215之间三个氨基酸E、P、E的插入突变，G339D、S371L、S373P、S375F、K417N、N440K、G446S、S477N、T478K、E484A、Q493R、G496S、Q498R、N501Y、Y505H、T547K、D614G、H655Y、N679K、P681H、N764K、D796Y、N856K、Q954H、N969K和L981F；
2)S1/S2切割位点RRAR经突变以失去被弗林样蛋白酶和溶酶体蛋白酶切割的能力，优选地，S1/S2切割位点突变为GGSG；
3)S2切割位点KR经突变以失去被弗林样蛋白酶和溶酶体蛋白酶切割的能力，优选地，S2切割位点突变为AN；
4)K986P和/或V987P突变。
在一些实施方案中，所述信号序列包含免疫球蛋白重链可变区(IGHV)信号序列。例如，所述信号序列具有SEQ ID NO.17所示的氨基酸序列。
在一些实施方案中，所述三聚体结构域为T4噬菌体fibritin三聚体基序。在一些实施方案中，所述T4噬菌体fibritin三聚体基序具有SEQ ID NO.18所示的氨基酸序列。
在优选的实施方案中，重组S蛋白从N端到C端由信号序列、胞外结构域、跨膜结构域和胞质结构域组成，所述胞外结构域相对于野生型序列具有以下突变：A67V、H69缺失突变、V70缺失突变、T95I、G142缺失突变、V143缺失突变、Y144缺失突变、Y145D、N211缺失突变、L212I、在R214和D215之间三个氨基酸E、P、E的插入突变，G339D、S371L、S373P、S375F、K417N、N440K、G446S、S477N、T478K、E484A、Q493R、G496S、Q498R、N501Y、Y505H、T547K、D614G、H655Y、N679K、P681H、N764K、D796Y、N856K、Q954H、N969K和L981F。例如，重组S蛋白具有如SEQ ID NO.20所示的氨基酸序列。
在另一优选的实施方案中，重组S蛋白从N端到C端由信号序列、胞外结构域、跨膜结构域和胞质结构域组成，所述胞外结构域相对于野生型序列具有以下突变：A67V、H69缺失突变、V70缺失突变、T95I、G142缺失突变、V143缺失突变、Y144缺失突变、Y145D、N211缺失突变、L212I、在R214和D215之间三个氨基酸E、P、E的插入突变，G339D、S371L、S373P、S375F、K417N、N440K、G446S、S477N、T478K、E484A、Q493R、G496S、Q498R、N501Y、Y505H、T547K、D614G、H655Y、N679K、P681H、N764K、D796Y、N856K、Q954H、N969K和L981F；S1/S2切割位点RRAR突变为GGSG；和S2切割位点KR突变为AN。例如，重组S蛋白具有如SEQ ID NO.21所示的氨基酸序列。
在另一优选的实施方案中，重组S蛋白从N端到C端由信号序列、胞外结构域、跨膜结构域和胞质结构域组成，所述胞外结构域相对于野生型序列具有以下突变：A67V、H69缺失突变、V70缺失突变、T95I、G142缺失突变、V143缺失突变、Y144缺失突变、Y145D、N211缺失突变、L212I、在R214和D215之间三个氨基酸E、P、E的插入突变，G339D、S371L、S373P、S375F、K417N、N440K、G446S、S477N、T478K、E484A、Q493R、G496S、Q498R、N501Y、Y505H、T547K、D614G、H655Y、N679K、P681H、N764K、D796Y、N856K、Q954H、N969K和L981F；K986P和V987P。例如，重组S蛋白具有如SEQ ID NO.22所示的氨基酸序列。
在另一优选的实施方案中，重组S蛋白从N端到C端由信号序列、胞外结构域、跨膜结构域和胞质结构域组成，所述胞外结构域相对于野生型序列具有以下突变：A67V、H69缺失突变、V70缺失突变、T95I、G142缺失突变、V143缺失突变、Y144缺失突变、Y145D、N211缺失突变、L212I、在R214和D215之间三个氨基酸E、P、E的插入突变，G339D、S371L、S373P、S375F、K417N、N440K、G446S、S477N、T478K、E484A、Q493R、G496S、Q498R、N501Y、Y505H、T547K、D614G、H655Y、N679K、P681H、N764K、D796Y、N856K、Q954H、N969K和L981F；K986P和V987P；S1/S2切割位点RRAR突变为GGSG。例如，重组S蛋白具有如SEQ ID NO.23所示的氨基酸序列。
在另一优选的实施方案中，重组S蛋白从N端到C端由信号序列、胞外结构域、跨膜结构域和胞质结构域组成，所述胞外结构域相对于野生型序列具有以下突变：A67V、H69缺失突变、V70缺失突变、T95I、G142缺失突变、V143缺失突变、Y144缺失突变、Y145D、N211缺失突变、L212I、在R214和D215之间三个氨基酸E、P、E的插入突变，G339D、S371L、S373P、S375F、K417N、N440K、G446S、S477N、T478K、E484A、Q493R、G496S、Q498R、N501Y、Y505H、T547K、D614G、H655Y、N679K、P681H、N764K、D796Y、N856K、Q954H、N969K和L981F；K986P和V987P；S2切割位点KR突变为AN。例如，重组S蛋白具有如SEQ ID NO.24所示的氨基酸序列。
在另一优选的实施方案中，重组S蛋白从N端到C端由信号序列、胞外结构域、跨膜结构域和胞质结构域组成，所述胞外结构域相对于野生型序列具有以下突变：A67V、H69缺失突变、V70缺失突变、T95I、G142缺失突变、V143缺失突变、Y144缺失突变、Y145D、N211缺失突变、L212I、在R214和D215之间三个氨基酸E、P、E的插入突变，G339D、S371L、S373P、S375F、K417N、N440K、G446S、S477N、T478K、E484A、Q493R、G496S、Q498R、N501Y、Y505H、T547K、D614G、H655Y、N679K、P681H、N764K、D796Y、N856K、Q954H、N969K和L981F；K986P和V987P；S1/S2切割位点RRAR突变为GGSG；和S2切割位点KR突变为AN。例如，重组S蛋白具有如SEQ IDNO.25所示的氨基酸序列。
在一些实施方案中，重组S蛋白具有选自SEQ ID NO.20-25中任一项所示的氨基酸序列。在优选的实施方案中，重组S蛋白具有选自SEQ ID NO.23-25中任一项所示的氨基酸序列。在最优选的实施方案中，重组S蛋白具有如SEQ ID NO.25所示的氨基酸序列
在第四方面，本发明提供了mRNA，其编码本发明第三方面的重组S蛋白。
在一些实施方案中，mRNA从5’到3’方向包含帽结构、5’-UTR、编码本发明的重组S蛋白的开放阅读框(ORF)、3’-UTR和polyA尾。
在一些实施方案中，帽结构可以具有m7G5’ppp5’(2’-OMe)NpG，其中m7G为N7-甲基鸟嘌呤核苷；p为磷酸；ppp为三磷酸；2’-OMe为2’-甲氧基修饰；N为任意核苷，例如腺嘌呤核苷(A)、鸟嘌呤核苷(G)、胞嘧啶核苷(C)和尿嘧啶核苷(U)，或天然存在的其它核苷或经修饰的核苷。
在一些实施方案中，5’-UTR可以包含选自下组的基因的5’-UTR或其同源物、片段或变体：β-珠蛋白(HBB)基因、热休克蛋白70(Hsp70)基因、轴丝动力蛋白重链2(DNAH2)基因、17β-羟基类固醇脱氢酶4(HSD17B4)基因。例如，变体序列可以与相应基因的野生型5’-UTR序列具有至少80％，至少85％，至少90％，至少95％，至少98％，或至少99％的序列同一性。
在一些实施方案中，5’-UTR包含衍生自17β-羟基类固醇脱氢酶4(HSD17B4)基因的5’-UTR或其同源物、片段或变体。在一些实施方案中，5’-UTR包含KOZAK序列。在一些实施方案中，5’-UTR包含衍生自17β-羟基类固醇脱氢酶4(HSD17B4)基因的5’-UTR或其同源物、片段或变体，和KOZAK序列。在一些实施方案中，5’-UTR包含如SEQ ID NO.8和/或SEQ ID NO.9所示的序列。
在一些实施方案中，3’-UTR可以包含选自下组的基因的3’-UTR或其同源物、片段或变体：白蛋白(ALB)基因、α-珠蛋白基因、β-珠蛋白(HBB)基因、酪氨酸羟化酶基因、热休克蛋白70(Hsp70)基因，脂氧合酶基因和胶原蛋白α基因。例如，变体序列可以与相应基因的野生型3’-UTR序列具有至少80％，至少85％，至少90％，至少95％，至少98％，或至少99％的序列同一性。在一些实施方案中，3’-UTR包含衍生自白蛋白(ALB)基因的3’-UTR或其同源物、片段或变体。优选地，3’-UTR包含如SEQ ID NO.10所示的序列。
在一些实施方案中，poly A尾的长度可以为100-200个核苷酸，例如约100个核苷酸，约110个核苷酸，约120个核苷酸，约130个核苷酸，约140个核苷酸，约150个核苷酸，约160个核苷酸，约170个核苷酸，约180个核苷酸，约190个核苷酸，或约200个核苷酸。在一些实施方案中，polyA尾的长度可以为100-150个核苷酸。在一些实施方案中，所述poly A尾的长度可以为约120个核苷酸。
在一些实施方案中，本发明的mRNA包含如SEQ ID NO.27所示的序列，或由SEQ IDNO.27所示的序列组成。
在一些实施方案中，mRNA中的一个或多个核苷酸可以是经修饰的。例如，mRNA中的一个或多个核苷酸(例如所有核苷酸)可以各自独立替换为天然存在的核苷酸类似物或人工合成的核苷酸类似物。
在一些实施方案中，天然存在的核苷酸类似物选自假尿苷、2-硫尿苷、5-甲基尿苷、5-甲基胞苷和N6-甲基腺苷。在一些实施方案中，人工合成的核苷酸类似物选自N1-甲基假尿苷和5-乙炔基尿苷。
在一些实施方案中，mRNA中的一个或多个尿苷三磷酸可以各自独立地替换为假尿苷三磷酸、2-硫尿苷三磷酸、5-甲基尿苷三磷酸、N1-甲基假尿苷三磷酸或5-乙炔基尿苷三磷酸，和/或一个或多个胞苷三磷酸可以各自独立地替换为5-甲基胞苷三磷酸，和/或一个或多个腺苷三磷酸可以各自独立地替换为N6-甲基腺苷三磷酸。
在一些实施方案中，mRNA中的一个或多个尿苷三磷酸可以各自独立替换为假尿苷三磷酸、1-甲基-假尿苷三磷酸或5-乙炔基尿苷三磷酸。在一些实施方案中，mRNA中的一个或多个胞苷三磷酸可以各自独立替换为5-甲基胞苷三磷酸。
在第五方面，本发明提供了组合物，其包含本发明第一方面的重组S蛋白或本发明第二方面的mRNA，和本发明第三方面的重组S蛋白或本发明第四方法的mRNA。
在本发明组合物的一些实施方案中，组合物包含本发明第一方面的重组S蛋白和本发明第三方面的重组S蛋白。在一些实施方案中，组合物包含本发明第一方面的重组S蛋白和本发明第四方法的mRNA。在一些实施方案中，组合物包含本发明第二方面的mRNA和本发明第三方面的重组S蛋白。在一些实施方案中，组合物包含本发明第二方面的mRNA和本发明第四方面的mRNA。
在一些实施方案中，组合物包含具有SEQ ID NO.14-16中任一项所示的序列的mRNA和具有如SEQ ID NO.27所示的序列的mRNA。在优选的实施方案中，组合物包含具有如SEQ ID NO.14所示的序列的mRNA和具有如SEQ ID NO.27所示的序列的mRNA。
在一些实施方案中，组合物中两种重组S蛋白或两种mRNA的摩尔比例为1-3:1-3，例如1:1、1:1.5、1:2、1:2.5、1:3、1.5:2.5、2:1.5、2:2.5、2:3、2.5:3，优选1:1。在优选的实施方案中，组合物中本发明第一方面的重组S蛋白与本发明第三方面的重组S蛋白的摩尔比例为1-3:1-3，例如1:1、1:1.5、1:2、1:2.5、1:3、1.5:1、1.5:2、1.5:2.5、2:1、2:1.5、2:2.5、2:3、2.5:1、2.5:1.5、2.5:2、2.5:3、3:1、3:2、3:2.5，优选1:1。在优选的实施方案中，组合物中本发明第二方面的mRNA与本发明第四方面的mRNA的摩尔比例为1-3:1-3，例如1:1、1:1.5、1:2、1:2.5、1:3、1.5:1、1.5:2、1.5:2.5、2:1、2:1.5、2:2.5、2:3、2.5:1、2.5:1.5、2.5:2、2.5:3、3:1、3:2、3:2.5，优选1:1。
在一些实施方案中，组合物还包含以下重组S蛋白或编码其的mRNA：
(a)相对于野生型S蛋白包括以下突变的重组S蛋白：K986P和V987P；和/或
(b)相对于野生型S蛋白包括以下突变的重组S蛋白：G75V、T76I、R246缺失、S247缺失、Y248缺失、L249缺失、T250缺失、P251缺失、G252缺失、D253N、L452Q、F490S、D614G、T859N；K986P；和V987P；
和/或
(c)相对于野生型S蛋白包括以下突变的重组S蛋白：S1/S2切割位点突变为GGSG；K986P；和V987P；
和/或
(d)相对于野生型S蛋白包括以下突变的重组S蛋白：S2切割位点突变为AN；K986P；和V987P；
和/或
(e)相对于野生型S蛋白包括以下突变的重组S蛋白：S1/S2切割位点突变为GGSG；S2切割位点突变为AN；K986P；和V987P；
和/或
(f)相对于野生型S蛋白包括以下突变的重组S蛋白：G75V、T76I、R246缺失、S247缺失、Y248缺失、L249缺失、T250缺失、P251缺失、G252缺失、D253N、L452Q、F490S、D614G、T859N；S1/S2切割位点突变为GGSG；K986P；和V987P；
和/或
(g)相对于野生型S蛋白包括以下突变的重组S蛋白：G75V、T76I、R246缺失、S247缺失、Y248缺失、L249缺失、T250缺失、P251缺失、G252缺失、D253N、L452Q、F490S、D614G、T859N；S2切割位点突变为AN；K986P；和V987P；
和/或
(h)相对于野生型S蛋白包括以下突变的重组S蛋白：G75V、T76I、R246缺失、S247缺失、Y248缺失、L249缺失、T250缺失、P251缺失、G252缺失、D253N、L452Q、F490S、D614G、T859N；S1/S2切割位点突变为GGSG；S2切割位点突变为AN；K986P；和V987P。
在一些实施方案中，重组S蛋白(a)中的S1/S2切割位点RRAR可以经突变以失去被蛋白酶例如弗林样蛋白酶和溶酶体蛋白酶切割的能力；优选地，S1/S2切割位点RRAR突变为GGSG。
在一些实施方案中，重组S蛋白(a)中的S2切割位点KR可以经突变以失去被蛋白酶例如弗林样蛋白酶和溶酶体蛋白酶切割的能力；优选地，S2切割位点KR突变为AN。
在一些实施方案中，重组S蛋白(a)还包含三聚体结构域，所述三聚体结构域当表达时促进所述重组S蛋白(a)形成三聚体。在一些实施方案中，重组S蛋白(a)中的三聚体结构域可以包含T4噬菌体fibritin三聚体基序。在一些实施方案中，所述T4噬菌体fibritin三聚体基序可以具有SEQ ID NO.18所示的氨基酸序列。
在一些实施方案中，重组S蛋白(a)中的三聚体结构域可以与重组S蛋白(a)直接融合。在另一些实施方案中，三聚体结构域可以通过接头与重组S蛋白(a)融合。在一些实施方案中，三聚体结构域可以与重组S蛋白(a)的N端融合。在另一些实施方案中，三聚体结构域可以与重组S蛋白(a)的C端融合。例如，三聚体结构域可以通过接头与重组S蛋白(a)的C端融合。在一些实施方案中，所述接头序列可以包含SEQ ID NO.19所示的序列。
在一些实施方案中，重组S蛋白(a)可以不包含功能性融合肽结构域(FP结构域。例如，重组S蛋白(a)可以包含经突变的融合肽结构域，例如通过一个或多个氨基酸残基的取代、缺失、插入和/或添加，导致融合肽结构域失去其天然功能，例如介导病毒与宿主细胞膜进行融合的功能。或者，在一些实施方案中，重组S蛋白(a)可以不包含融合肽结构域。
在一些实施方案中，重组S蛋白(a)可以不包含跨膜结构域和/或胞质结构域。在一些实施方案中，重组S蛋白(a)可以不包含胞质结构域。在一些实施方案中，重组S蛋白(a)可以不包含跨膜结构域和胞质结构域。
在一些实施方案中，重组S蛋白(a)还包含信号序列；优选地，所述信号序列包含免疫球蛋白重链可变区(IGHV)信号序列。例如，所述信号序列可以具有SEQ ID NO.17所示的氨基酸序列。
在一些实施方案中，重组S蛋白(a)从N端到C端由以下中的任一项组成：
i)胞外结构域和三聚体结构域；
ii)胞外结构域、跨膜结构域和三聚体结构域；
iii)信号序列、胞外结构域和三聚体结构域；和
iv)信号序列、胞外结构域、跨膜结构域和三聚体结构域。
在一些实施方案中，所述胞外结构域相对于野生型序列具有以下突变中的一种或多种：
1)S1/S2切割位点RRAR经突变以失去被弗林样蛋白酶和溶酶体蛋白酶切割的能力，优选地，S1/S2切割位点突变为GGSG；
2)S2切割位点KR经突变以失去被弗林样蛋白酶和溶酶体蛋白酶切割的能力，优选地，S2切割位点突变为AN；
3)K986P和/或V987P突变；
4)融合肽结构域经突变以失去介导病毒与宿主细胞膜进行融合的功能；优选融合肽结构域缺失突变。
在优选的实施方案中，重组S蛋白(a)从N端到C端由以下中的任一项组成：
i)胞外结构域和三聚体结构域；
ii)胞外结构域、跨膜结构域和三聚体结构域；
iii)信号序列、胞外结构域和三聚体结构域；和
iv)信号序列、胞外结构域、跨膜结构域和三聚体结构域；
其中所述胞外结构域的氨基酸序列为SEQ ID NO.29所示的氨基酸序列的氨基酸位置14-1213并进行了以下突变获得的序列：在SEQ ID NO.29所示的氨基酸序列的氨基酸位置986和987处具有K986P和V987P取代且在氨基酸位置817-987中不具有其它突变，且所述胞外结构域中的S2切割位点KR经突变以失去被弗林样蛋白酶和溶酶体蛋白酶切割的能力，并且所述胞外结构域中的S1/S2切割位点RRAR经突变以失去被弗林样蛋白酶和溶酶体蛋白酶切割的能力，
其中所述三聚体结构域当表达时促进所述重组S蛋白(a)形成三聚体，其中所述三聚体结构域为T4噬菌体fibritin三聚体基序，并且所述三聚体结构域通过任选的接头序列融合在所述胞外结构域或跨膜结构域的C端。
在优选的实施方案中，重组S蛋白(a)具有选自SEQ ID NO.30-33中任一项所示的氨基酸序列。在优选的实施方案中，编码重组S蛋白(a)的mRNA具有选自SEQ ID NO.34-37中任一项所示的序列。
重组S蛋白(a)的结构和编码其的mRNA可参见例如中国专利申请号202011369776.2，其通过引用整体并入本文。
在其他实施方案中，重组S蛋白(b)中的S1/S2切割位点RRAR可以经突变以失去被蛋白酶例如弗林样蛋白酶和溶酶体蛋白酶切割的能力；优选地，S1/S2切割位点RRAR突变为GGSG。
在一些实施方案中，重组S蛋白(b)中的S2切割位点KR可以经突变以失去被蛋白酶例如弗林样蛋白酶和溶酶体蛋白酶切割的能力；优选地，S2切割位点KR突变为AN。
在一些实施方案中，重组S蛋白(b)可以不包含跨膜结构域和/或胞质结构域。在一些实施方案中，重组S蛋白(b)可以不包含胞质结构域。在一些实施方案中，重组S蛋白(b)可以不包含跨膜结构域和胞质结构域。
在一些实施方案中，重组S蛋白(b)从N端到C端由以下中的任一项组成：i)胞外结构域、任选的跨膜结构域和任选的胞质结构域；和ii)信号序列、胞外结构域、任选的跨膜结构域和任选的胞质结构域。
在一些实施方案中，所述胞外结构域相对于野生型序列具有以下突变中的一种或多种：
1)G75V、T76I、R246缺失、S247缺失、Y248缺失、L249缺失、T250缺失、P251缺失、G252缺失、D253N、L452Q、F490S、D614G、T859N；
2)S1/S2切割位点RRAR经突变以失去被弗林样蛋白酶和溶酶体蛋白酶切割的能力，优选地，S1/S2切割位点突变为GGSG；
3)S2切割位点KR经突变以失去被弗林样蛋白酶和溶酶体蛋白酶切割的能力，优选地，S2切割位点突变为AN；
4)K986P和/或V987P突变。
在优选的实施方案中，重组S蛋白(b)从N端到C端由信号序列、胞外结构域、跨膜结构域和胞质结构域组成，所述胞外结构域相对于野生型序列具有以下突变：G75V、T76I、R246缺失、S247缺失、Y248缺失、L249缺失、T250缺失、P251缺失、G252缺失、D253N、L452Q、F490S、D614G、T859N。
在另一优选的实施方案中，重组S蛋白(b)从N端到C端由信号序列、胞外结构域、跨膜结构域和胞质结构域组成，所述胞外结构域相对于野生型序列具有以下突变：G75V、T76I、R246缺失、S247缺失、Y248缺失、L249缺失、T250缺失、P251缺失、G252缺失、D253N、L452Q、F490S、D614G、T859N、S1/S2切割位点RRAR突变为GGSG和S2切割位点KR突变为AN。例如，重组S蛋白(b)具有如SEQ ID NO.38所示的氨基酸序列。
在另一优选的实施方案中，重组S蛋白(b)从N端到C端由信号序列、胞外结构域、跨膜结构域和胞质结构域组成，所述胞外结构域相对于野生型序列具有以下突变：G75V、T76I、R246缺失、S247缺失、Y248缺失、L249缺失、T250缺失、P251缺失、G252缺失、D253N、L452Q、F490S、D614G、T859N、K986P和V987P。例如，重组S蛋白(b)具有如SEQ ID NO.39所示的氨基酸序列。
在又一优选的实施方案中，重组S蛋白(b)从N端到C端由信号序列、胞外结构域、跨膜结构域和胞质结构域组成，所述胞外结构域相对于野生型序列具有以下突变：G75V、T76I、R246缺失、S247缺失、Y248缺失、L249缺失、T250缺失、P251缺失、G252缺失、D253N、L452Q、F490S、D614G、T859N、K986P、V987P和S1/S2切割位点RRAR突变为GGSG。例如，重组S蛋白(b)具有如SEQ ID NO.40所示的氨基酸序列。
在另一优选的实施方案中，重组S蛋白(b)从N端到C端由信号序列、胞外结构域、跨膜结构域和胞质结构域组成，所述胞外结构域相对于野生型序列具有以下突变：G75V、T76I、R246缺失、S247缺失、Y248缺失、L249缺失、T250缺失、P251缺失、G252缺失、D253N、L452Q、F490S、D614G、T859N、K986P、V987P和S2切割位点KR突变为AN。例如，重组S蛋白(b)具有如SEQ ID NO.41所示的氨基酸序列。
在又一优选的实施方案中，重组S蛋白(b)从N端到C端由信号序列、胞外结构域、跨膜结构域和胞质结构域组成，所述胞外结构域相对于野生型序列具有以下突变：G75V、T76I、R246缺失、S247缺失、Y248缺失、L249缺失、T250缺失、P251缺失、G252缺失、D253N、L452Q、F490S、D614G、T859N、K986P、V987P、S1/S2切割位点RRAR突变为GGSG和S2切割位点KR突变为AN。例如，重组S蛋白(b)具有如SEQ ID NO.42所示的氨基酸序列。
在优选的实施方案中，重组S蛋白(b)具有选自SEQ ID NO.38-42中任一项所示的氨基酸序列。在优选的实施方案中，编码重组S蛋白(b)的mRNA具有如SEQ ID NO.43所示的序列。
重组S蛋白(b)的结构和编码其的mRNA可参见例如中国专利申请号202210159238.3，其通过引用整体并入本文。
在一些实施方案中，本发明的组合物还包含一种或多种药学上可接受的载体、赋形剂或稀释剂。
如本文所用的术语“药学上可接受的”是指在合理的医学判断的范围内适合用于与人和动物的组织接触而没有过度毒性、刺激性、变应性应答或其它问题或并发症，与合理的益处/风险比相称的载体、赋形剂或稀释剂。
用于本发明的组合物中的示例性载体包括盐水、缓冲盐水、葡萄糖和水。用于本发明的组合物中的示例性赋形剂包括填充剂、粘合剂、崩解剂、包衣剂、吸附剂、抗粘附剂、助流剂、防腐剂、抗氧化剂、调味剂、着色剂、甜味剂、溶剂、共溶剂、缓冲剂、螯合剂、粘度赋予剂、表面活性剂、稀释剂、润湿剂、载体、稀释剂、防腐剂、乳化剂、稳定剂和张力调节剂。本领域技术人员已知选择合适的赋形剂以制备本发明的组合物。通常，合适的赋形剂的选择尤其取决于所使用的活性剂、待治疗的疾病和组合物的期望剂型。
根据所采用的活性剂(如mRNA)，可将本公开的组合物制备成各种形式，如固态、液态、气态或冻干形式，特别可以是软膏剂、乳膏剂、透皮贴剂、凝胶剂、粉剂、片剂、溶液剂、气雾剂、颗粒剂、丸剂、混悬剂、乳剂、胶囊剂、糖浆剂、液体剂、酏剂、浸膏剂、酊剂或流浸膏提取物的形式，或者是特别适用于所需施用方法的形式。本发明已知的用于生产药物的过程在第22版的Remington’s Pharmaceutical Sciences(Ed.MaackPublishing Co，Easton，Pa.，2012)中显示，并可包括例如常规混合、溶解、制粒、制糖衣、研磨、乳化、包封、包埋或冻干过程。
在一些实施方案中，组合物可以是疫苗组合物，任选地，疫苗组合物还包含一种或多种佐剂。
如本文中使用的，术语“疫苗组合物”指诱导或改进对特定疾病的免疫的生物制剂。用疫苗组合物激发个体的免疫系统诱导免疫细胞的形成和/或增殖，该免疫细胞特异性识别由疫苗包含的化合物。所述免疫细胞的至少一部分保持存活一段时间，其可以延长至疫苗接种后10,20或30年。如果个体的免疫系统在前述时间段内遇到衍生能引发免疫应答的化合物的病原体，那么通过疫苗接种生成的免疫细胞被再活化，且相比于尚未用疫苗激发和首次遇到病原体的免疫原性化合物的个体的免疫应答，增强针对该病原体的免疫应答。
在本文中，“疫苗接种”、“免疫接种”、“免疫”或“接种”是指向受试者施用疫苗，其目的是防止受试者发展出疾病的一种或多种症状。原则上，疫苗接种包括初次疫苗接种和任选地一次或多次加强疫苗接种。初次疫苗接种或初次免疫定义为施用本文公开的组合物或单位剂量以建立保护性免疫应答的初次施用计划。加强疫苗接种或加强免疫是指在初次疫苗接种之后，例如在初次疫苗接种计划的最后一次施用后至少一周、至少2周、至少一个月、至少6个月、至少1年或甚至5或10年之后进行的施用或施用计划。加强施用试图增强或重建初次疫苗接种的免疫应答。
对本发明的组合物或疫苗组合物的免疫应答是受试者中形成对组合物中存在的抗原蛋白的体液和/或细胞免疫应答。出于本发明的目的，“体液免疫应答”指由抗体分子，包括分泌性(IgA)或IgG分子介导的免疫应答，而“细胞免疫应答”指由T-淋巴细胞和/或其它白细胞介导的免疫应答。细胞免疫的一个重要方面牵涉细胞裂解T细胞(“CTL”)的抗原特异性应答。CTL对由主要组织相容性复合物(MHC)编码并且在细胞表面上表达的蛋白质结合呈递的肽抗原具有特异性。CTL帮助诱导并促进对胞内微生物的破坏，或者此类微生物感染的细胞的裂解。细胞免疫的另一个方面牵涉辅助T细胞的抗原特异性应答。辅助T细胞作用为帮助刺激功能，并且聚焦非特异性效应细胞针对在其表面上展示与MHC分子结合的肽抗原的细胞的活性。细胞免疫应答还涉及生成由活化的T细胞和/或其它白细胞，包括那些自CD4+和CD8+T细胞衍生的细胞生成的细胞因子、趋化因子和其它此类分子。
因此，免疫应答可以是刺激CTL，和/或辅助T细胞生成或活化的应答。也可以刺激趋化因子和/或细胞因子的生成。本发明的组合物或疫苗组合物也可以引发抗体介导的免疫应答。因此，免疫应答可以包括下列一种或多种效应：B细胞的抗体(例如IgA或IgG)生成；和/或抑制物、细胞毒性或辅助T细胞和/或特异性针对疫苗中存在的蛋白的T细胞的活化。这些应答可以用来中和感染性，和/或介导抗体-补体，或抗体依赖性细胞细胞毒性(ADCC)以对经免疫个体提供保护。可以使用本领域中公知的标准免疫测定法和中和测定法测定此类应答。
如本文中使用的，术语“佐剂”指在细胞水平或体液水平增加、刺激、活化、加强或调节针对组合物的活性成分的免疫应答的试剂，例如，免疫佐剂刺激免疫系统针对实际抗原的应答但其本身没有免疫效应。这样的佐剂的实例包括但不限于无机佐剂(例如，无机金属盐例如磷酸铝或氢氧化铝)、有机佐剂(例如，皂角苷或鲨烯)、油基佐剂(如弗氏完全佐剂和弗氏不完全佐剂)、细胞因子(如IL-1β、IL-2、IL-7、IL-12、IL-18、GM-CFS和INF-γ)、颗粒状佐剂(例如，免疫刺激性复合物(ISCOMS)、脂质体或可生物降解的微球)、病毒颗粒、细菌佐剂(例如单磷酰脂质A或胞壁肽)、合成佐剂(例如，非离子型嵌段共聚物、胞壁肽类似物或合成脂质A)或合成多核苷酸佐剂(例如，多聚精氨酸或多聚赖氨酸)。优选地，佐剂选自铝佐剂(例如氢氧化铝、磷酸铝、硫酸铝、明矾)、MF59、AS03、病毒体(例如肝炎病毒病毒体和流感病毒病毒体)、AS04、热可逆水包油乳液、ISA51、弗氏佐剂、IL-12、CpG基序、甘露糖或其任意组合。
在一些实施方案中，所述组合物或疫苗组合物还包含一种或多种另外的治疗剂。例如，所述治疗剂可以选自另外的抗原性蛋白或多肽、抗体、激素或激素类似物，和小分子药物。
在第六方面，本发明提供了DNA，其编码本发明第二方面的mRNA和/或本发明第四方法的mRNA。在一些实施方案中，本发明的DNA编码本发明第二方面的mRNA。在一些实施方案中，本发明的DNA编码本发明第四方面的mRNA。在一些实施方案中，本发明的DNA编码本发明第二方面的mRNA和本发明第四方法的mRNA。在一些实施方案中，本发明的DNA可以用于在体外通过转录制备本发明的mRNA。
在一些实施方案中，本发明的DNA包含如SEQ ID NO.11-13和26中任一项所示的序列，或由SEQ ID NO.11-13和26中任一项所示的序列组成。
在第七方面，本发明提供了重组质粒，其包含本发明第六方面的DNA。
在一些实施方案中，重组质粒为pT7TS质粒。
在一些实施方案中，重组质粒还包含起始序列(Ori)、T7启动子、5’-UTR和3’-UTR。
在一些实施方案中，起始序列为ColE1型起始序列。优选地，起始序列包含如SEQID NO.6所示的序列，或由SEQ ID NO.6所示的序列组成。
在一些实施方案中，T7启动子包含如SEQ ID NO.7(TAATACGACTCACTATAATG)所示的序列，或由SEQ ID NO.7所示的序列组成。
在一些实施方案中，5’-UTR可以包含选自下组的基因的5’-UTR或其同源物、片段或变体：β-珠蛋白(HBB)基因、热休克蛋白70(Hsp70)基因、轴丝动力蛋白重链2(DNAH2)基因、17β-羟基类固醇脱氢酶4(HSD17B4)基因。例如，变体序列可以与相应基因的野生型5’-UTR序列具有至少80％，至少85％，至少90％，至少95％，至少98％，或至少99％的序列同一性。
在一些实施方案中，5’-UTR包含衍生自17β-羟基类固醇脱氢酶4(HSD17B4)基因的5’-UTR或其同源物、片段或变体。在一些实施方案中，5’-UTR包含KOZAK序列。在一些实施方案中，5’-UTR包含衍生自17β-羟基类固醇脱氢酶4(HSD17B4)基因的5’-UTR或其同源物、片段或变体，和KOZAK序列。在一些实施方案中，5’-UTR包含如SEQ ID NO.8和/或SEQ ID NO.9所示的序列。
在一些实施方案中，3’-UTR可以包含选自下组的基因的3’-UTR或其同源物、片段或变体：白蛋白(ALB)基因、α-珠蛋白基因、β-珠蛋白(HBB)基因、酪氨酸羟化酶基因、热休克蛋白70(Hsp70)基因，脂氧合酶基因和胶原蛋白α基因。例如，变体序列可以与相应基因的野生型3’-UTR序列具有至少80％，至少85％，至少90％，至少95％，至少98％，或至少99％的序列同一性。
在一些实施方案中，3’-UTR包含衍生自白蛋白(ALB)基因的3’-UTR或其同源物、片段或变体。优选地，3’-UTR包含如SEQ ID NO.10所示的序列。
在一些实施方案中，重组质粒还包含polyA、抗性基因启动子和抗性基因。
在一些实施方案中，poly A尾的长度可以为100-200个核苷酸，例如约100个核苷酸，约110个核苷酸，约120个核苷酸，约130个核苷酸，约140个核苷酸，约150个核苷酸，约160个核苷酸，约170个核苷酸，约180个核苷酸，约190个核苷酸，或约200个核苷酸。在一些实施方案中，polyA尾的长度可以为100-150个核苷酸。在一些实施方案中，所述poly A尾的长度可以为约120个核苷酸。
在一些实施方案中，抗性基因启动子为氨苄青霉素抗性基因启动子。
在一些实施方案中，抗性基因为硫酸卡那霉素抗性基因。
在优选的实施方案中，重组质粒包含如SEQ ID NO.28所示的核苷酸序列，或由SEQID NO.28所示的核苷酸序列组成。
在第八方面，本发明提供了mRNA-载体颗粒，其包含本发明第二方面的mRNA和/或本发明第四方法的mRNA，以及包封所述mRNA的载体材料。
在一些实施方案中，载体材料可以选自鱼精蛋白、脂质纳米颗粒(LNP)、高分子材料和无机纳米颗粒。在优选的实施方案中，所述载体材料是LNP。
在一些实施方案中，LNP可以包含离子化脂质、聚乙二醇化脂质、胆固醇及衍生物和磷脂中的一种或多种。例如，LNP可以包含离子化脂质、聚乙二醇化脂质、胆固醇及衍生物和磷脂中的任一种，其中任意两种、任意三种或全部四种的组合。
在第九方面，本发明提供了预防和/或治疗受试者中与SARS-CoV-2感染相关的疾病或病症的方法，其包括向受试者施用有效量的本发明的重组S蛋白、mRNA、组合物、重组质粒、或mRNA-载体颗粒。
如本文所用的术语“预防”或“治疗”是指疾病或病症的获得或发展的风险的降低，即，使疾病的临床症状的至少一种在疾病发作之前易受疾病感染、或未暴露于致病原的受试者中不发展。例如，治疗可以包括：(i)在可能易患疾病、障碍和/或病症但尚未诊断患有所述疾病、障碍和/或病症的患者中预防所述疾病、障碍和/或病症；(ii)抑制所述疾病、障碍和/或病症，即阻止其发展；或(iii)减轻所述疾病、障碍和/或病症，即引起所述疾病、障碍和/或病症的消退。
如本文所用的术语“有效量”意指当施用到受试者用于治疗或预防疾病时足以实现这样的治疗或预防的化合物的量。“有效量”可取决于化合物、疾病及其严重度、以及待治疗的受试者的年龄、体重等改变。“治疗有效量”是指用于治疗性治疗的有效量。“预防有效量”是指用于预防性治疗的有效量。
如本文所用的术语“施用”是指使用本领域技术人员已知的任何各种方法和递送系统将药剂物理导入受试者。示例性施用途径包括静脉内、肌肉内、皮下、腹膜内、脊柱或其他肠胃外施用途径，例如通过注射或输液。
如本文中使用的，术语“受试者”、“个体”、和“患者”是本领域中公知的，并且在本文中可互换使用，指需要治疗的任何受试者，特别是哺乳动物受试者。实例包括但不限于人和其它灵长类，包括非人灵长类，诸如黑猩猩及其它猿和猴物种。术语个体、受试者和患者本身不表示特定年龄、性别、人种等。
在本发明的方法的实施方案中，所述疾病或病症为SARS-CoV-2变异株，例如Delta变异株、Omicron变异株或Lambda变异株感染引起的疾病或病症。
在第十方面，本发明提供了本发明的重组S蛋白、mRNA、组合物、重组质粒、或mRNA-载体颗粒在制备用于在受试者中预防和/或治疗与SARS-CoV-2感染相关的疾病或病症的药物中的用途。
在第十一方面，本发明提供了本发明的重组S蛋白、mRNA、组合物、重组质粒、或mRNA-载体颗粒，其用于预防和/或治疗受试者中与SARS-CoV-2感染相关的疾病或病症。
在本发明的用途的实施方案中，所述疾病或病症为SARS-CoV-2变异株，例如Delta变异株、Omicron变异株或Lambda变异株感染引起的疾病或病症。
附图说明
图1显示了RBMRNA-Delta质粒结构示意图。
图2显示了通过RBMRNA-Delta质粒转录得到的核苷酸序列为SEQ ID NO:14的mRNA的电泳结果图。
图3显示了检测从RBMRNA-Delta mRNA表达的重组S蛋白的蛋白免疫印迹结果图。
图4显示了通过ELISPOT法检测的RBMRNA-Delta 1疫苗在小鼠体内诱导T淋巴细胞分泌IFN-γ、IL-2、IL-4和IL-5的结果。*：P<0.05，**：P<0.01。
图5显示了通过ELISPOT法检测的RBMRNA-Delta 2疫苗在小鼠体内诱导T淋巴细胞分泌IFN-γ、IL-2、IL-4和IL-5的结果。*：P<0.05，**：P<0.01。
图6显示了通过ELISPOT法检测的RBMRNA-Delta 3疫苗在小鼠体内诱导T淋巴细胞分泌IFN-γ、IL-2、IL-4和IL-5的结果。*：P<0.05，**：P<0.01。
图7显示了通过流式细胞术检测的RBMRNA-Delta 1疫苗在小鼠体内诱导细胞内因子IFN-γ、IL-2、IL-4和IL-5分泌的结果。*：P<0.05，**：P<0.01。
图8显示了通过流式细胞术检测的RBMRNA-Delta 2疫苗在小鼠体内诱导细胞内因子IFN-γ、IL-2、IL-4和IL-5分泌的结果。*：P<0.05，**：P<0.01。
图9显示了通过流式细胞术检测的RBMRNA-Delta 3疫苗在小鼠体内诱导细胞内因子IFN-γ、IL-2、IL-4和IL-5分泌的结果。*：P<0.05，**：P<0.01。
图10显示了通过流式细胞术检测的RBMRNA-Delta 1疫苗在小鼠体内诱导免疫应答的T细胞亚群结果图。*：P<0.05，**：P<0.01，***：P<0.001。
图11显示了通过流式细胞术检测的RBMRNA-Delta 2疫苗在小鼠体内诱导免疫应答的T细胞亚群结果图。*：P<0.05，**：P<0.01。
图12显示了通过流式细胞术检测的RBMRNA-Delta 3疫苗在小鼠体内诱导免疫应答的T细胞亚群结果图。*：P<0.05，**：P<0.01，***：P<0.001。
图13显示了接种RBMRNA-Delta 1疫苗后小鼠血清中IgG抗体水平的结果。
图14显示了接种RBMRNA-Delta 2疫苗后小鼠血清中IgG抗体水平的结果。
图15显示了接种RBMRNA-Delta 3疫苗后小鼠血清中IgG抗体水平的结果。
图16显示了接种RBMRNA-Delta 1疫苗后小鼠血清对野生型假病毒和德尔塔型假病毒的中和效果。
图17显示了接种RBMRNA-Delta 3疫苗后小鼠血清对野生型假病毒和德尔塔型假病毒的中和效果。
图18显示了RBMRNA-Omicron质粒的结构示意图。
图19显示了由RBMRNA-Omicron质粒转录得到的RBMRNA-Omicron mRNA的电泳结果图。
图20显示了检测由RBMRNA-Omicron mRNA表达的重组S蛋白的蛋白免疫印迹结果图。
图21显示了通过ELISPOT法检测的RBMRNA-Omicron疫苗在小鼠体内诱导T淋巴细胞分泌细胞因子IFN-γ、IL-2、IL-4和IL-5的结果图。*：P<0.05，**：P<0.01，***：P<0.001，****：P<0.0001。
图22显示了通过流式细胞术检测的RBMRNA-Omicron疫苗在小鼠体内诱导细胞因子IFN-γ、IL-2、IL-4和IL-5分泌的结果图。*：P<0.05，**：P<0.01。
图23显示了通过流式细胞术检测的RBMRNA-Omicron疫苗在小鼠体内诱导免疫应答的T细胞亚群结果图。*：P<0.05，**：P<0.01，***：P<0.001，****：P<0.0001。
图24显示了接种RBMRNA-Omicron疫苗或RBMRNA-组合疫苗后小鼠血清中IgG抗体水平的结果图。
图25显示了接种RBMRNA-组合疫苗后小鼠对Omicron型真病毒的中和抗体效价(NAb)的结果图。*：P<0.05，**：P<0.01，***：P<0.001，****：P<0.0001。
图26显示了接种RBMRNA-Omicron疫苗或者RBMRNA-组合疫苗后小鼠对Delta型病毒的TCID50结果图。*：P<0.05，**：P<0.01，***：P<0.001，****：P<0.0001。
图27显示了接种RBMRNA-Omicron疫苗或者RBMRNA-组合疫苗后小鼠对Omicron型病毒的TCID50结果图。*：P<0.05，**：P<0.01，***：P<0.001，****：P<0.0001。
具体实施方式
下面结合具体实施例来进一步描述本发明，本发明的优点和特点将会随着描述而更为清楚。应当理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。以下实施例中未注明具体条件的实验方法，通过按照本领域的常规条件，例如，Sambrook和Russeii等人，分子克隆：实验室手册(第三版)(2001)，CSHL出版社中所述的条件，或按照制造商所建议的条件。除非另有说明，否则以下实施例中所用的实验材料和试剂均可商购获得。
实施例1.mRNA的制备
RBMRNA-Delta mRNA的制备
在野生型SARS-CoV-2 S蛋白的基础上经过以下突变得到重组S蛋白(SEQ IDNO.3)：T19R、G142D、E156G、F157和R158缺失突变、A222V、L452R、T478K、D614G、P681R、D950N；682-685位(S1/S2切割位点)RRAR突变为GGSG；814-815位(S2切割位点)KR突变为AN；986-987位的KV序列突变为两个脯氨酸PP。
以该重组S蛋白的序列为基础，设计DNA编码序列(SEQ ID NO.11)。在DNA编码序列的基础上增加例如5’-UTR、3’-UTR、polyA序列后，通过同源重组插入至pT7TS质粒中进行构建，形成重组载体pT7TS-2.0，获得最终的重组质粒命名为RBMRNA-Delta质粒。
RBMRNA-Delta质粒包含的元件包括起始序列(SEQ ID NO.6)，T7启动子序列(SEQID NO.7)，5’-UTR序列(SEQ ID NO.8)，3’-UTR序列(SEQ ID NO.10)，3’端聚腺苷酸(polyA)序列，氨苄青霉素抗性基因启动子、硫酸卡那霉素抗性基因。通过这些非编码结构来调节RBMRNA-Delta质粒经转录得到的mRNA的稳定性、翻译效率和免疫原性。
对RBMRNA-Delta质粒的编码区及polyA区域进行测序，插入的目的基因测序与参考序列完全一致，构建成功的整个质粒结构如图1所示。RBMRNA-Delta质粒经体外转录后得到mRNA(命名为RBMRNA-Delta mRNA)，mRNA经翻译得到蛋白质(命名为RBMRNA-Delta蛋白质)。
通过重组质粒转录得到mRNA的序列如SEQ ID NO.14所示。用Agilent 2200Tapestation电泳系统对转录得到的mRNA大小及完整性进行分析。结果如图2所示，转录mRNA条带单一，无降解。
使用上述方法还得到了如SEQ ID NO.15和SEQ ID NO.16所示的mRNA，其DNA编码序列分别如SEQ ID NO.12和SEQ ID NO.13所示，编码的重组S蛋白的氨基酸序列分别如SEQID NO.5和SEQ ID NO.4所示。得到的重组S蛋白相对于野生型S蛋白包含以下突变：
(1)T19R、G142D、E156G、F157和R158缺失突变、A222V、L452R、T478K、D614G、P681R、D950N；682-685位(S1/S2切割位点)RRAR突变为GGSG；814-815位(S2切割位点)KR突变为AN；去除跨膜结构域和胞质结构域；去除融合肽结构域；将T4噬菌体fibritin基序连接至胞外结构域的C端；将信号肽序列替换成免疫球蛋白重链可变区(IGHV)信号序列(SEQ IDNO.4)；
(2)在上述(1)的基础上将986-987位的KV序列突变为两个脯氨酸PP(SEQ IDNO.5)。
RBMRNA-Omicron mRNA的制备
在野生型SARS-CoV-2 S蛋白的基础上经过以下突变得到重组S蛋白(SEQ IDNO.25)：A67V、H69del、V70del、T95I、G142del、V143del、Y144del、Y145D、N211del、L212I以及在R214和D215之间三个氨基酸E、P、E的插入突变、G339D、S371L、S373P、S375F、K417N、N440K、G446S、S477N、T478K、E484A、Q493R、G496S、Q498R、N501Y、Y505H、T547K、D614G、H655Y、N679K、P681H、N764K、D796Y、N856K、Q954H、N969K、L981F突变；682-685位(S1/S2切割位点)RRAR突变为GGSG；814-815位(S2切割位点)KR突变为AN；986-987位的KV序列突变为两个脯氨酸PP。
以该重组S蛋白的序列为基础，设计DNA编码序列(SEQ ID NO.26)。在DNA编码序列的基础上增加例如5’-UTR、3’-UTR、polyA序列后，通过同源重组插入至pT7TS质粒中进行构建，形成重组载体pT7TS-2.0，获得最终的重组质粒(命名为RBMRNA-Omicron质粒，其核苷酸序列为SEQ ID NO.28)。
RBMRNA-Omicron质粒包含的元件包括起始序列(SEQ ID NO.6)，T7启动子序列(SEQ ID NO.7)，5’-UTR序列(SEQ ID NO.8)，3’-UTR序列(SEQ ID NO.10)，3’端聚腺苷酸(polyA)序列，氨苄青霉素抗性基因启动子，硫酸卡那霉素抗性基因。通过这些非编码结构来调节RBMRNA-Omicron质粒经转录得到的mRNA的稳定性、翻译效率和免疫原性。
对RBMRNA-Omicron质粒的编码区及polyA区域进行测序，插入的目的基因测序与参考序列完全一致，构建成功的整个质粒结构如图18所示。RBMRNA-Omicron质粒(SEQ IDNO.28)经体外转录后得到mRNA(命名为RBMRNA-Omicron mRNA)(SEQ ID NO.27)，mRNA经翻译得到蛋白质(命名为RBMRNA-Omicron蛋白质)(SEQ ID NO.25)。
由重组质粒RBMRNA-Omicron质粒转录得到的mRNA大小及完整性用Agilent2200Tapestation电泳系统进行分析，结果显示转录的mRNA条带单一，无降解(图19)。
实施例2.mRNA的表达验证
参照Lipofectamine MessagerMAX(ThermoFisher Scientific)的说明书，分别用2.5μg实施例1中得到的RBMRNA-Delta mRNA(SEQ ID NO.14)和RBMRNA-Omicron mRNA(SEQID NO.27)转染293T细胞，未转染mRNA的细胞作为阴性对照。转染24小时后，通过蛋白免疫印迹检测预融合德尔塔型或奥密克戎型SARS-CoV-2 S蛋白的表达，其中用兔抗SARS-CoV-2S蛋白抗体(GeneTex，GTX632604)进行免疫印迹，使用羊抗兔-HRP二抗进行标记。结果分别如图3(德尔塔型)和图20(奥密克戎型)所示，成功在180kDa处检测到大小与预期一致的预融合S蛋白的表达。
实施例3.mRNA疫苗诱导的Th1或Th2免疫应答
mRNA疫苗的制备
使用实施例1中的3种RBMRNA-Delta mRNA序列(SEQ ID No.14-16)和RBMRNA-Omicron mRNA序列(SEQ ID NO.27)分别制备mRNA疫苗，分别称为RBMRNA-Delta 1疫苗、RBMRNA-Delta 2疫苗和RBMRNA-Delta3疫苗以及RBMRNA-Omicron疫苗。使用包括以下组分的脂质纳米颗粒包封mRNA：8-(3-羟基丙基)(9,12-二烯基-十八烷基-1)-胺基-辛酸十七烷-9-醇酯、二硬脂酰基磷脂酰胆碱(DSPC)、1,2-二肉豆蔻酰-rac-甘油-3-甲氧基聚乙二醇2000(DMG-PEG2000)和胆固醇。制备方法包括将上述组分溶解在乙醇溶液中，通过微流控混合器将所述脂质乙醇溶液与mRNA水溶液混合得到脂质纳米颗粒，以及对混合物进行透析、超滤和微米膜过滤从而获得mRNA的LNP疫苗制剂。具体的疫苗制备方法参见例如中国专利申请号202011369776.2，其通过引用整体并入本文。
RBMRNA-Delta疫苗诱导的Th1或Th2免疫应答
将得到的3种RBMRNA-Delta mRNA疫苗用于BALB/c小鼠免疫试验。免疫小鼠采用6-8周龄的雌性SPF级别健康BALB/c小鼠。实验根据小鼠体重均衡随机分为溶剂对照组(PBS)，疫苗低剂量组(1μg/只)，疫苗中剂量组(5μg/只)和疫苗高剂量组(20μg/只)，每组12只。分组后按照设定剂量通过肌肉注射分别于第0天和第14天接种2次疫苗制剂以进行初次和加强免疫，溶剂对照组给予等体积的PBS。
在加强免疫7天后，取小鼠脾脏分离脾脏淋巴细胞，通过ELISPOT法检测分泌IFN-γ、IL-2、IL-4和IL-5的T淋巴细胞。RBMRNA-Delta 1疫苗、RBMRNA-Delta 2疫苗和RBMRNA-Delta 3疫苗的结果分别如图4-6所示。结果显示，在接种低、中、高剂量的3种疫苗后，分泌Th1型细胞因子INFγ、IL2的T淋巴细胞显著多于分泌Th2型细胞因子IL4、IL5的T淋巴细胞。
在加强免疫9天后，使用抗IFN-γ抗体(Biolegend，货号505808)、抗IL-2抗体(Biolegend，货号503808)、抗IL-4抗体(Biolegend，货号504104)和抗IL-5抗体(Biolegend，货号504304)，通过流式细胞术分别检测细胞内因子IFN-γ、IL-2、IL-4和IL-5的水平，以进一步评估mRNA疫苗诱导的Th1或Th2免疫反应。RBMRNA-Delta 1疫苗、RBMRNA-Delta 2疫苗和RBMRNA-Delta 3疫苗的结果分别如图7-9所示。结果显示，3种疫苗均导致CD4+T细胞内Th1型细胞因子IFN-γ和IL-2的水平呈剂量依赖性增加，而CD8+T细胞内细胞因子的水平无明显变化。
这些结果表明3种RBMRNA-Delta疫苗诱导的免疫反应为Th1型偏倚的免疫反应。
RBMRNA-Omicron疫苗诱导的Th1或Th2免疫应答
将RBMRNA-Omicron疫苗用于BALB/c小鼠免疫试验，免疫小鼠采用SPF级别健康的BALB/c小鼠(6-8周龄，雌性)。实验根据小鼠体重均衡随机分为溶剂对照(PBS)组，RBMRNA-Omicron疫苗低剂量组(1μg/只)和RBMRNA-Omicron疫苗高剂量组(20μg/只)，每组3只。分组后分别于第0天和第21天，按照设定剂量通过肌肉注射向小鼠接种2次疫苗以进行初次免疫和加强免疫，溶剂对照组给予等体积的PBS。
在加强免疫后7天，以SARS-CoV-2 S蛋白作为刺激物，刺激分离的小鼠脾脏淋巴细胞，通过Elispot法检测分泌IFN-γ、IL-2、IL-4和IL-5细胞因子的T淋巴细胞数。如图21所示，与溶剂对照(PBS)组相比，低、高剂量RBMRNA-Omicron疫苗均可使分泌Th1型细胞因子IFN-γ、IL-2和Th2型细胞因子IL-4的T淋巴细胞数显著升高，而分泌Th2型细胞因子IL-5的T淋巴细胞数无明显变化。
在加强免疫后9天，使用抗IFN-γ抗体(Biolegend，货号505808)、抗IL-2抗体(Biolegend，货号503808)、抗IL-4抗体(Biolegend，货号504104)和抗IL-5抗体(Biolegend，货号504304)，通过流式细胞术分别检测细胞内因子IFN-γ、IL-2、IL-4和IL-5的水平，以进一步评估RBMRNA-Omicron疫苗诱导的Th1或Th2免疫反应。如图22所示,RBMRNA-Omicron疫苗导致CD4+T细胞内Th1型细胞因子IFN-γ和IL-2的水平呈剂量依赖性增加，而CD8+T细胞内细胞因子的水平无明显变化。
这些结果表明，RBMRNA-Omicron疫苗诱导的免疫反应为Th1型偏倚的免疫反应。
实施例4.T细胞亚群检测
采用与实施例3中相同的实验方法用3种RBMRNA-Delta疫苗和RBMRNA-Omicron疫苗免疫小鼠，每组3只小鼠，取分离的小鼠脾脏淋巴细胞进行免疫实验。在加强免疫9天后，通过流式细胞术检测T细胞亚群，以评价RBMRNA-Delta疫苗和RBMRNA-Omicron疫苗疫苗诱导的T淋巴细胞、CD4+ T细胞、CD8+ T细胞、效应记忆T(Tem)细胞的免疫应答水平。RBMRNA-Delta 1疫苗、RBMRNA-Delta 2疫苗和RBMRNA-Delta 3疫苗的结果分别如图10-12所示；RBMRNA-Omicron疫苗的结果如图23所示。
结果表明，与溶剂对照(PBS)组相比，3种RBMRNA-Delta疫苗和RBMRNA-Omicron疫苗均可诱导机体产生CD4+ T细胞和CD8+ T细胞介导的细胞免疫应答，并且能产生特异性效应记忆T细胞，使机体获得免疫记忆保护。
实施例5.mRNA疫苗诱导产生的IgG抗体效价的评估
RBMRNA-Delta疫苗诱导产生的IgG抗体效价
采用与实施例3中类似的实验方法将3种RBMRNA-Delta mRNA疫苗用于BALB/c小鼠免疫试验，每组取6只小鼠的血清进行实验。在加强免疫后14天，通过间接ELISA法检测小鼠血清中的特异性IgG抗体水平。RBMRNA-Delta 1疫苗、RBMRNA-Delta 2疫苗和RBMRNA-Delta3疫苗的结果分别如图13-15所示。
IgG抗体水平检测结果表明，3种疫苗在低、中、高剂量下，均能够在小鼠体内诱导产生高滴度的IgG抗体。
RBMRNA-Omicron疫苗和RBMRNA-组合疫苗诱导产生的IgG抗体效价
参考实施例3的方法制备RBMRNA-组合疫苗，RBMRNA-组合疫苗包含RBMRNA-Omicron mRNA(SEQ ID NO.27)和RBMRNA-Delta mRNA(SEQ ID NO.14)，这2种mRNA的摩尔比例为1:1。
将制备得到的RBMRNA-组合疫苗和RBMRNA-Omicro疫苗用于BALB/c小鼠免疫试验。实验根据小鼠体重均衡随机分为溶剂对照(PBS)组、RBMRNA-Omicron疫苗低剂量组(1μg/只)、RBMRNA-Omicron疫苗高剂量组(20μg/只)、RBMRNA-组合疫苗低剂量组(1μg/只)、RBMRNA-组合疫苗高剂量组(20μg/只)，每组6只。分组后分别于第0天和第21天，按照设定剂量通过肌肉注射向小鼠接种2次疫苗以进行初次免疫和加强免疫，溶剂对照组给予等体积的PBS，取小鼠的血清进行实验。在加强免疫后14天，通过间接ELISA法检测小鼠血清中特异性IgG抗体的效价。
如图24所示，RBMRNA-Omicron疫苗和RBMRNA-组合疫苗均能针对野生型、德尔塔型和奥密克戎型SARS-CoV-2在小鼠体内诱导产生高滴度的IgG抗体。
实施例6.RBMRNA-Delta mRNA疫苗诱导产生的中和抗体的评估
采用与实施例3中类似的实验方法将3种RBMRNA-Delta mRNA疫苗用于BALB/c小鼠免疫试验，每组取6只小鼠的血清进行实验。疫苗组仅采用高剂量(20μg/只)。分别在初次免疫后14天和加强免疫后14天采集小鼠血清，通过假病毒中和试验进行S蛋白特异性中和抗体检测。具体操作如下：
将血清水浴灭活后离心，收集上清液。用不含血清的DMEM培养基对灭活的血清进行稀释。向96孔板中加入稀释的血清和假病毒，在37℃下共孵育1小时。孵育结束后，向96孔板中加入293T-ACE2-p2A-mTagBFP2细胞(该细胞是通过CRISPR技术向293T细胞定点敲入ACE2-p2A-mTagBFP2而获得)。在37℃、5％CO2下培养48小时。培养结束后，洗掉培养基，然后加入PBS和萤火虫萤光素酶底物，避光震荡后通过多功能酶标仪检测萤光素酶化学发光值。采用Reed-muench法计算中和效价。
RBMRNA-Delta 1疫苗和RBMRNA-Delta 3疫苗诱导产生的抗体的中和效果分别如图16和17所示。
结果表明，经2种疫苗免疫的小鼠血清均可在初次免疫后14天基本上完全中和德尔塔型假病毒，并在加强免疫后14天基本上完全中和野生型假病毒。对于RBMRNA-Delta 1疫苗，经免疫的小鼠血清在初次免疫后14天，对野生型和德尔塔型假病毒的中和效价分别为282和966；在加强免疫后14天，对野生型和德尔塔型假病毒的中和效价分别为4007和6903(图16)。对于RBMRNA-Delta 3疫苗，经免疫的小鼠血清在初次免疫后14天，对野生型和德尔塔型假病毒的中和效价分别为271和874；在加强免疫后14天，对野生型和德尔塔型假病毒的中和效价分别为4232和4624(图17)。
实施例7.RBMRNA-Delta 1、RBMRNA-Omicron和RBMRNA-组合mRNA疫苗诱导产生的中和抗体的评估
采用与实施例3中类似的实验方法将RBMRNA-Delta 1mRNA疫苗、RBMRNA-Omicron疫苗和RBMRNA-组合疫苗用于BALB/c小鼠免疫试验，每组取6只小鼠的血清进行实验。疫苗组仅采用高剂量(20μg/只)。加强免疫后14天采集小鼠血清。不同疫苗被用于针对不同变异类型的假病毒中和实验中以评估不同疫苗针对不同假病毒的中和效价。具体操作如下：
将血清水浴灭活后离心，收集上清液。用不含血清的DMEM培养基对灭活的血清进行稀释。向96孔板中加入稀释的血清和假病毒，在37℃下共孵育1小时。孵育结束后，向96孔板中加入293T-ACE2-p2A-mTagBFP2细胞(该细胞是通过CRISPR技术向293T细胞定点敲入ACE2-p2A-mTagBFP2而获得)。在37℃、5％CO2下培养48小时。培养结束后，洗掉培养基，然后加入PBS和萤火虫萤光素酶底物，避光震荡后通过多功能酶标仪检测萤光素酶化学发光值。具体步骤参考例如中国专利申请号202210019169.6，其通过引用整体并入本文。
采用Reed-muench法计算中和效价。
RBMRNA-Delta 1疫苗、RBMRNA-Omicron疫苗和RBMRNA-组合疫苗针对不同假病毒诱导产生的抗体的中和效价如表1所示。
表1疫苗针对不同假病毒的中和效价
Figure BDA0003747844720000371
如表1所示，RBMRNA-Delta 1疫苗针对Delta型假病毒有强大的抑制作用，而RBMRNA-Omicron疫苗针对Omicron型假病毒有强大的抑制作用。此外，RBMRNA-组合疫苗针对所有的野生型、Beta型、Gamma型、Alpha型、Delta型、Omicron型和Deltacron型假病毒均有强大的抑制作用。
实施例8.真病毒实验
采用与实施例3中类似的实验方法将RBMRNA-组合疫苗用于BALB/c小鼠免疫试验，每组取6只小鼠的血清进行实验。疫苗组仅采用高剂量(20μg/只)。加强免疫后14天采集小鼠血清。自经免疫的小鼠采集的血清样本置于56℃中灭活30min，并以二倍稀释法用DMEM培养基(GBCO)连续稀释。经稀释的血清在96孔板中与100 TCID50 SARS-CoV-2真病毒(Omicron，B.1.1.529)以1:1(体积比)混合并于37℃下培养1小时。之后将病毒/血清混合物一式四份加入至96孔板中的单层Vero-E6细胞，且该板置于37℃、5％CO2的培养箱中培养3-5天。在显微镜下记录每个孔的细胞病变效应(CPE)，并计算50％中和抗体(NAb)效价。
如图25所示，相比于PBS对照组，经RBMRNA-组合疫苗免疫的小鼠血清显著提高针对SARS-CoV-2 Omicron型真病毒的中和效果，提示RBMRNA-组合疫苗能够有效地抑制Omicron型真病毒。
实施例9.病毒TCID50实验
RBMRNA-Omicron疫苗和RBMRNA-组合疫苗被用于TCID50实验，每组5只K18-hACE2小鼠。在第0天(作为初次免疫)和第21天(作为加强免疫)，对小鼠以5μg剂量的RBMRNA-Omicron疫苗或者RBMRNA-组合疫苗进行2次肌肉注射接种。对照组则注射等同体积的PBS。加强免疫后11天，用1×103蚀菌斑形成单位(PFU)的Delta(B.1.617.2)真病毒攻击小鼠。感染后5天，小鼠右肺的病毒滴度通过TCID50实验来测量，且结果如图26所示。加强免疫后31天，用1×104PFU的Omicron(B.1.1.529)真病毒攻击小鼠。感染后5天，小鼠右肺的病毒滴度通过TCID50实验来测量，且结果如图27所示。
如图26所示，相比于对照组，RBMRNA-Omicron疫苗和RBMRNA-组合疫苗均能减少Delta真病毒的病毒滴度。如图27所示，相比于对照组，RBMRNA-Omicron疫苗和RBMRNA-组合疫苗均能减少Omicron真病毒的病毒滴度。
这些结果说明，相比于对照组，RBMRNA-Omicron疫苗和RBMRNA-组合疫苗均能有效保护小鼠免受Delta型和Omicron型SARS-CoV-2的感染。
本申请参考了各种发行的专利、公开的专利申请、期刊文章和其他出版物，将所有这些引入本申请作为参考。若任何引入的参考文献和本说明书有冲突，则以本说明书为准。此外，落入现有技术范围的本发明的任何具体实施方案可以明确地从任何一个或多个权利要求中排除。因为所述实施方案被认为是本领域技术人员已知的，它们可以被排除，即使所述排除没有在本申请中明确列出。本发明的任何具体实施方案可从任何权利要求中以任何理由排除，不管是否与现有技术的存在有关。
虽然已经参考其特定实施方案描述本发明，本领域的技术人员应当理解可以进行各种改变且可以替换等同物而不脱离本发明的真正的精神和范围。另外，可作出许多修改以使特定的情况，材料，组合物，方法，方法步骤适于本发明的目的，精神和范围。所有这些修改都旨在权利要求的范围内。
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<110> 广州市锐博生物科技有限公司
阿格纳生物制药有限公司
<120> 基于SARS-COV-2的S蛋白的疫苗和组合物
<130> C21W0740
<160> 43
<170> PatentIn version 3.5
<210> 1
<211> 1271
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 1
Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val
1 5 10 15
Asn Leu Arg Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe
20 25 30
Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu
35 40 45
His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp
50 55 60
Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp
65 70 75 80
Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu
85 90 95
Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser
100 105 110
Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile
115 120 125
Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Asp Val Tyr
130 135 140
Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Gly Val Tyr Ser Ser
145 150 155 160
Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp
165 170 175
Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe
180 185 190
Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile
195 200 205
Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Val Leu Glu Pro Leu
210 215 220
Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu
225 230 235 240
Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser Gly Trp
245 250 255
Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr
260 265 270
Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp
275 280 285
Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe
290 295 300
Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro
305 310 315 320
Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe
325 330 335
Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn
