CN115960922A - 自复制rna分子设计及其应用 - Google Patents

Abstract

本申请涉及自复制RNA分子设计及其应用，其中，自复制RNA分子包括：第一RNA序列，编码N蛋白或其功能片段；第二RNA序列，编码P蛋白或其功能片段；和第三RNA序列，编码L蛋白或其功能片段，其中，所述自复制RNA分子在动物细胞内适于产生N蛋白、P蛋白、L蛋白或其功能片段。由此，这些第一、第二和第三RNA序列构成了自复制RNA分子的“核心区”区域，可以作为能够在动物细胞内实现自我复制和翻译的最小区域，进一步可以携带载货区，用于搭载其他的编码或非编码序列来表达或者复制靶分子表达小分子、多种肽、酶、抗体、通道蛋白以及受体蛋白等，从而达到不同的治疗目的。

Description

自复制RNA分子设计及其应用

技术领域

本申请属于生物医药领域，具体的，本申请涉及自复制RNA分子设计及其应用。

背景技术

基于mRNA的疫苗和治疗方法的开发近年来成为人们关注的热点之一。无论是用作疫苗还是治疗，mRNA的作用模式是刺激免疫系统作出反应或通过表达蛋白质进行治疗。因此，蛋白表达水平与治疗效果密切相关。但是，mRNA的缺点之一是它们不是很稳定，在细胞内几天内就会降解，导致不可持续的蛋白质表达水平。如果长期用于疾病治疗，可能需要患者注射大量的mRNA，这可能会增加mRNA治疗的毒副作用。

因此，目前基于mRNA的疫苗或者治疗手段仍有待改进。

发明内容

本申请旨在至少在一定程度上解决相关技术中的技术问题之一。

发明人通过对各种RNA病毒的RNA在细胞内的翻译及自我扩增机制进行了深入研究，发现通过采用编码来自弹状病毒的N蛋白，P蛋白及L蛋白的RNA分子作为核心区域，可以实现RNA在动物细胞内的自我复制和翻译，并且该核心区域作为强大的“引擎”，可以提供高效转录扩增和启动大分子蛋白的“动能”，能够进一步搭载“货物区”来复制或者翻译靶分子，这些靶分子几乎涵盖了目前市场上所有的蛋白质药物。根据本发明的实施例，“货物区”可设计不同的蛋白编码盒使机体在细胞内生产多种肽、酶、抗体、通道蛋白以及受体蛋白等，从而达到不同的治疗目的，覆盖肿瘤管线、疫苗管线、罕见病管线及前瞻性通用型产品管线。由此，本发明的发明人提出了一种新型的自复制RNA分子，并将其命名为reRNA^TM，如前所述，reRNA^TM包含可以实现RNA翻译及自我扩增的元件编码序列及搭载的编码靶分子的基因序列。

由此，在本发明的一个方面，本发明提出了一种自复制RNA分子，其包括：第一RNA序列，所述第一RNA序列编码N蛋白或其功能片段；第二RNA序列，所述第二RNA序列编码P蛋白或其功能片段；和第三RNA序列，所述第三RNA序列编码L蛋白或其功能片段，其中，所述自复制RNA分子在动物细胞内适于产生所述N蛋白、所述P蛋白、所述L蛋白或其功能片段。参考图1，由此，这些第一、第二和第三RNA序列构成了自复制RNA分子的“核心区”区域，也称为“核心生命区”，可以作为能够在动物细胞内实现自我复制和翻译的最小区域，进一步可以携带载货区，用于搭载其他的编码或非编码序列来表达或者复制靶分子，表达小分子、多种肽、酶、抗体、通道蛋白以及受体蛋白等，从而达到不同的治疗目的，覆盖肿瘤管线、疫苗管线、罕见病管线及前瞻性通用型产品管线。

在本发明的第二方面，本发明提出了一种DNA分子，其编码前面任一项所述的自复制RNA分子。

在本发明的第三方面，本发明提出了一种表达载体，其携带前面所述的DNA分子。

在本发明的第四方面，本发明提出了一种制备前面所述自复制RNA分子的方法，其包括：使前面所述的DNA分子或者表达载体在宿主细胞中表达所述自复制RNA分子；和收集所述自复制RNA分子。

在本发明的第五方面，本发明提出了一种RNA-蛋白质复合物，其包括：

前面所述自复制RNA分子；和

蛋白质，所述蛋白质包括N蛋白或其功能片段、P蛋白或其功能片段和L蛋白或其功能片段。

在本发明的第六方面，本发明提出了一种药物组合物，其特征在于，包括前面所述的自复制RNA分子、DNA分子、表达载体或者RNA-蛋白质复合物。

在本发明的第七方面，本发明提出了一种在个体中表达靶分子的方法，其包括：

为个体给予前面所述的自复制RNA分子、DNA分子、表达载体、RNA-蛋白质复合物或者药物组合物。

本申请的附加方面和优点将在下面的描述中部分给出，部分将从下面的描述中变得明显，或通过本申请的实践了解到。

附图说明

本申请的上述和/或附加的方面和优点从结合下面附图对实施例的描述中将变得明显和容易理解，其中：

图1显示了根据本发明一个实施例的自复制RNA分子(reRNA^TM)的结构示意图；

图2显示了根据本发明一个实施例的含有所述自复制RNA分子的蛋白质-RNA复合物的结构示意图；

图3、5、6分别显示了根据本发明实施例1、3和4的绿色荧光蛋白表达结果；

图4显示了根据本发明实施例2的绿色荧光蛋白RNA自复制结果；

图7显示了根据本申请实施例8中利用流式细胞检测技术对抗原呈递细胞进行OVA抗原呈递检测的检测结果。

具体实施方式

下面详细描述本申请的实施例，所述实施例的示例在附图中示出。下面通过参考附图描述的实施例是示例性的，旨在用于解释本申请，而不能理解为对本申请的限制。

在本文中，术语“包含”或“包括”为开放式表达，即包括本发明所指明的内容，但并不排除其他方面的内容。

在本文中，术语“任选地”、“任选的”或“任选”通常是指随后所述的事件或状况可以但未必发生，并且该描述包括其中发生该事件或状况的情况，以及其中未发生该事件或状况的情况。

自复制RNA分子

在本文中所使用的术语“自复制RNA分子”也可以称为“自我扩增RNA”，与普通mRNA相比，自复制RNA分子的重要的区别在于它可以使用自己的RNA序列作为模板进行自我复制。根据本申请的实施例，自复制RNA分子还可以在细胞质进行翻译和复制工作，不进入细胞核，可以避免与基因组发生整合所带来的潜在风险。通常mRNA编码需要表达的蛋白质，细胞内的核糖体用来完成翻译和蛋白质生产。根据本申请的实施例，自复制RNA分子会携带一个能够表达RNA聚合酶(RNA依赖RNA聚合酶)的序列，在该RNA分子在细胞质中通过翻译产生RNA聚合酶后，它可以用该自复制RNA分子作为模板来产生更多的自复制RNA分子。

发明人通过对各种RNA病毒的RNA在细胞内的翻译及自我扩增机制进行了深入研究，发现通过采用编码来自弹状病毒的N蛋白，P蛋白及L蛋白的RNA分子作为核心区域，或者以编码正链RNA病毒来源的NSP1、NSP2、NSP3和NSP4蛋白的RNA分子作为核心区域，可以实现RNA在动物细胞内的自我复制和翻译，并且该核心区域作为强大的“引擎”，可以提供高效转录扩增和启动大分子蛋白的“动能”，能够进一步搭载“货物区”来复制或者翻译靶分子，这些靶分子几乎涵盖了目前市场上所有的蛋白质药物。根据本发明的实施例，“货物区”可设计不同的蛋白编码盒使机体在细胞内生产多种肽、酶、抗体、通道蛋白以及受体蛋白等，从而达到不同的治疗目的，覆盖肿瘤管线、疫苗管线、罕见病管线及前瞻性通用型产品管线。由此，本发明的发明人提出了一种新型的自复制RNA分子，并将其命名为reRNA^TM，如前所述，reRNA^TM包含可以实现RNA翻译及自我扩增的元件编码序列及搭载的编码靶分子的基因序列。根据本申请的实施例，reRNA^TM技术借鉴了RNA病毒的特性，在体内实现从RNA到mRNA，再从mRNA到蛋白质的放大扩增，从而高效长效的表达治疗性药物以达到更好的临床疗效。

mRNA通常是通过刺激免疫系统作出反应或通过表达蛋白质进行治疗。因此，蛋白表达水平与治疗效果密切相关。然而，如前所述，传统mRNA不是很稳定，在细胞内几天内就会降解，导致不可持续的蛋白质表达水平。如果长期用于疾病治疗，可能需要患者注射大量的mRNA，这可能会增加mRNA治疗的毒副作用。自扩增RNA因为其可以在细胞质内进行自我复制，因此可以在很低的剂量下达到与传统mRNA相同的蛋白表达水平。例如，自复制RNA分子比传统信使RNA的用量可以小数百倍甚至数千倍，同样能够发挥相同的免疫刺激作用和蛋白质生物活性。可以减少药物治疗或者免疫刺激中使用的注射剂量和次数，从而延长治疗效果，减少药物和给药载体可能产生的毒副作用。另外，自复制RNA分子在复制过程中会形成双链RNA，这与病毒RNA在复制过程中非常相似，可能会刺激细胞的固有免疫反应，进一步增强疫苗的效果。

发明人在研究过程中发现，由于自复制RNA分子将至少编码RNA聚合酶的序列与表达靶蛋白的序列连接起来，因此整个mRNA分子的分子量比传统mRNA的分子量大得多，分子量过大可能导致递送效率、翻译效率以及复制效率显著降低。为了对这些效率进行改善，发明人进行了深入研究，希望能够寻找到最短的核酸片段，可以正常发挥自我复制和翻译的功能。

在本发明的一个方面，本发明提出了一种自复制RNA分子，其包括：第一RNA序列、第二RNA序列和第三RNA序列，其中，所述第一RNA序列编码N蛋白或其功能片段，所述第二RNA序列编码P蛋白或其功能片段，所述第三RNA序列编码L蛋白或其功能片段；或者，所述自复制RNA分子包括第四RNA序列，所述第四RNA序列编码Nsp蛋白或其功能片段；其中，所述自复制RNA分子在动物细胞内适于产生所述N蛋白、所述P蛋白、所述L蛋白或其功能片段，或者所述自复制RNA分子在动物细胞内适于产生所述Nsp蛋白或其功能片段。由此，这些第一、第二和第三RNA序列，或者所述第四RNA序列构成了自复制RNA分子的“核心区”区域，也称为“核心生命区”，可以作为能够在动物细胞内实现自我复制和翻译的最小区域，进一步可以搭载其他的编码序列来表达或者复制靶分子，从而发挥药物或者疫苗的功能。

在本文中所使用的术语“功能片段”是指蛋白质的全长序列的一部分，但仍能够发挥与RNA分子自我复制相关的功能，例如可以是全长序列的截断型的，也可以是蛋白质全长序列的氨基酸序列发生替换、突变或者删除等改变后的蛋白质。根据本申请的实施例，对于N蛋白的功能片段，可以结合RNA分子，保护RNA不受核酸酶的影响，对于P蛋白的功能片段，能够结合N蛋白，在模板上定位L聚合酶，同时也能够作为RNA聚合酶转录和复制复合体的基本组成部分，进一步L蛋白的功能片段能够发挥RNA聚合酶的功能，与RNA的转录及复制有关。

根据本申请的实施例，如何实现RNA在细胞内的翻译同时具备自我扩增特点是最大的技术难点，目前已知的mRNA技术可以实现RNA在细胞内的翻译，但无法实现RNA的自我扩增，为了解决这一问题，发明人研究各种RNA病毒RNA在细胞内的翻译及自我扩增机制，试图找到一种蛋白或蛋白复合体，可以实现RNA在细胞内的翻译及自我扩增，经过大量的研究工作，最终确定弹状病毒的核衣壳蛋白复合体为最佳的候选组合。弹状病毒核衣壳蛋白复合体包含N蛋白，P蛋白及L蛋白，为了确定最小的功能单位，发明人使用水泡性口炎病毒(VSV，Indiana)的核衣壳蛋白复合体蛋白进行了筛选，结果发现，单独的N蛋白、P蛋白、L蛋白不能实现reRNA^TM在细胞内的翻译及自我扩增，而两蛋白的组合NP、NL、PL也不能实现reRNA^TM在细胞内的翻译及自我扩增，三蛋白复合体NPL可以实现reRNA^TM在细胞内的翻译及自我扩增。需要说明的是，对于这种组合包括但不限于弹状病毒科的水泡性病毒属、狂犬病病毒属、短暂热病毒属、非毒粒弹状病毒属等的NPL蛋白组合，以及交叉的组合方式。在此组合中N蛋白的功能为RNA结合蛋白，保护RNA不受核酸酶的影响；P蛋白的功能为连接蛋白，结合N蛋白，在模板上定位L聚合酶，同时也是RNA聚合酶转录和复制复合体的基本组成部分；L蛋白的功能为RNA聚合酶的功能，与RNA的转录及复制有关。本领域技术人员能够理解的是，对于N蛋白，P蛋白和L蛋白，其可以来自于不同的病毒类型。