340 345 350
Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn
355 360 365
Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys
370 375 380
Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile
385 390 395 400
Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile
405 410 415
Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile
420 425 430
Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn
435 440 445
Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg
450 455 460
Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Lys Pro Cys Asn Gly
465 470 475 480
Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln
485 490 495
Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser
500 505 510
Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser
515 520 525
Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu
530 535 540
Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe
545 550 555 560
Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp
565 570 575
Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly
580 585 590
Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val
595 600 605
Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile His Ala
610 615 620
Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val
625 630 635 640
Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val Asn Asn
645 650 655
Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr
660 665 670
Gln Thr Gln Thr Asn Ser Arg Arg Arg Ala Arg Ser Val Ala Ser Gln
675 680 685
Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala
690 695 700
Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val
705 710 715 720
Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys
725 730 735
Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu
740 745 750
Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile
755 760 765
Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys
770 775 780
Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe Asn Phe
785 790 795 800
Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile
805 810 815
Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile
820 825 830
Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile
835 840 845
Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr
850 855 860
Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile
865 870 875 880
Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe
885 890 895
Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn
900 905 910
Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala
915 920 925
Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly
930 935 940
Lys Leu Gln Asn Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu
945 950 955 960
Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn
965 970 975
Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln Ile Asp
980 985 990
Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln
995 1000 1005
Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala
1010 1015 1020
Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val
1025 1030 1035
Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser
1040 1045 1050
Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala
1055 1060 1065
Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly
1070 1075 1080
Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr
1085 1090 1095
His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile
1100 1105 1110
Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile
1115 1120 1125
Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu
1130 1135 1140
Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr
1145 1150 1155
Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser
1160 1165 1170
Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala
1175 1180 1185
Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys
1190 1195 1200
Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe
1205 1210 1215
Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys
1220 1225 1230
Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys
1235 1240 1245
Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu
1250 1255 1260
Lys Gly Val Lys Leu His Tyr Thr
1265 1270
<210> 2
<211> 1271
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 2
Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val
1 5 10 15
Asn Leu Arg Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe
20 25 30
Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu
35 40 45
His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp
50 55 60
Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp
65 70 75 80
Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu
85 90 95
Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser
100 105 110
Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile
115 120 125
Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Asp Val Tyr
130 135 140
Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Gly Val Tyr Ser Ser
145 150 155 160
Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp
165 170 175
Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe
180 185 190
Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile
195 200 205
Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Val Leu Glu Pro Leu
210 215 220
Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu
225 230 235 240
Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser Gly Trp
245 250 255
Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr
260 265 270
Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp
275 280 285
Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe
290 295 300
Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro
305 310 315 320
Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe
325 330 335
Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn
340 345 350
Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn
355 360 365
Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys
370 375 380
Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile
385 390 395 400
Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile
405 410 415
Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile
420 425 430
Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn
435 440 445
Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg
450 455 460
Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Lys Pro Cys Asn Gly
465 470 475 480
Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln
485 490 495
Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser
500 505 510
Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser
515 520 525
Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu
530 535 540
Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe
545 550 555 560
Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp
565 570 575
Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly
580 585 590
Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val
595 600 605
Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile His Ala
610 615 620
Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val
625 630 635 640
Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val Asn Asn
645 650 655
Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr
660 665 670
Gln Thr Gln Thr Asn Ser Arg Gly Gly Ser Gly Ser Val Ala Ser Gln
675 680 685
Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala
690 695 700
Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val
705 710 715 720
Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys
725 730 735
Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu
740 745 750
Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile
755 760 765
Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys
770 775 780
Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe Asn Phe
785 790 795 800
Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Ala Asn Ser Phe Ile
805 810 815
Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile
820 825 830
Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile
835 840 845
Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr
850 855 860
Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile
865 870 875 880
Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe
885 890 895
Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn
900 905 910
Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala
915 920 925
Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly
930 935 940
Lys Leu Gln Asn Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu
945 950 955 960
Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn
965 970 975
Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln Ile Asp
980 985 990
Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln
995 1000 1005
Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala
1010 1015 1020
Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val
1025 1030 1035
Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser
1040 1045 1050
Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala
1055 1060 1065
Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly
1070 1075 1080
Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr
1085 1090 1095
His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile
1100 1105 1110
Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile
1115 1120 1125
Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu
1130 1135 1140
Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr
1145 1150 1155
Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser
1160 1165 1170
Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala
1175 1180 1185
Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys
1190 1195 1200
Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe
1205 1210 1215
Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys
1220 1225 1230
Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys
1235 1240 1245
Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu
1250 1255 1260
Lys Gly Val Lys Leu His Tyr Thr
1265 1270
<210> 3
<211> 1271
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 3
Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val
1 5 10 15
Asn Leu Arg Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe
20 25 30
Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu
35 40 45
His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp
50 55 60
Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp
65 70 75 80
Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu
85 90 95
Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser
100 105 110
Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile
115 120 125
Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Asp Val Tyr
130 135 140
Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Gly Val Tyr Ser Ser
145 150 155 160
Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp
165 170 175
Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe
180 185 190
Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile
195 200 205
Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Val Leu Glu Pro Leu
210 215 220
Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu
225 230 235 240
Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser Gly Trp
245 250 255
Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr
260 265 270
Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp
275 280 285
Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe
290 295 300
Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro
305 310 315 320
Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe
325 330 335
Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn
340 345 350
Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn
355 360 365
Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys
370 375 380
Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile
385 390 395 400
Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile
405 410 415
Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile
420 425 430
Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn
435 440 445
Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg
450 455 460
Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Lys Pro Cys Asn Gly
465 470 475 480
Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln
485 490 495
Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser
500 505 510
Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser
515 520 525
Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu
530 535 540
Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe
545 550 555 560
Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp
565 570 575
Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly
580 585 590
Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val
595 600 605
Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile His Ala
610 615 620
Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val
625 630 635 640
Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val Asn Asn
645 650 655
Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr
660 665 670
Gln Thr Gln Thr Asn Ser Arg Gly Gly Ser Gly Ser Val Ala Ser Gln
675 680 685
Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala
690 695 700
Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val
705 710 715 720
Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys
725 730 735
Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu
740 745 750
Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile
755 760 765
Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys
770 775 780
Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe Asn Phe
785 790 795 800
Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Ala Asn Ser Phe Ile
805 810 815
Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile
820 825 830
Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile
835 840 845
Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr
850 855 860
Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile
865 870 875 880
Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe
885 890 895
Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn
900 905 910
Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala
915 920 925
Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly
930 935 940
Lys Leu Gln Asn Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu
945 950 955 960
Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn
965 970 975
Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp
980 985 990
Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln
995 1000 1005
Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala
1010 1015 1020
Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val
1025 1030 1035
Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser
1040 1045 1050
Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala
1055 1060 1065
Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly
1070 1075 1080
Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr
1085 1090 1095
His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile
1100 1105 1110
Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile
1115 1120 1125
Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu
1130 1135 1140
Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr
1145 1150 1155
Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser
1160 1165 1170
Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala
1175 1180 1185
Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys
1190 1195 1200
Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe
1205 1210 1215
Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys
1220 1225 1230
Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys
1235 1240 1245
Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu
1250 1255 1260
Lys Gly Val Lys Leu His Tyr Thr
1265 1270
<210> 4
<211> 1229
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 4
Met Asp Trp Ile Trp Arg Ile Leu Phe Leu Val Gly Ala Ala Thr Gly
1 5 10 15
Ala His Ser Gln Cys Val Asn Leu Arg Thr Arg Thr Gln Leu Pro Pro
20 25 30
Ala Tyr Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val
35 40 45
Phe Arg Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe
50 55 60
Phe Ser Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn
65 70 75 80
Gly Thr Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val
85 90 95
Tyr Phe Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe
100 105 110
Gly Thr Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn
115 120 125
Ala Thr Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp
130 135 140
Pro Phe Leu Asp Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu
145 150 155 160
Ser Gly Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser
165 170 175
Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn
180 185 190
Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr
195 200 205
Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe
210 215 220
Ser Val Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr
225 230 235 240
Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly
245 250 255
Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly
260 265 270
Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr
275 280 285
Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys
290 295 300
Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser
305 310 315 320
Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile
325 330 335
Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala
340 345 350
Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp
355 360 365
Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr
370 375 380
Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr
385 390 395 400
Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro
405 410 415
Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp
420 425 430
Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys
435 440 445
Val Gly Gly Asn Tyr Asn Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn
450 455 460
Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly
465 470 475 480
Ser Lys Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu
485 490 495
Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr
500 505 510
Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val
515 520 525
Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn
530 535 540
Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn
545 550 555 560
Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr
565 570 575
Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr
580 585 590
Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr
595 600 605
Ser Asn Gln Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val
610 615 620
Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr
625 630 635 640
Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly
645 650 655
Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala
660 665 670
Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gly Gly Ser
675 680 685
Gly Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly
690 695 700
Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr
705 710 715 720
Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr
725 730 735
Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu
740 745 750
Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn
755 760 765
Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu
770 775 780
Val Phe Ala Gln Val Lys Gln Pro Asp Pro Ser Lys Pro Ser Ala Asn
785 790 795 800
Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala
805 810 815
Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg
820 825 830
Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro
835 840 845
Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala
850 855 860
Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln
865 870 875 880
Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val
885 890 895
Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe
900 905 910
Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser
915 920 925
Ala Leu Gly Lys Leu Gln Asn Val Val Asn Gln Asn Ala Gln Ala Leu
930 935 940
Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser
945 950 955 960
Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val
965 970 975
Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr
980 985 990
Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn
995 1000 1005
Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys
1010 1015 1020
Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro
1025 1030 1035
Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val
1040 1045 1050
Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His
1055 1060 1065
Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn
1070 1075 1080
Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln
1085 1090 1095
Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val
1100 1105 1110
Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro
1115 1120 1125
Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn
1130 1135 1140
His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn
1145 1150 1155
Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu
1160 1165 1170
Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu
1175 1180 1185
Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Ser Ala Ile Gly Gly
1190 1195 1200
Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg Lys
1205 1210 1215
Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu
1220 1225
<210> 5
<211> 1230
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 5
Met Asp Trp Ile Trp Arg Ile Leu Phe Leu Val Gly Ala Ala Thr Gly
1 5 10 15
Ala His Ser Gln Cys Val Asn Leu Arg Thr Arg Thr Gln Leu Pro Pro
20 25 30
Ala Tyr Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val
35 40 45
Phe Arg Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe
50 55 60
Phe Ser Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn
65 70 75 80
Gly Thr Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val
85 90 95
Tyr Phe Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe
100 105 110
Gly Thr Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn
115 120 125
Ala Thr Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp
130 135 140
Pro Phe Leu Asp Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu
145 150 155 160
Ser Gly Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser
165 170 175
Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn
180 185 190
Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr
195 200 205
Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe
210 215 220
Ser Val Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr
225 230 235 240
Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly
245 250 255
Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly
260 265 270
Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr
275 280 285
Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys
290 295 300
Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser
305 310 315 320
Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile
325 330 335
Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala
340 345 350
Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp
355 360 365
Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr
370 375 380
Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr
385 390 395 400
Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro
405 410 415
Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp
420 425 430
Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys
435 440 445
Val Gly Gly Asn Tyr Asn Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn
450 455 460
Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly
465 470 475 480
Ser Lys Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu
485 490 495
Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr
500 505 510
Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val
515 520 525
Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn
530 535 540
Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn
545 550 555 560
Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr
565 570 575
Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr
580 585 590
Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr
595 600 605
Ser Asn Gln Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val
610 615 620
Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr
625 630 635 640
Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly
645 650 655
Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala
660 665 670
Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gly Gly Ser
675 680 685
Gly Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly
690 695 700
Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr
705 710 715 720
Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr
725 730 735
Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu
740 745 750
Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn
755 760 765
Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu
770 775 780
Val Phe Ala Gln Val Lys Gln Pro Asp Pro Ser Lys Pro Ser Ala Asn
785 790 795 800
Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala
805 810 815
Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg
820 825 830
Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro
835 840 845
Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala
850 855 860
Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln
865 870 875 880
Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val
885 890 895
Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe
900 905 910
Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser
915 920 925
Ala Leu Gly Lys Leu Gln Asn Val Val Asn Gln Asn Ala Gln Ala Leu
930 935 940
Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser
945 950 955 960
Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Val Glu Ala Glu
965 970 975
Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr
980 985 990
Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala
995 1000 1005
Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser
1010 1015 1020
Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe
1025 1030 1035
Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr
1040 1045 1050
Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys
1055 1060 1065
His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser
1070 1075 1080
Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro
1085 1090 1095
Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp
1100 1105 1110
Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln
1115 1120 1125
Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys
1130 1135 1140
Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile
1145 1150 1155
Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn
1160 1165 1170
Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu
1175 1180 1185
Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Ser Ala Ile Gly
1190 1195 1200
Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg
1205 1210 1215
Lys Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu
1220 1225 1230
<210> 6
<211> 802
<212> DNA
<213> 人工序列
<220>
<223> 起始序列
<400> 6
agtactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 60
tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 120
gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 180
taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 240
aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 300
ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 360
catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 420
ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 480
ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 540
agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 600
taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 660
atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 720
cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggtccctgg 780
ccttttgctg gccttttgct ca 802
<210> 7
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> T7启动子
<400> 7
taatacgact cactataatg 20
<210> 8
<211> 62
<212> DNA
<213> 人工序列
<220>
<223> 5'UTR
<400> 8
gtcccgcagt cggcgtccag cggctctgct tgttcgtgtg tgtgtcgttg caggccttat 60
tc 62
<210> 9
<211> 24
<212> DNA
<213> 人工序列
<220>
<223> kozak序列
<400> 9
agatctaccg gtggtaccgc cacc 24