根据本发明的实施例，所述N蛋白、所述P蛋白、所述L蛋白的至少之一分别独立地来自弹状病毒科病毒。根据本发明的实施例，所述弹状病毒科病毒包括选自水泡性病毒属病毒、狂犬病病毒属病毒和短暂热病毒属病毒的至少之一。其中，水泡性病毒属包括但不限于Vesicular stomatitis New Jersey virus,Maraba virus,Vesicular stomatitisAlagoas virus Indiana 3,Cocal virus Indiana 2,Isfahan virus,Chandipura virus,Spring viraemia of carp virus,American bat vesiculovirus,Carajasvesiculovirus,Jurona vesiculovirus,Malpais Spring vesiculovirus,Morretonvesiculovirus,Perinet vesiculovirus,Piry vesiculovirus,Radi vesiculovirus,YugBogdanovac vesiculovirus。狂犬病病毒属包括但不限于West Caucasian bat virus,Aravan lyssavirus,Khujand lyssavirus,Rabies virus,Australian bat lyssavirus,European bat lyssavirus 1,European bat lyssavirus 2,Irkut virus,Shimoni batvirus,Lagos bat virus,Mokola virus,Lleida bat lyssavirus,Ikoma lyssavirus，Gannoruwa bat lyssavirus,Bokeloh bat lyssavirus,Duvenhage lyssavirus。短暂热病毒属包括但不限于Bovine fever ephemerovirus,Adelaide River ephemerovirus,Berrimah ephemerovirus,Kimberley ephemerovirus

由此，根据本发明的实施例，所述弹状病毒科病毒包括选自水疱性口炎新泽西病毒、马拉巴病毒、水疱性口炎阿拉戈斯病毒印第安纳3号、科卡尔病毒印第安纳2号、伊斯法罕病毒、钱迪普拉病毒、鲤鱼春季病毒血症病毒、美国蝙蝠水疱病毒、卡拉哈斯水疱病毒、朱罗纳水疱病毒、马尔帕斯春季水疱病毒、莫雷顿水疱病毒、佩里特水疱病毒、皮里水泡病毒、拉迪水泡病毒、尤格·博格达诺瓦克水泡病毒、西高加索蝙蝠病毒、Aravan狂犬病毒、Khujand狂犬病毒、狂犬病病毒、澳大利亚蝙蝠狂犬病毒、欧洲蝙蝠狂犬病毒1、欧洲蝙蝠狂犬病毒2、伊尔库特病毒、希莫尼蝙蝠病毒、拉各斯蝙蝠病毒、莫科拉病毒、莱伊达蝙蝠狂犬病毒、伊科马狂犬病毒、甘诺鲁瓦蝙蝠狂犬病毒、博克洛蝙蝠狂犬病毒、杜文哈格狂犬病毒、热蜉蝣病毒、阿德莱德河蜉蝣病毒、伯里玛蜉蝣病毒和金伯利蜉蝣病毒的至少之一。

根据本发明的实施例，N蛋白、P蛋白和L蛋白也可以分别独立地来自相同病毒种的不同的病毒血清型，例如包括但不限于水泡性口炎病毒(VSV)印第安那株、新泽西株和可卡尔株。

根据本发明的实施例，所述N蛋白具有SEQ ID NO：1、7、13、19或25所示氨基酸序列。

MSVTVKRIIDNTVVVPKLPANEDPVEYPADYFRKSKEIPLYINTTKSLSDLRGYVYQGLKSGNVSIIHVNSYLYGALKDIRGKLDKDWSSFGINIGKAGDTIGIFDLVSLKALDGVLPDGVSDASRTSADDKWLPLYLLGLYRVGRTQMPEYRKKLMDGLTNQCKMINEQFEPLVPEGRDIFDVWGNDSNYTKIVAAVDMFFHMFKKHECASFRYGTIVSRFKDCAALATFGHLCKITGMSTEDVTTWILNREVADEMVQMMLPGQEIDKADSYMPYLIDFGLSSKSPYSSVKNPAFHFWGQLTALLLRSTRARNARQPDDIEYTSLTTAGLLYAYAVGSSADLAQQFCVGDNKYTPDDSTGGLTTNAPPQGRDVVEWLGWFEDQNRKPTPDMMQYAKRAVMSLQGLREKTIGKYAKSEFDK(SEQ ID NO：1)。

MSVTVKRIIDNTVIVPKLPANEDPVEYPADYFRKSKEIPLYINTTKSLSDLRGYVYQGLKSGNVSIIHVNSYLYGALKDIRGKLDKDWSSFGINIGKAGDTIGIFDLVSLKALDGVLPDGVSDASRTSADDKWLPLYLLGLYRVGRTQMPEYRKKLMDGLTNQCKMINEQFEPLVPEGRDIFDVWGNDSNYTKIVAAVDMFFHMFKKHECASFRYGTIVSRFKDCAALATFGHLCKITGMSTEDVTTWILNREVADEMVQMMLPGQEIDKADSYMPYLIDFGLSSKSPYSSVKNPAFHFWGQLTALLLRSTRARNARQPDDIEYTSLTTAGLLYAYAVGSSADLAQQFCVGDNKYTPDDSTGGLTTNAPPQGRDVVEWLGWFEDQNRKPTPDMMQYAKRAVMSLQGLREKTIGKYAKSEFDK(SEQ ID NO:7)。

MAPTVKRIINDSIIQPKLPANEDPVEYPADYFKNNTNIVLYVSTKVALNDLRAYVYQGIKSGNPSILHINAYLYAALKGVEGTLDRDWVSFGRTIGKREENVKIFDLVKVEELKTALPDGKSDPDRSAEDDKWLPIYILGLYRVGRSKVTDYRKKLLDGLENQCRVASTRFESLVEDGLDFFDIWENDPNFTKIVAAVDMFFHMFKKHERAPIRYGTIVSRFKDCAALATFGHLSKVSGLSIEDLTTWVLNREVADELCQMMYPGQEIDKADSYMPYMIDFGLSQKSPYSSVKNPAFHFWGQLAALLLRSTRAKNARQPDDIEYTSLTCASLLLSFAVGSSADIEQQFYIGEDKYTTEKDDSLKKSDVPPKGRNVVDWLGWYDDNGGKPTPDMLNFARRAVSSLQSLREKTIGKYAKVEFDK(SEQ ID NO:13)。

MSSQVFCISTGQTVSVCLPANEDPVEFPGAFFTPNARKPTVYIKKETDLSLLRSHVYDGIKDGSVTVSQINSYLYMVLKDIREKPDKNWTSFGVELGKKNEPMGIFDLLNVEDVKGKELDKKGQDTRLPGDDLWLPTLILGLYRVSRATQVEYKKTLMTNLYAQCKLRTKDAEEIVDETAEFFNAWANDSNFTKIVAAVDMYFHHFKKSDHAPIRFGTIVSRFKDCAALSTLSHLQKVTGLPIEEVFTWVFNKSVQDDLLRMMTPGQEIDQADSYMPYLIDMGLSTKSPYSSTKNPSFHFWGQLTAFLVKSARAKNALVPVDIAYHELTTAALLFAYAIGRSSELEQRFVLNGKKFTKEKDSRDDNDTTPPSERNVVVWLAWWEDIKHEITPDMKAFAKRAVERVGDIRVNSVAEYARKLFA(SEQ ID NO:19)。

MDADKIVFKVNNQVVSLKPEIIVDQYEYKYPAIKDLKKPCITLGKAPDLNKAYKSVLSGMSAAKLNPDDVCSYLAAAMQFFEGTCPEDWTSYGIVIARKGDKITPGSLVEIKRTDVEGNWALTGGMELTRDPTVPEHASLVGLLLSLYRLSKISGQNTGNYKTNIADRIEQIFETAPFVKIVEHHTLMTTHKMCANWSTIPNFRFLAGTYDMFFSRIEHLYSAIRVGTVVTAYEDCSGLVSFTGFIKQINLTAREAILYFFHKNFEEEIRRMFEPGQETAVPHSYFIHFRSLGLSGKSPYSSNAVGHVFNLIHFVGCYMGQVRSLNATVIAACAPHEMSVLGGYLGEEFFGKGTFERRFFRDEKELQEYEAAELTKTDVALADDGTVNSDDEDYFSGETRSPEAVYTRIMMNGGRLKRSHIRRYVSVSSNHQARPNSFAEFLNKTYSSDS(SEQ ID NO:25)。

根据本发明的实施例，所述P蛋白具有SEQ ID NO：2、8、14、20或26所示氨基酸序列。

MDNLTKVREYLKSYSRLDQAVGEIDEIEAQRAEKSNYELFQEDGVEEHTKPSYFQAADDSDTESEPEIEDNQGLYAPDPEAEQVEGFIQGPLDDYADEEVDVVFTSDWKQPELESDEHGKTLRLTSPEGLSGEQKSQWLSTIKAVVQSAKYWNLAECTFEASGEGVIMKERQITPDVYKVTPVMNTHPSQSEAVSDVWSLSKTSMTFQPKKASLQPLTISLDELFSSRGEFISVGGDGRMSHKEAILLGLRYKKLYNQARVKYSL(SEQ ID NO：2)。

MDNLTKVREYLKSYSRLDQAVGEIDEIEAQRAEKSNYELFQEDGVEEHTKPSYFQAADDSDTESEPEIEDNQGLYAQDPEAEQVEGFIQGPLDDYADEEVDVVFTSDWKPPELESDEHGKTLRLTSPEGLSGEQKSQWLSTIKAVVQSAKYWNLAECTFEASGEGVIMKERQITPDVYKVTPVMNTHPSQSEAVSDVWSLSKTSMTFQPKKASLQPLTISLDELFSSRGEFISVGGDGRMSHKEAILLGLRYKKLYNQARVKYSL(SEQ ID NO:8)。

MDSVDRLKTYLATYDNLDSALQDANESEERREDKYLQDLFIEDQGDKPTPSYYQEEESSDSDTDYNAEHLTMLSPDERIDKWEEDLPELEKIDDDIPVTFSDWTQPVMKENGGEKSLSLFPPVGLTKIQTEQWKKTIEAVCESSKYWNLSECQILNLEDSLTLKGRLMTPDCSSSVKSQNSVRRSEPLYSSHSPGPPLKVSESINLWDLKSTEVQLISKRAGVKDMTVKLTDFFGSEEEYYSVCPEGAPDLMGAIIMGLKYKKLFNQARMKYRL(SEQ ID NO:14)。

MEDSQLYQALKNYPKLQDTLDSIENLEDDTKSEPSECGSPTERGIPSYYLAEELDECEEEDSEDDDDNLPTEIPDPPTVDMLEAIMEDEIDDTAYQVHFEAKQTWKPVIETGGNERGKFTLSVPQNLSALQLLQWETGIHALAERLGGCRLLQISTRGTRDGIEFTVRETPCVSPASDPIPSTSRSSSIASNVSTRQTESPGSKSNTSLGIPEVPANLIDIGAIDKEFILAAISPSDPPYKNTLRNLFGSGDSFEQYNQTGIYSLKELVVAGLKRKGIYNRIRIRCHLEPQFN(SEQID NO:20)。

MSKIFVNPSAIRAGLADLEMAEETVDLINRNIEDNQAHLQGEPIEVDNLPEDMGRLHLDDGKSPNHGEIAKVGEGKYREDFQMDEGEDPSFLFQSYLENVGVQIVRQMRSGERFLKIWSQTVEEIISYVAVNFPNPPGKSSEDKSTQTTGRELKKETTPTPSQRESQSSKARMAAQIASGPPALEWSATNEEDDLSVEAEIAHQIAESFSKKYKFPSRSSGILLYNFEQLKMNLDDIVKEAKNVPGVTRLAHDGSKLPLRCVLGWVALANSKKFQLLVESDKLSKIMQDDLNRYTSC(SEQ ID NO:26)。

根据本发明的实施例，所述L蛋白具有SEQ ID NO：3、9、15、21或27所示氨基酸序列。

MEVHDFETDEFNDFNEDDYATREFLNPDERMTYLNHADYNLNSPLISDDIDNLIRKFNSLPIPSMWDSKNWDGVLEMLTSCQANPIPTSQMHKWMGSWLMSDNHDASQGYSFLHEVDKEAEITFDVVETFIRGWGNKPIEYIKKERWTDSFKILAYLCQKFLDLHKLTLILNAVSEVELLNLARTFKGKVRRSSHGTNICRIRVPSLGPTFISEGWAYFKKLDILMDRNFLLMVKDVIIGRMQTVLSMVCRIDNLFSEQDIFSLLNIYRIGDKIVERQGNFSYDLIKMVEPICNLKLMKLARESRPLVPQFPHFENHIKTSVDEGAKIDRGIRFLHDQIMSVKTVDLTLVIYGSFRHWGHPFIDYYTGLEKLHSQVTMKKDIDVSYAKALASDLARIVLFQQFNDHKKWFVNGDLLPHDHPFKSHVKENTWPTAAQVQDFGDKWHELPLIKCFEIPDLLDPSIIYSDKSHSMNRSEVLKHVRMNPNTPIPSKKVLQTMLDTKATNWKEFLKEIDEKGLDDDDLIIGLKGKERELKLAGRFFSLMSWKLREYFVITEYLIKTHFVPMFKGLTMADDLTAVIKKMLDSSSGQGLKSYEAICIANHIDYEKWNNHQRKLSNGPVFRVMGQFLGYPSLIERTHEFFEKSLIYYNGRPDLMRVHNNTLINSTSQRVCWQGQEGGLEGLRQKGWSILNLLVIQREAKIRNTAVKVLAQGDNQVICTQYKTKKSRNVVELQGALNQMVSNNEKIMTAIKIGTGKLGLLINDDETMQSADYLNYGKIPIFRGVIRGLETKRWSRVTCVTNDQIPTCANIMSSVSTNALTVAHFAENPINAMIQYNYFGTFARLLLMMHDPALRQSLYEVQDKIPGLHSSTFKYAMLYLDPSIGGVSGMSLSRFLIRAFPDPVTESLSFWRFIHVHARSEHLKEMSAVFGNPEIAKFRITHIDKLVEDPTSLNIAMGMSPANLLKTEVKKCLIESRQTIRNQVIKDATIYLYHEEDRLRSFLWSINPLFPRFLSEFKSGTFLGVADGLISLFQNSRTIRNSFKKKYHRELDDLIVRSEVSSLTHLGKLHLRRGSCKMWTCSATHADTLRYKSWGRTVIGTTVPHPLEMLGPQHRKETPCAPCNTSGFNYVSVHCPDGIHDVFSSRGPLPAYLGSKTSESTSILQPWERESKVPLIKRATRLRDAISWFVEPDSKLAMTILSNIHSLTGEEWTKRQHGFKRTGSALHRFSTSRMSHGGFASQSTAALTRLMATTDTMRDLGDQNFDFLFQATLLYAQITTTVARDGWITSCTDHYHIACKSCLRPIEEITLDSSMDYTPPDVSHVLKTWRNGEGSWGQEIKQIYPLEGNWKNLAPAEQSYQVGRCIGFLYGDLAYRKSTHAEDSSLFPLSIQGRIRGRGFLKGLLDGLMRASCCQVIHRRSLAHLKRPANAVYGGLIYLIDKLSVSPPFLSLTRSGPIRDELETIPHKIPTSYPTSNRDMGVIVRNYFKYQCRLIEKGKYRSHYSQLWLFSDVLSIDFIGPFSISTTLLQILYKPFLSGKDKNELRELANLSSLLRSGEGWEDIHVKFFTKDILLCPEEIRHACKFGIAKDNNKDMSYPPWGRESRGTITTIPVYYTTTPYPKMLEMPPRIQNPLLSGIRLGQLPTGAHYKIRSILHGMGIHYRDFLSCGDGSGGMTAALLRENVHSRGIFNSLLELSGSVMRGASPEPPSALETLGGDKSRCVNGETCWEYPSDLCDPRTWDYFLRLKAGLGLQIDLIVMDMEVRDSSTSLKIETNVRNYVHRILDEQGVLIYKTYGTYICESEKNAVTILGPMFKTVDLVQTEFSSSQTSEVYMVCKGLKKLIDEPNPDWSSINESWKNLYAFQSSEQEFARAKKVSTYFTLTGIPSQFIPDPFVNIETMLQIFGVPTGVSHAAALKSSDRPADLLTISLFYMAIISYYNINHIRVGPIPPNPPSDGIAQNVGIAITGISFWLSLMEKDIPLYQQCLAVIQQSFPIRWEAVSVKGGYKQKWSTRGDGLPKDTRISDSLAPIGNWIRSLELVRNQVRLNPFNEILFNQLCRTVDNHLKWSNLRRNTGMIEWINRRISKEDRSILMLKSDLHEENSWRD(SEQ ID NO：3)。

MEVHDFETDEFNDFNEDDYATREFLNPDERMTYLNHADYNLNSPLISDDIDNLIRKFNSLPIPSMWDSKNWDGVLEMLTSCQANPISTSQMHKWMGSWLMSDNHDASQGYSFLHEVDKEAEITFDVVETFIRGWGNKPIEYIKKERWTDSFKILAYLCQKFLDLHKLTLILNAVSEVELLNLARTFKGKVRRSSHGTNICRIRVPSLGPTFISEGWAYFKKLDILMDRNFLLMVKDVIIGRMQTVLSMVCRIDNLFSEQDIFSLLNIYRIGDKIVERQGNFSYDLIKMVEPICNLKLMKLARESRPLVPQFPHFENHIKTSVDEGAKIDRGIRFLHDQIMSVKTVDLTLVIYGSFRHWGHPFIDYYTGLEKLHSQVTMKKDIDVSYAKALASDLARIVLFQQFNDHKKWFVNGDLLPHDHPFKSHVKENTWPTAAQVQDFGDKWHELPLIKCFEIPDLLDPSIIYSDKSHSMNRSEVLKHVRMNPNTPIPSKKVLQTMLDTKATNWKEFLKEIDEKGLDDDDLIIGLKGKERELKLAGRFFSLMSWKLREYFVITEYLIKTHFVPMFKGLTMADDLTAVIKKMLDSSSGQGLKSYEAICIANHIDYEKWNNHQRKLSNGPVFRVMGQFLGYPSLIERTHEFFEKSLIYYNGRPDLMRVHNNTLINSTSQRVCWQGQEGGLEGLRQKGWTILNLLVIQREAKIRNTAVKVLAQGDNQVICTQYKTKKSRNVVELQGALNQMVSNNEKIMTAIKIGTGKLGLLINDDETMQSADYLNYGKIPIFRGVIRGLETKRWSRVTCVTNDQIPTCANIMSSVSTNALTVAHFAENPINAMIQYNYFGTFARLLLMMHDPALRQSLYEVQDKIPGLHSSTFKYAMLYLDPSIGGVSGMSLSRFLIRAFPDPVTESLSFWRFIHVHARSEHLKEMSAVFGNPEIAKFRITHIDKLVEDPTSLNIAMGMSPANLLKTEVKKCLIESRQTIRNQVIKDATIYLYHEEDRLRSFLWSINPLFPRFLSEFKSGTFLGVADGLISLFQNSRTIRNSFKKKYHRELDDLIVRSEVSSLTHLGKLHLRRGSCKMWTCSATHADTLRYKSWGRTVIGTTVPHPLEMLGPQHRKETPCAPCNTSGFNYVSVHCPDGIHDVFSSRGPLPAYLGSKTSESTSILQPWERESKVPLIKRATRLRDAISWFVEPDSKLAMTILSNIHSLTGEEWTKRQHGFKRTGSALHRFSTSRMSHGGFASQSTAALTRLMATTDTMRDLGDQNFDFLFQATLLYAQITTTVARDGWITSCTDHYHIACKSCLRPIEEITLDSSMDYTPPDVSHVLKTWRNGEGSWGQEIKQIYPLEGNWKNLAPAEQSYQVGRCIGFLYGDLAYRKSTHAEDSSLFPLSIQGRIRGRGFLKGLLDGLMRASCCQVIHRRSLAHLKRPANAVYGGLIYLIDKLSVSPPFLSLTRSGPIRDELETIPHKIPTSYPTSNRDMGVIVRNYFKYQCRLIEKGKYRSHYSQLWLFSDVLSIDFIGPFSISTTLLQILYKPFLSGKDKNELRELANLSSLLRSGEGWEDIHVKFFTKDILLCPEEIRHACKFGIAKDNNKDMSYPPWGRESRGTITTIPVYYTTTPYPKMLEMPPRIQNPLLSGIRLGQLPTGAHYKIRSILHGMGIHYRDFLSCGDGSGGMTAALLRENVHSRGIFNSLLELSGSVMRGASPEPPSALETLGGDKSRCVNGETCWEYPSDLCDPRTWDYFLRLKAGLGLQIDLIVMDMEVRDSSTSLKIETNVRNYVHRILDEQGVLIYKTYGTYICESEKNAVTILGPMFKTVDLVQTEFSSSQTSEVYMVCKGLKKLIDEPNPDWSSINESWKNLYAFQSSEQEFARAKKVSTYFTLTGIPSQFIPDPFVNIETMLQIFGVPTGVSHAAALKSSDRPADLLTISLFYMAIISYYNINHIRVGPIPPNPPSDGIAQNVGIAITGISFWLSLMEKDIPLYQQCLAVIQQSFPIRWEAVSVKGGYKQKWSTRGDGLPKDTRTSDSLAPIGNWIRSLELVRNQVRLNPFNEILFNQLCRTVDNHLKWSNLRRNTGMIEWINRRISKEDRSILMLKSDLHEENSWRD(SEQ ID NO:9)。

MDFDLIEDSANWEDDESDFFLRDILSQEDQMSYLNTADYNLNSPLISDDMVYIIKRMNHEEVPPIWRSKEWDSPLDMLRGCQAQPMSHQEMHNWFGTWIQNVQHDSAQPFTFLKEVDKESEMTYDLVSTFLKGWVGKDYPFKSKNKEIDSMALVGPLCQKFLDLHKITLILNAVSLGETKELLTTFKGKYRMSCENIPIARLRLPSLGPVFMCKGWTYIHKERVLMDRTSFSCVKDVIIGRMQTFLSMIGRSDNKFSPDQIYTLANVYRIGDKILEQCGNKAYDLIKMIEPICNLKMMELARLHRPKIPKFPHFEEHVKGSVQELTQRSNRIQTLYDLIMSMKDVDLVLVVYGSFRHWGHPFIDYFQGLEKLHTQVNMEKHIDKEYPQQLASDLARLVLNKQFSESKKWFVDPSKMSPKHPFYEHVINKTWPTAAKIQDFGDNWHKLPLIQCFEIPDLIDPSVIYSDKSHSMNKKEVIQHVRSKPNIPIPSNKVLQTMLTNRATNWKAFLKDIDENGLDDDDLIIGLKGKERELKIAGRFFSLMSWRLREYFVITEYLIKTYYVPLFKGLTMADDLTSVIKKMMDSSSGQGLDDYSSVCLANHIDYEKWNNHQRKESNGPIFRVMGQFLGYPSLIERTHEFFEKSLIYYNGRPDLMTIRNGTLCNSTKHRVCWNGQKGGLEGLRQKGWSIVNLLVIQREAKIRNTAVKVLAQGDNQVICTQYKTKKTRSELELRAVLHQMAGNNNKIMEEIKRGTEKLGLIINDDETMQSADYLNYGKIPIFRGVIRGLETKRWSRVTCVTNDQIPTCANLMSSVSTNALTVAHFAENPINAMIQYNYFGTFARLLLFMHDPAIRQSLYKVQEKIPGLHTRTFKYAMLYLDPSIGGVCGMALSRFLIRAFPDPVTESLSFWKFIYEHASEPHLKKMEVMFETPQGAKFRIEHINKLLEDPTSLNISMGMSPANLLKSEVKKCLIESRSSIKNEIIKDATIYMHQEEEKLRGFLWSIKPLFPRFLSEFKAGTFLGVSEGFINLFQNSRTIRNSFKKRYHKDLDELIIKREISSLSHLGSMHYRLGDNQIWSCSASRADILRYKSWTRKVVGTTVPHPLEMHGPPSKKERPCQLCNSSGLTYISVHCPKGIIDVFNRRGPLPAYLGSNTSESTSILQPWEKESKIPIIKRATRLRDAISWFIPPESPLSTCILNNIQALTGEDWSSKQHGFKRTGSALHRFSTSRMSNGGFASQSPATLTRMIATTDTMRDFGTKNYDFMFQASLLYGQMTTSISRYGTPGSCTDHYHIRCKGCIREIEEVELNTSLEYKTPDVSHILEKWRNNTGSWGHQIKQLKPAEGNWESLSPVEQSYQVARCIGFLYGELTHKKSRQADDSSLFPLSIQLKVRGRGFLRGLLDGLMRSSCCQVIHRRSVSTLKRPANAVYGGLIYLIDKLSASSPFLSLVRTGPIRQELEQVPHKMSTSYPTNIRDLGSIVRNYFKYQCRPVERGNYKTYYNQIWLFSDVLSTEFIGPMRISSSLLRLLYRPSLTKKDREELRELAALSSNLRSGEDWDDLHIKFFSNDLLFCSQEIRHACKFGIKKDNEDITFYPNWGTEYIGNVTDIPVFYRAQNVKKDIRVPPRIQNPLMSGLRLGQLPTGAHYKMRAIVFRLKIPYHDFLDCGDGSGGMTAALLRYNRTSRGIFNSLLDLSDTMLRGSSPEPPSALETLGGERVRCVNGDSCWEHPSDLSDENTWKYFLHLKKGCGMSINLITMDMEVQDSVISYKIESLVRQYVPVLLESDGCLIYKTYGTYIATQEDNSLTLIGSLFHSVQLVQTDLSSSNTSELYLVCRRLKDYVDTPFVDWIELYDNWEKQYAFRSFKDEFQRAQSLTPETTLIGIPPQFVPDPGVNLETLFQIAGVPTGVAHGITHHILQSKDKLISNAIGSMCVISHFHINTIRTTDSMPGPPSDGDVNKMCSALIGACFWLDWMESDLNLYKHCLRSIMKSMPVRWFRTLKNEKWSQKWDCKGDAIPKDSRLGDSLANIGNWIRAWELIRNGNKSEPFDSMVAEALTKSVDKSLSWRKISKSTGIPRLLNSDIDLVDQSILNVQIDIVENQAWQN(SEQ ID NO:15)。