<210> 10
<211> 89
<212> DNA
<213> 人工序列
<220>
<223> 3'UTR
<400> 10
agccaacacc ctgtctaaaa aacataaatt tctttaatca ttttgcctct tttctctgtg 60
cttcaattaa taaaaaatgg aaagaacct 89
<210> 11
<211> 3816
<212> DNA
<213> 人工序列
<220>
<223> RBMRNA-Delta 1疫苗的DNA序列
<400> 11
atgttcgtgt tcctggtgct gctgcctctg gtgagcagcc agtgcgtgaa tctgagaacc 60
agaacccagc tgcctcctgc ctacaccaat agcttcacca gaggagttta ttatcccgat 120
aaggtgttca gaagtagtgt attacatagt acccaggacc tgttcctacc tttcttcagt 180
aacgtgacct ggttccacgc catccacgtg agcggcacca atggcaccaa gagattcgac 240
aatcctgtgc tgcctttcaa tgacggcgtg tacttcgcca gcaccgagaa gagcaatatc 300
atcagaggct ggatcttcgg caccaccttg gattccaaga ctcagagcct gctgattgta 360
aacaacgcta caaatgtggt gatcaaggtg tgcgagttcc agttctgcaa tgaccctttc 420
ctggacgttt attatcataa gaacaacaag agctggatgg agagcggagt atattcgtcg 480
gctaataatt gcaccttcga gtacgtgagc cagcctttcc tgatggacct ggagggcaag 540
cagggcaatt tcaagaatct gagagagttc gtgttcaaga atatcgacgg ctacttcaag 600
atctacagca agcacacacc cattaatctg gtgagagacc tgcctcaggg cttcagcgtc 660
ctggagcctc tggtggacct gcctatcggc atcaatatca ccagattcca gaccctgctg 720
gccctgcaca gatcatatct tacaccaggc gattcgtcaa gcggttggac cgctggagct 780
gcggcatatt acgtgggcta cctgcagcct agaaccttcc tgctgaagta caatgagaat 840
ggtacgataa ccgacgcagt tgattgtgcc ctggaccctc tgagcgagac caagtgcacc 900
ctgaagagct tcaccgtgga gaagggcatc taccagacca gcaatttcag agtgcagcct 960
accgagagca tcgtgagatt ccctaatatc accaatctgt gccctttcgg cgaggtgttc 1020
aatgccacca gattcgccag cgtgtacgca tggaaccgca agcggataag caattgcgtg 1080
gccgactaca gcgtgctgta caatagcgcc agcttcagca ccttcaaatg ttatggtgtt 1140
tcgccaacaa agctgaatga cctgtgcttc accaatgtgt acgccgacag cttcgtgatc 1200
agaggcgacg aggtgagaca gatcgcgcca gggcagaccg gcaagatcgc cgactacaat 1260
tacaagctgc ctgacgactt caccggctgc gtgatcgcgt ggaactctaa caacctggac 1320
tcgaaagttg gaggcaatta caattaccgc tacagactgt tcagaaagag caatctgaag 1380
cctttcgaga gagacatcag caccgagatc taccaggccg gcagcaagcc gtgtaatggc 1440
gtggagggct tcaattgcta cttccctctg cagagctacg gcttccagcc taccaatggc 1500
gtgggctacc agccttacag agtggtggtg ctgagcttcg agctgctgca cgctcccgct 1560
accgtgtgcg gccctaagaa gagcaccaat ctggtgaaga ataagtgcgt gaatttcaat 1620
ttcaatggtc taactggaac gggcgtgctg accgagagca ataagaagtt tcttcccttt 1680
caacaattcg gcagagacat cgccgacacc acagatgctg taagagaccc tcagaccctg 1740
gagatcctgg acatcactcc gtgtagcttc ggcggcgtga gcgtgatcac accgggcacc 1800
aataccagca atcaggtggc cgtgctgtac cagggcgtga attgcaccga ggtgcctgtg 1860
gccatccacg ccgaccagct gactcccact tggagggtat attccacggg aagcaatgtg 1920
ttccagacca gagccggctg cctgatcggc gccgagcacg tgaataatag ctacgagtgc 1980
gacatcccta tcggcgccgg catctgcgcc agctaccaga cccagaccaa tagccgcggc 2040
ggcagcggca gcgtggccag ccagagcatc atcgcctaca ccatgagcct gggcgccgag 2100
aatagcgtgg cctacagcaa taatagcatc gccatcccta ccaatttcac catcagcgtg 2160
accaccgaaa tattaccagt ctccatgacc aagaccagcg tggactgcac catgtacatc 2220
tgcggcgaca gcaccgagtg cagcaatctg ctgctgcagt acggcagctt ctgcacccag 2280
ctgaatagag ccctgaccgg catcgccgtg gagcaggaca agaataccca ggaggtgttc 2340
gcccaggtga agcagatcta caagactccg ccgatcaagg acttcggcgg cttcaatttc 2400
agccaaatac tcccagatcc aagcaagcct agcgccaaca gcttcatcga ggacctgctg 2460
ttcaataagg tgaccctggc cgacgccggc ttcatcaagc agtacggcga ctgcctaggt 2520
gatattgcgg caagagacct gatctgcgcc cagaagttta acggtttgac agtactacct 2580
cctctgctga ccgacgagat gatagcacaa tatacgtcgg cattgctcgc tggcacgatc 2640
acatcgggct ggactttcgg cgccggagca gcgttgcaaa tccctttcgc catgcagatg 2700
gcctacagat tcaatggcat cggcgtgacc cagaatgtgc tgtacgagaa tcagaagctg 2760
atcgccaatc agttcaatag cgccatcggc aagatccagg acagcctgag cagcaccgcc 2820
agcgccctgg gcaagctgca gaacgtggtg aatcagaatg cccaggccct gaataccctg 2880
gtgaagcagc tgagcagcaa tttcggcgcc atcagtagtg tactcaacga catcctgagc 2940
agactggacc caccagaggc cgaggtgcaa attgatcgtc ttattactgg cagactgcag 3000
agcctgcaga cctacgtgac ccagcagctg atcagagccg ccgagatcag agccagcgcc 3060
aatctggccg ccaccaagat gagcgagtgc gtgctgggcc agagcaagag agtggacttc 3120
tgcggcaagg gctaccacct gatgagcttc cctcagagcg ctccacatgg cgtggtgttc 3180
ctgcacgtga cctacgtgcc tgcccaggag aagaatttca ccaccgcacc cgcaatctgc 3240
cacgacggca aggcccactt ccctagagag ggcgtgttcg tgagcaatgg cacccactgg 3300
ttcgtgaccc agagaaattt ctacgagcct cagatcatca ccaccgacaa taccttcgtg 3360
agcggcaatt gcgacgtggt gatcgggata gtcaataata ctgtctacga ccctctgcag 3420
cctgagctgg acagcttcaa ggaggagctg gacaagtact tcaagaatca caccagccct 3480
gacgtggacc tcggtgatat ttcgggaatc aatgccagcg tggtgaatat ccagaaggaa 3540
attgatcggc tcaacgaagt ggccaagaat ctgaatgaga gcctgatcga cctgcaggag 3600
ctgggcaagt acgagcagta catcaagtgg ccttggtaca tctggctggg cttcatcgcc 3660
ggcctgatcg ccatcgtgat ggtgaccatc atgctgtgct gcatgacctc ctgttgttcc 3720
tgtttgaaag ggtgttgttc gtgtgggtcc tgctgcaagt tcgacgagga cgacagcgag 3780
cctgtgctga agggcgtgaa gctgcactac acctaa 3816
<210> 12
<211> 3690
<212> DNA
<213> 人工序列
<220>
<223> RBMRNA-Delta 2疫苗的DNA序列
<400> 12
atggactgga tctggagaat cctgttcctg gtgggggctg ccactggcgc ccatagccag 60
tgcgtgaatc tgagaaccag aacccagctg cctcctgcct acaccaatag cttcaccaga 120
ggagtttatt atcccgataa ggtgttcaga agtagtgtat tacatagtac ccaggacctg 180
ttcctacctt tcttcagtaa cgtgacctgg ttccacgcca tccacgtgag cggcaccaat 240
ggcaccaaga gattcgacaa tcctgtgctg cctttcaatg acggcgtgta cttcgccagc 300
accgagaaga gcaatatcat cagaggctgg atcttcggca ccaccttgga ttccaagact 360
cagagcctgc tgattgtaaa caacgctaca aatgtggtga tcaaggtgtg cgagttccag 420
ttctgcaatg accctttcct ggacgtttat tatcataaga acaacaagag ctggatggag 480
agcggagtat attcgtcggc taataattgc accttcgagt acgtgagcca gcctttcctg 540
atggacctgg agggcaagca gggcaatttc aagaatctga gagagttcgt gttcaagaat 600
atcgacggct acttcaagat ctacagcaag cacacaccca ttaatctggt gagagacctg 660
cctcagggct tcagcgtcct ggagcctctg gtggacctgc ctatcggcat caatatcacc 720
agattccaga ccctgctggc cctgcacaga tcatatctta caccaggcga ttcgtcaagc 780
ggttggaccg ctggagctgc ggcatattac gtgggctacc tgcagcctag aaccttcctg 840
ctgaagtaca atgagaatgg tacgataacc gacgcagttg attgtgccct ggaccctctg 900
agcgagacca agtgcaccct gaagagcttc accgtggaga agggcatcta ccagaccagc 960
aatttcagag tgcagcctac cgagagcatc gtgagattcc ctaatatcac caatctgtgc 1020
cctttcggcg aggtgttcaa tgccaccaga ttcgccagcg tgtacgcatg gaaccgcaag 1080
cggataagca attgcgtggc cgactacagc gtgctgtaca atagcgccag cttcagcacc 1140
ttcaaatgtt atggtgtttc gccaacaaag ctgaatgacc tgtgcttcac caatgtgtac 1200
gccgacagct tcgtgatcag aggcgacgag gtgagacaga tcgcgccagg gcagaccggc 1260
aagatcgccg actacaatta caagctgcct gacgacttca ccggctgcgt gatcgcgtgg 1320
aactctaaca acctggactc gaaagttgga ggcaattaca attaccgcta cagactgttc 1380
agaaagagca atctgaagcc tttcgagaga gacatcagca ccgagatcta ccaggccggc 1440
agcaagccgt gtaatggcgt ggagggcttc aattgctact tccctctgca gagctacggc 1500
ttccagccta ccaatggcgt gggctaccag ccttacagag tggtggtgct gagcttcgag 1560
ctgctgcacg ctcccgctac cgtgtgcggc cctaagaaga gcaccaatct ggtgaagaat 1620
aagtgcgtga atttcaattt caatggtcta actggaacgg gcgtgctgac cgagagcaat 1680
aagaagtttc ttccctttca acaattcggc agagacatcg ccgacaccac agatgctgta 1740
agagaccctc agaccctgga gatcctggac atcactccgt gtagcttcgg cggcgtgagc 1800
gtgatcacac cgggcaccaa taccagcaat caggtggccg tgctgtacca gggcgtgaat 1860
tgcaccgagg tgcctgtggc catccacgcc gaccagctga ctcccacttg gagggtatat 1920
tccacgggaa gcaatgtgtt ccagaccaga gccggctgcc tgatcggcgc cgagcacgtg 1980
aataatagct acgagtgcga catccctatc ggcgccggca tctgcgccag ctaccagacc 2040
cagaccaata gccctcgggg cagcggcagc gtggccagcc agagcatcat cgcctacacc 2100
atgagcctgg gcgccgagaa tagcgtggcc tacagcaata atagcatcgc catccctacc 2160
aatttcacca tcagcgtgac caccgaaata ttaccagtct ccatgaccaa gaccagcgtg 2220
gactgcacca tgtacatctg cggcgacagc accgagtgca gcaatctgct gctgcagtac 2280
ggcagcttct gcacccagct gaatagagcc ctgaccggca tcgccgtgga gcaggacaag 2340
aatacccagg aggtgttcgc ccaggtgaag cagccagatc caagcaagcc tagcgccaac 2400
agcttcatcg aggacctgct gttcaataag gtgaccctgg ccgacgccgg cttcatcaag 2460
cagtacggcg actgcctagg tgatattgcg gcaagagacc tgatctgcgc ccagaagttt 2520
aacggtttga cagtactacc tcctctgctg accgacgaga tgatagcaca atatacgtcg 2580
gcattgctcg ctggcacgat cacatcgggc tggactttcg gcgccggagc agcgttgcaa 2640
atccctttcg ccatgcagat ggcctacaga ttcaatggca tcggcgtgac ccagaatgtg 2700
ctgtacgaga atcagaagct gatcgccaat cagttcaata gcgccatcgg caagatccag 2760
gacagcctga gcagcaccgc cagcgccctg ggcaagctgc agaacgtggt gaatcagaat 2820
gcccaggccc tgaataccct ggtgaagcag ctgagcagca atttcggcgc catcagtagt 2880
gtactcaacg acatcctgag cagactggac ccgccggagg ccgaggtgca aattgatcgt 2940
cttattactg gcagactgca gagcctgcag acctacgtga cccagcagct gatcagagcc 3000
gccgagatca gagccagcgc caatctggcc gccaccaaga tgagcgagtg cgtgctgggc 3060
cagagcaaga gagtggactt ctgcggcaag ggctaccacc tgatgagctt ccctcagagc 3120
gctccacatg gcgtggtgtt cctgcacgtg acctacgtgc ctgcccagga gaagaatttc 3180
accaccgcac ccgcaatctg ccacgacggc aaggcccact tccctagaga gggcgtgttc 3240
gtgagcaatg gcacccactg gttcgtgacc cagagaaatt tctacgagcc tcagatcatc 3300
accaccgaca ataccttcgt gagcggcaat tgcgacgtgg tgatcgggat agtcaataat 3360
actgtctacg accctctgca gcctgagctg gacagcttca aggaggagct ggacaagtac 3420
ttcaagaatc acaccagccc tgacgtggac ctcggtgata tttcgggaat caatgccagc 3480
gtggtgaata tccagaagga aattgatcgg ctcaacgaag tggccaagaa tctgaatgag 3540
agcctgatcg acctgcagga gctgggcaag tacgagcagt acatcaagtg gcctagcgcc 3600
atcggcggct acatccccga ggcccccagg gacggccagg cctacgtgag gaaggacggc 3660
gagtgggtgc tgctgagcac cttcctgtga 3690
<210> 13
<211> 3816
<212> DNA
<213> 人工序列
<220>
<223> RBMRNA-Delta 3疫苗的DNA序列
<400> 13
atgttcgtgt tcctggtgct gctgcccctg gtgagcagcc agtgcgtgaa cctgagaacc 60
cggacccagc tgccaccagc ctacaccaac agcttcaccc ggggcgtcta ctaccccgac 120
aaggtgttcc ggagcagcgt cctgcacagc acccaggacc tgttcctgcc cttcttcagc 180
aacgtgacct ggttccacgc catccacgtg agcggcacca acggcaccaa gcggttcgac 240
aaccccgtgc tgcccttcaa cgacggcgtg tacttcgcca gcaccgagaa gagcaacatc 300
atccggggct ggatcttcgg caccaccctg gacagcaaga cccagagcct gctgatcgtg 360
aataacgcca ccaacgtggt gatcaaggtg tgcgagttcc agttctgcaa cgaccccttc 420
ctggacgtgt actaccacaa gaacaacaag agctggatgg agagcggagt gtacagcagc 480
gccaacaact gcaccttcga gtacgtgagc cagcccttcc tgatggacct ggagggcaag 540
cagggcaact tcaagaacct gcgggagttc gtgttcaaga acatcgacgg ctacttcaag 600
atctacagca agcacacccc aatcaacctg gtgcgggatc tgccccaggg cttctcagtc 660
ctggagcccc tggtggacct gcccatcggc atcaacatca cccggttcca gaccctgctg 720
gccctgcacc ggagctacct gaccccaggc gacagcagca gcgggtggac agcaggcgcg 780
gctgcttact acgtgggcta cctgcagccc cggaccttcc tgctgaagta caacgagaac 840
ggcaccatca ccgacgccgt ggactgcgcc ctggaccctc tgagcgagac caagtgcacc 900
ctgaagagct tcaccgtgga gaagggcatc taccagacca gcaacttccg ggtgcagccc 960
accgagagca tcgtgcggtt ccccaacatc accaacctgt gccccttcgg cgaggtgttc 1020
aacgccaccc ggttcgccag cgtgtacgcc tggaaccgga agcggatcag caactgcgtg 1080
gccgactaca gcgtgctgta caacagcgcc agcttcagca ccttcaagtg ctacggcgtg 1140
agccccacca agctgaacga cctgtgcttc accaacgtgt acgccgacag cttcgtgatc 1200
cgtggcgacg aggtgcggca gatcgcaccc ggccagacag gcaagatcgc cgactacaac 1260
tacaagctgc ccgacgactt caccggctgc gtgatcgcct ggaacagcaa caacctcgac 1320
agcaaggtgg gcggcaacta caactaccgc taccggctgt tccggaagag caacctgaag 1380
cccttcgagc gggacatcag caccgagatc taccaagccg gctccaagcc ttgcaacggc 1440
gtggagggct tcaactgcta cttccctctg cagagctacg gcttccagcc caccaacggc 1500
gtgggctacc agccctaccg ggtggtggtg ctgagcttcg agctgctgca cgccccagcc 1560
accgtgtgtg gccccaagaa gagcaccaac ctggtgaaga acaagtgcgt gaacttcaac 1620
ttcaacggcc ttaccggcac cggcgtgctg accgagagca acaagaaatt cctgcccttt 1680
cagcagttcg gccgggacat cgccgacacc accgacgctg tgcgggatcc ccagaccctg 1740
gagatcctgg acatcacccc ttgcagcttc ggcggcgtga gcgtgatcac cccaggcacc 1800
aacaccagca accaggtggc cgtgctgtac cagggcgtga actgcaccga ggtgcccgtg 1860
gccatccacg ccgaccagct gacacccacc tggcgggtct acagcaccgg cagcaacgtg 1920
ttccagaccc gggccggttg cctgatcggc gccgagcacg tgaacaacag ctacgagtgc 1980
gacatcccca tcggcgccgg catctgtgcc agctaccaga cccagaccaa ttcacgcggc 2040
ggcagcggca gcgtggccag ccagagcatc atcgcctaca ccatgagcct gggcgccgag 2100
aacagcgtgg cctacagcaa caacagcatc gccatcccca ccaacttcac catcagcgtg 2160
accaccgaga ttctgcccgt gagcatgacc aagaccagcg tggactgcac catgtacatc 2220
tgcggcgaca gcaccgagtg cagcaacctg ctgctgcagt acggcagctt ctgcacccag 2280
ctgaaccggg ccctgaccgg catcgccgtg gagcaggaca agaacaccca ggaggtgttc 2340
gcccaggtga agcagatcta caagacccct cccatcaagg acttcggcgg cttcaacttc 2400
agccagatcc tgcccgaccc cagcaagccc agcgccaaca gcttcatcga ggacctgctg 2460
ttcaacaagg tgaccctagc cgacgccggc ttcatcaagc agtacggcga ctgcctcggc 2520
gacatagccg cccgggacct gatctgcgcc cagaagttca acggcctgac cgtgctgcct 2580
cccctgctga ccgacgagat gatcgcccag tacaccagcg ccctgttagc cggaaccatc 2640
accagcggct ggactttcgg cgctggagcc gctctgcaga tccccttcgc catgcagatg 2700
gcctaccggt tcaacggcat cggcgtgacc cagaacgtgc tgtacgagaa ccagaagctg 2760
atcgccaacc agttcaacag cgccatcggc aagatccagg acagcctgag cagcaccgct 2820
agcgccctgg gcaagctgca gaacgtggtg aaccagaacg cccaggccct gaacaccctg 2880
gtgaagcagc tgagcagcaa cttcggcgcc atcagcagcg tgctgaacga catcctgagc 2940
cggctggacc ctcccgaggc cgaggtgcag atcgaccggc tgatcactgg ccggctgcag 3000
agcctgcaga cctacgtgac ccagcagctg atccgggccg ccgagattcg ggccagcgcc 3060
aacctggccg ccaccaagat gagcgagtgc gtgctgggcc agagcaagcg ggtggacttc 3120
tgcggcaagg gctaccacct gatgagcttt ccccagagcg caccccacgg agtggtgttc 3180
ctgcacgtga cctacgtgcc cgcccaggag aagaacttca ccaccgcccc agccatctgc 3240
cacgacggca aggcccactt tccccgggag ggcgtgttcg tgagcaacgg cacccactgg 3300
ttcgtgaccc agcggaactt ctacgagccc cagatcatca ccaccgacaa caccttcgtg 3360
agcggcaact gcgacgtggt gatcggcatc gtgaacaaca ccgtgtacga tcccctgcag 3420
cccgagctgg acagcttcaa ggaggagctg gacaagtact tcaagaatca caccagcccc 3480
gacgtggacc tgggcgacat cagcggcatc aacgccagcg tggtgaacat ccagaaggag 3540
atcgatcggc tgaacgaggt ggccaagaac ctgaacgaga gcctgatcga cctgcaggag 3600
ctgggcaagt acgagcagta catcaagtgg ccctggtaca tctggctggg cttcatcgcc 3660
ggcctgatcg ccatcgtgat ggtgaccatc atgctgtgct gcatgaccag ctgctgcagc 3720
tgcctgaagg gctgttgcag ctgcggcagc tgctgcaagt tcgacgagga cgacagcgag 3780
cccgtgctga agggcgtgaa gctgcactac acctga 3816
<210> 14
<211> 4130
<212> RNA
<213> 人工序列
<220>
<223> RBMRNA-Delta 1疫苗的mRNA序列
<400> 14
augagacaag cuugucccgc agucggcguc cagcggcucu gcuuguucgu gugugugucg 60
uugcaggccu uauucagauc uaccgguggu accgccacca uguucguguu ccuggugcug 120
cugccucugg ugagcagcca gugcgugaau cugagaacca gaacccagcu gccuccugcc 180
uacaccaaua gcuucaccag aggaguuuau uaucccgaua agguguucag aaguagugua 240
uuacauagua cccaggaccu guuccuaccu uucuucagua acgugaccug guuccacgcc 300
auccacguga gcggcaccaa uggcaccaag agauucgaca auccugugcu gccuuucaau 360
gacggcgugu acuucgccag caccgagaag agcaauauca ucagaggcug gaucuucggc 420
accaccuugg auuccaagac ucagagccug cugauuguaa acaacgcuac aaauguggug 480
aucaaggugu gcgaguucca guucugcaau gacccuuucc uggacguuua uuaucauaag 540
aacaacaaga gcuggaugga gagcggagua uauucgucgg cuaauaauug caccuucgag 600
uacgugagcc agccuuuccu gauggaccug gagggcaagc agggcaauuu caagaaucug 660
agagaguucg uguucaagaa uaucgacggc uacuucaaga ucuacagcaa gcacacaccc 720
auuaaucugg ugagagaccu gccucagggc uucagcgucc uggagccucu gguggaccug 780
ccuaucggca ucaauaucac cagauuccag acccugcugg cccugcacag aucauaucuu 840
acaccaggcg auucgucaag cgguuggacc gcuggagcug cggcauauua cgugggcuac 900
cugcagccua gaaccuuccu gcugaaguac aaugagaaug guacgauaac cgacgcaguu 960
gauugugccc uggacccucu gagcgagacc aagugcaccc ugaagagcuu caccguggag 1020
aagggcaucu accagaccag caauuucaga gugcagccua ccgagagcau cgugagauuc 1080
ccuaauauca ccaaucugug cccuuucggc gagguguuca augccaccag auucgccagc 1140
guguacgcau ggaaccgcaa gcggauaagc aauugcgugg ccgacuacag cgugcuguac 1200
aauagcgcca gcuucagcac cuucaaaugu uaugguguuu cgccaacaaa gcugaaugac 1260
cugugcuuca ccaaugugua cgccgacagc uucgugauca gaggcgacga ggugagacag 1320
aucgcgccag ggcagaccgg caagaucgcc gacuacaauu acaagcugcc ugacgacuuc 1380
accggcugcg ugaucgcgug gaacucuaac aaccuggacu cgaaaguugg aggcaauuac 1440
aauuaccgcu acagacuguu cagaaagagc aaucugaagc cuuucgagag agacaucagc 1500
accgagaucu accaggccgg cagcaagccg uguaauggcg uggagggcuu caauugcuac 1560
uucccucugc agagcuacgg cuuccagccu accaauggcg ugggcuacca gccuuacaga 1620
gugguggugc ugagcuucga gcugcugcac gcucccgcua ccgugugcgg cccuaagaag 1680
agcaccaauc uggugaagaa uaagugcgug aauuucaauu ucaauggucu aacuggaacg 1740
ggcgugcuga ccgagagcaa uaagaaguuu cuucccuuuc aacaauucgg cagagacauc 1800
gccgacacca cagaugcugu aagagacccu cagacccugg agauccugga caucacuccg 1860
uguagcuucg gcggcgugag cgugaucaca ccgggcacca auaccagcaa ucagguggcc 1920
gugcuguacc agggcgugaa uugcaccgag gugccugugg ccauccacgc cgaccagcug 1980
acucccacuu ggaggguaua uuccacggga agcaaugugu uccagaccag agccggcugc 2040
cugaucggcg ccgagcacgu gaauaauagc uacgagugcg acaucccuau cggcgccggc 2100
aucugcgcca gcuaccagac ccagaccaau agccgcggcg gcagcggcag cguggccagc 2160
cagagcauca ucgccuacac caugagccug ggcgccgaga auagcguggc cuacagcaau 2220
aauagcaucg ccaucccuac caauuucacc aucagcguga ccaccgaaau auuaccaguc 2280
uccaugacca agaccagcgu ggacugcacc auguacaucu gcggcgacag caccgagugc 2340
agcaaucugc ugcugcagua cggcagcuuc ugcacccagc ugaauagagc ccugaccggc 2400
aucgccgugg agcaggacaa gaauacccag gagguguucg cccaggugaa gcagaucuac 2460
aagacuccgc cgaucaagga cuucggcggc uucaauuuca gccaaauacu cccagaucca 2520
agcaagccua gcgccaacag cuucaucgag gaccugcugu ucaauaaggu gacccuggcc 2580
gacgccggcu ucaucaagca guacggcgac ugccuaggug auauugcggc aagagaccug 2640
aucugcgccc agaaguuuaa cgguuugaca guacuaccuc cucugcugac cgacgagaug 2700
auagcacaau auacgucggc auugcucgcu ggcacgauca caucgggcug gacuuucggc 2760
gccggagcag cguugcaaau cccuuucgcc augcagaugg ccuacagauu caauggcauc 2820
ggcgugaccc agaaugugcu guacgagaau cagaagcuga ucgccaauca guucaauagc 2880
gccaucggca agauccagga cagccugagc agcaccgcca gcgcccuggg caagcugcag 2940
aacgugguga aucagaaugc ccaggcccug aauacccugg ugaagcagcu gagcagcaau 3000
uucggcgcca ucaguagugu acucaacgac auccugagca gacuggaccc accagaggcc 3060
gaggugcaaa uugaucgucu uauuacuggc agacugcaga gccugcagac cuacgugacc 3120
cagcagcuga ucagagccgc cgagaucaga gccagcgcca aucuggccgc caccaagaug 3180
agcgagugcg ugcugggcca gagcaagaga guggacuucu gcggcaaggg cuaccaccug 3240
augagcuucc cucagagcgc uccacauggc gugguguucc ugcacgugac cuacgugccu 3300
gcccaggaga agaauuucac caccgcaccc gcaaucugcc acgacggcaa ggcccacuuc 3360
ccuagagagg gcguguucgu gagcaauggc acccacuggu ucgugaccca gagaaauuuc 3420
uacgagccuc agaucaucac caccgacaau accuucguga gcggcaauug cgacguggug 3480
aucgggauag ucaauaauac ugucuacgac ccucugcagc cugagcugga cagcuucaag 3540
gaggagcugg acaaguacuu caagaaucac accagcccug acguggaccu cggugauauu 3600
ucgggaauca augccagcgu ggugaauauc cagaaggaaa uugaucggcu caacgaagug 3660
gccaagaauc ugaaugagag ccugaucgac cugcaggagc ugggcaagua cgagcaguac 3720
aucaaguggc cuugguacau cuggcugggc uucaucgccg gccugaucgc caucgugaug 3780
gugaccauca ugcugugcug caugaccucc uguuguuccu guuugaaagg guguuguucg 3840
uguggguccu gcugcaaguu cgacgaggac gacagcgagc cugugcugaa gggcgugaag 3900
cugcacuaca ccuaacucga gacuaguagc caacacccug ucuaaaaaac auaaauuucu 3960
uuaaucauuu ugccucuuuu cucugugcuu caauuaauaa aaaauggaaa gaaccuaaaa 4020
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 4080
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 4130
<210> 15
<211> 4004
<212> RNA
<213> 人工序列
<220>
<223> RBMRNA-Delta 2疫苗的mRNA序列
<400> 15
augagacaag cuugucccgc agucggcguc cagcggcucu gcuuguucgu gugugugucg 60
uugcaggccu uauucagauc uaccgguggu accgccacca uggacuggau cuggagaauc 120
cuguuccugg ugggggcugc cacuggcgcc cauagccagu gcgugaaucu gagaaccaga 180
acccagcugc cuccugccua caccaauagc uucaccagag gaguuuauua ucccgauaag 240
guguucagaa guaguguauu acauaguacc caggaccugu uccuaccuuu cuucaguaac 300
gugaccuggu uccacgccau ccacgugagc ggcaccaaug gcaccaagag auucgacaau 360
ccugugcugc cuuucaauga cggcguguac uucgccagca ccgagaagag caauaucauc 420
agaggcugga ucuucggcac caccuuggau uccaagacuc agagccugcu gauuguaaac 480
aacgcuacaa auguggugau caaggugugc gaguuccagu ucugcaauga cccuuuccug 540
gacguuuauu aucauaagaa caacaagagc uggauggaga gcggaguaua uucgucggcu 600
aauaauugca ccuucgagua cgugagccag ccuuuccuga uggaccugga gggcaagcag 660
ggcaauuuca agaaucugag agaguucgug uucaagaaua ucgacggcua cuucaagauc 720
uacagcaagc acacacccau uaaucuggug agagaccugc cucagggcuu cagcguccug 780
gagccucugg uggaccugcc uaucggcauc aauaucacca gauuccagac ccugcuggcc 840
cugcacagau cauaucuuac accaggcgau ucgucaagcg guuggaccgc uggagcugcg 900
gcauauuacg ugggcuaccu gcagccuaga accuuccugc ugaaguacaa ugagaauggu 960
acgauaaccg acgcaguuga uugugcccug gacccucuga gcgagaccaa gugcacccug 1020
aagagcuuca ccguggagaa gggcaucuac cagaccagca auuucagagu gcagccuacc 1080
gagagcaucg ugagauuccc uaauaucacc aaucugugcc cuuucggcga gguguucaau 1140
gccaccagau ucgccagcgu guacgcaugg aaccgcaagc ggauaagcaa uugcguggcc 1200
gacuacagcg ugcuguacaa uagcgccagc uucagcaccu ucaaauguua ugguguuucg 1260
ccaacaaagc ugaaugaccu gugcuucacc aauguguacg ccgacagcuu cgugaucaga 1320
ggcgacgagg ugagacagau cgcgccaggg cagaccggca agaucgccga cuacaauuac 1380
aagcugccug acgacuucac cggcugcgug aucgcgugga acucuaacaa ccuggacucg 1440
aaaguuggag gcaauuacaa uuaccgcuac agacuguuca gaaagagcaa ucugaagccu 1500
uucgagagag acaucagcac cgagaucuac caggccggca gcaagccgug uaauggcgug 1560
gagggcuuca auugcuacuu cccucugcag agcuacggcu uccagccuac caauggcgug 1620
ggcuaccagc cuuacagagu gguggugcug agcuucgagc ugcugcacgc ucccgcuacc 1680
gugugcggcc cuaagaagag caccaaucug gugaagaaua agugcgugaa uuucaauuuc 1740
aauggucuaa cuggaacggg cgugcugacc gagagcaaua agaaguuucu ucccuuucaa 1800
caauucggca gagacaucgc cgacaccaca gaugcuguaa gagacccuca gacccuggag 1860
auccuggaca ucacuccgug uagcuucggc ggcgugagcg ugaucacacc gggcaccaau 1920
accagcaauc agguggccgu gcuguaccag ggcgugaauu gcaccgaggu gccuguggcc 1980
auccacgccg accagcugac ucccacuugg aggguauauu ccacgggaag caauguguuc 2040
cagaccagag ccggcugccu gaucggcgcc gagcacguga auaauagcua cgagugcgac 2100
aucccuaucg gcgccggcau cugcgccagc uaccagaccc agaccaauag cccucggggc 2160
agcggcagcg uggccagcca gagcaucauc gccuacacca ugagccuggg cgccgagaau 2220
agcguggccu acagcaauaa uagcaucgcc aucccuacca auuucaccau cagcgugacc 2280
accgaaauau uaccagucuc caugaccaag accagcgugg acugcaccau guacaucugc 2340
ggcgacagca ccgagugcag caaucugcug cugcaguacg gcagcuucug cacccagcug 2400
aauagagccc ugaccggcau cgccguggag caggacaaga auacccagga gguguucgcc 2460
caggugaagc agccagaucc aagcaagccu agcgccaaca gcuucaucga ggaccugcug 2520
uucaauaagg ugacccuggc cgacgccggc uucaucaagc aguacggcga cugccuaggu 2580
gauauugcgg caagagaccu gaucugcgcc cagaaguuua acgguuugac aguacuaccu 2640
ccucugcuga ccgacgagau gauagcacaa uauacgucgg cauugcucgc uggcacgauc 2700
acaucgggcu ggacuuucgg cgccggagca gcguugcaaa ucccuuucgc caugcagaug 2760
gccuacagau ucaauggcau cggcgugacc cagaaugugc uguacgagaa ucagaagcug 2820
aucgccaauc aguucaauag cgccaucggc aagauccagg acagccugag cagcaccgcc 2880
agcgcccugg gcaagcugca gaacguggug aaucagaaug cccaggcccu gaauacccug 2940
gugaagcagc ugagcagcaa uuucggcgcc aucaguagug uacucaacga cauccugagc 3000
agacuggacc cgccggaggc cgaggugcaa auugaucguc uuauuacugg cagacugcag 3060
agccugcaga ccuacgugac ccagcagcug aucagagccg ccgagaucag agccagcgcc 3120
aaucuggccg ccaccaagau gagcgagugc gugcugggcc agagcaagag aguggacuuc 3180
ugcggcaagg gcuaccaccu gaugagcuuc ccucagagcg cuccacaugg cgugguguuc 3240
cugcacguga ccuacgugcc ugcccaggag aagaauuuca ccaccgcacc cgcaaucugc 3300
cacgacggca aggcccacuu cccuagagag ggcguguucg ugagcaaugg cacccacugg 3360
uucgugaccc agagaaauuu cuacgagccu cagaucauca ccaccgacaa uaccuucgug 3420
agcggcaauu gcgacguggu gaucgggaua gucaauaaua cugucuacga cccucugcag 3480
ccugagcugg acagcuucaa ggaggagcug gacaaguacu ucaagaauca caccagcccu 3540
gacguggacc ucggugauau uucgggaauc aaugccagcg uggugaauau ccagaaggaa 3600
auugaucggc ucaacgaagu ggccaagaau cugaaugaga gccugaucga ccugcaggag 3660
cugggcaagu acgagcagua caucaagugg ccuagcgcca ucggcggcua cauccccgag 3720
gcccccaggg acggccaggc cuacgugagg aaggacggcg agugggugcu gcugagcacc 3780
uuccugugac ucgagacuag uagccaacac ccugucuaaa aaacauaaau uucuuuaauc 3840
auuuugccuc uuuucucugu gcuucaauua auaaaaaaug gaaagaaccu aaaaaaaaaa 3900
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 3960
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaa 4004
<210> 16
<211> 4130
<212> RNA
<213> 人工序列
<220>
<223> RBMRNA-Delta 3疫苗的mRNA序列
<400> 16
augagacaag cuugucccgc agucggcguc cagcggcucu gcuuguucgu gugugugucg 60
uugcaggccu uauucagauc uaccgguggu accgccacca uguucguguu ccuggugcug 120
cugccccugg ugagcagcca gugcgugaac cugagaaccc ggacccagcu gccaccagcc 180
uacaccaaca gcuucacccg gggcgucuac uaccccgaca agguguuccg gagcagcguc 240
cugcacagca cccaggaccu guuccugccc uucuucagca acgugaccug guuccacgcc 300
auccacguga gcggcaccaa cggcaccaag cgguucgaca accccgugcu gcccuucaac 360
gacggcgugu acuucgccag caccgagaag agcaacauca uccggggcug gaucuucggc 420
accacccugg acagcaagac ccagagccug cugaucguga auaacgccac caacguggug 480
aucaaggugu gcgaguucca guucugcaac gaccccuucc uggacgugua cuaccacaag 540
aacaacaaga gcuggaugga gagcggagug uacagcagcg ccaacaacug caccuucgag 600
uacgugagcc agcccuuccu gauggaccug gagggcaagc agggcaacuu caagaaccug 660
cgggaguucg uguucaagaa caucgacggc uacuucaaga ucuacagcaa gcacacccca 720
aucaaccugg ugcgggaucu gccccagggc uucucagucc uggagccccu gguggaccug 780
cccaucggca ucaacaucac ccgguuccag acccugcugg cccugcaccg gagcuaccug 840
accccaggcg acagcagcag cggguggaca gcaggcgcgg cugcuuacua cgugggcuac 900
cugcagcccc ggaccuuccu gcugaaguac aacgagaacg gcaccaucac cgacgccgug 960
gacugcgccc uggacccucu gagcgagacc aagugcaccc ugaagagcuu caccguggag 1020
aagggcaucu accagaccag caacuuccgg gugcagccca ccgagagcau cgugcgguuc 1080
cccaacauca ccaaccugug ccccuucggc gagguguuca acgccacccg guucgccagc 1140
guguacgccu ggaaccggaa gcggaucagc aacugcgugg ccgacuacag cgugcuguac 1200
aacagcgcca gcuucagcac cuucaagugc uacggcguga gccccaccaa gcugaacgac 1260
cugugcuuca ccaacgugua cgccgacagc uucgugaucc guggcgacga ggugcggcag 1320
aucgcacccg gccagacagg caagaucgcc gacuacaacu acaagcugcc cgacgacuuc 1380
accggcugcg ugaucgccug gaacagcaac aaccucgaca gcaagguggg cggcaacuac 1440
aacuaccgcu accggcuguu ccggaagagc aaccugaagc ccuucgagcg ggacaucagc 1500
accgagaucu accaagccgg cuccaagccu ugcaacggcg uggagggcuu caacugcuac 1560
uucccucugc agagcuacgg cuuccagccc accaacggcg ugggcuacca gcccuaccgg 1620
gugguggugc ugagcuucga gcugcugcac gccccagcca ccgugugugg ccccaagaag 1680
agcaccaacc uggugaagaa caagugcgug aacuucaacu ucaacggccu uaccggcacc 1740
ggcgugcuga ccgagagcaa caagaaauuc cugcccuuuc agcaguucgg ccgggacauc 1800
gccgacacca ccgacgcugu gcgggauccc cagacccugg agauccugga caucaccccu 1860
ugcagcuucg gcggcgugag cgugaucacc ccaggcacca acaccagcaa ccagguggcc 1920
gugcuguacc agggcgugaa cugcaccgag gugcccgugg ccauccacgc cgaccagcug 1980
acacccaccu ggcgggucua cagcaccggc agcaacgugu uccagacccg ggccgguugc 2040
cugaucggcg ccgagcacgu gaacaacagc uacgagugcg acauccccau cggcgccggc 2100
aucugugcca gcuaccagac ccagaccaau ucacgcggcg gcagcggcag cguggccagc 2160
cagagcauca ucgccuacac caugagccug ggcgccgaga acagcguggc cuacagcaac 2220
aacagcaucg ccauccccac caacuucacc aucagcguga ccaccgagau ucugcccgug 2280
agcaugacca agaccagcgu ggacugcacc auguacaucu gcggcgacag caccgagugc 2340
agcaaccugc ugcugcagua cggcagcuuc ugcacccagc ugaaccgggc ccugaccggc 2400
aucgccgugg agcaggacaa gaacacccag gagguguucg cccaggugaa gcagaucuac 2460
aagaccccuc ccaucaagga cuucggcggc uucaacuuca gccagauccu gcccgacccc 2520
agcaagccca gcgccaacag cuucaucgag gaccugcugu ucaacaaggu gacccuagcc 2580
gacgccggcu ucaucaagca guacggcgac ugccucggcg acauagccgc ccgggaccug 2640
aucugcgccc agaaguucaa cggccugacc gugcugccuc cccugcugac cgacgagaug 2700
aucgcccagu acaccagcgc ccuguuagcc ggaaccauca ccagcggcug gacuuucggc 2760
gcuggagccg cucugcagau ccccuucgcc augcagaugg ccuaccgguu caacggcauc 2820
ggcgugaccc agaacgugcu guacgagaac cagaagcuga ucgccaacca guucaacagc 2880
gccaucggca agauccagga cagccugagc agcaccgcua gcgcccuggg caagcugcag 2940
aacgugguga accagaacgc ccaggcccug aacacccugg ugaagcagcu gagcagcaac 3000
uucggcgcca ucagcagcgu gcugaacgac auccugagcc ggcuggaccc ucccgaggcc 3060
gaggugcaga ucgaccggcu gaucacuggc cggcugcaga gccugcagac cuacgugacc 3120
cagcagcuga uccgggccgc cgagauucgg gccagcgcca accuggccgc caccaagaug 3180
agcgagugcg ugcugggcca gagcaagcgg guggacuucu gcggcaaggg cuaccaccug 3240
augagcuuuc cccagagcgc accccacgga gugguguucc ugcacgugac cuacgugccc 3300
gcccaggaga agaacuucac caccgcccca gccaucugcc acgacggcaa ggcccacuuu 3360
ccccgggagg gcguguucgu gagcaacggc acccacuggu ucgugaccca gcggaacuuc 3420
uacgagcccc agaucaucac caccgacaac accuucguga gcggcaacug cgacguggug 3480
aucggcaucg ugaacaacac cguguacgau ccccugcagc ccgagcugga cagcuucaag 3540
gaggagcugg acaaguacuu caagaaucac accagccccg acguggaccu gggcgacauc 3600
agcggcauca acgccagcgu ggugaacauc cagaaggaga ucgaucggcu gaacgaggug 3660
gccaagaacc ugaacgagag ccugaucgac cugcaggagc ugggcaagua cgagcaguac 3720
aucaaguggc ccugguacau cuggcugggc uucaucgccg gccugaucgc caucgugaug 3780
gugaccauca ugcugugcug caugaccagc ugcugcagcu gccugaaggg cuguugcagc 3840
ugcggcagcu gcugcaaguu cgacgaggac gacagcgagc ccgugcugaa gggcgugaag 3900