MDLNPVDDAAELSEENFFSGKLSKECRIRGLNSVDYNLNSPLVSDDLTYLLNKFKGKPVPIRWKMKKWDSILDQLRKHDLEYLRPSDLHQWFAEWMLYSKHGSKQGEDFLKTVDEEASDTFEVVRSFIRGWTGGEINFVRKSGKHMGYCAELCQKFLDLHKLTLLGNAATDNELLQLSKTFGDDKIYKKRLIKLPSLGRVIFDSGFFIVLDQRVLMDRNFMLMMKDVIIGRMQTVLSMISRCDDKFSSKDIDFLLKVYSTGDKIIRKLGNDGYELIKTVEPMCNLRLSDLARRFRPLVPPFPHFRRHIESTVDELSAKTPLIRELFSLIDTSPNVDSTLVVYGSFRHWGHPFINYFEGLEKLHKQVTMEKEIDTNYSEALASDLARIVLTKEFNEKKQWAVDYHRVPTSHPFKNHIRDNTWPTAAVIQDFGDHWHELPLIQCFDIPDLIDPSIIYSDKSHSMNRSEVLNHVRTKPHTPIPSKKVLESMIDKPATNWLEFLEEIDKNGLSDEDLVIGLKGKERELKIAGRFFSLMSWKLREYFVVTEYLIKTHFVPLFHGLTMADDMTAVIKKMLESSSGQGLTNYDSVCIANHIDYEKWNNHQRKLSNGPVFKVMGQFLGYPNLIYRTHEFFEKSLIYYNERPDLMKVRNGILENSTHQRVCWNGQAGGLEGLRQKGWSILNLLVIQREAKIRNTAVKVLAQGDNQVICTQYKTKQYRNDIELRQALNQMAANNDVIMKAIESGTNKLGLLINQDETMQSADYLNYGKVPIFRGVIRGLETKRWSRVTCVTNDQLPTCANLMSSVSTNALTVAHFDVHPLNAMIQFNFFGNFARLLLIMHDPAIRQSLNQLKGPNINVHSYGFKVAMLYLDPSIGGVCGTALSRFLIRSFPDPVTESLSFWKLIHHSTSDIRLKNLSEQFGNPKIAVFRESHIEKLLEDPTSLNISMGMSAANLLKTEIKKNLLQKKSSIGNQIVKDAVYYIHSEDEKLRTFLWSITPLFPRFLSEFKAGTFMGVASSIVSLFQNSRTIRNVFRDYMSQTIDDLIVKSELTSLEHLSNYTDRKGSGGIWSCSAEQADKLRRMSWKRPVLGTTVPHPLEMHGRGTLKSPLSKCCKESRMDYISVHIPEGLNKVLDGRGSLPAYLGSKTSESTSILQPWEKESKIPIIRRATRLRDAIHWFVDPDSNLARSILNNIESLTGEKWEGALKGYKRTGSALHRFSTSRVSHGGFSSQSPACLTRMMATTDTMRDYAQLNYDFMFQASLLYSQMTSSVILMGTTVSNTIHFHVTCRKCIREITEPMLESPREYRGKDVHLVLAKWKNSSNGWGETLQLLKPVEGDWDTIPPVEKSYHVGRILGFLYGDLKSQNSSRADDSSIFPLSIQMRLRGRGFLRGILDGLVRASACQVIHRRSVALLSKPANAIYGGLIYLIDKISASTSFTTLCRDGPIREELSSIPHKIPTSYPTSNSDMGLHIRNYLKFQCKTVELGKYQSDIKDLWLFSDVMTSNIAGPFALSTKILKCLYKPALSQKDRNNIRKISNFSKMMRSQLSWDPTSSEFITSQILVCNEEIRHACKFGIPKLSLKFDDPVWGPEDYGLIWSIPVDYSSQSVPKNLKPCPRIQNPSISGFRLGQLPTGAHYKLRSILRKKNIHYRDALCGGDGSGGMTAAVLRYNLKARAIFNSILDFDGSTMKGASPDPPSALETVVNGRTRCVNAESCWENPSDLSEQRTWDYFKFLKTHHGLKIDLIVLDMEVRDFAISASIEKCVRNNVSSILEEDGVLIYKTYGSTIAAESSNAVVNIGVLFESVELIQTEYSSTSTSEVYMYCRKIKKFVDAPHPDWISLDYYWSKLFCFRSYKEEFFRSYEVSRKESLKGIPNSFIPDPLVNLETLLQIAGVPSGISHQLAIDIKESQLTQITAAMVLCGMIANYTLDVTKKRDSYNPPSDGRLIRMSAALVGISFWISVKYYDKELNFELEQILSRSFPIRWMLSRNYLFWTTKKGFRNAKDVRLSGNMANIGNWIRCMELLHLPPGSLSKDEVTTTCGKYIRSLKYSVILQQTGIIDLWKSRVASDDRSLMEVKTEFIESEHWVD(SEQ ID NO:21)。

MLDPGEVYDDPIDPIELEAEPRGTPIVPNILRNSDYNLNSPLIEDPARLMLEWLKTGNRPYRMTLTDNCSRSFRVLKDYFKKVDLGSLKVGGMAAQSMISLWLYGAHSESNRSRRCITDLAHFYSKSSPIEKLLNLTLGNRGLRIPPEGVLSCLERVDYDNAFGRYLANTYSSYLFFHVITLYMNALDWDEEKTILALWKDLTSVDIGKDLVKFKDQIWGLLIVTKDFVYSQSSNCLFDRNYTLMLKDLFLSRFNSLMVLLSPPEPRYSDDLISQLCQLYIAGDQVLSMCGNSGYEVIKILEPYVVNSLVQRAEKFRPLIHSLGDFPVFIKDKVSQLEETFGPCARRFFRALDQFDNIHDLVFVFGCYRHWGHPYIDYRKGLSKLYDQVHLKKVIDKSYQECLASDLARRILRWGFDKYSKWYLDSRFLARDHPLTPYIKTQTWPPKHIVDLVGDTWHKLPITQIFEIPESMDPSEILDDKSHSFTRTRLASWLSENRGGPVPSEKVIITALSKPPVNPREFLRSIDLGGLPDEDLIIGLKPKERELKIEGRFFALMSWNLRLYFVITEKLLANYILPLFDALTMTDNLNKVFKKLIDRVTGQGLLDYSRVTYAFHLDYEKWNNHQRLESTEDVFSVLDQVFGLKRVFSRTHEFFQKAWIYYSDRSDLIGLREDQIYCLDASNGPTCWNGQDGGLEGLRQKGWSLVSLLMIDRESQIRNTRTKILAQGDNQVLCPTYMLSPGLSQEGLLYELERISRNALSIYRAVEEGASKLGLIIKKEETMCSYDFLIYGKTPLFRGNILVPESKRWARVSCVSNDQIVNLANIMSTVSTNALTVAQHSQSLIKPMRDFLLMSVQAVFHYLLFSPILKGRVYKILSAEGESFLLAMSRIIYLDPSLGGISGMSLGRFHIRQFSDPVSEGLSFWREIWLSSQESWIHALCQEAGNPDLGERTLESFTRLLEDPTTLNIRGGASPTILLKDAIRKALYDEVDKVENSEFREAILLSKTHRDNFILFLISVEPLFPRFLSELFSSSFLGIPESIIGLIQNSRTIRRQFRKSLSKTLEESFYNSEIHGISRMTQTPQRVGGVWPCSSERADQLREISWGRKVVGTTVPHPSEMLGLLPKSSISCTCGATGGGNPRVSVSVLPSFDQSFFSRGPLKGYLGSSTSMSTQLFHAWEKVTNVHVVKRALSLKESINWFITRDSNLAQALIRNIMSLTGPDFPLEEAPVFKRTGSALHRFKSARYSEGGYSSVCPNLLSHISVSTDTMSDLTQDGKNYDFMFQPLMLYAQTWTSELVQRDTRLRDSTFHWHLRCNRCVRPIDDVTLETSQIFEFPDVSKRISRMVSGAVPHFQRLPDIRLRPGDFESLSGREKSHHIGSAQGLLYSILVAIHDSGYNDGTIFPVNIYGKVSPRDYLRGLARGVLIGSSICFLTRMTNININRPLELVSGVISYILLRLDNHPSLYIMLREPSLRGEIFSIPQKIPAAYPTTMKEGNRSILCYLQHVLRYEREIITASPENDWLWIFSDFRSAKMTYLSLITYQSHLLLQRVERNLSKSMRDNLRQLSSLMRQVLGGHGEDTLESDDNIQRLLKDSLRRTRWVDQEVRHAARTMTGDYSPNKKVSRKVGCSEWVCSAQQVAVSTSANPAPVSELDIRALSKRFQNPLISGLRVVQWATGAHYKLKPILDDLNVFPSLCLVVGDGSGGISRAVLNMFPDAKLVFNSLLEVNDLMASGTHPLPPSAIMRGGNDIVSRVIDLDSIWEKPSDLRNLATWKYFQSVQKQVNMSYDLIICDAEVTDIASINRITLLMSDFALSIDGPLYLVFKTYGTMLVNPNYKAIQHLSRAFPSVTGFITQVTSSFSSELYLRFSKRGKFFRDAEYLTSSTLREMSLVLFNCSSPKSEMQRARSLNYQDLVRGFPEEIISNPYNEMIITLIDSDVESFLVHKMVDDLELQRGTLSKVAIIIAIMIVFSNRVFNVSKPLTDPSFYPPSDPKILRHFNICCSTMMYLSTALGDVPSFARLHDLYNRPITYYFRKQVIRGNVYLSWSWSNDTSVFKRVACNSSLSLSSHWIRLIYKIVKTTRLVGSIKDLSREVERHLHRYNRWITLEDIRSRSSLLDYSCL(SEQ ID NO:27)。

根据本发明的实施例，所述Nsp蛋白具有SEQ ID NO:31～34所示氨基酸序列中的至少之一。发明人发现，正链RNA病毒来源的自复制RNA序列的核心复合物为非结构蛋白(Nsp)。

MEKPVVNVDVDPQSPFVVQLQKSFPQFEVVAQQVTPNDHANARAFSHLASKLIELEVPTTATILDIGSAPARRMFSEHQYHCVCPMRSPEDPDRMMKYASKLAEKACKITNKNLHEKIKDLRTVLDTPDAETPSLCFHNDVTCNMRAEYSVMQDVYINAPGTIYHQAMKGVRTLYWIGFDTTQFMFSAMAGSYPAYNTNWADEKVLEARNIGLCSTKLSEGRTGKLSIMRKKELKPGSRVYFSVGSTLYPEHRASLQSWHLPSVFHLNGKQSYTCRCDTVVSCEGYVVKKITISPGITGETVGYAVTHNSEGFLLCKVTDTVKGERVSFPVCTYIPATICDQMTGIMATDISPDDAQKLLVGLNQRIVINGRTNRNTNTMQNYLLPIIAQGFSKWAKERKDDLDNEKMLGTRERKLTYGCLWAFRTKKVHSFYRPPGTQTCVKVPASFSAFPMSSVWTTSLPMSLRQKLKLALQPKKEEKLLQVSEELVMEAKAAFEDAQEEARAEKLREALPPLVADKGIEAAAEVVCEVEGLQADIGA(SEQ ID NO:31)。

ALVETPRGHVRIIPQANDRMIGQYIVVSPNSVLKNAKLAPAHPLADQVKIITHSGRSGRYAVEPYDAKVLMPAGGAVPWPEFLALSESATLVYNEREFVNRKLYHIAMHGPAKNTEEEQYKVTKAELAETEYVFDVDKKRCVKKEEASGLVLSGELTNPPYHELALEGLKTRPAVPYKVETIGVIGTPGSGKSAIIKSTVTARDLVTSGKKENCREIEADVLRLRGMQITSKTVDSVMLNGCHKAVEVLYVDEAFACHAGALLALIAIVRPRKKVVLCGDPMQCGFFNMMQLKVHFNHPEKDICTKTFYKYISRRCTQPVTAIVSTLHYDGKMKTTNPCKKNIEIDITGATKPKPGDIILTCFRGWVKQLQIDYPGHEVMTAAASQGLTRKGVYAVRQKVNENPLYAITSEHVNVLLTRTEDRLVWKTLQGDPWIKQPTNIPKGNFQATIEDWEAEHKGIIAAINSPTPRANPFSCKTNVCWAKALEPILATAGIVLTGCQWSELFPQFADDKPHSAIYALDVICIKFFGMDLTSGLFSKQSIPLTYHPADSARPVAHWDNSPGTRKYGYDHAIAAELSRRFPVFQLAGKGTQLDLQTGRTRVISAQHNLVPVNRNLPHALVPEYKEKQPGPVKKFLNQFKHHSVLVVSEEKIEAPRKRIEWIAPIGIAGADKNYNLAFGFPPQARYDLVFINIGTKYRNHHFQQCEDHAATLKTLSRSALNCLNPGGTLVVKSYGYADRNSEDVVTALARKFVRVSAARPDCVSSNTEMYLIFRQLDNSRTRQFTPHHLNCVISSVYEGTRDGVGA(SEQ ID NO:32)。

APSYRTKRENIADCQEEAVVNAANPLGRPGEGVCRAIYKRWPTSFTDSATETGTARMTVCLGKKVIHAVGPDFRKHPEAEALKLLQNAYHAVADLVNEHNIKSVAIPLLSTGIYAAGKDRLEVSLNCLTTALDRTDADVTIYCLDKKWKERIDAALQLKESVTELKDEDMEIDDELVWIHPDSCLKGRKGFSTTKGKLYSYFEGTKFHQAAKDMAEIKVLFPNDQESNEQLCAYILGETMEAIREKCPVDHNPSSSPPKTLPCLCMYAMTPERVHRLRSNNVKEVTVCSSTPLPKHKIKNVQKVQCTKVVLFNPHTPAFVPARKYIEVPEQPTAPPAQAEEAPEVVATPSPSTADNTSLDVTDISLDMDDSSEGSLFSSFSGSDNSITSMDSWSSGPSSLEIVDRRQVVVADVHAVQEPAPIPPPRLKKMARLAAARKEPTPPASNSSESLHLSFGGVSMSLGSIFDGETARQAAVQPLATGPTDVPMSFGSFSDGEIDELSRRVTESEPVLFGSFEPGEVNSIISSRSAVSFPLRKQRRRRRSRRTEY(SEQ ID NO:33)。

MMPTEANKSRYQSRKVENQKAITTERLLSGLRLYNSATDQPECYKITYPKPLYSSSVPANYSDPQFAVAVCNNYLHENYPTVASYQITDEYDAYLDMVDGTVACLDTATFCPAKLRSYPKKHEYRAPNIRSAVPSAMQNTLQNVLIAATKRNCNVTQMRELPTLDSATFNVECFRKYACNDEYWEEFARKPIRITTEFVTAYVARLKGPKAAALFAKTYNLVPLQEVPMDRFVMDMKRDVKVTPGTKHTEERPKVQVIQAAEPLATAYLCGIHRELVRRLTAVLLPNIHTLFDMSAEDFDAIIAEHFKQGDPVLETDIASFDKSQDDAMALTGLMILEDLGVDQPLLDLIECAFGEISSTHLPTGTRFKFGAMMKSGMFLTLFVNTVLNVVIASRVLEERLKTSRCAAFIGDDNIIHGVVSDKEMAERCATWLNMEVKIIDAVIGERPPYFCGGFILQDSVTSTACRVADPLKRLFKLGKPLPADDEQDEDRRRALLDETKAWFRVGITGTLAVAVTTRYEVDNITPVLLALRTFAQSKRAFQAIRGEIKHLYGGPK(SEQ ID NO:34)。

根据本发明的一些具体的实施例，所述自复制核心序列包括编码以下6种蛋白质的碱基序列组合中的一种：

所述N蛋白具有SEQ ID NO：1所示氨基酸序列，所述P蛋白具有SEQ ID NO：2所示氨基酸序列，所述L蛋白具有SEQ ID NO：3所示氨基酸序列。

所述N蛋白具有SEQ ID NO：7所示氨基酸序列，所述P蛋白具有SEQ ID NO：8所示氨基酸序列，所述L蛋白具有SEQ ID NO：9所示氨基酸序列。

所述N蛋白具有SEQ ID NO：13所示氨基酸序列，所述P蛋白具有SEQ ID NO：14所示氨基酸序列的，所述L蛋白具有SEQ ID NO：15所示氨基酸序列。

所述N蛋白具有SEQ ID NO：19所示氨基酸序列，所述P蛋白具有SEQ ID NO：20所示氨基酸序列，所述L蛋白具有SEQ ID NO：21所示氨基酸序列。

所述N蛋白具有SEQ ID NO：25所示氨基酸序列，所述P蛋白具有SEQ ID NO：26所示氨基酸序列，所述L蛋白具有SEQ ID NO：27所示氨基酸序列。

所述Nsp蛋白具有SEQ ID NO：31～34所示氨基酸序列。发明人发现，正链RNA病毒来源的自复制RNA序列的核心复合物为NSP1(SEQ ID NO：31)、NSP2(SEQ ID NO：32)、NSP3(SEQ ID NO：33)和NSP4(SEQ ID NO：34)。

根据本发明的实施例，所述自复制RNA分子中的所述第一RNA序列具有SEQ ID NO：4、10、16、22或28所示碱基序列。

AUGUCUGUUACAGUCAAGAGAAUCAUUGACAACACAGUCGUAGUUCCAAAACUUCCUGCAAAUGAGGAUCCAGUGGAAUACCCGGCAGAUUACUUCAGAAAAUCAAAGGAGAUUCCUCUUUACAUCAAUACUACAAAAAGUUUGUCAGAUCUAAGAGGAUAUGUCUACCAAGGCCUCAAAUCCGGAAAUGUAUCAAUCAUACAUGUCAACAGCUACUUGUAUGGAGCAUUAAAGGACAUCCGGGGUAAGUUGGAUAAAGAUUGGUCAAGUUUCGGAAUAAACAUCGGGAAAGCAGGGGAUACAAUCGGAAUAUUUGACCUUGUAUCCUUGAAAGCCCUGGACGGCGUACUUCCAGAUGGAGUAUCGGAUGCUUCCAGAACCAGCGCAGAUGACAAAUGGUUGCCUUUGUAUCUACUUGGCUUAUACAGAGUGGGCAGAACACAAAUGCCUGAAUACAGAAAAAAGCUCAUGGAUGGGCUGACAAAUCAAUGCAAAAUGAUCAAUGAACAGUUUGAACCUCUUGUGCCAGAAGGUCGUGACAUUUUUGAUGUGUGGGGAAAUGACAGUAAUUACACAAAAAUUGUCGCUGCAGUGGACAUGUUCUUCCACAUGUUCAAAAAACAUGAAUGUGCCUCGUUCAGAUACGGAACUAUUGUUUCCAGAUUCAAAGAUUGUGCUGCAUUGGCAACAUUUGGACACCUCUGCAAAAUAACCGGAAUGUCUACAGAAGAUGUAACGACCUGGAUCUUGAACCGAGAAGUUGCAGAUGAAAUGGUCCAAAUGAUGCUUCCAGGCCAAGAAAUUGACAAGGCCGAUUCAUACAUGCCUUAUUUGAUCGACUUUGGAUUGUCUUCUAAGUCUCCAUAUUCUUCCGUCAAAAACCCUGCCUUCCACUUCUGGGGGCAAUUGACAGCUCUUCUGCUCAGAUCCACCAGAGCAAGGAAUGCCCGACAGCCUGAUGACAUUGAGUAUACAUCUCUUACUACAGCAGGUUUGUUGUACGCUUAUGCAGUAGGAUCCUCUGCCGACUUGGCACAACAGUUUUGUGUUGGAGAUAACAAAUACACUCCAGAUGAUAGUACCGGAGGAUUGACGACUAAUGCACCGCCACAAGGCAGAGAUGUGGUCGAAUGGCUCGGAUGGUUUGAAGAUCAAAACAGAAAACCGACUCCUGAUAUGAUGCAGUAUGCGAAAAGAGCAGUCAUGUCACUGCAAGGCCUAAGAGAGAAGACAAUUGGCAAGUAUGCUAAGUCAGAAUUUGACAAAUGA(SEQ ID NO：4)。

AUGUCUGUUACAGUCAAGAGAAUCAUUGACAACACAGUCAUAGUUCCAAAACUUCCUGCAAAUGAGGAUCCAGUGGAAUACCCGGCAGAUUACUUCAGAAAAUCAAAGGAGAUUCCUCUUUACAUCAAUACUACAAAAAGUUUGUCAGAUCUAAGAGGAUAUGUCUACCAAGGCCUCAAAUCCGGAAAUGUAUCAAUCAUACAUGUCAACAGCUACUUGUAUGGAGCAUUAAAGGACAUCCGGGGUAAGUUGGAUAAAGAUUGGUCAAGUUUCGGAAUAAACAUCGGGAAAGCAGGGGAUACAAUCGGAAUAUUUGACCUUGUAUCCUUGAAAGCCCUGGACGGCGUACUUCCAGAUGGAGUAUCGGAUGCUUCCAGAACCAGCGCAGAUGACAAAUGGUUGCCUUUGUAUCUACUUGGCUUAUACAGAGUGGGCAGAACACAAAUGCCUGAAUACAGAAAAAAGCUCAUGGAUGGGCUGACAAAUCAAUGCAAAAUGAUCAAUGAACAGUUUGAACCUCUUGUGCCAGAAGGUCGUGACAUUUUUGAUGUGUGGGGAAAUGACAGUAAUUACACAAAAAUUGUCGCUGCAGUGGACAUGUUCUUCCACAUGUUCAAAAAACAUGAAUGUGCCUCGUUCAGAUACGGAACUAUUGUUUCCAGAUUCAAAGAUUGUGCUGCAUUGGCAACAUUUGGACACCUCUGCAAAAUAACCGGAAUGUCUACAGAAGAUGUAACGACCUGGAUCUUGAACCGAGAAGUUGCAGAUGAAAUGGUCCAAAUGAUGCUUCCAGGCCAAGAAAUUGACAAGGCCGAUUCAUACAUGCCUUAUUUGAUCGACUUUGGAUUGUCUUCUAAGUCUCCAUAUUCUUCCGUCAAAAACCCUGCCUUCCACUUCUGGGGGCAAUUGACAGCUCUUCUGCUCAGAUCCACCAGAGCAAGGAAUGCCCGACAGCCUGAUGACAUUGAGUAUACAUCUCUUACUACAGCAGGUUUGUUGUACGCUUAUGCAGUAGGAUCCUCUGCCGACUUGGCACAACAGUUUUGUGUUGGAGAUAACAAAUACACUCCAGAUGAUAGUACCGGAGGAUUGACGACUAAUGCACCGCCACAAGGCAGAGAUGUGGUCGAAUGGCUCGGAUGGUUUGAAGAUCAAAACAGAAAACCGACUCCUGAUAUGAUGCAGUAUGCGAAAAGAGCAGUCAUGUCACUGCAAGGCCUAAGAGAGAAGACAAUUGGCAAGUAUGCUAAGUCAGAAUUUGACAAAUGA(SEQ ID NO:10)。

AUGGCUCCUACAGUUAAGAGAAUCAUUAAUGACUCCAUAAUUCAGCCUAAACUACCGGCCAAUGAGGAUCCGGUAGAGUAUCCAGCUGAUUACUUCAAAAAUAACACCAACAUAGUAUUAUAUGUGAGCACUAAAGUAGCCCUAAAUGAUUUAAGGGCCUAUGUCUACCAGGGAAUCAAAUCCGGUAAUCCAUCAAUACUCCACAUAAAUGCUUAUCUUUAUGCCGCGUUAAAGGGAGUAGAAGGAACAUUAGACAGAGACUGGGUUAGCUUUGGAAGAACGAUCGGAAAGAGGGAAGAGAAUGUCAAAAUCUUCGAUCUAGUCAAAGUUGAGGAACUGAAGACAGCUCUUCCUGAUGGGAAAUCGGAUCCAGAUCGAUCAGCUGAAGAUGAUAAAUGGCUUCCUAUUUACAUCCUAGGCCUAUAUCGAGUGGGUAGAUCCAAAGUUACAGAUUACAGAAAGAAACUCCUGGACGGUCUAGAAAAUCAGUGUAGAGUGGCAUCAACCAGAUUUGAAAGCCUUGUUGAAGAUGGUCUUGACUUCUUUGACAUCUGGGAAAAUGAUCCCAACUUCACCAAAAUAGUUGCUGCAGUGGAUAUGUUUUUUCACAUGUUCAAGAAGCAUGAACGUGCUCCAAUCAGAUAUGGAACCAUAGUCUCACGAUUCAAAGACUGUGCAGCACUUGCUACAUUUGGACAUCUCAGCAAGGUUAGCGGACUUUCAAUUGAAGACCUAACAACAUGGGUUCUGAAUCGAGAAGUCGCAGACGAACUAUGCCAAAUGAUGUACCCAGGCCAAGAAAUUGACAAGGCAGAUUCAUACAUGCCAUAUAUGAUUGAUUUCGGGUUAUCCCAAAAAUCUCCUUAUUCAUCAGUGAAGAAUCCAGCUUUCCACUUCUGGGGUCAACUGGCCGCACUCUUGCUAAGAUCAACUCGAGCAAAAAAUGCUCGGCAACCCGAUGACAUUGAGUACACUUCGCUGACUUGUGCAAGUUUACUACUGUCAUUUGCAGUUGGAUCUUCAGCAGACAUUGAACAACAAUUCUACAUCGGAGAGGACAAAUACACAACAGAGAAAGAUGACAGUCUCAAGAAAUCUGAUGUCCCACCGAAAGGAAGAAAUGUUGUAGACUGGCUCGGUUGGUAUGAUGACAAUGGAGGGAAACCCACACCGGAUAUGCUCAACUUCGCACGAAGAGCAGUCAGUUCGUUGCAGUCACUCCGCGAGAAAACAAUUGGCAAAUACGCAAAAGUAGAAUUUGACAAAUGA(SEQ ID NO:16)。

AUGAGUUCUCAAGUAUUCUGCAUUUCUACAGGUCAAACAGUCUCCGUAUGCCUUCCAGCAAAUGAAGACCCAGUAGAGUUUCCAGGGGCGUUCUUCACUCCAAAUGCUAGGAAACCGACGGUGUACAUAAAGAAGGAGACAGAUCUGAGUCUUUUGAGGAGUCAUGUUUAUGACGGGAUUAAGGAUGGUUCUGUGACAGUGUCCCAGAUAAAUUCUUAUCUAUACAUGGUUUUGAAAGAUAUCCGGGAGAAACCAGAUAAAAAUUGGACUUCUUUCGGAGUCGAAUUGGGAAAAAAGAAUGAGCCAAUGGGAAUUUUCGAUCUGUUGAAUGUUGAAGAUGUUAAGGGGAAAGAAUUGGACAAAAAAGGGCAAGACACCCGACUGCCCGGAGAUGAUCUAUGGCUUCCAACGCUGAUCCUCGGUCUUUAUCGCGUGUCCAGGGCGACACAGGUCGAAUACAAGAAGACUCUGAUGACCAAUCUAUAUGCUCAGUGUAAAUUGCGCACGAAGGAUGCAGAGGAGAUUGUGGAUGAAACAGCAGAAUUCUUCAAUGCUUGGGCAAAUGACUCCAACUUCACAAAAAUCGUAGCUGCUGUGGAUAUGUACUUCCACCACUUCAAGAAGAGUGAUCAUGCUCCUAUUCGUUUCGGAACAAUUGUGUCUAGAUUCAAAGACUGUGCAGCAUUGUCCACUCUGUCGCACCUCCAAAAGGUCACAGGGCUCCCCAUUGAGGAGGUCUUCACCUGGGUUUUCAAUAAAUCAGUACAGGAUGAUUUGUUGCGGAUGAUGACUCCUGGCCAGGAAAUUGACCAAGCUGAUUCCUAUAUGCCCUAUCUGAUCGACAUGGGCUUGUCCACGAAGUCUCCUUACUCUUCAACAAAAAAUCCCAGUUUCCAUUUCUGGGGGCAACUGACCGCAUUCUUGGUAAAAUCCGCAAGAGCAAAGAAUGCGUUAGUCCCAGUUGACAUCGCAUACCAUGAACUGACCACUGCUGCACUGCUUUUUGCCUAUGCAAUUGGCAGAUCGUCUGAGCUGGAGCAGAGAUUCGUGCUUAACGGGAAGAAGUUCACCAAGGAGAAGGACUCUCGAGAUGACAAUGACACUACUCCGCCAUCGGAGAGAAAUGUUGUUGUUUGGCUGGCCUGGUGGGAGGACAUUAAGCAUGAAAUUACUCCGGACAUGAAGGCCUUUGCCAAGAGAGCAGUGGAAAGGGUAGGAGAUAUUCGAGUGAACUCAGUUGCAGAGUAUGCCAGGAAACUCUUUGCAUGA(SEQ ID NO:22)。