cugcacuaca ccugacucga gacuaguagc caacacccug ucuaaaaaac auaaauuucu 3960
uuaaucauuu ugccucuuuu cucugugcuu caauuaauaa aaaauggaaa gaaccuaaaa 4020
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 4080
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 4130
<210> 17
<211> 19
<212> PRT
<213> 人工序列
<220>
<223> 信号序列
<400> 17
Met Asp Trp Ile Trp Arg Ile Leu Phe Leu Val Gly Ala Ala Thr Gly
1 5 10 15
Ala His Ser
<210> 18
<211> 27
<212> PRT
<213> 人工序列
<220>
<223> 三聚体结构域的氨基酸序列
<400> 18
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> 19
<211> 4
<212> PRT
<213> 人工序列
<220>
<223> 接头序列
<400> 19
Ser Ala Ile Gly
1
<210> 20
<211> 1270
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 20
Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val
1 5 10 15
Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe
20 25 30
Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu
35 40 45
His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp
50 55 60
Phe His Val Ile Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp Asn Pro
65 70 75 80
Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Ile Glu Lys Ser
85 90 95
Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser Lys Thr
100 105 110
Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile Lys Val
115 120 125
Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Asp His Lys Asn Asn
130 135 140
Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr Ser Ser Ala Asn Asn
145 150 155 160
Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp Leu Glu Gly
165 170 175
Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe Lys Asn Ile
180 185 190
Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile Ile Val Arg
195 200 205
Glu Pro Glu Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu Pro Leu Val
210 215 220
Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu Ala
225 230 235 240
Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser Gly Trp Thr
245 250 255
Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr Phe
260 265 270
Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys
275 280 285
Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe Thr
290 295 300
Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro Thr
305 310 315 320
Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Asp
325 330 335
Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg
340 345 350
Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Leu
355 360 365
Ala Pro Phe Phe Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu
370 375 380
Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg
385 390 395 400
Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile Ala
405 410 415
Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala
420 425 430
Trp Asn Ser Asn Lys Leu Asp Ser Lys Val Ser Gly Asn Tyr Asn Tyr
435 440 445
Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp
450 455 460
Ile Ser Thr Glu Ile Tyr Gln Ala Gly Asn Lys Pro Cys Asn Gly Val
465 470 475 480
Ala Gly Phe Asn Cys Tyr Phe Pro Leu Arg Ser Tyr Ser Phe Arg Pro
485 490 495
Thr Tyr Gly Val Gly His Gln Pro Tyr Arg Val Val Val Leu Ser Phe
500 505 510
Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr
515 520 525
Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Lys
530 535 540
Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln
545 550 555 560
Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp Pro
565 570 575
Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val
580 585 590
Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu
595 600 605
Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile His Ala Asp
610 615 620
Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val Phe
625 630 635 640
Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu Tyr Val Asn Asn Ser
645 650 655
Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr Gln
660 665 670
Thr Gln Thr Lys Ser His Arg Arg Ala Arg Ser Val Ala Ser Gln Ser
675 680 685
Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala Tyr
690 695 700
Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val Thr
705 710 715 720
Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr
725 730 735
Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln
740 745 750
Tyr Gly Ser Phe Cys Thr Gln Leu Lys Arg Ala Leu Thr Gly Ile Ala
755 760 765
Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln
770 775 780
Ile Tyr Lys Thr Pro Pro Ile Lys Tyr Phe Gly Gly Phe Asn Phe Ser
785 790 795 800
Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile Glu
805 810 815
Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys
820 825 830
Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys
835 840 845
Ala Gln Lys Phe Lys Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp
850 855 860
Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr
865 870 875 880
Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala
885 890 895
Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val
900 905 910
Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile
915 920 925
Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys
930 935 940
Leu Gln Asp Val Val Asn His Asn Ala Gln Ala Leu Asn Thr Leu Val
945 950 955 960
Lys Gln Leu Ser Ser Lys Phe Gly Ala Ile Ser Ser Val Leu Asn Asp
965 970 975
Ile Phe Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln Ile Asp Arg
980 985 990
Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln
995 1000 1005
Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala
1010 1015 1020
Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp
1025 1030 1035
Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala
1040 1045 1050
Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala Gln
1055 1060 1065
Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys
1070 1075 1080
Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His
1085 1090 1095
Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr
1100 1105 1110
Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly
1115 1120 1125
Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp
1130 1135 1140
Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser
1145 1150 1155
Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val
1160 1165 1170
Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys
1175 1180 1185
Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr
1190 1195 1200
Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile
1205 1210 1215
Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys Cys
1220 1225 1230
Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly
1235 1240 1245
Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys
1250 1255 1260
Gly Val Lys Leu His Tyr Thr
1265 1270
<210> 21
<211> 1270
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 21
Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val
1 5 10 15
Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe
20 25 30
Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu
35 40 45
His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp
50 55 60
Phe His Val Ile Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp Asn Pro
65 70 75 80
Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Ile Glu Lys Ser
85 90 95
Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser Lys Thr
100 105 110
Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile Lys Val
115 120 125
Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Asp His Lys Asn Asn
130 135 140
Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr Ser Ser Ala Asn Asn
145 150 155 160
Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp Leu Glu Gly
165 170 175
Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe Lys Asn Ile
180 185 190
Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile Ile Val Arg
195 200 205
Glu Pro Glu Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu Pro Leu Val
210 215 220
Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu Ala
225 230 235 240
Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser Gly Trp Thr
245 250 255
Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr Phe
260 265 270
Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys
275 280 285
Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe Thr
290 295 300
Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro Thr
305 310 315 320
Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Asp
325 330 335
Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg
340 345 350
Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Leu
355 360 365
Ala Pro Phe Phe Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu
370 375 380
Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg
385 390 395 400
Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile Ala
405 410 415
Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala
420 425 430
Trp Asn Ser Asn Lys Leu Asp Ser Lys Val Ser Gly Asn Tyr Asn Tyr
435 440 445
Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp
450 455 460
Ile Ser Thr Glu Ile Tyr Gln Ala Gly Asn Lys Pro Cys Asn Gly Val
465 470 475 480
Ala Gly Phe Asn Cys Tyr Phe Pro Leu Arg Ser Tyr Ser Phe Arg Pro
485 490 495
Thr Tyr Gly Val Gly His Gln Pro Tyr Arg Val Val Val Leu Ser Phe
500 505 510
Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr
515 520 525
Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Lys
530 535 540
Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln
545 550 555 560
Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp Pro
565 570 575
Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val
580 585 590
Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu
595 600 605
Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile His Ala Asp
610 615 620
Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val Phe
625 630 635 640
Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu Tyr Val Asn Asn Ser
645 650 655
Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr Gln
660 665 670
Thr Gln Thr Lys Ser His Gly Gly Ser Gly Ser Val Ala Ser Gln Ser
675 680 685
Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala Tyr
690 695 700
Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val Thr
705 710 715 720
Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr
725 730 735
Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln
740 745 750
Tyr Gly Ser Phe Cys Thr Gln Leu Lys Arg Ala Leu Thr Gly Ile Ala
755 760 765
Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln
770 775 780
Ile Tyr Lys Thr Pro Pro Ile Lys Tyr Phe Gly Gly Phe Asn Phe Ser
785 790 795 800
Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Ala Asn Ser Phe Ile Glu
805 810 815
Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys
820 825 830
Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys
835 840 845
Ala Gln Lys Phe Lys Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp
850 855 860
Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr
865 870 875 880
Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala
885 890 895
Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val
900 905 910
Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile
915 920 925
Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys
930 935 940
Leu Gln Asp Val Val Asn His Asn Ala Gln Ala Leu Asn Thr Leu Val
945 950 955 960
Lys Gln Leu Ser Ser Lys Phe Gly Ala Ile Ser Ser Val Leu Asn Asp
965 970 975
Ile Phe Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln Ile Asp Arg
980 985 990
Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln
995 1000 1005
Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala
1010 1015 1020
Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp
1025 1030 1035
Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala
1040 1045 1050
Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala Gln
1055 1060 1065
Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys
1070 1075 1080
Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His
1085 1090 1095
Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr
1100 1105 1110
Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly
1115 1120 1125
Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp
1130 1135 1140
Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser
1145 1150 1155
Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val
1160 1165 1170
Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys
1175 1180 1185
Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr
1190 1195 1200
Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile
1205 1210 1215
Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys Cys
1220 1225 1230
Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly
1235 1240 1245
Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys
1250 1255 1260
Gly Val Lys Leu His Tyr Thr
1265 1270
<210> 22
<211> 1270
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 22
Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val
1 5 10 15
Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe
20 25 30
Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu
35 40 45
His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp
50 55 60
Phe His Val Ile Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp Asn Pro
65 70 75 80
Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Ile Glu Lys Ser
85 90 95
Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser Lys Thr
100 105 110
Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile Lys Val
115 120 125
Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Asp His Lys Asn Asn
130 135 140
Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr Ser Ser Ala Asn Asn
145 150 155 160
Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp Leu Glu Gly
165 170 175
Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe Lys Asn Ile
180 185 190
Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile Ile Val Arg
195 200 205
Glu Pro Glu Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu Pro Leu Val
210 215 220
Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu Ala
225 230 235 240
Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser Gly Trp Thr
245 250 255
Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr Phe
260 265 270
Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys
275 280 285
Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe Thr
290 295 300
Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro Thr
305 310 315 320
Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Asp
325 330 335
Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg
340 345 350
Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Leu
355 360 365
Ala Pro Phe Phe Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu
370 375 380
Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg
385 390 395 400
Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile Ala
405 410 415
Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala
420 425 430
Trp Asn Ser Asn Lys Leu Asp Ser Lys Val Ser Gly Asn Tyr Asn Tyr
435 440 445
Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp
450 455 460
Ile Ser Thr Glu Ile Tyr Gln Ala Gly Asn Lys Pro Cys Asn Gly Val
465 470 475 480
Ala Gly Phe Asn Cys Tyr Phe Pro Leu Arg Ser Tyr Ser Phe Arg Pro
485 490 495
Thr Tyr Gly Val Gly His Gln Pro Tyr Arg Val Val Val Leu Ser Phe
500 505 510
Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr
515 520 525
Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Lys
530 535 540
Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln
545 550 555 560
Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp Pro
565 570 575
Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val
580 585 590
Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu
595 600 605
Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile His Ala Asp
610 615 620
Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val Phe
625 630 635 640
Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu Tyr Val Asn Asn Ser
645 650 655
Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr Gln
660 665 670
Thr Gln Thr Lys Ser His Arg Arg Ala Arg Ser Val Ala Ser Gln Ser
675 680 685
Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala Tyr
690 695 700
Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val Thr
705 710 715 720
Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr
725 730 735
Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln
740 745 750
Tyr Gly Ser Phe Cys Thr Gln Leu Lys Arg Ala Leu Thr Gly Ile Ala
755 760 765
Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln
770 775 780
Ile Tyr Lys Thr Pro Pro Ile Lys Tyr Phe Gly Gly Phe Asn Phe Ser
785 790 795 800
Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile Glu
805 810 815
Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys
820 825 830
Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys
835 840 845
Ala Gln Lys Phe Lys Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp
850 855 860
Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr
865 870 875 880
Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala
885 890 895
Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val
900 905 910
Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile
915 920 925
Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys
930 935 940
Leu Gln Asp Val Val Asn His Asn Ala Gln Ala Leu Asn Thr Leu Val
945 950 955 960
Lys Gln Leu Ser Ser Lys Phe Gly Ala Ile Ser Ser Val Leu Asn Asp
965 970 975
Ile Phe Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp Arg
980 985 990
Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln
995 1000 1005
Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala
1010 1015 1020
Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp
1025 1030 1035
Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala
1040 1045 1050
Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala Gln
1055 1060 1065
Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys
1070 1075 1080
Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His
1085 1090 1095
Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr
1100 1105 1110
Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly
1115 1120 1125
Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp
1130 1135 1140
Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser
1145 1150 1155
Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val
1160 1165 1170
Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys
1175 1180 1185
Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr
1190 1195 1200
Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile
1205 1210 1215
Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys Cys
1220 1225 1230
Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly
1235 1240 1245
Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys
1250 1255 1260
Gly Val Lys Leu His Tyr Thr
1265 1270
<210> 23
<211> 1270
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 23
Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val
1 5 10 15
Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe
20 25 30
Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu
35 40 45
His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp
50 55 60
Phe His Val Ile Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp Asn Pro
65 70 75 80
Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Ile Glu Lys Ser
85 90 95
Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser Lys Thr
100 105 110
Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile Lys Val
115 120 125
Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Asp His Lys Asn Asn
130 135 140
Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr Ser Ser Ala Asn Asn
145 150 155 160
Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp Leu Glu Gly
165 170 175
Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe Lys Asn Ile
180 185 190
Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile Ile Val Arg
195 200 205
Glu Pro Glu Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu Pro Leu Val
210 215 220
Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu Ala
225 230 235 240
Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser Gly Trp Thr
245 250 255
Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr Phe
260 265 270
Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys
275 280 285
Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe Thr
290 295 300
Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro Thr
305 310 315 320
Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Asp
325 330 335
Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg
340 345 350
Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Leu
355 360 365
Ala Pro Phe Phe Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu
370 375 380
Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg
385 390 395 400
Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile Ala
405 410 415
Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala
420 425 430
Trp Asn Ser Asn Lys Leu Asp Ser Lys Val Ser Gly Asn Tyr Asn Tyr
435 440 445
Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp
450 455 460
Ile Ser Thr Glu Ile Tyr Gln Ala Gly Asn Lys Pro Cys Asn Gly Val
465 470 475 480
Ala Gly Phe Asn Cys Tyr Phe Pro Leu Arg Ser Tyr Ser Phe Arg Pro
485 490 495
Thr Tyr Gly Val Gly His Gln Pro Tyr Arg Val Val Val Leu Ser Phe
500 505 510
Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr
515 520 525
Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Lys
530 535 540
Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln
545 550 555 560
Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp Pro
565 570 575
Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val
580 585 590
Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu
595 600 605
Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile His Ala Asp
610 615 620
Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val Phe
625 630 635 640
Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu Tyr Val Asn Asn Ser
645 650 655
Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr Gln
660 665 670
Thr Gln Thr Lys Ser His Gly Gly Ser Gly Ser Val Ala Ser Gln Ser
675 680 685
Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala Tyr
690 695 700
Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val Thr
705 710 715 720
Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr
725 730 735
Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln
740 745 750
Tyr Gly Ser Phe Cys Thr Gln Leu Lys Arg Ala Leu Thr Gly Ile Ala
755 760 765
Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln
770 775 780
Ile Tyr Lys Thr Pro Pro Ile Lys Tyr Phe Gly Gly Phe Asn Phe Ser
785 790 795 800
Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile Glu
805 810 815
Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys
820 825 830
Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys
835 840 845
Ala Gln Lys Phe Lys Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp
850 855 860
Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr
865 870 875 880
Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala
885 890 895
Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val
900 905 910
Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile
915 920 925
Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys
930 935 940
Leu Gln Asp Val Val Asn His Asn Ala Gln Ala Leu Asn Thr Leu Val
945 950 955 960
Lys Gln Leu Ser Ser Lys Phe Gly Ala Ile Ser Ser Val Leu Asn Asp
965 970 975
Ile Phe Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp Arg
980 985 990
Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln
995 1000 1005
Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala
1010 1015 1020
Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp
1025 1030 1035
Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala
1040 1045 1050
Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala Gln
1055 1060 1065
Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys
1070 1075 1080
Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His
1085 1090 1095
Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr
1100 1105 1110
Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly
1115 1120 1125
Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp
1130 1135 1140
Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser
1145 1150 1155
Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val
1160 1165 1170
Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys
1175 1180 1185
Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr
1190 1195 1200
Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile
1205 1210 1215
Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys Cys
1220 1225 1230
Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly
1235 1240 1245
Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys
1250 1255 1260
Gly Val Lys Leu His Tyr Thr
1265 1270
<210> 24
<211> 1270
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 24
Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val
1 5 10 15
Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe
20 25 30
Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu
35 40 45
His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp
50 55 60
Phe His Val Ile Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp Asn Pro
65 70 75 80
Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Ile Glu Lys Ser
85 90 95
Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser Lys Thr
100 105 110
Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile Lys Val
115 120 125
Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Asp His Lys Asn Asn
130 135 140
Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr Ser Ser Ala Asn Asn
145 150 155 160
Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp Leu Glu Gly
165 170 175
Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe Lys Asn Ile
180 185 190
Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile Ile Val Arg
195 200 205
Glu Pro Glu Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu Pro Leu Val
210 215 220
Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu Ala
225 230 235 240
Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser Gly Trp Thr
245 250 255
Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr Phe
260 265 270
Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys
275 280 285
Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe Thr
290 295 300
Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro Thr
305 310 315 320
Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Asp
325 330 335
Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg
340 345 350
Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Leu
355 360 365
Ala Pro Phe Phe Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu
370 375 380
Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg
385 390 395 400
Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile Ala
405 410 415
Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala
420 425 430
Trp Asn Ser Asn Lys Leu Asp Ser Lys Val Ser Gly Asn Tyr Asn Tyr
435 440 445
Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp
450 455 460
Ile Ser Thr Glu Ile Tyr Gln Ala Gly Asn Lys Pro Cys Asn Gly Val
465 470 475 480
Ala Gly Phe Asn Cys Tyr Phe Pro Leu Arg Ser Tyr Ser Phe Arg Pro
485 490 495
Thr Tyr Gly Val Gly His Gln Pro Tyr Arg Val Val Val Leu Ser Phe
500 505 510
Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr
515 520 525
Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Lys
530 535 540
Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln
545 550 555 560
Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp Pro
565 570 575
Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val
580 585 590
Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu
595 600 605
Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile His Ala Asp
610 615 620
Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val Phe
625 630 635 640
Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu Tyr Val Asn Asn Ser
645 650 655
Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr Gln
660 665 670
Thr Gln Thr Lys Ser His Arg Arg Ala Arg Ser Val Ala Ser Gln Ser
675 680 685
Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala Tyr
690 695 700
Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val Thr
705 710 715 720
Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr
725 730 735
Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln
740 745 750