AUGGAUGCCGACAAGAUUGUAUUCAAAGUCAAUAAUCAGGUGGUCUCUUUGAAGCCUGAGAUUAUCGUGGAUCAAUAUGAGUACAAGUACCCUGCCAUCAAAGAUUUGAAAAAGCCCUGUAUAACCCUAGGAAAGGCUCCCGAUUUAAAUAAAGCAUACAAGUCAGUUUUGUCAGGCAUGAGCGCCGCCAAACUUAAUCCUGACGAUGUAUGUUCCUAUUUGGCAGCGGCAAUGCAGUUUUUUGAGGGGACAUGUCCGGAAGACUGGACCAGCUAUGGAAUUGUGAUUGCACGAAAAGGAGAUAAGAUCACCCCAGGUUCUCUGGUGGAGAUAAAACGUACUGAUGUAGAAGGGAAUUGGGCUCUGACAGGAGGCAUGGAACUGACAAGAGACCCCACUGUCCCUGAGCAUGCGUCCUUAGUCGGUCUUCUCUUGAGUCUGUAUAGGUUGAGCAAAAUAUCCGGGCAAAACACUGGUAACUAUAAGACAAACAUUGCAGACAGGAUAGAGCAGAUUUUUGAGACAGCCCCUUUUGUUAAAAUCGUGGAACACCAUACUCUAAUGACAACUCACAAAAUGUGUGCUAAUUGGAGUACUAUACCAAACUUCAGAUUUUUGGCCGGAACCUAUGACAUGUUUUUCUCCCGGAUUGAGCAUCUAUAUUCAGCAAUCAGAGUGGGCACAGUUGUCACUGCUUAUGAAGACUGUUCAGGACUGGUAUCAUUUACUGGGUUCAUAAAACAAAUCAAUCUCACCGCUAGAGAGGCAAUACUAUAUUUCUUCCACAAGAACUUUGAGGAAGAGAUAAGAAGAAUGUUUGAGCCAGGGCAGGAGACAGCUGUUCCUCACUCUUAUUUCAUCCACUUCCGUUCACUAGGCUUGAGUGGGAAAUCUCCUUAUUCAUCAAAUGCUGUUGGUCACGUGUUCAAUCUCAUUCACUUUGUAGGAUGCUAUAUGGGUCAAGUCAGAUCCCUAAAUGCAACGGUUAUUGCUGCAUGUGCUCCUCAUGAAAUGUCUGUUCUAGGGGGCUAUCUGGGAGAGGAAUUCUUCGGGAAAGGGACAUUUGAAAGAAGAUUCUUCAGAGAUGAGAAAGAACUUCAAGAAUACGAGGCGGCUGAACUGACAAAGACUGACGUAGCACUGGCAGAUGAUGGAACUGUCAACUCUGACGACGAGGACUACUUUUCAGGUGAAACCAGAAGUCCGGAGGCUGUUUAUACUCGAAUCAUGAUGAAUGGAGGUCGACUAAAGAGAUCUCACAUACGGAGAUAUGUCUCAGUCAGUUCCAAUCAUCAAGCCCGUCCAAACUCAUUCGCCGAGUUUCUAAACAAGACAUAUUCGAGUGACUCAUAA(SEQ IDNO:28)。

根据本发明的实施例，所述第二RNA序列具有SEQ ID NO：5、11、17、23或29所示碱基序列。

AUGGAUAAUCUCACAAAAGUUCGUGAGUAUCUCAAGUCCUAUUCUCGUCUGGAUCAGGCGGUAGGAGAGAUAGAUGAGAUCGAAGCACAACGAGCUGAAAAGUCCAAUUAUGAGUUGUUCCAAGAGGAUGGAGUGGAAGAGCAUACUAAGCCCUCUUAUUUUCAGGCAGCAGAUGAUUCUGACACAGAAUCUGAACCAGAAAUUGAAGACAAUCAAGGCUUGUAUGCACCAGAUCCAGAAGCUGAGCAAGUUGAAGGCUUUAUACAGGGGCCUUUAGAUGACUAUGCAGAUGAGGAAGUGGAUGUUGUAUUUACUUCGGACUGGAAACAGCCUGAGCUUGAAUCUGACGAGCAUGGAAAGACCUUACGGUUGACAUCGCCAGAGGGUUUAAGUGGAGAGCAGAAAUCCCAGUGGCUUUCGACGAUUAAAGCAGUCGUGCAAAGUGCCAAAUACUGGAAUCUGGCAGAGUGCACAUUUGAAGCAUCGGGAGAAGGGGUCAUUAUGAAGGAGCGCCAGAUAACUCCGGAUGUAUAUAAGGUCACUCCAGUGAUGAACACACAUCCGUCCCAAUCAGAAGCAGUAUCAGAUGUUUGGUCUCUCUCAAAGACAUCCAUGACUUUCCAACCCAAGAAAGCAAGUCUUCAGCCUCUCACCAUAUCCUUGGAUGAAUUGUUCUCAUCUAGAGGAGAGUUCAUCUCUGUCGGAGGUGACGGACGAAUGUCUCAUAAAGAGGCCAUCCUGCUCGGCCUGAGAUACAAAAAGUUGUACAAUCAGGCGAGAGUCAAAUAUUCUCUGUAG(SEQ ID NO：5)。

AUGGAUAAUCUCACAAAAGUUCGUGAGUAUCUCAAGUCCUAUUCUCGUCUGGAUCAGGCGGUAGGAGAGAUAGAUGAGAUCGAAGCACAACGAGCUGAAAAGUCCAAUUAUGAGUUGUUCCAAGAGGAUGGAGUGGAAGAGCAUACUAAGCCCUCUUAUUUUCAGGCAGCAGAUGAUUCUGACACAGAAUCUGAACCAGAAAUUGAAGACAAUCAAGGUUUGUAUGCACAGGAUCCAGAAGCUGAGCAAGUUGAAGGCUUUAUACAGGGGCCUUUAGAUGACUAUGCAGAUGAGGAAGUGGAUGUUGUAUUUACUUCGGACUGGAAACCACCUGAGCUUGAAUCUGACGAGCAUGGAAAGACCUUACGGUUGACAUCGCCAGAGGGUUUAAGUGGAGAGCAGAAAUCCCAGUGGCUUUCGACGAUUAAAGCAGUCGUGCAAAGUGCCAAAUACUGGAAUCUGGCAGAGUGCACAUUUGAAGCAUCGGGAGAAGGGGUCAUUAUGAAGGAGCGCCAGAUAACUCCGGAUGUAUAUAAGGUCACUCCAGUGAUGAACACACAUCCGUCCCAAUCAGAAGCAGUAUCAGAUGUUUGGUCUCUCUCAAAGACAUCCAUGACUUUCCAACCCAAGAAAGCAAGUCUUCAGCCUCUCACCAUAUCCUUGGAUGAAUUGUUCUCAUCUAGAGGAGAGUUCAUCUCUGUCGGAGGUGACGGACGAAUGUCUCAUAAAGAGGCCAUCCUGCUCGGCCUGAGAUACAAAAAGUUGUACAAUCAGGCGAGAGUCAAAUAUUCUCUGUAG(SEQ ID NO:11)。

AUGGACAGUGUUGAUAGGCUCAAGACUUACUUAGCCACUUAUGAUAAUUUGGAUUCUGCCUUGCAGGAUGCCAAUGAAUCUGAGGAAAGACGAGAGGAUAAAUAUCUCCAAGACCUCUUCAUCGAAGAUCAAGGAGAUAAACCAACUCCGUCAUAUUAUCAGGAAGAAGAAUCGUCAGAUUCAGAUACUGAUUAUAAUGCUGAACAUCUUACGAUGCUGUCACCGGAUGAAAGAAUAGACAAGUGGGAAGAAGAUUUGCCUGAAUUAGAAAAGAUUGAUGAUGAUAUACCGGUGACCUUUUCUGAUUGGACACAGCCUGUAAUGAAGGAAAAUGGGGGAGAGAAAUCAUUGUCUCUGUUCCCUCCAGUCGGGUUAACAAAGAUUCAAACAGAACAAUGGAAAAAAACCAUUGAGGCGGUUUGUGAGAGUUCAAAAUAUUGGAAUUUAUCAGAAUGCCAAAUUCUUAACUUGGAAGACAGCCUCACUCUCAAAGGCCGAUUGAUGACUCCUGAUUGUAGUUCUUCAGUAAAAUCUCAAAAUUCUGUCCGGAGGUCAGAACCUCUCUACUCCUCUCAUUCUCCAGGUCCCCCACUCAAGGUAUCAGAGUCCAUCAAUUUAUGGGAUUUAAAGUCCACUGAAGUACAAUUGAUCUCCAAGAGAGCCGGAGUUAAGGACAUGACAGUCAAAUUGACAGACUUCUUUGGAAGUGAGGAAGAGUAUUAUUCAGUAUGCCCAGAAGGGGCGCCAGACUUGAUGGGAGCUAUCAUCAUGGGACUGAAGUACAAGAAACUCUUCAAUCAGGCAAGAAUGAAAUAUCGUCUUUAA(SEQID NO:17)。

AUGGAAGACUCUCAACUAUAUCAAGCUCUCAAGAAUUACCCAAAGCUUCAGGACACUCUCGACUCAAUUGAGAAUCUUGAAGAUGACACAAAGUCAGAGCCAUCUGAGUGUGGUUCCCCCACAGAGAGAGGAAUUCCCAGCUAUUACCUGGCUGAAGAAUUGGAUGAAUGUGAGGAAGAAGAUUCGGAAGAUGAUGAUGACAAUUUACCUACUGAGAUACCGGAUCCUCCUACUGUUGACAUGCUGGAGGCAAUUAUGGAAGAUGAGAUUGAUGAUACGGCUUACCAGGUACAUUUUGAGGCCAAACAGACUUGGAAACCGGUGAUAGAGACAGGGGGAAAUGAAAGAGGUAAGUUCACCCUUUCAGUCCCUCAGAAUCUGAGCGCUCUACAACUUCUCCAGUGGGAGACAGGGAUUCAUGCAUUGGCGGAGAGACUUGGGGGAUGCAGACUGCUGCAGAUCAGCACACGGGGAACGAGGGAUGGGAUAGAGUUCACUGUAAGAGAAACGCCUUGCGUUUCACCUGCUUCUGAUCCAAUCCCUUCCACAUCCAGAUCCAGUUCAAUUGCUUCCAAUGUCUCUACCAGACAAACCGAAUCACCUGGCUCCAAAUCCAAUACAAGUCUCGGGAUACCAGAAGUCCCGGCUAAUUUGAUCGACAUAGGAGCAAUUGAUAAAGAAUUCAUCCUGGCAGCUAUCUCUCCGUCUGACCCACCUUAUAAGAACACUCUGAGAAACCUCUUCGGGUCCGGAGAUUCCUUUGAACAGUACAACCAGACAGGAAUUUACUCUCUUAAGGAACUCGUCGUUGCUGGAUUGAAGAGAAAAGGAAUUUAUAAUAGAAUCAGAAUCAGAUGUCAUCUUGAGCCCCAGUUCAAUUGA(SEQ ID NO:23)。

AUGAGCAAGAUCUUUGUCAAUCCUAGUGCUAUUAGAGCCGGUCUGGCCGAUCUUGAGAUGGCUGAAGAAACUGUUGAUCUGAUCAAUAGAAAUAUCGAAGACAAUCAGGCUCAUCUCCAAGGGGAACCCAUAGAGGUGGACAAUCUCCCUGAGGAUAUGGGGCGACUUCACCUGGAUGAUGGAAAAUCGCCCAACCAUGGUGAGAUAGCCAAGGUGGGAGAAGGCAAGUAUCGAGAGGACUUUCAGAUGGAUGAAGGAGAGGAUCCUAGCUUCCUGUUCCAGUCAUACCUGGAAAAUGUUGGAGUCCAAAUAGUCAGACAAAUGAGGUCAGGAGAGAGAUUUCUCAAGAUAUGGUCACAGACCGUAGAAGAGAUUAUAUCCUAUGUCGCGGUCAACUUUCCCAACCCUCCAGGAAAGUCUUCAGAGGAUAAAUCAACCCAGACUACUGGCCGAGAGCUCAAGAAGGAGACAACACCCACUCCUUCUCAGAGAGAAAGCCAAUCAUCGAAAGCCAGGAUGGCGGCUCAAAUUGCUUCUGGCCCUCCAGCCCUUGAAUGGUCGGCUACCAAUGAAGAGGAUGAUCUAUCAGUGGAGGCUGAGAUCGCUCACCAGAUUGCAGAAAGUUUCUCCAAAAAAUAUAAGUUUCCCUCUCGAUCCUCAGGGAUACUCUUGUAUAAUUUUGAGCAAUUGAAAAUGAACCUUGAUGAUAUAGUUAAAGAGGCAAAAAAUGUACCAGGUGUGACCCGUUUAGCCCAUGACGGGUCCAAACUCCCCCUAAGAUGUGUACUGGGAUGGGUCGCUUUGGCCAACUCUAAGAAAUUCCAGUUGUUAGUCGAAUCCGACAAGCUGAGUAAAAUCAUGCAAGAUGACUUGAAUCGCUAUACAUCUUGCUAA(SEQ ID NO:29)。

根据本发明的实施例，所述第三RNA序列具有SEQ ID NO：6、12、18、24或30所示碱基序列。

AUGGAAGUCCACGAUUUUGAGACCGACGAGUUCAAUGAUUUCAAUGAAGAUGACUAUGCCACAAGAGAAUUCCUGAAUCCCGAUGAGCGCAUGACGUACUUGAAUCAUGCUGAUUACAACCUGAAUUCUCCUCUAAUUAGUGAUGAUAUUGACAAUUUAAUCAGGAAAUUCAAUUCUCUUCCAAUUCCCUCGAUGUGGGAUAGUAAGAACUGGGAUGGAGUUCUUGAGAUGUUAACGUCAUGUCAAGCCAAUCCCAUCCCAACAUCUCAGAUGCAUAAAUGGAUGGGAAGUUGGUUAAUGUCUGAUAAUCAUGAUGCCAGUCAAGGGUAUAGUUUUUUACAUGAAGUGGACAAAGAGGCAGAAAUAACAUUUGACGUGGUGGAGACCUUCAUCCGCGGCUGGGGCAACAAACCAAUUGAAUACAUCAAAAAGGAAAGAUGGACUGACUCAUUCAAAAUUCUCGCUUAUUUGUGUCAAAAGUUUUUGGACUUACACAAGUUGACAUUAAUCUUAAAUGCUGUCUCUGAGGUGGAAUUGCUCAACUUGGCGAGGACUUUCAAAGGCAAAGUCAGAAGAAGUUCUCAUGGAACGAACAUAUGCAGGAUUAGGGUUCCCAGCUUGGGUCCUACUUUUAUUUCAGAAGGAUGGGCUUACUUCAAGAAACUUGAUAUUCUAAUGGACCGAAACUUUCUGUUAAUGGUCAAAGAUGUGAUUAUAGGGAGGAUGCAAACGGUGCUAUCCAUGGUAUGUAGAAUAGACAACCUGUUCUCAGAGCAAGACAUCUUCUCCCUUCUAAAUAUCUACAGAAUUGGAGAUAAAAUUGUGGAGAGGCAGGGAAAUUUUUCUUAUGACUUGAUUAAAAUGGUGGAACCGAUAUGCAACUUGAAGCUGAUGAAAUUAGCAAGAGAAUCAAGGCCUUUAGUCCCACAAUUCCCUCAUUUUGAAAAUCAUAUCAAGACUUCUGUUGAUGAAGGGGCAAAAAUUGACCGAGGUAUAAGAUUCCUCCAUGAUCAGAUAAUGAGUGUGAAAACAGUGGAUCUCACACUGGUGAUUUAUGGAUCGUUCAGACAUUGGGGUCAUCCUUUUAUAGAUUAUUACACUGGACUAGAAAAAUUACAUUCCCAAGUAACCAUGAAGAAAGAUAUUGAUGUGUCAUAUGCAAAAGCACUUGCAAGUGAUUUAGCUCGGAUUGUUCUAUUUCAACAGUUCAAUGAUCAUAAAAAGUGGUUCGUGAAUGGAGACUUGCUCCCUCAUGAUCAUCCCUUUAAAAGUCAUGUUAAAGAAAAUACAUGGCCCACAGCUGCUCAAGUUCAAGAUUUUGGAGAUAAAUGGCAUGAACUUCCGCUGAUUAAAUGUUUUGAAAUACCCGACUUACUAGACCCAUCGAUAAUAUACUCUGACAAAAGUCAUUCAAUGAAUAGGUCAGAGGUGUUGAAACAUGUCCGAAUGAAUCCGAACACUCCUAUCCCUAGUAAAAAGGUGUUGCAGACUAUGUUGGACACAAAGGCUACCAAUUGGAAAGAAUUUCUUAAAGAGAUUGAUGAGAAGGGCUUAGAUGAUGAUGAUCUAAUUAUUGGUCUUAAAGGAAAGGAGAGGGAACUGAAGUUGGCAGGUAGAUUUUUCUCCCUAAUGUCUUGGAAAUUGCGAGAAUACUUUGUAAUUACCGAAUAUUUGAUAAAGACUCAUUUCGUCCCUAUGUUUAAAGGCCUGACAAUGGCGGACGAUCUAACUGCAGUCAUUAAAAAGAUGUUAGAUUCCUCAUCCGGCCAAGGAUUGAAGUCAUAUGAGGCAAUUUGCAUAGCCAAUCACAUUGAUUACGAAAAAUGGAAUAACCACCAAAGGAAGUUAUCAAACGGCCCAGUGUUCCGAGUUAUGGGCCAGUUCUUAGGUUAUCCAUCCUUAAUCGAGAGAACUCAUGAAUUUUUUGAGAAAAGUCUUAUAUACUACAAUGGAAGACCAGACUUGAUGCGUGUUCACAACAACACACUGAUCAAUUCAACCUCCCAACGAGUUUGUUGGCAAGGACAAGAGGGUGGACUGGAAGGUCUACGGCAAAAAGGAUGGAGUAUCCUCAAUCUACUGGUUAUUCAAAGAGAGGCUAAAAUCAGAAACACU

GCUGUCAAAGUCUUGGCACAAGGUGAUAAUCAAGUUAUUUGCACACAGUAUAAAACGAAGAAAUCGAGAAACGUUGUAGAAUUACAGGGUGCUCUCAAUCAAAUGGUUUCUAAUAAUGAGAAAAUUAUGACUGCAAUCAAAAUAGGGACAGGGAAGUUAGGACUUUUGAUAAAUGACGAUGAGACUAUGCAAUCUGCAGAUUACUUGAAUUAUGGAAAAAUACCGAUUUUCCGUGGAGUGAUUAGAGGGUUAGAGACCAAGAGAUGGUCACGAGUGACUUGUGUCACCAAUGACCAAAUACCCACUUGUGCUAAUAUAAUGAGCUCAGUUUCCACAAAUGCUCUCACCGUAGCUCAUUUUGCUGAGAACCCAAUCAAUGCCAUGAUACAGUACAAUUAUUUUGGGACAUUUGCUAGACUCUUGUUGAUGAUGCAUGAUCCUGCUCUUCGUCAAUCAUUGUAUGAAGUUCAAGAUAAGAUACCGGGCUUGCACAGUUCUACUUUCAAAUACGCCAUGUUGUAUUUGGACCCUUCCAUUGGAGGAGUGUCGGGCAUGUCUUUGUCCAGGUUUUUGAUUAGAGCCUUCCCAGAUCCCGUAACAGAAAGUCUCUCAUUCUGGAGAUUCAUCCAUGUACAUGCUCGAAGUGAGCAUCUGAAGGAGAUGAGUGCAGUAUUUGGAAACCCCGAGAUAGCCAAGUUUCGAAUAACUCACAUAGACAAGCUAGUAGAAGAUCCAACCUCUCUGAACAUCGCUAUGGGAAUGAGUCCAGCGAACUUGUUAAAGACUGAGGUUAAAAAAUGCUUAAUCGAAUCAAGACAAACCAUCAGGAACCAGGUGAUUAAGGAUGCAACCAUAUAUUUGUAUCAUGAAGAGGAUCGGCUCAGAAGUUUCUUAUGGUCAAUAAAUCCUCUGUUCCCUAGAUUUUUAAGUGAAUUCAAAUCAGGCACUUUUUUGGGAGUCGCAGACGGGCUCAUCAGUCUAUUUCAAAAUUCUCGUACUAUUCGGAACUCCUUUAAGAAAAAGUAUCAUAGGGAAUUGGAUGAUUUGAUUGUGAGGAGUGAGGUAUCCUCUUUGACACAUUUAGGGAAACUUCAUUUGAGAAGGGGAUCAUGUAAAAUGUGGACAUGUUCAGCUACUCAUGCUGACACAUUAAGAUACAAAUCCUGGGGCCGUACAGUUAUUGGGACAACUGUACCCCAUCCAUUAGAAAUGUUGGGUCCACAACAUCGAAAAGAGACUCCUUGUGCACCAUGUAACACAUCAGGGUUCAAUUAUGUUUCUGUGCAUUGUCCAGACGGGAUCCAUGACGUCUUUAGUUCACGGGGACCAUUGCCUGCUUAUCUAGGGUCUAAAACAUCUGAAUCUACAUCUAUUUUGCAGCCUUGGGAAAGGGAAAGCAAAGUCCCACUGAUUAAAAGAGCUACACGUCUUAGAGAUGCUAUCUCUUGGUUUGUUGAACCCGACUCUAAACUAGCAAUGACUAUACUUUCUAACAUCCACUCUUUAACAGGCGAAGAAUGGACCAAAAGGCAGCAUGGGUUCAAAAGAACAGGGUCUGCCCUUCAUAGGUUUUCGACAUCUCGGAUGAGCCAUGGUGGGUUCGCAUCUCAGAGCACUGCAGCAUUGACCAGGUUGAUGGCAACUACAGACACCAUGAGGGAUCUGGGAGAUCAGAAUUUCGACUUUUUAUUCCAAGCAACGUUGCUCUAUGCUCAAAUUACCACCACUGUUGCAAGAGACGGAUGGAUCACCAGUUGUACAGAUCAUUAUCAUAUUGCCUGUAAGUCCUGUUUGAGACCCAUAGAAGAGAUCACCCUGGACUCAAGUAUGGACUACACGCCCCCAGAUGUAUCCCAUGUGCUGAAGACAUGGAGGAAUGGGGAAGGUUCGUGGGGACAAGAGAUAAAACAGAUCUAUCCUUUAGAAGGGAAUUGGAAGAAUUUAGCACCUGCUGAGCAAUCCUAUCAAGUCGGCAGAUGUAUAGGUUUUCUAUAUGGAGACUUGGCGUAUAGAAAAUCUACUCAUGCCGAGGACAGUUCUCUAUUUCCUCUAUCUAUACAAGGUCGUAUUAGAGGUCGAGGUUUCUUAAAAGGGUUGCUAGACGGAUUAAUGAGAGCAAGUUGCUGCCAAGUAAUACACCGGAGAAGUCUGGCUCAUUUGAAGAGGCCGGCCAACGCAGUGUACGGAGGUUUGAUUUACUUGAUUGAUAAAUUGAGUGUAUCACCUCCAUUCCUUUCUCUUACUAGAUCAGGACCUAUUAGAGACGAAUUAGAAACGAUUCCCCACAAGAUCCCAACCUCCUAUCCGACAAGCAACCGUGAUAUGGGGGUGAUUGUCAGAAAUUACUUCAAAUACCAAUGCCGUCUAAUUGAAAAGGGAAAAUACAGAUCACAUUAUUCACAAUUAUGGUUAUUCUCAGAUGUCUUAUCCAUAGACUUCAUUGGACCAUUCUCUAUUUCCACCACCCUCUUGCAAAUCCUAUACAAGCCAUUUUUAUCUGGGAAAGAUAAGAAUGAGUUGAGAGAGCUGGCAAAUCUUUCUUCAUUGCUAAGAUCAGGAGAGGGGUGGGAAGACAUACAUGUGAAAUUCUUCACCAAGGACAUAUUAUUGUGUCCAGAGGAAAUCAGACAUGCUUGCAAGUUCGGGAUUGCUAAGGAUAAUAAUAAAGACAUGAGCUAUCCCCCUUGGGGAAGGGAAUCCAGAGGGACAAUUACAACAAUCCCUGUUUAUUAUACGACCACCCCUUACCCAAAGAUGCUAGAGAUGCCUCCAAGAAUCCAAAAUCCCCUGCUGUCCGGAAUCAGGUUGGGCCAAUUACCAACUGGCGCUCAUUAUAAAAUUCGGAGUAUAUUACAUGGAAUGGGAAUCCAUUACAGGGACUUCUUGAGUUGUGGAGACGGCUCCGGAGGGAUGACUGCUGCAUUACUACGAGAAAAUGUGCAUAGCAGAGGAAUAUUCAAUAGUCUGUUAGAAUUAUCAGGGUCAGUCAUGCGAGGCGCCUCUCCUGAGCCCCCCAGUGCCCUAGAAACUUUAGGAGGAGAUAAAUCGAGAUGUGUAAAUGGUGAAACAUGUUGGGAAUAUCCAUCUGACUUAUGUGACCCAAGGACUUGGGACUAUUUCCUCCGACUCAAAGCAGGCUUGGGGCUUCAAAUUGAUUUAAUUGUAAUGGAUAUGGAAGUUCGGGAUUCUUCUACUAGCCUGAAAAUUGAGACGAAUGUUAGAAAUUAUGUGCACCGGAUUUUGGAUGAGCAAGGAGUUUUAAUCUACAAGACUUAUGGAACAUAUAUUUGUGAGAGCGAAAAGAAUGCAGUAACAAUCCUUGGUCCCAUGUUCAAGACGGUCGACUUAGUUCAAACAGAAUUUAGUAGUUCUCAAACGUCUGAAGUAUAUAUGGUAUGUAAAGGUUUGAAGAAAUUAAUCGAUGAACCCAAUCCCGAUUGGUCUUCCAUCAAUGAAUCCUGGAAAAACCUGUACGCAUUCCAGUCAUCAGAACAGGAAUUUGCCAGAGCAAAGAAGGUUAGUACAUACUUUACCUUGACAGGUAUUCCCUCCCAAUUCAUUCCUGAUCCUUUUGUAAACAUUGAGACUAUGCUACAAAUAUUCGGAGUACCCACGGGUGUGUCUCAUGCGGCUGCCUUAAAAUCAUCUGAUAGACCUGCAGAUUUAUUGACCAUUAGCCUUUUUUAUAUGGCGAUUAUAUCGUAUUAUAACAUCAAUCAUAUCAGAGUAGGACCGAUACCUCCGAACCCCCCAUCAGAUGGAAUUGCACAAAAUGUGGGGAUCGCUAUAACUGGUAUAAGCUUUUGGCUGAGUUUGAUGGAGAAAGACAUUCCACUAUAUCAACAGUGUUUAGCAGUUAUCCAGCAAUCAUUCCCGAUUAGGUGGGAGGCUGUUUCAGUAAAAGGAGGAUACAAGCAGAAGUGGAGUACUAGAGGUGAUGGGCUCCCAAAAGAUACCCGAAUUUCAGACUCCUUGGCCCCAAUCGGGAACUGGAUCAGAUCUCUGGAAUUGGUCCGAAACCAAGUUCGUCUAAAUCCAUUCAAUGAGAUCUUGUUCAAUCAGCUAUGUCGUACAGUGGAUAAUCAUUUGAAAUGGUCAAAUUUGCGAAGAAACACAGGAAUGAUUGAAUGGAUCAAUAGACGAAUUUCAAAAGAAGACCGGUCUAUACUGAUGUUGAAGAGUGACCUACACGAGGAAAACUCUUGGAGAGAUUAA(SEQ ID NO：6)。