Tyr Gly Ser Phe Cys Thr Gln Leu Lys Arg Ala Leu Thr Gly Ile Ala
755 760 765
Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln
770 775 780
Ile Tyr Lys Thr Pro Pro Ile Lys Tyr Phe Gly Gly Phe Asn Phe Ser
785 790 795 800
Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Ala Asn Ser Phe Ile Glu
805 810 815
Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys
820 825 830
Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys
835 840 845
Ala Gln Lys Phe Lys Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp
850 855 860
Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr
865 870 875 880
Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala
885 890 895
Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val
900 905 910
Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile
915 920 925
Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys
930 935 940
Leu Gln Asp Val Val Asn His Asn Ala Gln Ala Leu Asn Thr Leu Val
945 950 955 960
Lys Gln Leu Ser Ser Lys Phe Gly Ala Ile Ser Ser Val Leu Asn Asp
965 970 975
Ile Phe Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp Arg
980 985 990
Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln
995 1000 1005
Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala
1010 1015 1020
Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp
1025 1030 1035
Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala
1040 1045 1050
Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala Gln
1055 1060 1065
Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys
1070 1075 1080
Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His
1085 1090 1095
Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr
1100 1105 1110
Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly
1115 1120 1125
Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp
1130 1135 1140
Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser
1145 1150 1155
Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val
1160 1165 1170
Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys
1175 1180 1185
Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr
1190 1195 1200
Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile
1205 1210 1215
Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys Cys
1220 1225 1230
Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly
1235 1240 1245
Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys
1250 1255 1260
Gly Val Lys Leu His Tyr Thr
1265 1270
<210> 25
<211> 1270
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 25
Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val
1 5 10 15
Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe
20 25 30
Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu
35 40 45
His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp
50 55 60
Phe His Val Ile Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp Asn Pro
65 70 75 80
Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Ile Glu Lys Ser
85 90 95
Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser Lys Thr
100 105 110
Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile Lys Val
115 120 125
Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Asp His Lys Asn Asn
130 135 140
Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr Ser Ser Ala Asn Asn
145 150 155 160
Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp Leu Glu Gly
165 170 175
Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe Lys Asn Ile
180 185 190
Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile Ile Val Arg
195 200 205
Glu Pro Glu Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu Pro Leu Val
210 215 220
Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu Ala
225 230 235 240
Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser Gly Trp Thr
245 250 255
Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr Phe
260 265 270
Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys
275 280 285
Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe Thr
290 295 300
Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro Thr
305 310 315 320
Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Asp
325 330 335
Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg
340 345 350
Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Leu
355 360 365
Ala Pro Phe Phe Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu
370 375 380
Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg
385 390 395 400
Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile Ala
405 410 415
Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala
420 425 430
Trp Asn Ser Asn Lys Leu Asp Ser Lys Val Ser Gly Asn Tyr Asn Tyr
435 440 445
Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp
450 455 460
Ile Ser Thr Glu Ile Tyr Gln Ala Gly Asn Lys Pro Cys Asn Gly Val
465 470 475 480
Ala Gly Phe Asn Cys Tyr Phe Pro Leu Arg Ser Tyr Ser Phe Arg Pro
485 490 495
Thr Tyr Gly Val Gly His Gln Pro Tyr Arg Val Val Val Leu Ser Phe
500 505 510
Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr
515 520 525
Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Lys
530 535 540
Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln
545 550 555 560
Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp Pro
565 570 575
Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val
580 585 590
Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu
595 600 605
Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile His Ala Asp
610 615 620
Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val Phe
625 630 635 640
Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu Tyr Val Asn Asn Ser
645 650 655
Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr Gln
660 665 670
Thr Gln Thr Lys Ser His Gly Gly Ser Gly Ser Val Ala Ser Gln Ser
675 680 685
Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala Tyr
690 695 700
Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val Thr
705 710 715 720
Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr
725 730 735
Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln
740 745 750
Tyr Gly Ser Phe Cys Thr Gln Leu Lys Arg Ala Leu Thr Gly Ile Ala
755 760 765
Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln
770 775 780
Ile Tyr Lys Thr Pro Pro Ile Lys Tyr Phe Gly Gly Phe Asn Phe Ser
785 790 795 800
Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Ala Asn Ser Phe Ile Glu
805 810 815
Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys
820 825 830
Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys
835 840 845
Ala Gln Lys Phe Lys Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp
850 855 860
Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr
865 870 875 880
Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala
885 890 895
Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val
900 905 910
Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile
915 920 925
Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys
930 935 940
Leu Gln Asp Val Val Asn His Asn Ala Gln Ala Leu Asn Thr Leu Val
945 950 955 960
Lys Gln Leu Ser Ser Lys Phe Gly Ala Ile Ser Ser Val Leu Asn Asp
965 970 975
Ile Phe Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp Arg
980 985 990
Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln
995 1000 1005
Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala
1010 1015 1020
Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp
1025 1030 1035
Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala
1040 1045 1050
Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala Gln
1055 1060 1065
Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys
1070 1075 1080
Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His
1085 1090 1095
Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr
1100 1105 1110
Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly
1115 1120 1125
Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp
1130 1135 1140
Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser
1145 1150 1155
Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val
1160 1165 1170
Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys
1175 1180 1185
Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr
1190 1195 1200
Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile
1205 1210 1215
Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys Cys
1220 1225 1230
Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly
1235 1240 1245
Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys
1250 1255 1260
Gly Val Lys Leu His Tyr Thr
1265 1270
<210> 26
<211> 3813
<212> DNA
<213> 人工序列
<220>
<223> RBMRNA-Omicron疫苗的DNA序列
<400> 26
atgttcgtgt tcctggtgct gctgcctctg gtgagcagcc agtgcgtgaa tctgaccacc 60
agaacccagc tgcctcctgc ctacaccaat agcttcacca gaggagttta ttatcccgat 120
aaggtgttca gaagtagtgt attacatagt acccaggacc tgttcctacc tttcttcagt 180
aacgtgacct ggttccacgt catcagcggc accaatggca ccaagagatt cgacaatcct 240
gtgctgcctt tcaatgacgg cgtgtacttc gccagcatcg agaagagcaa tatcatcaga 300
ggctggatct tcggcaccac cttggattcc aagactcaga gcctgctgat tgtaaacaac 360
gctacaaatg tggtgatcaa ggtgtgcgag ttccagttct gcaatgaccc tttcctggac 420
cataagaaca acaagagctg gatggagagc gagttccgcg tatattcgtc ggctaataat 480
tgcaccttcg agtacgtgag ccagcctttc ctgatggacc tggagggcaa gcagggcaat 540
ttcaagaatc tgagagagtt cgtgttcaag aatatcgacg gctacttcaa gatctacagc 600
aagcacacac ccattatcgt gagagagccg gaggacctgc ctcagggctt cagcgccctg 660
gagcctctgg tggacctgcc tatcggcatc aatatcacca gattccagac cctgctggcc 720
ctgcacagat catatcttac accaggcgat tcgtcaagcg gttggaccgc tggagctgcg 780
gcatattacg tgggctacct gcagcctaga accttcctgc tgaagtacaa tgagaatggt 840
acgataaccg acgcagttga ttgtgccctg gaccctctga gcgagaccaa gtgcaccctg 900
aagagcttca ccgtggagaa gggcatctac cagaccagca atttcagagt gcagcctacc 960
gagagcatcg tgagattccc taatatcacc aatctgtgcc ctttcgacga ggtgttcaat 1020
gccaccagat tcgccagcgt gtacgcatgg aaccgcaagc ggataagcaa ttgcgtggcc 1080
gactacagcg tgctgtacaa tctggcccca ttcttcacct tcaaatgtta tggtgtttcg 1140
ccaacaaagc tgaatgacct gtgcttcacc aatgtgtacg ccgacagctt cgtgatcaga 1200
ggcgacgagg tgagacagat cgcgccaggg cagaccggca acatcgccga ctacaattac 1260
aagctgcctg acgacttcac cggctgcgtg atcgcgtgga actctaacaa gctggactcg 1320
aaagtttcag gcaattacaa ttacctgtac agactgttca gaaagagcaa tctgaagcct 1380
ttcgagagag acatcagcac cgagatctac caggccggca acaagccgtg taatggcgtg 1440
gcgggcttca attgctactt ccctctgcgc agctacagct tccggcctac ctacggcgtg 1500
ggccaccagc cttacagagt ggtggtgctg agcttcgagc tgctgcacgc tcccgctacc 1560
gtgtgcggcc ctaagaagag caccaatctg gtgaagaata agtgcgtgaa tttcaatttc 1620
aatggtctaa agggaacggg cgtgctgacc gagagcaata agaagtttct tccctttcaa 1680
caattcggca gagacatcgc cgacaccaca gatgctgtaa gagaccctca gaccctggag 1740
atcctggaca tcactccgtg tagcttcggc ggcgtgagcg tgatcacacc gggcaccaat 1800
accagcaatc aggtggccgt gctgtaccag ggcgtgaatt gcaccgaggt gcctgtggcc 1860
atccacgccg accagctgac tcccacttgg agggtatatt ccacgggaag caatgtgttc 1920
cagaccagag ccggctgcct gatcggcgcc gagtacgtga ataatagcta cgagtgcgac 1980
atccctatcg gcgccggcat ctgcgccagc taccagaccc agaccaagag ccacggcggc 2040
agcggcagcg tggccagcca gagcatcatc gcctacacca tgagcctggg cgccgagaat 2100
agcgtggcct acagcaataa tagcatcgcc atccctacca atttcaccat cagcgtgacc 2160
accgaaatat taccagtctc catgaccaag accagcgtgg actgcaccat gtacatctgc 2220
ggcgacagca ccgagtgcag caatctgctg ctgcagtacg gcagcttctg cacccagctg 2280
aagagagccc tgaccggcat cgccgtggag caggacaaga atacccagga ggtgttcgcc 2340
caggtgaagc agatctacaa gactccgccg atcaagtact tcggcggctt caatttcagc 2400
caaatactcc cagatccaag caagcctagc gccaacagct tcatcgagga cctgctgttc 2460
aataaggtga ccctggccga cgccggcttc atcaagcagt acggcgactg cctaggtgat 2520
attgcggcaa gagacctgat ctgcgcccag aagtttaagg gtttgacagt actacctcct 2580
ctgctgaccg acgagatgat agcacaatat acgtcggcat tgctcgctgg cacgatcaca 2640
tcgggctgga ctttcggcgc cggagcagcg ttgcaaatcc ctttcgccat gcagatggcc 2700
tacagattca atggcatcgg cgtgacccag aatgtgctgt acgagaatca gaagctgatc 2760
gccaatcagt tcaatagcgc catcggcaag atccaggaca gcctgagcag caccgccagc 2820
gccctgggca agctgcagga cgtggtgaat cacaatgccc aggccctgaa taccctggtg 2880
aagcagctga gcagcaagtt cggcgccatc agtagtgtac tcaacgacat cttcagcaga 2940
ctggacccac cagaggccga ggtgcaaatt gatcgtctta ttactggcag actgcagagc 3000
ctgcagacct acgtgaccca gcagctgatc agagccgccg agatcagagc cagcgccaat 3060
ctggccgcca ccaagatgag cgagtgcgtg ctgggccaga gcaagagagt ggacttctgc 3120
ggcaagggct accacctgat gagcttccct cagagcgctc cacatggcgt ggtgttcctg 3180
cacgtgacct acgtgcctgc ccaggagaag aatttcacca ccgcacccgc aatctgccac 3240
gacggcaagg cccacttccc tagagagggc gtgttcgtga gcaatggcac ccactggttc 3300
gtgacccaga gaaatttcta cgagcctcag atcatcacca ccgacaatac cttcgtgagc 3360
ggcaattgcg acgtggtgat cgggatagtc aataatactg tctacgaccc tctgcagcct 3420
gagctggaca gcttcaagga ggagctggac aagtacttca agaatcacac cagccctgac 3480
gtggacctcg gtgatatttc gggaatcaat gccagcgtgg tgaatatcca gaaggaaatt 3540
gatcggctca acgaagtggc caagaatctg aatgagagcc tgatcgacct gcaggagctg 3600
ggcaagtacg agcagtacat caagtggcct tggtacatct ggctgggctt catcgccggc 3660
ctgatcgcca tcgtgatggt gaccatcatg ctgtgctgca tgacctcctg ttgttcctgt 3720
ttgaaagggt gttgttcgtg tgggtcctgc tgcaagttcg acgaggacga cagcgagcct 3780
gtgctgaagg gcgtgaagct gcactacacc taa 3813
<210> 27
<211> 4127
<212> RNA
<213> 人工序列
<220>
<223> RBMRNA-Omicron疫苗的mRNA序列
<400> 27
augagacaag cuugucccgc agucggcguc cagcggcucu gcuuguucgu gugugugucg 60
uugcaggccu uauucagauc uaccgguggu accgccacca uguucguguu ccuggugcug 120
cugccucugg ugagcagcca gugcgugaau cugaccacca gaacccagcu gccuccugcc 180
uacaccaaua gcuucaccag aggaguuuau uaucccgaua agguguucag aaguagugua 240
uuacauagua cccaggaccu guuccuaccu uucuucagua acgugaccug guuccacguc 300
aucagcggca ccaauggcac caagagauuc gacaauccug ugcugccuuu caaugacggc 360
guguacuucg ccagcaucga gaagagcaau aucaucagag gcuggaucuu cggcaccacc 420
uuggauucca agacucagag ccugcugauu guaaacaacg cuacaaaugu ggugaucaag 480
gugugcgagu uccaguucug caaugacccu uuccuggacc auaagaacaa caagagcugg 540
auggagagcg aguuccgcgu auauucgucg gcuaauaauu gcaccuucga guacgugagc 600
cagccuuucc ugauggaccu ggagggcaag cagggcaauu ucaagaaucu gagagaguuc 660
guguucaaga auaucgacgg cuacuucaag aucuacagca agcacacacc cauuaucgug 720
agagagccgg aggaccugcc ucagggcuuc agcgcccugg agccucuggu ggaccugccu 780
aucggcauca auaucaccag auuccagacc cugcuggccc ugcacagauc auaucuuaca 840
ccaggcgauu cgucaagcgg uuggaccgcu ggagcugcgg cauauuacgu gggcuaccug 900
cagccuagaa ccuuccugcu gaaguacaau gagaauggua cgauaaccga cgcaguugau 960
ugugcccugg acccucugag cgagaccaag ugcacccuga agagcuucac cguggagaag 1020
ggcaucuacc agaccagcaa uuucagagug cagccuaccg agagcaucgu gagauucccu 1080
aauaucacca aucugugccc uuucgacgag guguucaaug ccaccagauu cgccagcgug 1140
uacgcaugga accgcaagcg gauaagcaau ugcguggccg acuacagcgu gcuguacaau 1200
cuggccccau ucuucaccuu caaauguuau gguguuucgc caacaaagcu gaaugaccug 1260
ugcuucacca auguguacgc cgacagcuuc gugaucagag gcgacgaggu gagacagauc 1320
gcgccagggc agaccggcaa caucgccgac uacaauuaca agcugccuga cgacuucacc 1380
ggcugcguga ucgcguggaa cucuaacaag cuggacucga aaguuucagg caauuacaau 1440
uaccuguaca gacuguucag aaagagcaau cugaagccuu ucgagagaga caucagcacc 1500
gagaucuacc aggccggcaa caagccgugu aauggcgugg cgggcuucaa uugcuacuuc 1560
ccucugcgca gcuacagcuu ccggccuacc uacggcgugg gccaccagcc uuacagagug 1620
guggugcuga gcuucgagcu gcugcacgcu cccgcuaccg ugugcggccc uaagaagagc 1680
accaaucugg ugaagaauaa gugcgugaau uucaauuuca auggucuaaa gggaacgggc 1740
gugcugaccg agagcaauaa gaaguuucuu cccuuucaac aauucggcag agacaucgcc 1800
gacaccacag augcuguaag agacccucag acccuggaga uccuggacau cacuccgugu 1860
agcuucggcg gcgugagcgu gaucacaccg ggcaccaaua ccagcaauca gguggccgug 1920
cuguaccagg gcgugaauug caccgaggug ccuguggcca uccacgccga ccagcugacu 1980
cccacuugga ggguauauuc cacgggaagc aauguguucc agaccagagc cggcugccug 2040
aucggcgccg aguacgugaa uaauagcuac gagugcgaca ucccuaucgg cgccggcauc 2100
ugcgccagcu accagaccca gaccaagagc cacggcggca gcggcagcgu ggccagccag 2160
agcaucaucg ccuacaccau gagccugggc gccgagaaua gcguggccua cagcaauaau 2220
agcaucgcca ucccuaccaa uuucaccauc agcgugacca ccgaaauauu accagucucc 2280
augaccaaga ccagcgugga cugcaccaug uacaucugcg gcgacagcac cgagugcagc 2340
aaucugcugc ugcaguacgg cagcuucugc acccagcuga agagagcccu gaccggcauc 2400
gccguggagc aggacaagaa uacccaggag guguucgccc aggugaagca gaucuacaag 2460
acuccgccga ucaaguacuu cggcggcuuc aauuucagcc aaauacuccc agauccaagc 2520
aagccuagcg ccaacagcuu caucgaggac cugcuguuca auaaggugac ccuggccgac 2580
gccggcuuca ucaagcagua cggcgacugc cuaggugaua uugcggcaag agaccugauc 2640
ugcgcccaga aguuuaaggg uuugacagua cuaccuccuc ugcugaccga cgagaugaua 2700
gcacaauaua cgucggcauu gcucgcuggc acgaucacau cgggcuggac uuucggcgcc 2760
ggagcagcgu ugcaaauccc uuucgccaug cagauggccu acagauucaa uggcaucggc 2820
gugacccaga augugcugua cgagaaucag aagcugaucg ccaaucaguu caauagcgcc 2880
aucggcaaga uccaggacag ccugagcagc accgccagcg cccugggcaa gcugcaggac 2940
guggugaauc acaaugccca ggcccugaau acccugguga agcagcugag cagcaaguuc 3000
ggcgccauca guaguguacu caacgacauc uucagcagac uggacccacc agaggccgag 3060
gugcaaauug aucgucuuau uacuggcaga cugcagagcc ugcagaccua cgugacccag 3120
cagcugauca gagccgccga gaucagagcc agcgccaauc uggccgccac caagaugagc 3180
gagugcgugc ugggccagag caagagagug gacuucugcg gcaagggcua ccaccugaug 3240
agcuucccuc agagcgcucc acauggcgug guguuccugc acgugaccua cgugccugcc 3300
caggagaaga auuucaccac cgcacccgca aucugccacg acggcaaggc ccacuucccu 3360
agagagggcg uguucgugag caauggcacc cacugguucg ugacccagag aaauuucuac 3420
gagccucaga ucaucaccac cgacaauacc uucgugagcg gcaauugcga cguggugauc 3480
gggauaguca auaauacugu cuacgacccu cugcagccug agcuggacag cuucaaggag 3540
gagcuggaca aguacuucaa gaaucacacc agcccugacg uggaccucgg ugauauuucg 3600
ggaaucaaug ccagcguggu gaauauccag aaggaaauug aucggcucaa cgaaguggcc 3660
aagaaucuga augagagccu gaucgaccug caggagcugg gcaaguacga gcaguacauc 3720
aaguggccuu gguacaucug gcugggcuuc aucgccggcc ugaucgccau cgugauggug 3780
accaucaugc ugugcugcau gaccuccugu uguuccuguu ugaaagggug uuguucgugu 3840
ggguccugcu gcaaguucga cgaggacgac agcgagccug ugcugaaggg cgugaagcug 3900
cacuacaccu aacucgagac uaguagccaa cacccugucu aaaaaacaua aauuucuuua 3960
aucauuuugc cucuuuucuc ugugcuucaa uuaauaaaaa auggaaagaa ccuaaaaaaa 4020
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 4080
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaa 4127
<210> 28
<211> 6800
<212> DNA
<213> 人工序列
<220>
<223> RBMRNA-Omicron疫苗的质粒模板的DNA序列
<400> 28
tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 60
cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 120
agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 180
gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 240
aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 300
tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag 360
ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg 420
aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct 480
ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt 540
agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg 600
gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc 660
tctaatacga ctcactataa tgagacaagc ttgtcccgca gtcggcgtcc agcggctctg 720
cttgttcgtg tgtgtgtcgt tgcaggcctt attcagatct accggtggta ccgccaccat 780
gttcgtgttc ctggtgctgc tgcctctggt gagcagccag tgcgtgaatc tgaccaccag 840
aacccagctg cctcctgcct acaccaatag cttcaccaga ggagtttatt atcccgataa 900
ggtgttcaga agtagtgtat tacatagtac ccaggacctg ttcctacctt tcttcagtaa 960
cgtgacctgg ttccacgtca tcagcggcac caatggcacc aagagattcg acaatcctgt 1020
gctgcctttc aatgacggcg tgtacttcgc cagcatcgag aagagcaata tcatcagagg 1080
ctggatcttc ggcaccacct tggattccaa gactcagagc ctgctgattg taaacaacgc 1140
tacaaatgtg gtgatcaagg tgtgcgagtt ccagttctgc aatgaccctt tcctggacca 1200
taagaacaac aagagctgga tggagagcga gttccgcgta tattcgtcgg ctaataattg 1260
caccttcgag tacgtgagcc agcctttcct gatggacctg gagggcaagc agggcaattt 1320
caagaatctg agagagttcg tgttcaagaa tatcgacggc tacttcaaga tctacagcaa 1380
gcacacaccc attatcgtga gagagccgga ggacctgcct cagggcttca gcgccctgga 1440
gcctctggtg gacctgccta tcggcatcaa tatcaccaga ttccagaccc tgctggccct 1500
gcacagatca tatcttacac caggcgattc gtcaagcggt tggaccgctg gagctgcggc 1560
atattacgtg ggctacctgc agcctagaac cttcctgctg aagtacaatg agaatggtac 1620
gataaccgac gcagttgatt gtgccctgga ccctctgagc gagaccaagt gcaccctgaa 1680
gagcttcacc gtggagaagg gcatctacca gaccagcaat ttcagagtgc agcctaccga 1740
gagcatcgtg agattcccta atatcaccaa tctgtgccct ttcgacgagg tgttcaatgc 1800
caccagattc gccagcgtgt acgcatggaa ccgcaagcgg ataagcaatt gcgtggccga 1860
ctacagcgtg ctgtacaatc tggccccatt cttcaccttc aaatgttatg gtgtttcgcc 1920
aacaaagctg aatgacctgt gcttcaccaa tgtgtacgcc gacagcttcg tgatcagagg 1980
cgacgaggtg agacagatcg cgccagggca gaccggcaac atcgccgact acaattacaa 2040
gctgcctgac gacttcaccg gctgcgtgat cgcgtggaac tctaacaagc tggactcgaa 2100
agtttcaggc aattacaatt acctgtacag actgttcaga aagagcaatc tgaagccttt 2160
cgagagagac atcagcaccg agatctacca ggccggcaac aagccgtgta atggcgtggc 2220
gggcttcaat tgctacttcc ctctgcgcag ctacagcttc cggcctacct acggcgtggg 2280
ccaccagcct tacagagtgg tggtgctgag cttcgagctg ctgcacgctc ccgctaccgt 2340
gtgcggccct aagaagagca ccaatctggt gaagaataag tgcgtgaatt tcaatttcaa 2400
tggtctaaag ggaacgggcg tgctgaccga gagcaataag aagtttcttc cctttcaaca 2460
attcggcaga gacatcgccg acaccacaga tgctgtaaga gaccctcaga ccctggagat 2520
cctggacatc actccgtgta gcttcggcgg cgtgagcgtg atcacaccgg gcaccaatac 2580
cagcaatcag gtggccgtgc tgtaccaggg cgtgaattgc accgaggtgc ctgtggccat 2640
ccacgccgac cagctgactc ccacttggag ggtatattcc acgggaagca atgtgttcca 2700
gaccagagcc ggctgcctga tcggcgccga gtacgtgaat aatagctacg agtgcgacat 2760
ccctatcggc gccggcatct gcgccagcta ccagacccag accaagagcc acggcggcag 2820
cggcagcgtg gccagccaga gcatcatcgc ctacaccatg agcctgggcg ccgagaatag 2880
cgtggcctac agcaataata gcatcgccat ccctaccaat ttcaccatca gcgtgaccac 2940
cgaaatatta ccagtctcca tgaccaagac cagcgtggac tgcaccatgt acatctgcgg 3000
cgacagcacc gagtgcagca atctgctgct gcagtacggc agcttctgca cccagctgaa 3060
gagagccctg accggcatcg ccgtggagca ggacaagaat acccaggagg tgttcgccca 3120
ggtgaagcag atctacaaga ctccgccgat caagtacttc ggcggcttca atttcagcca 3180
aatactccca gatccaagca agcctagcgc caacagcttc atcgaggacc tgctgttcaa 3240
taaggtgacc ctggccgacg ccggcttcat caagcagtac ggcgactgcc taggtgatat 3300
tgcggcaaga gacctgatct gcgcccagaa gtttaagggt ttgacagtac tacctcctct 3360
gctgaccgac gagatgatag cacaatatac gtcggcattg ctcgctggca cgatcacatc 3420
gggctggact ttcggcgccg gagcagcgtt gcaaatccct ttcgccatgc agatggccta 3480
cagattcaat ggcatcggcg tgacccagaa tgtgctgtac gagaatcaga agctgatcgc 3540
caatcagttc aatagcgcca tcggcaagat ccaggacagc ctgagcagca ccgccagcgc 3600
cctgggcaag ctgcaggacg tggtgaatca caatgcccag gccctgaata ccctggtgaa 3660
gcagctgagc agcaagttcg gcgccatcag tagtgtactc aacgacatct tcagcagact 3720
ggacccacca gaggccgagg tgcaaattga tcgtcttatt actggcagac tgcagagcct 3780
gcagacctac gtgacccagc agctgatcag agccgccgag atcagagcca gcgccaatct 3840
ggccgccacc aagatgagcg agtgcgtgct gggccagagc aagagagtgg acttctgcgg 3900
caagggctac cacctgatga gcttccctca gagcgctcca catggcgtgg tgttcctgca 3960
cgtgacctac gtgcctgccc aggagaagaa tttcaccacc gcacccgcaa tctgccacga 4020
cggcaaggcc cacttcccta gagagggcgt gttcgtgagc aatggcaccc actggttcgt 4080
gacccagaga aatttctacg agcctcagat catcaccacc gacaatacct tcgtgagcgg 4140
caattgcgac gtggtgatcg ggatagtcaa taatactgtc tacgaccctc tgcagcctga 4200
gctggacagc ttcaaggagg agctggacaa gtacttcaag aatcacacca gccctgacgt 4260
ggacctcggt gatatttcgg gaatcaatgc cagcgtggtg aatatccaga aggaaattga 4320
tcggctcaac gaagtggcca agaatctgaa tgagagcctg atcgacctgc aggagctggg 4380
caagtacgag cagtacatca agtggccttg gtacatctgg ctgggcttca tcgccggcct 4440
gatcgccatc gtgatggtga ccatcatgct gtgctgcatg acctcctgtt gttcctgttt 4500
gaaagggtgt tgttcgtgtg ggtcctgctg caagttcgac gaggacgaca gcgagcctgt 4560
gctgaagggc gtgaagctgc actacaccta actcgagact agtagccaac accctgtcta 4620
aaaaacataa atttctttaa tcattttgcc tcttttctct gtgcttcaat taataaaaaa 4680
tggaaagaac ctaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 4740
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 4800
aaaaaaggat cctctagact gcaggagctc gatatcgaat tcgtattcta tagtgtcacc 4860
taaatccaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac 4920
ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc 4980
ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg 5040
gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggggca ctctcagtac 5100
aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc 5160
gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg 5220
gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct 5280
cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg 5340
tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc 5400
aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag 5460
gaagagtatg agccatattc aacgggaaac gtcttgctct aggccgcgat taaattccaa 5520
catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc 5580
gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa 5640
aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt 5700
tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac 5760
cactgcgatc cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga 5820
aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa 5880
ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa 5940
cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt 6000
ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga 6060
tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg 6120
acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga 6180
gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat 6240
gaataaattg cagtttcatt tgatgctcga tgagtttttc taactgtcag accaagttta 6300
ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa 6360
gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 6420
gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 6480
ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 6540
gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 6600
tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 6660
cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 6720
cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 6780
ttcgtgcaca cagcccagct 6800
<210> 29
<211> 1273
<212> PRT
<213> 未知
<220>
<223> SARS-CoV-2野生型S蛋白的氨基酸序列
<400> 29
Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val
1 5 10 15
Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe
20 25 30
Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu
35 40 45
His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp
50 55 60
Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp
65 70 75 80
Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu
85 90 95
Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser
100 105 110
Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile
115 120 125
Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr
130 135 140
Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr
145 150 155 160
Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu
165 170 175
Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe
180 185 190
Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr
195 200 205
Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu
210 215 220
Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr
225 230 235 240
Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser
245 250 255
Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro
260 265 270
Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala
275 280 285
Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys
290 295 300
Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val
305 310 315 320
Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys
325 330 335
Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala
340 345 350
Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu
355 360 365
Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro
370 375 380
Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe
385 390 395 400
Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly
405 410 415
Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys
420 425 430
Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn
435 440 445
Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe
450 455 460
Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys
465 470 475 480
Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly
485 490 495
Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val
500 505 510
Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys
515 520 525
Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn
530 535 540
Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu
545 550 555 560
Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val
565 570 575
Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe
580 585 590
Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val
595 600 605
Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile
610 615 620
His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser
625 630 635 640
Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val
645 650 655
Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala
660 665 670
Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser Val Ala
675 680 685
Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser
690 695 700
Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile
705 710 715 720
Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val
725 730 735
Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu
740 745 750
Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr
755 760 765
Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln
770 775 780
Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe
785 790 795 800
Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser
805 810 815
Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly
820 825 830
Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp
835 840 845
Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu
850 855 860
Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly
865 870 875 880
Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile
885 890 895
Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr
900 905 910
Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn
915 920 925
Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala
930 935 940
Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn
945 950 955 960
Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val
965 970 975
Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln
980 985 990
Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val
995 1000 1005
Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn
1010 1015 1020
Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys
1025 1030 1035
Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro
1040 1045 1050
Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val
1055 1060 1065
Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His
1070 1075 1080
Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn
1085 1090 1095
Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln
1100 1105 1110
Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val
1115 1120 1125
Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro
1130 1135 1140
Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn
1145 1150 1155
His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn
1160 1165 1170
Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu
1175 1180 1185
Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu
1190 1195 1200
Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu
1205 1210 1215
Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met
1220 1225 1230
Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys
1235 1240 1245
Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro
1250 1255 1260
Val Leu Lys Gly Val Lys Leu His Tyr Thr
1265 1270
<210> 30
<211> 1250
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 30
Met Asp Trp Ile Trp Arg Ile Leu Phe Leu Val Gly Ala Ala Thr Gly
1 5 10 15
Ala His Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro
20 25 30
Ala Tyr Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val
35 40 45
Phe Arg Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe
50 55 60
Phe Ser Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn
65 70 75 80
Gly Thr Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val
85 90 95
Tyr Phe Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe
100 105 110
Gly Thr Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn
115 120 125
Ala Thr Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp
130 135 140
Pro Phe Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu
145 150 155 160
Ser Glu Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr
165 170 175
Val Ser Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe
180 185 190
Lys Asn Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys
195 200 205
Ile Tyr Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln
210 215 220
Gly Phe Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn
225 230 235 240
Ile Thr Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr
245 250 255
Pro Gly Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr
260 265 270
Val Gly Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn
275 280 285
Gly Thr Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu
290 295 300
Thr Lys Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln
305 310 315 320
Thr Ser Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro
325 330 335
Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg
340 345 350
Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val
355 360 365
Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys
370 375 380
Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn
385 390 395 400
Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile
405 410 415
Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro
420 425 430
Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp
435 440 445
Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys
450 455 460
Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln
465 470 475 480
Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe
485 490 495
Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln
500 505 510
Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala
515 520 525
Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys
530 535 540
Val Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu
545 550 555 560
Ser Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala
565 570 575
Asp Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp
580 585 590
Ile Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr
595 600 605
Asn Thr Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr
610 615 620
Glu Val Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg
625 630 635 640
Val Tyr Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu
645 650 655
Ile Gly Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile
660 665 670
Gly Ala Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gly
675 680 685
Gly Ser Gly Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser
690 695 700
Leu Gly Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile
705 710 715 720
Pro Thr Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser
725 730 735
Met Thr Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser
740 745 750
Thr Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln
755 760 765
Leu Asn Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr
770 775 780
Gln Glu Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile
785 790 795 800
Lys Asp Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser
805 810 815
Lys Pro Ser Ala Asn Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val
820 825 830
Thr Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly
835 840 845
Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu
850 855 860
Thr Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr
865 870 875 880
Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala
885 890 895
Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe
900 905 910
Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu
915 920 925
Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu
930 935 940
Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln
945 950 955 960
Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe
965 970 975
Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro
980 985 990
Pro Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln
995 1000 1005
Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu
1010 1015 1020
Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys
1025 1030 1035
Val Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr
1040 1045 1050
His Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe
1055 1060 1065
Leu His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr
1070 1075 1080
Ala Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu
1085 1090 1095
Gly Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg
1100 1105 1110
Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val
1115 1120 1125
Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val
1130 1135 1140
Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu
1145 1150 1155
Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly
1160 1165 1170
Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu
1175 1180 1185
Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu
1190 1195 1200
Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp
1205 1210 1215
Pro Ser Ala Ile Gly Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly
1220 1225 1230
Gln Ala Tyr Val Arg Lys Asp Gly Glu Trp Val Leu Leu Ser Thr
1235 1240 1245
Phe Leu
1250
<210> 31
<211> 1231
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 31
Met Asp Trp Ile Trp Arg Ile Leu Phe Leu Val Gly Ala Ala Thr Gly
1 5 10 15
Ala His Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro
20 25 30
Ala Tyr Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val
35 40 45
Phe Arg Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe
50 55 60
Phe Ser Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn
65 70 75 80
Gly Thr Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val
85 90 95
Tyr Phe Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe
100 105 110
Gly Thr Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn
115 120 125
Ala Thr Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp
130 135 140
Pro Phe Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu
145 150 155 160
Ser Glu Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr
165 170 175
Val Ser Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe
180 185 190
Lys Asn Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys
195 200 205
Ile Tyr Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln
210 215 220
Gly Phe Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn
225 230 235 240
Ile Thr Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr
245 250 255
Pro Gly Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr
260 265 270
Val Gly Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn
275 280 285
Gly Thr Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu
290 295 300
Thr Lys Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln
305 310 315 320
Thr Ser Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro
325 330 335
Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg
340 345 350
Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val
355 360 365
Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys
370 375 380
Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn
385 390 395 400
Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile
405 410 415
Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro
420 425 430
Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp
435 440 445
Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys
450 455 460
Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln
465 470 475 480
Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe
485 490 495
Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln
500 505 510
Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala
515 520 525
Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys
530 535 540
Val Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu
545 550 555 560
Ser Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala
565 570 575
Asp Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp
580 585 590
Ile Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr
595 600 605
Asn Thr Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr
610 615 620
Glu Val Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg
625 630 635 640
Val Tyr Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu
645 650 655
Ile Gly Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile
660 665 670
Gly Ala Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gly
675 680 685
Gly Ser Gly Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser
690 695 700
Leu Gly Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile
705 710 715 720
Pro Thr Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser
725 730 735
Met Thr Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser
740 745 750
Thr Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln
755 760 765
Leu Asn Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr
770 775 780
Gln Glu Val Phe Ala Gln Val Lys Gln Pro Asp Pro Ser Lys Pro Ser
785 790 795 800
Ala Asn Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala
805 810 815
Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala
820 825 830
Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu
835 840 845
Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu
850 855 860
Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala
865 870 875 880
Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile
885 890 895
Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn
900 905 910
Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr
915 920 925
Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln
930 935 940
Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile
945 950 955 960
Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala
965 970 975
Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln
980 985 990
Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser
995 1000 1005
Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln
1010 1015 1020
Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser
1025 1030 1035
Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr
1040 1045 1050
Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile
1055 1060 1065
Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val
1070 1075 1080
Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu
1085 1090 1095
Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys
1100 1105 1110
Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu
1115 1120 1125
Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe
1130 1135 1140
Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly
1145 1150 1155
Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu
1160 1165 1170
Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln
1175 1180 1185
Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Ser Ala Ile
1190 1195 1200
Gly Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val
1205 1210 1215
Arg Lys Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu
1220 1225 1230
<210> 32
<211> 1274
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 32
Met Asp Trp Ile Trp Arg Ile Leu Phe Leu Val Gly Ala Ala Thr Gly
1 5 10 15
Ala His Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro
20 25 30
Ala Tyr Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val
35 40 45
Phe Arg Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe
50 55 60
Phe Ser Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn
65 70 75 80
Gly Thr Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val
85 90 95
Tyr Phe Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe
100 105 110
Gly Thr Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn
115 120 125
Ala Thr Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp
130 135 140
Pro Phe Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu
145 150 155 160
Ser Glu Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr
165 170 175
Val Ser Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe
180 185 190
Lys Asn Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys
195 200 205
Ile Tyr Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln
210 215 220
Gly Phe Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn
225 230 235 240
Ile Thr Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr
245 250 255
Pro Gly Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr
260 265 270
Val Gly Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn
275 280 285
Gly Thr Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu
290 295 300
Thr Lys Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln
305 310 315 320
Thr Ser Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro
325 330 335
Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg
340 345 350
Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val
355 360 365
Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys
370 375 380
Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn
385 390 395 400
Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile
405 410 415
Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro
420 425 430
Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp
435 440 445
Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys
450 455 460
Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln
465 470 475 480
Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe
485 490 495
Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln
500 505 510
Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala
515 520 525
Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys
530 535 540
Val Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu
545 550 555 560
Ser Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala
565 570 575
Asp Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp
580 585 590
Ile Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr
595 600 605
Asn Thr Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr
610 615 620
Glu Val Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg
625 630 635 640
Val Tyr Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu
645 650 655
Ile Gly Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile
660 665 670
Gly Ala Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gly
675 680 685
Gly Ser Gly Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser
690 695 700
Leu Gly Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile
705 710 715 720
Pro Thr Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser
725 730 735
Met Thr Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser
740 745 750
Thr Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln
755 760 765
Leu Asn Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr
770 775 780
Gln Glu Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile
785 790 795 800
Lys Asp Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser
805 810 815
Lys Pro Ser Ala Asn Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val
820 825 830
Thr Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly
835 840 845
Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu
850 855 860
Thr Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr
865 870 875 880
Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala
885 890 895
Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe
900 905 910
Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu
915 920 925
Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu
930 935 940
Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln
945 950 955 960
Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe
965 970 975
Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro
980 985 990
Pro Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln
995 1000 1005
Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu
1010 1015 1020
Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys
1025 1030 1035
Val Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr
1040 1045 1050
His Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe
1055 1060 1065
Leu His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr
1070 1075 1080
Ala Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu
1085 1090 1095
Gly Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg
1100 1105 1110
Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val
1115 1120 1125
Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val
1130 1135 1140
Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu
1145 1150 1155
Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly
1160 1165 1170
Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu
1175 1180 1185
Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu
1190 1195 1200
Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp
1205 1210 1215
Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile
1220 1225 1230
Val Met Val Thr Ile Met Leu Cys Cys Met Ser Ala Ile Gly Gly
1235 1240 1245
Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg Lys
1250 1255 1260
Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu
1265 1270
<210> 33
<211> 1255
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 33
Met Asp Trp Ile Trp Arg Ile Leu Phe Leu Val Gly Ala Ala Thr Gly
1 5 10 15
Ala His Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro
20 25 30
Ala Tyr Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val
35 40 45
Phe Arg Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe
50 55 60
Phe Ser Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn
65 70 75 80
Gly Thr Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val
85 90 95
Tyr Phe Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe
100 105 110
Gly Thr Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn
115 120 125
Ala Thr Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp
130 135 140
Pro Phe Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu
145 150 155 160
Ser Glu Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr
165 170 175
Val Ser Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe
180 185 190
Lys Asn Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys
195 200 205
Ile Tyr Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln
210 215 220
Gly Phe Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn
225 230 235 240
Ile Thr Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr
245 250 255
Pro Gly Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr
260 265 270
Val Gly Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn
275 280 285
Gly Thr Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu
290 295 300
Thr Lys Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln
305 310 315 320
Thr Ser Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro
325 330 335
Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg
340 345 350
Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val
355 360 365
Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys
370 375 380
Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn
385 390 395 400
Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile
405 410 415
Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro
420 425 430
Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp
435 440 445
Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys
450 455 460
Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln
465 470 475 480
Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe
485 490 495
Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln
500 505 510
Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala
515 520 525
Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys
530 535 540
Val Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu
545 550 555 560
Ser Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala
565 570 575
Asp Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp
580 585 590
Ile Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr
595 600 605
Asn Thr Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr
610 615 620
Glu Val Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg
625 630 635 640
Val Tyr Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu
645 650 655
Ile Gly Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile
660 665 670
Gly Ala Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gly
675 680 685
Gly Ser Gly Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser
690 695 700
Leu Gly Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile
705 710 715 720
Pro Thr Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser
725 730 735
Met Thr Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser
740 745 750
Thr Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln
755 760 765
Leu Asn Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr
770 775 780
Gln Glu Val Phe Ala Gln Val Lys Gln Pro Asp Pro Ser Lys Pro Ser
785 790 795 800
Ala Asn Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala
805 810 815
Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala
820 825 830
Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu
835 840 845
Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu
850 855 860
Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala
865 870 875 880
Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile
885 890 895
Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn
900 905 910
Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr
915 920 925
Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln
930 935 940
Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile
945 950 955 960
Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala
965 970 975
Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln
980 985 990
Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser
995 1000 1005
Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln
1010 1015 1020
Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser
1025 1030 1035
Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr
1040 1045 1050
Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile
1055 1060 1065
Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val
1070 1075 1080
Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu
1085 1090 1095
Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys
1100 1105 1110
Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu
1115 1120 1125
Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe
1130 1135 1140
Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly
1145 1150 1155
Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu
1160 1165 1170
Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln
1175 1180 1185
Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile
1190 1195 1200
Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr
1205 1210 1215
Ile Met Leu Cys Cys Met Ser Ala Ile Gly Gly Tyr Ile Pro Glu
1220 1225 1230
Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg Lys Asp Gly Glu Trp
1235 1240 1245
Val Leu Leu Ser Thr Phe Leu
1250 1255
<210> 34
<211> 4085
<212> RNA
<213> 人工序列
<220>
<223> 重组S蛋白的mRNA序列
<400> 34
aggagacaag cuugucccgc agucggcguc cagcggcucu gcuuguucgu gugugugucg 60
uugcaggccu uauucagauc uaccgguggu accgccacca uggacuggau cuggagaauc 120
cuguuccugg ugggggcugc cacuggcgcc cauagccagu gcgugaaucu gaccaccaga 180
acccagcugc cuccugccua caccaauagc uucaccagag gaguuuauua ucccgauaag 240
guguucagaa guaguguauu acauaguacc caggaccugu uccuaccuuu cuucaguaac 300
gugaccuggu uccacgccau ccacgugagc ggcaccaaug gcaccaagag auucgacaau 360
ccugugcugc cuuucaauga cggcguguac uucgccagca ccgagaagag caauaucauc 420
agaggcugga ucuucggcac caccuuggau uccaagacuc agagccugcu gauuguaaac 480
aacgcuacaa auguggugau caaggugugc gaguuccagu ucugcaauga cccuuuccug 540
gguguuuauu aucauaagaa caacaagagc uggauggaga gcgaguuccg cguauauucg 600
ucggcuaaua auugcaccuu cgaguacgug agccagccuu uccugaugga ccuggagggc 660
aagcagggca auuucaagaa ucugagagag uucguguuca agaauaucga cggcuacuuc 720
aagaucuaca gcaagcacac acccauuaau cuggugagag accugccuca gggcuucagc 780
gcccuggagc cucuggugga ccugccuauc ggcaucaaua ucaccagauu ccagacccug 840
cuggcccugc acagaucaua ucuuacacca ggcgauucgu caagcgguug gaccgcugga 900
gcugcggcau auuacguggg cuaccugcag ccuagaaccu uccugcugaa guacaaugag 960
aaugguacga uaaccgacgc aguugauugu gcccuggacc cucugagcga gaccaagugc 1020
acccugaaga gcuucaccgu ggagaagggc aucuaccaga ccagcaauuu cagagugcag 1080
ccuaccgaga gcaucgugag auucccuaau aucaccaauc ugugcccuuu cggcgaggug 1140
uucaaugcca ccagauucgc cagcguguac gcauggaacc gcaagcggau aagcaauugc 1200
guggccgacu acagcgugcu guacaauagc gccagcuuca gcaccuucaa auguuauggu 1260
guuucgccaa caaagcugaa ugaccugugc uucaccaaug uguacgccga cagcuucgug 1320
aucagaggcg acgaggugag acagaucgcg ccagggcaga ccggcaagau cgccgacuac 1380
aauuacaagc ugccugacga cuucaccggc ugcgugaucg cguggaacuc uaacaaccug 1440
gacucgaaag uuggaggcaa uuacaauuac cuguacagac uguucagaaa gagcaaucug 1500
aagccuuucg agagagacau cagcaccgag aucuaccagg ccggcagcac accguguaau 1560
ggcguggagg gcuucaauug cuacuucccu cugcagagcu acggcuucca gccuaccaau 1620
ggcgugggcu accagccuua cagaguggug gugcugagcu ucgagcugcu gcacgcuccc 1680
gcuaccgugu gcggcccuaa gaagagcacc aaucugguga agaauaagug cgugaauuuc 1740
aauuucaaug gucuaacugg aacgggcgug cugaccgaga gcaauaagaa guuucuuccc 1800
uuucaacaau ucggcagaga caucgccgac accacagaug cuguaagaga cccucagacc 1860
cuggagaucc uggacaucac uccguguagc uucggcggcg ugagcgugau cacaccgggc 1920
accaauacca gcaaucaggu ggccgugcug uaccaggacg ugaauugcac cgaggugccu 1980
guggccaucc acgccgacca gcugacuccc acuuggaggg uauauuccac gggaagcaau 2040
guguuccaga ccagagccgg cugccugauc ggcgccgagc acgugaauaa uagcuacgag 2100
ugcgacaucc cuaucggcgc cggcaucugc gccagcuacc agacccagac caauagcccu 2160
ggcggcagcg gcagcguggc cagccagagc aucaucgccu acaccaugag ccugggcgcc 2220
gagaauagcg uggccuacag caauaauagc aucgccaucc cuaccaauuu caccaucagc 2280
gugaccaccg aaauauuacc agucuccaug accaagacca gcguggacug caccauguac 2340
aucugcggcg acagcaccga gugcagcaau cugcugcugc aguacggcag cuucugcacc 2400
cagcugaaua gagcccugac cggcaucgcc guggagcagg acaagaauac ccaggaggug 2460
uucgcccagg ugaagcagau cuacaagacu ccgccgauca aggacuucgg cggcuucaau 2520
uucagccaaa uacucccaga uccaagcaag ccuagcgcca acagcuucau cgaggaccug 2580
cuguucaaua aggugacccu ggccgacgcc ggcuucauca agcaguacgg cgacugccua 2640
ggugauauug cggcaagaga ccugaucugc gcccagaagu uuaacgguuu gacaguacua 2700
ccuccucugc ugaccgacga gaugauagca caauauacgu cggcauugcu cgcuggcacg 2760
aucacaucgg gcuggacuuu cggcgccgga gcagcguugc aaaucccuuu cgccaugcag 2820
auggccuaca gauucaaugg caucggcgug acccagaaug ugcuguacga gaaucagaag 2880
cugaucgcca aucaguucaa uagcgccauc ggcaagaucc aggacagccu gagcagcacc 2940
gccagcgccc ugggcaagcu gcaggacgug gugaaucaga augcccaggc ccugaauacc 3000
cuggugaagc agcugagcag caauuucggc gccaucagua guguacucaa cgacauccug 3060
agcagacugg acccgccgga ggccgaggug caaauugauc gucuuauuac uggcagacug 3120
cagagccugc agaccuacgu gacccagcag cugaucagag ccgccgagau cagagccagc 3180
gccaaucugg ccgccaccaa gaugagcgag ugcgugcugg gccagagcaa gagaguggac 3240
uucugcggca agggcuacca ccugaugagc uucccucaga gcgcuccaca uggcguggug 3300
uuccugcacg ugaccuacgu gccugcccag gagaagaauu ucaccaccgc acccgcaauc 3360
ugccacgacg gcaaggccca cuucccuaga gagggcgugu ucgugagcaa uggcacccac 3420
ugguucguga cccagagaaa uuucuacgag ccucagauca ucaccaccga caauaccuuc 3480
gugagcggca auugcgacgu ggugaucggg auagucaaua auacugucua cgacccucug 3540
cagccugagc uggacagcuu caaggaggag cuggacaagu acuucaagaa ucacaccagc 3600
ccugacgugg accucgguga uauuucggga aucaaugcca gcguggugaa uauccagaag 3660
gaaauugauc ggcucaacga aguggccaag aaucugaaug agagccugau cgaccugcag 3720
gagcugggca aguacgagca guacaucaag uggccuagcg ccaucggcgg cuacaucccc 3780
gaggccccca gggacggcca ggccuacgug aggaaggacg gcgagugggu gcugcugagc 3840
accuuccugu gacucgagac uaguagccaa cacccugucu aaaaaacaua aauuucuuua 3900
aucauuuugc cucuuuucuc ugugcuucaa uuaauaaaaa auggaaagaa ccuggauccg 3960
ucgacaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 4020
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 4080
aaaaa 4085
<210> 35
<211> 3973
<212> RNA
<213> 人工序列
<220>
<223> 重组S蛋白的mRNA序列
<400> 35
gucccgcagu cggcguccag cggcucugcu uguucgugug ugugucguug caggccuuau 60
ucgccaccau ggacuggauc uggagaaucc uguuccuggu gggggcugcc acuggcgccc 120
auagccagug cgugaaucug accaccagaa cccagcugcc uccugccuac accaauagcu 180
ucaccagagg aguuuauuau cccgauaagg uguucagaag uaguguauua cauaguaccc 240
aggaccuguu ccuaccuuuc uucaguaacg ugaccugguu ccacgccauc cacgugagcg 300
gcaccaaugg caccaagaga uucgacaauc cugugcugcc uuucaaugac ggcguguacu 360
ucgccagcac cgagaagagc aauaucauca gaggcuggau cuucggcacc accuuggauu 420
ccaagacuca gagccugcug auuguaaaca acgcuacaaa uguggugauc aaggugugcg 480
aguuccaguu cugcaaugac ccuuuccugg guguuuauua ucauaagaac aacaagagcu 540
ggauggagag cgaguuccgc guauauucgu cggcuaauaa uugcaccuuc gaguacguga 600
gccagccuuu ccugauggac cuggagggca agcagggcaa uuucaagaau cugagagagu 660
ucguguucaa gaauaucgac ggcuacuuca agaucuacag caagcacaca cccauuaauc 720
uggugagaga ccugccucag ggcuucagcg cccuggagcc ucugguggac cugccuaucg 780
gcaucaauau caccagauuc cagacccugc uggcccugca cagaucauau cuuacaccag 840
gcgauucguc aagcgguugg accgcuggag cugcggcaua uuacgugggc uaccugcagc 900
cuagaaccuu ccugcugaag uacaaugaga augguacgau aaccgacgca guugauugug 960
cccuggaccc ucugagcgag accaagugca cccugaagag cuucaccgug gagaagggca 1020
ucuaccagac cagcaauuuc agagugcagc cuaccgagag caucgugaga uucccuaaua 1080
ucaccaaucu gugcccuuuc ggcgaggugu ucaaugccac cagauucgcc agcguguacg 1140
cauggaaccg caagcggaua agcaauugcg uggccgacua cagcgugcug uacaauagcg 1200
ccagcuucag caccuucaaa uguuauggug uuucgccaac aaagcugaau gaccugugcu 1260
ucaccaaugu guacgccgac agcuucguga ucagaggcga cgaggugaga cagaucgcgc 1320
cagggcagac cggcaagauc gccgacuaca auuacaagcu gccugacgac uucaccggcu 1380
gcgugaucgc guggaacucu aacaaccugg acucgaaagu uggaggcaau uacaauuacc 1440
uguacagacu guucagaaag agcaaucuga agccuuucga gagagacauc agcaccgaga 1500
ucuaccaggc cggcagcaca ccguguaaug gcguggaggg cuucaauugc uacuucccuc 1560
ugcagagcua cggcuuccag ccuaccaaug gcgugggcua ccagccuuac agaguggugg 1620
ugcugagcuu cgagcugcug cacgcucccg cuaccgugug cggcccuaag aagagcacca 1680
aucuggugaa gaauaagugc gugaauuuca auuucaaugg ucuaacugga acgggcgugc 1740
ugaccgagag caauaagaag uuucuucccu uucaacaauu cggcagagac aucgccgaca 1800
ccacagaugc uguaagagac ccucagaccc uggagauccu ggacaucacu ccguguagcu 1860
ucggcggcgu gagcgugauc acaccgggca ccaauaccag caaucaggug gccgugcugu 1920
accaggacgu gaauugcacc gaggugccug uggccaucca cgccgaccag cugacuccca 1980
cuuggagggu auauuccacg ggaagcaaug uguuccagac cagagccggc ugccugaucg 2040
gcgccgagca cgugaauaau agcuacgagu gcgacauccc uaucggcgcc ggcaucugcg 2100
ccagcuacca gacccagacc aauagcccug gcggcagcgg cagcguggcc agccagagca 2160
ucaucgccua caccaugagc cugggcgccg agaauagcgu ggccuacagc aauaauagca 2220
ucgccauccc uaccaauuuc accaucagcg ugaccaccga aauauuacca gucuccauga 2280
ccaagaccag cguggacugc accauguaca ucugcggcga cagcaccgag ugcagcaauc 2340
ugcugcugca guacggcagc uucugcaccc agcugaauag agcccugacc ggcaucgccg 2400
uggagcagga caagaauacc caggaggugu ucgcccaggu gaagcagcca gauccaagca 2460
agccuagcgc caacagcuuc aucgaggacc ugcuguucaa uaaggugacc cuggccgacg 2520
ccggcuucau caagcaguac ggcgacugcc uaggugauau ugcggcaaga gaccugaucu 2580
gcgcccagaa guuuaacggu uugacaguac uaccuccucu gcugaccgac gagaugauag 2640
cacaauauac gucggcauug cucgcuggca cgaucacauc gggcuggacu uucggcgccg 2700
gagcagcguu gcaaaucccu uucgccaugc agauggccua cagauucaau ggcaucggcg 2760
ugacccagaa ugugcuguac gagaaucaga agcugaucgc caaucaguuc aauagcgcca 2820
ucggcaagau ccaggacagc cugagcagca ccgccagcgc ccugggcaag cugcaggacg 2880
uggugaauca gaaugcccag gcccugaaua cccuggugaa gcagcugagc agcaauuucg 2940
gcgccaucag uaguguacuc aacgacaucc ugagcagacu ggacccgccg gaggccgagg 3000
ugcaaauuga ucgucuuauu acuggcagac ugcagagccu gcagaccuac gugacccagc 3060
agcugaucag agccgccgag aucagagcca gcgccaaucu ggccgccacc aagaugagcg 3120
agugcgugcu gggccagagc aagagagugg acuucugcgg caagggcuac caccugauga 3180
gcuucccuca gagcgcucca cauggcgugg uguuccugca cgugaccuac gugccugccc 3240
aggagaagaa uuucaccacc gcacccgcaa ucugccacga cggcaaggcc cacuucccua 3300
gagagggcgu guucgugagc aauggcaccc acugguucgu gacccagaga aauuucuacg 3360
agccucagau caucaccacc gacaauaccu ucgugagcgg caauugcgac guggugaucg 3420
ggauagucaa uaauacuguc uacgacccuc ugcagccuga gcuggacagc uucaaggagg 3480
agcuggacaa guacuucaag aaucacacca gcccugacgu ggaccucggu gauauuucgg 3540
gaaucaaugc cagcguggug aauauccaga aggaaauuga ucggcucaac gaaguggcca 3600
agaaucugaa ugagagccug aucgaccugc aggagcuggg caaguacgag caguacauca 3660
aguggccuag cgccaucggc ggcuacaucc ccgaggcccc cagggacggc caggccuacg 3720
ugaggaagga cggcgagugg gugcugcuga gcaccuuccu gugaagccaa cacccugucu 3780
aaaaaacaua aauuucuuua aucauuuugc cucuuuucuc ugugcuucaa uuaauaaaaa 3840
auggaaagaa ccuaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 3900
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 3960
aaaaaaaaaa aaa 3973
<210> 36
<211> 4157
<212> RNA
<213> 人工序列
<220>
<223> 重组S蛋白的mRNA序列
<400> 36
aggagacaag cuugucccgc agucggcguc cagcggcucu gcuuguucgu gugugugucg 60
uugcaggccu uauucagauc uaccgguggu accgccacca uggacuggau cuggagaauc 120
cuguuccugg ugggggcugc cacuggcgcc cauagccagu gcgugaaucu gaccaccaga 180
acccagcugc cuccugccua caccaauagc uucaccagag gaguuuauua ucccgauaag 240
guguucagaa guaguguauu acauaguacc caggaccugu uccuaccuuu cuucaguaac 300
gugaccuggu uccacgccau ccacgugagc ggcaccaaug gcaccaagag auucgacaau 360
ccugugcugc cuuucaauga cggcguguac uucgccagca ccgagaagag caauaucauc 420
agaggcugga ucuucggcac caccuuggau uccaagacuc agagccugcu gauuguaaac 480
aacgcuacaa auguggugau caaggugugc gaguuccagu ucugcaauga cccuuuccug 540
gguguuuauu aucauaagaa caacaagagc uggauggaga gcgaguuccg cguauauucg 600
ucggcuaaua auugcaccuu cgaguacgug agccagccuu uccugaugga ccuggagggc 660
aagcagggca auuucaagaa ucugagagag uucguguuca agaauaucga cggcuacuuc 720
aagaucuaca gcaagcacac acccauuaau cuggugagag accugccuca gggcuucagc 780
gcccuggagc cucuggugga ccugccuauc ggcaucaaua ucaccagauu ccagacccug 840
cuggcccugc acagaucaua ucuuacacca ggcgauucgu caagcgguug gaccgcugga 900
gcugcggcau auuacguggg cuaccugcag ccuagaaccu uccugcugaa guacaaugag 960
aaugguacga uaaccgacgc aguugauugu gcccuggacc cucugagcga gaccaagugc 1020
acccugaaga gcuucaccgu ggagaagggc aucuaccaga ccagcaauuu cagagugcag 1080
ccuaccgaga gcaucgugag auucccuaau aucaccaauc ugugcccuuu cggcgaggug 1140
uucaaugcca ccagauucgc cagcguguac gcauggaacc gcaagcggau aagcaauugc 1200
guggccgacu acagcgugcu guacaauagc gccagcuuca gcaccuucaa auguuauggu 1260
guuucgccaa caaagcugaa ugaccugugc uucaccaaug uguacgccga cagcuucgug 1320
aucagaggcg acgaggugag acagaucgcg ccagggcaga ccggcaagau cgccgacuac 1380
aauuacaagc ugccugacga cuucaccggc ugcgugaucg cguggaacuc uaacaaccug 1440
gacucgaaag uuggaggcaa uuacaauuac cuguacagac uguucagaaa gagcaaucug 1500
aagccuuucg agagagacau cagcaccgag aucuaccagg ccggcagcac accguguaau 1560
ggcguggagg gcuucaauug cuacuucccu cugcagagcu acggcuucca gccuaccaau 1620
ggcgugggcu accagccuua cagaguggug gugcugagcu ucgagcugcu gcacgcuccc 1680
gcuaccgugu gcggcccuaa gaagagcacc aaucugguga agaauaagug cgugaauuuc 1740
aauuucaaug gucuaacugg aacgggcgug cugaccgaga gcaauaagaa guuucuuccc 1800
uuucaacaau ucggcagaga caucgccgac accacagaug cuguaagaga cccucagacc 1860
cuggagaucc uggacaucac uccguguagc uucggcggcg ugagcgugau cacaccgggc 1920
accaauacca gcaaucaggu ggccgugcug uaccaggacg ugaauugcac cgaggugccu 1980
guggccaucc acgccgacca gcugacuccc acuuggaggg uauauuccac gggaagcaau 2040
guguuccaga ccagagccgg cugccugauc ggcgccgagc acgugaauaa uagcuacgag 2100
ugcgacaucc cuaucggcgc cggcaucugc gccagcuacc agacccagac caauagcccu 2160
ggcggcagcg gcagcguggc cagccagagc aucaucgccu acaccaugag ccugggcgcc 2220
gagaauagcg uggccuacag caauaauagc aucgccaucc cuaccaauuu caccaucagc 2280
gugaccaccg aaauauuacc agucuccaug accaagacca gcguggacug caccauguac 2340
aucugcggcg acagcaccga gugcagcaau cugcugcugc aguacggcag cuucugcacc 2400
cagcugaaua gagcccugac cggcaucgcc guggagcagg acaagaauac ccaggaggug 2460
uucgcccagg ugaagcagau cuacaagacu ccgccgauca aggacuucgg cggcuucaau 2520
uucagccaaa uacucccaga uccaagcaag ccuagcgcca acagcuucau cgaggaccug 2580
cuguucaaua aggugacccu ggccgacgcc ggcuucauca agcaguacgg cgacugccua 2640
ggugauauug cggcaagaga ccugaucugc gcccagaagu uuaacgguuu gacaguacua 2700
ccuccucugc ugaccgacga gaugauagca caauauacgu cggcauugcu cgcuggcacg 2760
aucacaucgg gcuggacuuu cggcgccgga gcagcguugc aaaucccuuu cgccaugcag 2820
auggccuaca gauucaaugg caucggcgug acccagaaug ugcuguacga gaaucagaag 2880
cugaucgcca aucaguucaa uagcgccauc ggcaagaucc aggacagccu gagcagcacc 2940
gccagcgccc ugggcaagcu gcaggacgug gugaaucaga augcccaggc ccugaauacc 3000
cuggugaagc agcugagcag caauuucggc gccaucagua guguacucaa cgacauccug 3060
agcagacugg acccgccgga ggccgaggug caaauugauc gucuuauuac uggcagacug 3120
cagagccugc agaccuacgu gacccagcag cugaucagag ccgccgagau cagagccagc 3180
gccaaucugg ccgccaccaa gaugagcgag ugcgugcugg gccagagcaa gagaguggac 3240
uucugcggca agggcuacca ccugaugagc uucccucaga gcgcuccaca uggcguggug 3300
uuccugcacg ugaccuacgu gccugcccag gagaagaauu ucaccaccgc acccgcaauc 3360
ugccacgacg gcaaggccca cuucccuaga gagggcgugu ucgugagcaa uggcacccac 3420
ugguucguga cccagagaaa uuucuacgag ccucagauca ucaccaccga caauaccuuc 3480
gugagcggca auugcgacgu ggugaucggg auagucaaua auacugucua cgacccucug 3540
cagccugagc uggacagcuu caaggaggag cuggacaagu acuucaagaa ucacaccagc 3600
ccugacgugg accucgguga uauuucggga aucaaugcca gcguggugaa uauccagaag 3660
gaaauugauc ggcucaacga aguggccaag aaucugaaug agagccugau cgaccugcag 3720
gagcugggca aguacgagca guacaucaag uggccuuggu acaucuggcu gggcuucauc 3780
gccggccuga ucgccaucgu gauggugacc aucaugcugu gcugcaugag cgccaucggc 3840
ggcuacaucc ccgaggcccc cagggacggc caggccuacg ugaggaagga cggcgagugg 3900
gugcugcuga gcaccuuccu gugacucgag acuaguagcc aacacccugu cuaaaaaaca 3960
uaaauuucuu uaaucauuuu gccucuuuuc ucugugcuuc aauuaauaaa aaauggaaag 4020
aaccuggauc cgucgacaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 4080
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 4140
aaaaaaaaaa aaaaaaa 4157
<210> 37
<211> 4100
<212> RNA
<213> 人工序列
<220>
<223> 重组S蛋白的mRNA序列
<400> 37
aggagacaag cuugucccgc agucggcguc cagcggcucu gcuuguucgu gugugugucg 60
uugcaggccu uauucagauc uaccgguggu accgccacca uggacuggau cuggagaauc 120
cuguuccugg ugggggcugc cacuggcgcc cauagccagu gcgugaaucu gaccaccaga 180
acccagcugc cuccugccua caccaauagc uucaccagag gaguuuauua ucccgauaag 240
guguucagaa guaguguauu acauaguacc caggaccugu uccuaccuuu cuucaguaac 300
gugaccuggu uccacgccau ccacgugagc ggcaccaaug gcaccaagag auucgacaau 360
ccugugcugc cuuucaauga cggcguguac uucgccagca ccgagaagag caauaucauc 420
agaggcugga ucuucggcac caccuuggau uccaagacuc agagccugcu gauuguaaac 480
aacgcuacaa auguggugau caaggugugc gaguuccagu ucugcaauga cccuuuccug 540
gguguuuauu aucauaagaa caacaagagc uggauggaga gcgaguuccg cguauauucg 600
ucggcuaaua auugcaccuu cgaguacgug agccagccuu uccugaugga ccuggagggc 660
aagcagggca auuucaagaa ucugagagag uucguguuca agaauaucga cggcuacuuc 720
aagaucuaca gcaagcacac acccauuaau cuggugagag accugccuca gggcuucagc 780
gcccuggagc cucuggugga ccugccuauc ggcaucaaua ucaccagauu ccagacccug 840
cuggcccugc acagaucaua ucuuacacca ggcgauucgu caagcgguug gaccgcugga 900
gcugcggcau auuacguggg cuaccugcag ccuagaaccu uccugcugaa guacaaugag 960
aaugguacga uaaccgacgc aguugauugu gcccuggacc cucugagcga gaccaagugc 1020
acccugaaga gcuucaccgu ggagaagggc aucuaccaga ccagcaauuu cagagugcag 1080
ccuaccgaga gcaucgugag auucccuaau aucaccaauc ugugcccuuu cggcgaggug 1140
uucaaugcca ccagauucgc cagcguguac gcauggaacc gcaagcggau aagcaauugc 1200
guggccgacu acagcgugcu guacaauagc gccagcuuca gcaccuucaa auguuauggu 1260
guuucgccaa caaagcugaa ugaccugugc uucaccaaug uguacgccga cagcuucgug 1320
aucagaggcg acgaggugag acagaucgcg ccagggcaga ccggcaagau cgccgacuac 1380
aauuacaagc ugccugacga cuucaccggc ugcgugaucg cguggaacuc uaacaaccug 1440
gacucgaaag uuggaggcaa uuacaauuac cuguacagac uguucagaaa gagcaaucug 1500
aagccuuucg agagagacau cagcaccgag aucuaccagg ccggcagcac accguguaau 1560
ggcguggagg gcuucaauug cuacuucccu cugcagagcu acggcuucca gccuaccaau 1620
ggcgugggcu accagccuua cagaguggug gugcugagcu ucgagcugcu gcacgcuccc 1680
gcuaccgugu gcggcccuaa gaagagcacc aaucugguga agaauaagug cgugaauuuc 1740
aauuucaaug gucuaacugg aacgggcgug cugaccgaga gcaauaagaa guuucuuccc 1800
uuucaacaau ucggcagaga caucgccgac accacagaug cuguaagaga cccucagacc 1860
cuggagaucc uggacaucac uccguguagc uucggcggcg ugagcgugau cacaccgggc 1920
accaauacca gcaaucaggu ggccgugcug uaccaggacg ugaauugcac cgaggugccu 1980
guggccaucc acgccgacca gcugacuccc acuuggaggg uauauuccac gggaagcaau 2040
guguuccaga ccagagccgg cugccugauc ggcgccgagc acgugaauaa uagcuacgag 2100
ugcgacaucc cuaucggcgc cggcaucugc gccagcuacc agacccagac caauagcccu 2160
ggcggcagcg gcagcguggc cagccagagc aucaucgccu acaccaugag ccugggcgcc 2220
gagaauagcg uggccuacag caauaauagc aucgccaucc cuaccaauuu caccaucagc 2280
gugaccaccg aaauauuacc agucuccaug accaagacca gcguggacug caccauguac 2340
aucugcggcg acagcaccga gugcagcaau cugcugcugc aguacggcag cuucugcacc 2400
cagcugaaua gagcccugac cggcaucgcc guggagcagg acaagaauac ccaggaggug 2460
uucgcccagg ugaagcagcc agauccaagc aagccuagcg ccaacagcuu caucgaggac 2520
cugcuguuca auaaggugac ccuggccgac gccggcuuca ucaagcagua cggcgacugc 2580
cuaggugaua uugcggcaag agaccugauc ugcgcccaga aguuuaacgg uuugacagua 2640
cuaccuccuc ugcugaccga cgagaugaua gcacaauaua cgucggcauu gcucgcuggc 2700
acgaucacau cgggcuggac uuucggcgcc ggagcagcgu ugcaaauccc uuucgccaug 2760
cagauggccu acagauucaa uggcaucggc gugacccaga augugcugua cgagaaucag 2820
aagcugaucg ccaaucaguu caauagcgcc aucggcaaga uccaggacag ccugagcagc 2880
accgccagcg cccugggcaa gcugcaggac guggugaauc agaaugccca ggcccugaau 2940
acccugguga agcagcugag cagcaauuuc ggcgccauca guaguguacu caacgacauc 3000
cugagcagac uggacccgcc ggaggccgag gugcaaauug aucgucuuau uacuggcaga 3060
cugcagagcc ugcagaccua cgugacccag cagcugauca gagccgccga gaucagagcc 3120
agcgccaauc uggccgccac caagaugagc gagugcgugc ugggccagag caagagagug 3180
gacuucugcg gcaagggcua ccaccugaug agcuucccuc agagcgcucc acauggcgug 3240
guguuccugc acgugaccua cgugccugcc caggagaaga auuucaccac cgcacccgca 3300
aucugccacg acggcaaggc ccacuucccu agagagggcg uguucgugag caauggcacc 3360
cacugguucg ugacccagag aaauuucuac gagccucaga ucaucaccac cgacaauacc 3420
uucgugagcg gcaauugcga cguggugauc gggauaguca auaauacugu cuacgacccu 3480
cugcagccug agcuggacag cuucaaggag gagcuggaca aguacuucaa gaaucacacc 3540
agcccugacg uggaccucgg ugauauuucg ggaaucaaug ccagcguggu gaauauccag 3600
aaggaaauug aucggcucaa cgaaguggcc aagaaucuga augagagccu gaucgaccug 3660
caggagcugg gcaaguacga gcaguacauc aaguggccuu gguacaucug gcugggcuuc 3720
aucgccggcc ugaucgccau cgugauggug accaucaugc ugugcugcau gagcgccauc 3780
ggcggcuaca uccccgaggc ccccagggac ggccaggccu acgugaggaa ggacggcgag 3840
ugggugcugc ugagcaccuu ccugugacuc gagacuagua gccaacaccc ugucuaaaaa 3900
acauaaauuu cuuuaaucau uuugccucuu uucucugugc uucaauuaau aaaaaaugga 3960
aagaaccugg auccgucgac aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 4020
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 4080
aaaaaaaaaa aaaaaaaaaa 4100
<210> 38
<211> 1266
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 38
Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val
1 5 10 15
Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe
20 25 30
Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu
35 40 45
His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp
50 55 60
Phe His Ala Ile His Val Ser Gly Thr Asn Val Ile Lys Arg Phe Asp
65 70 75 80
Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu
85 90 95
Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser
100 105 110
Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile
115 120 125
Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr
130 135 140
Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr
145 150 155 160
Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu
165 170 175
Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe
180 185 190
Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr
195 200 205
Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu
210 215 220
Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr
225 230 235 240
Leu Leu Ala Leu His Asn Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala
245 250 255
Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr
260 265 270
Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro
275 280 285
Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly
290 295 300
Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val
305 310 315 320
Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn
325 330 335
Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser
340 345 350
Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser
355 360 365
Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys
370 375 380
Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val
385 390 395 400
Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr
405 410 415
Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn
420 425 430
Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Gln Tyr Arg Leu
435 440 445
Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu
450 455 460
Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn
465 470 475 480
Cys Tyr Ser Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val
485 490 495
Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His
500 505 510
Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys
515 520 525
Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val
530 535 540
Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg
545 550 555 560
Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu
565 570 575
Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr
580 585 590
Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu Tyr Gln Gly Val
595 600 605
Asn Cys Thr Glu Val Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro
610 615 620
Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala
625 630 635 640
Gly Cys Leu Ile Gly Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp
645 650 655
Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn
660 665 670
Ser Pro Gly Gly Ser Gly Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr
675 680 685
Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser
690 695 700
Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu
705 710 715 720
Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys
725 730 735
Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe
740 745 750
Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp
755 760 765
Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr
770 775 780
Pro Pro Ile Lys Asp Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro
785 790 795 800
Asp Pro Ser Lys Pro Ser Ala Asn Ser Phe Ile Glu Asp Leu Leu Phe
805 810 815
Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp
820 825 830
Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe
835 840 845
Asn Gly Leu Asn Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala
850 855 860
Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr
865 870 875 880
Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala
885 890 895
Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn
900 905 910
Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln
915 920 925
Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val
930 935 940
Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser
945 950 955 960
Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg
965 970 975
Leu Asp Lys Val Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly
980 985 990
Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala
995 1000 1005
Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser
1010 1015 1020
Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys
1025 1030 1035
Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val
1040 1045 1050
Val Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe
1055 1060 1065
Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro
1070 1075 1080
Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr
1085 1090 1095
Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr
1100 1105 1110
Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn
1115 1120 1125
Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu
1130 1135 1140
Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp
1145 1150 1155
Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln
1160 1165 1170
Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu
1175 1180 1185
Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile
1190 1195 1200
Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile
1205 1210 1215
Ala Ile Val Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys
1220 1225 1230
Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys
1235 1240 1245
Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu
1250 1255 1260
His Tyr Thr
1265
<210> 39
<211> 1266
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 39
Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val
1 5 10 15
Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe
20 25 30
Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu
35 40 45
His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp
50 55 60
Phe His Ala Ile His Val Ser Gly Thr Asn Val Ile Lys Arg Phe Asp
65 70 75 80
Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu
85 90 95
Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser
100 105 110
Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile
115 120 125
Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr
130 135 140
Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr
145 150 155 160
Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu
165 170 175
Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe
180 185 190
Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr
195 200 205
Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu
210 215 220
Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr
225 230 235 240
Leu Leu Ala Leu His Asn Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala
245 250 255
Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr
260 265 270
Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro
275 280 285
Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly
290 295 300
Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val
305 310 315 320
Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn
325 330 335
Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser
340 345 350
Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser
355 360 365
Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys
370 375 380
Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val
385 390 395 400
Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr
405 410 415
Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn
420 425 430
Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Gln Tyr Arg Leu
435 440 445
Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu
450 455 460
Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn
465 470 475 480
Cys Tyr Ser Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val
485 490 495
Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His
500 505 510
Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys
515 520 525
Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val
530 535 540
Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg
545 550 555 560
Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu
565 570 575
Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr
580 585 590
Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu Tyr Gln Gly Val
595 600 605
Asn Cys Thr Glu Val Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro
610 615 620
Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala
625 630 635 640
Gly Cys Leu Ile Gly Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp
645 650 655
Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn
660 665 670
Ser Pro Arg Arg Ala Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr
675 680 685
Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser
690 695 700
Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu
705 710 715 720
Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys
725 730 735
Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe
740 745 750
Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp
755 760 765
Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr
770 775 780
Pro Pro Ile Lys Asp Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro
785 790 795 800
Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe
805 810 815
Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp
820 825 830
Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe
835 840 845
Asn Gly Leu Asn Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala
850 855 860
Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr
865 870 875 880
Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala
885 890 895
Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn
900 905 910
Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln
915 920 925
Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val
930 935 940
Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser
945 950 955 960
Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg
965 970 975
Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly
980 985 990
Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala
995 1000 1005
Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser
1010 1015 1020
Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys
1025 1030 1035
Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val
1040 1045 1050
Val Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe
1055 1060 1065
Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro
1070 1075 1080
Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr
1085 1090 1095
Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr
1100 1105 1110
Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn
1115 1120 1125
Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu
1130 1135 1140
Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp
1145 1150 1155
Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln
1160 1165 1170
Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu
1175 1180 1185
Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile
1190 1195 1200
Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile
1205 1210 1215
Ala Ile Val Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys
1220 1225 1230
Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys
1235 1240 1245
Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu
1250 1255 1260
His Tyr Thr
1265
<210> 40
<211> 1266
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 40
Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val
1 5 10 15
Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe
20 25 30
Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu
35 40 45
His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp
50 55 60
Phe His Ala Ile His Val Ser Gly Thr Asn Val Ile Lys Arg Phe Asp
65 70 75 80
Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu
85 90 95
Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser
100 105 110
Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile
115 120 125
Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr
130 135 140
Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr
145 150 155 160
Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu
165 170 175
Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe
180 185 190
Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr
195 200 205
Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu
210 215 220
Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr
225 230 235 240
Leu Leu Ala Leu His Asn Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala
245 250 255
Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr
260 265 270
Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro
275 280 285
Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly
290 295 300
Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val
305 310 315 320
Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn
325 330 335
Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser
340 345 350
Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser
355 360 365
Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys
370 375 380
Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val
385 390 395 400
Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr
405 410 415
Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn
420 425 430
Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Gln Tyr Arg Leu
435 440 445
Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu
450 455 460
Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn
465 470 475 480
Cys Tyr Ser Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val
485 490 495
Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His
500 505 510
Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys
515 520 525
Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val
530 535 540
Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg
545 550 555 560
Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu
565 570 575
Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr
580 585 590
Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu Tyr Gln Gly Val
595 600 605
Asn Cys Thr Glu Val Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro
610 615 620
Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala
625 630 635 640
Gly Cys Leu Ile Gly Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp
645 650 655
Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn
660 665 670
Ser Pro Gly Gly Ser Gly Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr
675 680 685
Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser
690 695 700
Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu
705 710 715 720
Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys
725 730 735
Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe
740 745 750
Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp
755 760 765
Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr
770 775 780
Pro Pro Ile Lys Asp Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro
785 790 795 800
Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe
805 810 815
Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp
820 825 830
Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe
835 840 845
Asn Gly Leu Asn Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala
850 855 860
Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr
865 870 875 880
Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala
885 890 895
Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn
900 905 910
Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln
915 920 925
Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val
930 935 940
Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser
945 950 955 960
Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg
965 970 975
Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly
980 985 990
Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala
995 1000 1005
Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser
1010 1015 1020
Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys
1025 1030 1035
Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val
1040 1045 1050
Val Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe
1055 1060 1065
Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro
1070 1075 1080
Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr
1085 1090 1095
Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr
1100 1105 1110
Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn
1115 1120 1125
Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu
1130 1135 1140
Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp
1145 1150 1155
Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln
1160 1165 1170
Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu
1175 1180 1185
Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile
1190 1195 1200
Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile
1205 1210 1215
Ala Ile Val Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys
1220 1225 1230
Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys
1235 1240 1245
Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu
1250 1255 1260
His Tyr Thr
1265
<210> 41
<211> 1266
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 41
Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val
1 5 10 15
Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe
20 25 30
Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu
35 40 45
His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp
50 55 60
Phe His Ala Ile His Val Ser Gly Thr Asn Val Ile Lys Arg Phe Asp
65 70 75 80
Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu
85 90 95
Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser
100 105 110
Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile
115 120 125
Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr
130 135 140
Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr
145 150 155 160
Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu
165 170 175
Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe
180 185 190
Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr
195 200 205
Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu
210 215 220
Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr
225 230 235 240
Leu Leu Ala Leu His Asn Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala
245 250 255
Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr
260 265 270
Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro
275 280 285
Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly
290 295 300
Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val
305 310 315 320
Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn
325 330 335
Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser
340 345 350
Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser
355 360 365
Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys
370 375 380
Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val
385 390 395 400
Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr
405 410 415
Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn
420 425 430
Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Gln Tyr Arg Leu
435 440 445
Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu
450 455 460
Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn
465 470 475 480
Cys Tyr Ser Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val
485 490 495
Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His
500 505 510
Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys
515 520 525
Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val
530 535 540
Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg
545 550 555 560
Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu
565 570 575
Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr
580 585 590
Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu Tyr Gln Gly Val
595 600 605
Asn Cys Thr Glu Val Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro
610 615 620
Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala
625 630 635 640
Gly Cys Leu Ile Gly Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp
645 650 655
Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn
660 665 670
Ser Pro Arg Arg Ala Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr
675 680 685
Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser
690 695 700
Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu
705 710 715 720
Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys
725 730 735
Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe
740 745 750
Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp
755 760 765
Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr
770 775 780
Pro Pro Ile Lys Asp Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro
785 790 795 800
Asp Pro Ser Lys Pro Ser Ala Asn Ser Phe Ile Glu Asp Leu Leu Phe
805 810 815
Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp
820 825 830
Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe
835 840 845
Asn Gly Leu Asn Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala
850 855 860
Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr
865 870 875 880
Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala
885 890 895
Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn
900 905 910
Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln
915 920 925
Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val
930 935 940
Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser
945 950 955 960
Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg
965 970 975
Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly
980 985 990
Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala
995 1000 1005
Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser
1010 1015 1020
Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys
1025 1030 1035
Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val
1040 1045 1050
Val Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe
1055 1060 1065
Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro
1070 1075 1080
Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr
1085 1090 1095
Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr
1100 1105 1110
Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn
1115 1120 1125
Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu
1130 1135 1140
Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp
1145 1150 1155
Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln
1160 1165 1170
Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu
1175 1180 1185
Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile
1190 1195 1200
Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile
1205 1210 1215
Ala Ile Val Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys
1220 1225 1230
Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys
1235 1240 1245
Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu
1250 1255 1260
His Tyr Thr
1265
<210> 42
<211> 1266
<212> PRT
<213> 人工序列
<220>
<223> 重组S蛋白的氨基酸序列
<400> 42
Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val
1 5 10 15
Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe
20 25 30
Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu
35 40 45
His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp
50 55 60
Phe His Ala Ile His Val Ser Gly Thr Asn Val Ile Lys Arg Phe Asp
65 70 75 80
Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu
85 90 95
Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser
100 105 110
Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile
115 120 125
Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr
130 135 140
Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr
145 150 155 160
Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu
165 170 175
Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe
180 185 190
Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr
195 200 205
Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu
210 215 220
Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr
225 230 235 240
Leu Leu Ala Leu His Asn Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala
245 250 255
Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr
260 265 270
Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro
275 280 285
Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly
290 295 300
Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val
305 310 315 320
Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn
325 330 335
Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser
340 345 350
Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser
355 360 365
Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys
370 375 380
Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val
385 390 395 400
Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr
405 410 415
Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn
420 425 430
Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Gln Tyr Arg Leu
435 440 445
Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu
450 455 460
Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn
465 470 475 480
Cys Tyr Ser Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val
485 490 495
Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His
500 505 510
Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys
515 520 525
Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val
530 535 540
Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg
545 550 555 560
Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu
565 570 575
Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr
580 585 590
Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu Tyr Gln Gly Val
595 600 605
Asn Cys Thr Glu Val Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro
610 615 620
Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala
625 630 635 640
Gly Cys Leu Ile Gly Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp
645 650 655
Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn
660 665 670
Ser Pro Gly Gly Ser Gly Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr
675 680 685
Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser
690 695 700
Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu
705 710 715 720
Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys
725 730 735
Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe
740 745 750
Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp
755 760 765
Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr
770 775 780
Pro Pro Ile Lys Asp Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro
785 790 795 800
Asp Pro Ser Lys Pro Ser Ala Asn Ser Phe Ile Glu Asp Leu Leu Phe
805 810 815
Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp
820 825 830
Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe
835 840 845
Asn Gly Leu Asn Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala
850 855 860
Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr
865 870 875 880
Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala
885 890 895
Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn
900 905 910
Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln
915 920 925
Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val
930 935 940
Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser
945 950 955 960
Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg
965 970 975
Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly
980 985 990
Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala
995 1000 1005
Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser
1010 1015 1020
Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys
1025 1030 1035
Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val
1040 1045 1050
Val Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe
1055 1060 1065
Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro
1070 1075 1080
Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr
1085 1090 1095
Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr
1100 1105 1110
Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn
1115 1120 1125
Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu
1130 1135 1140
Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp
1145 1150 1155
Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln
1160 1165 1170
Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu
1175 1180 1185
Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile
1190 1195 1200
Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile
1205 1210 1215
Ala Ile Val Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys
1220 1225 1230
Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys
1235 1240 1245
Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu
1250 1255 1260
His Tyr Thr
1265
<210> 43
<211> 4115
<212> RNA
<213> 人工序列
<220>
<223> RBMRNA-Lambda疫苗的mRNA序列
<400> 43
augagacaag cuugucccgc agucggcguc cagcggcucu gcuuguucgu gugugugucg 60
uugcaggccu uauucagauc uaccgguggu accgccacca uguucguguu ccuggugcug 120
cugccucugg ugagcagcca gugcgugaau cugaccacca gaacccagcu gccuccugcc 180
uacaccaaua gcuucaccag aggaguuuau uaucccgaua agguguucag aaguagugua 240
uuacauagua cccaggaccu guuccuaccu uucuucagua acgugaccug guuccacgcc 300
auccacguga gcggcaccaa ugucaucaag agauucgaca auccugugcu gccuuucaau 360
gacggcgugu acuucgccag caccgagaag agcaauauca ucagaggcug gaucuucggc 420
accaccuugg auuccaagac ucagagccug cugauuguaa acaacgcuac aaauguggug 480
aucaaggugu gcgaguucca guucugcaau gacccuuucc uggguguuua uuaucauaag 540
aacaacaaga gcuggaugga gagcgaguuc cgcguauauu cgucggcuaa uaauugcacc 600
uucgaguacg ugagccagcc uuuccugaug gaccuggagg gcaagcaggg caauuucaag 660
aaucugagag aguucguguu caagaauauc gacggcuacu ucaagaucua cagcaagcac 720
acacccauua aucuggugag agaccugccu cagggcuuca gcgcccugga gccucuggug 780
gaccugccua ucggcaucaa uaucaccaga uuccagaccc ugcuggcccu gcacaacucg 840
ucaagcgguu ggaccgcugg agcugcggca uauuacgugg gcuaccugca gccuagaacc 900
uuccugcuga aguacaauga gaaugguacg auaaccgacg caguugauug ugcccuggac 960
ccucugagcg agaccaagug cacccugaag agcuucaccg uggagaaggg caucuaccag 1020
accagcaauu ucagagugca gccuaccgag agcaucguga gauucccuaa uaucaccaau 1080
cugugcccuu ucggcgaggu guucaaugcc accagauucg ccagcgugua cgcauggaac 1140
cgcaagcgga uaagcaauug cguggccgac uacagcgugc uguacaauag cgccagcuuc 1200
agcaccuuca aauguuaugg uguuucgcca acaaagcuga augaccugug cuucaccaau 1260
guguacgccg acagcuucgu gaucagaggc gacgagguga gacagaucgc gccagggcag 1320
accggcaaga ucgccgacua caauuacaag cugccugacg acuucaccgg cugcgugauc 1380
gcguggaacu cuaacaaccu ggacucgaaa guuggaggca auuacaauua ccaguacaga 1440
cuguucagaa agagcaaucu gaagccuuuc gagagagaca ucagcaccga gaucuaccag 1500
gccggcagca caccguguaa uggcguggag ggcuucaauu gcuacucccc ucugcagagc 1560
uacggcuucc agccuaccaa uggcgugggc uaccagccuu acagaguggu ggugcugagc 1620
uucgagcugc ugcacgcucc cgcuaccgug ugcggcccua agaagagcac caaucuggug 1680
aagaauaagu gcgugaauuu caauuucaau ggucuaacug gaacgggcgu gcugaccgag 1740
agcaauaaga aguuucuucc cuuucaacaa uucggcagag acaucgccga caccacagau 1800
gcuguaagag acccucagac ccuggagauc cuggacauca cuccguguag cuucggcggc 1860
gugagcguga ucacaccggg caccaauacc agcaaucagg uggccgugcu guaccagggc 1920
gugaauugca ccgaggugcc uguggccauc cacgccgacc agcugacucc cacuuggagg 1980
guauauucca cgggaagcaa uguguuccag accagagccg gcugccugau cggcgccgag 2040
cacgugaaua auagcuacga gugcgacauc ccuaucggcg ccggcaucug cgccagcuac 2100
cagacccaga ccaauagccc uggcggcagc ggcagcgugg ccagccagag caucaucgcc 2160
uacaccauga gccugggcgc cgagaauagc guggccuaca gcaauaauag caucgccauc 2220
ccuaccaauu ucaccaucag cgugaccacc gaaauauuac cagucuccau gaccaagacc 2280
agcguggacu gcaccaugua caucugcggc gacagcaccg agugcagcaa ucugcugcug 2340
caguacggca gcuucugcac ccagcugaau agagcccuga ccggcaucgc cguggagcag 2400
gacaagaaua cccaggaggu guucgcccag gugaagcaga ucuacaagac uccgccgauc 2460
aaggacuucg gcggcuucaa uuucagccaa auacucccag auccaagcaa gccuagcgcc 2520
aacagcuuca ucgaggaccu gcuguucaau aaggugaccc uggccgacgc cggcuucauc 2580
aagcaguacg gcgacugccu aggugauauu gcggcaagag accugaucug cgcccagaag 2640
uuuaacgguu ugaacguacu accuccucug cugaccgacg agaugauagc acaauauacg 2700
ucggcauugc ucgcuggcac gaucacaucg ggcuggacuu ucggcgccgg agcagcguug 2760
caaaucccuu ucgccaugca gauggccuac agauucaaug gcaucggcgu gacccagaau 2820
gugcuguacg agaaucagaa gcugaucgcc aaucaguuca auagcgccau cggcaagauc 2880
caggacagcc ugagcagcac cgccagcgcc cugggcaagc ugcaggacgu ggugaaucag 2940
aaugcccagg cccugaauac ccuggugaag cagcugagca gcaauuucgg cgccaucagu 3000
aguguacuca acgacauccu gagcagacug gacccaccag aggccgaggu gcaaauugau 3060
cgucuuauua cuggcagacu gcagagccug cagaccuacg ugacccagca gcugaucaga 3120
gccgccgaga ucagagccag cgccaaucug gccgccacca agaugagcga gugcgugcug 3180
ggccagagca agagagugga cuucugcggc aagggcuacc accugaugag cuucccucag 3240
agcgcuccac auggcguggu guuccugcac gugaccuacg ugccugccca ggagaagaau 3300
uucaccaccg cacccgcaau cugccacgac ggcaaggccc acuucccuag agagggcgug 3360
uucgugagca auggcaccca cugguucgug acccagagaa auuucuacga gccucagauc 3420
aucaccaccg acaauaccuu cgugagcggc aauugcgacg uggugaucgg gauagucaau 3480
aauacugucu acgacccucu gcagccugag cuggacagcu ucaaggagga gcuggacaag 3540
uacuucaaga aucacaccag cccugacgug gaccucggug auauuucggg aaucaaugcc 3600
agcgugguga auauccagaa ggaaauugau cggcucaacg aaguggccaa gaaucugaau 3660
gagagccuga ucgaccugca ggagcugggc aaguacgagc aguacaucaa guggccuugg 3720
uacaucuggc ugggcuucau cgccggccug aucgccaucg ugauggugac caucaugcug 3780
ugcugcauga ccuccuguug uuccuguuug aaaggguguu guucgugugg guccugcugc 3840
aaguucgacg aggacgacag cgagccugug cugaagggcg ugaagcugca cuacaccuaa 3900
cucgagacua guagccaaca cccugucuaa aaaacauaaa uuucuuuaau cauuuugccu 3960
cuuuucucug ugcuucaauu aauaaaaaau ggaaagaacc uaaaaaaaaa aaaaaaaaaa 4020
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 4080
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaa 4115

Claims (36)

1.重组SARS-CoV-2刺突蛋白(S蛋白)，所述重组S蛋白相对于野生型S蛋白在胞外结构域中包括以下突变：
T19R、G142D、E156G、F157和R158缺失突变、A222V、L452R、T478K、D614G、P681R和D950N；
其中，氨基酸位置按照如SEQ ID NO.29所示的野生型S蛋白的氨基酸序列进行编号。
2.根据权利要求1所述的重组S蛋白，其中S1/S2切割位点RRAR和/或S2切割位点KR经突变以失去被弗林样蛋白酶和溶酶体蛋白酶切割的能力；优选地，所述S1/S2切割位点突变为GGSG，和/或所述S2切割位点突变为AN。
3.根据权利要求1或2所述的重组S蛋白，其还包括K986P和V987P突变。
4.根据权利要求1-3中任一项所述的重组S蛋白，其不包含融合肽结构域(FP)；任选地，所述重组S蛋白不包含跨膜结构域和胞质结构域；任选地，所述重组S蛋白还包含三聚体结构域，所述三聚体结构域当表达时促进所述重组S蛋白形成三聚体，优选地，所述三聚体结构域为T4噬菌体fibritin三聚体基序；任选地，所述重组S蛋白还包含信号序列，优选地，所述信号序列为免疫球蛋白重链可变区信号序列。
5.根据权利要求1所述的重组S蛋白，其具有选自SEQ ID NO.1-5中任一项所示的氨基酸序列，优选具有选自SEQ ID NO.3-5中任一项所示的氨基酸序列。
6.重组SARS-CoV-2刺突蛋白(S蛋白)，所述重组S蛋白相对于野生型S蛋白在胞外结构域中包括以下突变：
A67V、H69缺失突变、V70缺失突变、T95I、G142缺失突变、V143缺失突变、Y144缺失突变、Y145D、N211缺失突变、L212I、在R214和D215之间三个氨基酸E、P、E的插入突变，G339D、S371L、S373P、S375F、K417N、N440K、G446S、S477N、T478K、E484A、Q493R、G496S、Q498R、N501Y、Y505H、T547K、D614G、H655Y、N679K、P681H、N764K、D796Y、N856K、Q954H、N969K和L981F；
其中，氨基酸位置按照如SEQ ID NO.29所示的野生型S蛋白的氨基酸序列进行编号。
7.根据权利要求6所述的重组S蛋白，其中S1/S2切割位点RRAR和/或S2切割位点KR经突变以失去被弗林样蛋白酶和溶酶体蛋白酶切割的能力；优选地，所述S1/S2切割位点突变为GGSG，和/或所述S2切割位点突变为AN。
8.根据权利要求6或7所述的重组S蛋白，其还包括K986P和V987P突变。
9.根据权利要求6-8中任一项所述的重组S蛋白，其从N端到C端包含胞外结构域、跨膜结构域和胞质结构域；优选地，所述重组S蛋白还包含信号序列，所述信号序列优选为免疫球蛋白重链可变区信号序列。
10.根据权利要求6所述的重组S蛋白，其具有选自SEQ ID NO.20-25中任一项所示的氨基酸序列，优选具有SEQ ID NO.25所示的氨基酸序列。
11.mRNA，所述mRNA编码权利要求1-5中任一项所述的重组S蛋白。
12.根据权利要求11所述的mRNA，其从5’到3’方向包含帽结构、5’-UTR、开放阅读框(ORF)、3’-UTR和polyA尾。
13.根据权利要求12所述的mRNA，其中所述5’-UTR包含衍生自17β-羟基类固醇脱氢酶4(HSD17B4)基因的5’-UTR或其同源物、片段或变体，和/或KOZAK序列，优选地，所述5’-UTR包含如SEQ ID NO.8和/或SEQ ID NO.9所示的序列；
其中所述3’-UTR包含衍生自白蛋白(ALB)基因的3’-UTR或其同源物、片段或变体，优选地，所述3’-UTR包含如SEQ ID NO.10所示的序列；和/或
其中所述polyA尾的长度为100-150个核苷酸。
14.根据权利要求11所述的mRNA，其中所述mRNA的序列如SEQ ID NO.14-16中任一项所示。
15.根据权利要求11-14中任一项所述的mRNA，其中所述mRNA中的一个或多个核苷酸各自独立替换为天然存在的核苷酸类似物或人工合成的核苷酸类似物，其中所述天然存在的核苷酸类似物选自假尿苷、2-硫尿苷、5-甲基尿苷、5-甲基胞苷和N6-甲基腺苷，和所述人工合成的核苷酸类似物选自N1-甲基假尿苷和5-乙炔基尿苷；
优选地，所述mRNA中的一个或多个尿苷三磷酸各自独立替换为假尿苷三磷酸、1-甲基-假尿苷三磷酸或5-乙炔基尿苷三磷酸，和/或一个或多个胞苷三磷酸替换为5-甲基胞苷三磷酸。
16.mRNA，所述mRNA编码权利要求6-10中任一项所述的重组S蛋白。
17.根据权利要求16所述的mRNA，其从5’到3’方向包含帽结构、5’-UTR、开放阅读框(ORF)、3’-UTR和polyA尾。
18.根据权利要求17所述的mRNA，其中所述5’-UTR包含衍生自17β-羟基类固醇脱氢酶4(HSD17B4)基因的5’-UTR或其同源物、片段或变体，和/或KOZAK序列，优选地，所述5’-UTR包含如SEQ ID NO.8和/或SEQ ID NO.9所示的序列；
其中所述3’-UTR包含衍生自白蛋白(ALB)基因的3’-UTR或其同源物、片段或变体，优选地，所述3’-UTR包含如SEQ ID NO.10所示的序列；和/或
其中所述polyA尾的长度为100-150个核苷酸。
19.根据权利要求16所述的mRNA，其中所述mRNA的序列如SEQ ID NO.27所示。
20.根据权利要求16-19中任一项所述的mRNA，其中所述mRNA中的一个或多个核苷酸各自独立替换为天然存在的核苷酸类似物或人工合成的核苷酸类似物，其中所述天然存在的核苷酸类似物选自假尿苷、2-硫尿苷、5-甲基尿苷、5-甲基胞苷和N6-甲基腺苷，和所述人工合成的核苷酸类似物选自N1-甲基假尿苷和5-乙炔基尿苷；
优选地，所述mRNA中的一个或多个尿苷三磷酸各自独立替换为假尿苷三磷酸、1-甲基-假尿苷三磷酸或5-乙炔基尿苷三磷酸，和/或一个或多个胞苷三磷酸替换为5-甲基胞苷三磷酸。
21.组合物，其包含权利要求1-5中任一项所述的重组S蛋白或权利要求11-15中任一项所述的mRNA，和权利要求6-10中任一项所述的重组S蛋白或权利要求16-20中任一项所述的mRNA。
22.根据权利要求21所述的组合物，其中所述组合物包含权利要求11-15中任一项所述的mRNA和权利要求16-20中任一项所述的mRNA。
23.根据权利要求22所述的组合物，其中所述组合物包含具有SEQ ID NO.14-16中任一项所示的序列的mRNA和具有如SEQ ID NO.27所示的序列的mRNA。
24.根据权利要求21-23中任一项所述的组合物，其中所述组合物中两种重组S蛋白或两种mRNA的摩尔比例为1-3:1-3，优选1:1。
25.根据权利要求21-24中任一项所述的组合物，其还包含以下重组S蛋白或编码其的mRNA：
(a)相对于野生型S蛋白包括以下突变的重组S蛋白：K986P和V987P；和/或
(b)相对于野生型S蛋白包括以下突变的重组S蛋白：G75V、T76I、R246缺失、S247缺失、Y248缺失、L249缺失、T250缺失、P251缺失、G252缺失、D253N、L452Q、F490S、D614G、T859N；K986P；和V987P；
和/或
(c)相对于野生型S蛋白包括以下突变的重组S蛋白：S1/S2切割位点突变为GGSG；K986P；和V987P；
和/或
(d)相对于野生型S蛋白包括以下突变的重组S蛋白：S2切割位点突变为AN；K986P；和V987P；
和/或
(e)相对于野生型S蛋白包括以下突变的重组S蛋白：S1/S2切割位点突变为GGSG；S2切割位点突变为AN；K986P；和V987P；
和/或
(f)相对于野生型S蛋白包括以下突变的重组S蛋白：G75V、T76I、R246缺失、S247缺失、Y248缺失、L249缺失、T250缺失、P251缺失、G252缺失、D253N、L452Q、F490S、D614G、T859N；S1/S2切割位点突变为GGSG；K986P；和V987P；
和/或
(g)相对于野生型S蛋白包括以下突变的重组S蛋白：G75V、T76I、R246缺失、S247缺失、Y248缺失、L249缺失、T250缺失、P251缺失、G252缺失、D253N、L452Q、F490S、D614G、T859N；S2切割位点突变为AN；K986P；和V987P；
和/或
(h)相对于野生型S蛋白包括以下突变的重组S蛋白：G75V、T76I、R246缺失、S247缺失、Y248缺失、L249缺失、T250缺失、P251缺失、G252缺失、D253N、L452Q、F490S、D614G、T859N；S1/S2切割位点突变为GGSG；S2切割位点突变为AN；K986P；和V987P。
26.DNA，其编码权利要求11-14中任一项所述的mRNA和/或权利要求16-19中任一项所述的mRNA。
27.根据权利要求26所述的DNA，其中所述DNA的序列如SEQ ID NO.11-13和26中任一项所示。
28.重组质粒，其包含权利要求26或27所述的DNA。
29.根据权利要求28所述的重组质粒，其中所述质粒为pT7TS质粒。
30.根据权利要求29所述的重组质粒，其还包含起始序列(Ori)、T7启动子、5’-UTR和3’-UTR。
31.根据权利要求30所述的重组质粒，其中所述起始序列为ColE1型起始序列，优选地，所述起始序列为SEQ ID NO.6所示的起始序列；
其中所述T7启动子的序列如SEQ ID NO.7所示；
其中所述5’-UTR包含衍生自HSD17B4的5’-UTR或其同源物、片段或变体，和/或KOZAK序列，优选地，所述5’-UTR包含如SEQ ID NO.8和/或SEQ ID NO.9所示的序列；和/或
其中所述3’-UTR包含衍生自ALB的3’-UTR或其同源物、片段或变体，优选地，所述3’-UTR包含如SEQ ID NO.10所示的序列。
32.根据权利要求28-31中任一项所述的重组质粒，其中所述重组质粒还包含polyA、抗性基因启动子和抗性基因；
优选地，所述polyA的长度为100-150个核苷酸；所述抗性基因启动子为氨苄青霉素抗性基因启动子；和/或所述抗性基因为硫酸卡那霉素抗性基因。
33.根据权利要求28所述的重组质粒，其中所述重组质粒的核苷酸序列如SEQ IDNO.28所示。
34.mRNA-载体颗粒，其包含权利要求11-15中任一项所述的mRNA和/或权利要求16-20中任一项所述的mRNA，以及包封所述mRNA的载体材料。
35.用于在受试者中预防和/或治疗与SARS-CoV-2感染相关的疾病或病症的方法，其包括向所述受试者施用有效量的权利要求1-10中任一项所述的重组S蛋白、权利要求11-20中任一项所述的mRNA、权利要求21-25中任一项所述的组合物、权利要求28-33中任一项所述的重组质粒、或权利要求34所述的mRNA-载体颗粒。
36.根据权利要求35所述的方法，其中所述疾病或病症为SARS-CoV-2变异株，例如Delta变异株、Omicron变异株或Lambda变异株感染引起的疾病或病症。
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