AUGGAAGUCCACGAUUUUGAGACCGACGAGUUCAAUGAUUUCAAUGAAGAUGACUAUGCCACAAGAGAAUUCCUGAAUCCCGAUGAGCGCAUGACGUACUUGAAUCAUGCUGAUUACAAUUUGAAUUCUCCUCUAAUUAGUGAUGAUAUUGACAAUUUGAUCAGGAAAUUCAAUUCUCUUCCGAUUCCCUCGAUGUGGGAUAGUAAGAACUGGGAUGGAGUUCUUGAGAUGUUAACAUCAUGUCAAGCCAAUCCCAUCUCAACAUCUCAGAUGCAUAAAUGGAUGGGAAGUUGGUUAAUGUCUGAUAAUCAUGAUGCCAGUCAAGGGUAUAGUUUUUUACAUGAAGUGGACAAAGAGGCAGAAAUAACAUUUGACGUGGUGGAGACCUUCAUCCGCGGCUGGGGCAACAAACCAAUUGAAUACAUCAAAAAGGAAAGAUGGACUGACUCAUUCAAAAUUCUCGCUUAUUUGUGUCAAAAGUUUUUGGACUUACACAAGUUGACAUUAAUCUUAAAUGCUGUCUCUGAGGUGGAAUUGCUCAACUUGGCGAGGACUUUCAAAGGCAAAGUCAGAAGAAGUUCUCAUGGAACGAACAUAUGCAGGAUUAGGGUUCCCAGCUUGGGUCCUACUUUUAUUUCAGAAGGAUGGGCUUACUUCAAGAAACUUGAUAUUCUAAUGGACCGAAACUUUCUGUUAAUGGUCAAAGAUGUGAUUAUAGGGAGGAUGCAAACGGUGCUAUCCAUGGUAUGUAGAAUAGACAACCUGUUCUCAGAGCAAGACAUCUUCUCCCUUCUAAAUAUCUACAGAAUUGGAGAUAAAAUUGUGGAGAGGCAGGGAAAUUUUUCUUAUGACUUGAUUAAAAUGGUGGAACCGAUAUGCAACUUGAAGCUGAUGAAAUUAGCAAGAGAAUCAAGGCCUUUAGUCCCACAAUUCCCUCAUUUUGAAAAUCAUAUCAAGACUUCUGUUGAUGAAGGGGCAAAAAUUGACCGAGGUAUAAGAUUCCUCCAUGAUCAGAUAAUGAGUGUGAAAACAGUGGAUCUCACACUGGUGAUUUAUGGAUCGUUCAGACAUUGGGGUCAUCCUUUUAUAGAUUAUUACACUGGACUAGAAAAAUUACAUUCCCAAGUAACCAUGAAGAAAGAUAUUGAUGUGUCAUAUGCAAAAGCACUUGCAAGUGAUUUAGCUCGGAUUGUUCUAUUUCAACAGUUCAAUGAUCAUAAAAAGUGGUUCGUGAAUGGAGACUUGCUCCCUCAUGAUCAUCCCUUUAAAAGUCAUGUUAAAGAAAAUACAUGGCCCACAGCUGCUCAAGUUCAAGAUUUUGGAGAUAAAUGGCAUGAACUUCCGCUGAUUAAAUGUUUUGAAAUACCCGACUUACUAGACCCAUCGAUAAUAUACUCUGACAAAAGUCAUUCAAUGAAUAGGUCAGAGGUGUUGAAACAUGUCCGAAUGAAUCCGAACACUCCUAUCCCUAGUAAAAAGGUGUUGCAGACUAUGUUGGACACAAAGGCUACCAAUUGGAAAGAAUUUCUUAAAGAGAUUGAUGAGAAGGGCUUAGAUGAUGAUGAUCUAAUUAUUGGUCUUAAAGGAAAGGAGAGGGAACUGAAGUUGGCAGGUAGAUUUUUCUCCCUAAUGUCUUGGAAAUUGCGAGAAUACUUUGUAAUUACCGAAUAUUUGAUAAAGACUCAUUUCGUCCCUAUGUUUAAAGGCCUGACAAUGGCGGACGAUCUAACUGCAGUCAUUAAAAAGAUGUUAGAUUCCUCAUCCGGCCAAGGAUUGAAGUCAUAUGAGGCAAUUUGCAUAGCCAAUCACAUUGAUUACGAAAAAUGGAAUAACCACCAAAGGAAGUUAUCAAACGGCCCAGUGUUCCGAGUUAUGGGCCAGUUCUUAGGUUAUCCAUCCUUAAUCGAGAGAACUCAUGAAUUUUUUGAGAAAAGUCUUAUAUACUACAAUGGAAGACCAGACUUGAUGCGUGUUCACAACAACACACUGAUCAAUUCAACCUCCCAACGAGUUUGUUGGCAAGGACAAGAGGGUGGACUGGAAGGUCUACGGCAAAAAGGAUGGACUAUCCUCAAUCUACUGGUUAUUCAAAGAGAGGCUAAAAUCAGAAACACUGCUGUCAAAGUCUUGGCACAAGGUGAUAAUCAAGUUAUUUGCACACAGUAUAAAACGAAGAAAUCGAGAAACGUUGUAGAAUUACAGGGUGCUCUCAAUCAAAUGGUUUCUAAUAAUGAGAAAAUUAUGACUGCAAUCAAAAUAGGGACAGGGAAGUUAGGACUUUUGAUAAAUGACGAUGAGACUAUGCAAUCUGCAGAUUACUUGAAUUAUGGAAAAAUACCGAUUUUCCGUGGAGUGAUUAGAGGGUUAGAGACCAAGAGAUGGUCACGAGUGACUUGUGUCACCAAUGACCAAAUACCCACUUGUGCUAAUAUAAUGAGCUCAGUUUCCACAAAUGCUCUCACCGUAGCUCAUUUUGCUGA

GAACCCAAUCAAUGCCAUGAUACAGUACAAUUAUUUUGGGACAUUUGCUAGACUCUUGUUGAUGAUGCAUGAUCCUGCUCUUCGUCAAUCAUUGUAUGAAGUUCAAGAUAAGAUACCGGGCUUGCACAGUUCUACUUUCAAAUACGCCAUGUUGUAUUUGGACCCUUCCAUUGGAGGAGUGUCGGGCAUGUCUUUGUCCAGGUUUUUGAUUAGAGCCUUCCCAGAUCCCGUAACAGAAAGUCUCUCAUUCUGGAGAUUCAUCCAUGUACAUGCUCGAAGUGAGCAUCUGAAGGAGAUGAGUGCAGUAUUUGGAAACCCCGAGAUAGCCAAGUUUCGAAUAACUCACAUAGACAAGCUAGUAGAAGAUCCAACCUCUCUGAACAUCGCUAUGGGAAUGAGUCCAGCGAACUUGUUAAAGACUGAGGUUAAAAAAUGCUUAAUCGAAUCAAGACAAACCAUCAGGAACCAGGUGAUUAAGGAUGCAACCAUAUAUUUGUAUCAUGAAGAGGAUCGGCUCAGAAGUUUCUUAUGGUCAAUAAAUCCUCUGUUCCCUAGAUUUUUAAGUGAAUUCAAAUCAGGCACUUUUUUGGGAGUCGCAGACGGGCUCAUCAGUCUAUUUCAAAAUUCUCGUACUAUUCGGAACUCCUUUAAGAAAAAGUAUCAUAGGGAAUUGGAUGAUUUGAUUGUGAGGAGUGAGGUAUCCUCUUUGACACAUUUAGGGAAACUUCAUUUGAGAAGGGGAUCAUGUAAAAUGUGGACAUGUUCAGCUACUCAUGCUGACACAUUAAGAUACAAAUCCUGGGGCCGUACAGUUAUUGGGACAACUGUACCCCAUCCAUUAGAAAUGUUGGGUCCACAACAUCGAAAAGAGACUCCUUGUGCACCAUGUAACACAUCAGGGUUCAAUUAUGUUUCUGUGCAUUGUCCAGACGGGAUCCAUGACGUCUUUAGUUCACGGGGACCAUUGCCUGCUUAUCUAGGGUCUAAAACAUCUGAAUCUACAUCUAUUUUGCAGCCUUGGGAAAGGGAAAGCAAAGUCCCACUGAUUAAAAGAGCUACACGUCUUAGAGAUGCUAUCUCUUGGUUUGUUGAACCCGACUCUAAACUAGCAAUGACUAUACUUUCUAACAUCCACUCUUUAACAGGCGAAGAAUGGACCAAAAGGCAGCAUGGGUUCAAAAGAACAGGGUCUGCCCUUCAUAGGUUUUCGACAUCUCGGAUGAGCCAUGGUGGGUUCGCAUCUCAGAGCACUGCAGCAUUGACCAGGUUGAUGGCAACUACAGACACCAUGAGGGAUCUGGGAGAUCAGAAUUUCGACUUUUUAUUCCAAGCAACGUUGCUCUAUGCUCAAAUUACCACCACUGUUGCAAGAGACGGAUGGAUCACCAGUUGUACAGAUCAUUAUCAUAUUGCCUGUAAGUCCUGUUUGAGACCCAUAGAAGAGAUCACCCUGGACUCAAGUAUGGACUACACGCCCCCAGAUGUAUCCCAUGUGCUGAAGACAUGGAGGAAUGGGGAAGGUUCGUGGGGACAAGAGAUAAAACAGAUCUAUCCUUUAGAAGGGAAUUGGAAGAAUUUAGCACCUGCUGAGCAAUCCUAUCAAGUCGGCAGAUGUAUAGGUUUUCUAUAUGGAGACUUGGCGUAUAGAAAAUCUACUCAUGCCGAGGACAGUUCUCUAUUUCCUCUAUCUAUACAAGGUCGUAUUAGAGGUCGAGGUUUCUUAAAAGGGUUGCUAGACGGAUUAAUGAGAGCAAGUUGCUGCCAAGUAAUACACCGGAGAAGUCUGGCUCAUUUGAAGAGGCCGGCCAACGCAGUGUACGGAGGUUUGAUUUACUUGAUUGAUAAAUUGAGUGUAUCACCUCCAUUCCUUUCUCUUACUAGAUCAGGACCUAUUAGAGACGAAUUAGAAACGAUUCCCCACAAGAUCCCAACCUCCUAUCCGACAAGCAACCGUGAUAUGGGGGUGAUUGUCAGAAAUUACUUCAAAUACCAAUGCCGUCUAAUUGAAAAGGGAAAAUACAGAUCACAUUAUUCACAAUUAUGGUUAUUCUCAGAUGUCUUAUCCAUAGACUUCAUUGGACCAUUCUCUAUUUCCACCACCCUCUUGCAAAUCCUAUACAAGCCAUUUUUAUCUGGGAAAGAUAAGAAUGAGUUGAGAGAGCUGGCAAAUCUUUCUUCAUUGCUAAGAUCAGGAGAGGGGUGGGAAGACAUACAUGUGAAAUUCUUCACCAAGGACAUAUUAUUGUGUCCAGAGGAAAUCAGACAUGCUUGCAAGUUCGGGAUUGCUAAGGAUAAUAAUAAAGACAUGAGCUAUCCCCCUUGGGGAAGGGAAUCCAGAGGGACAAUUACAACAAUCCCUGUUUAUUAUACGACCACCCCUUACCCAAAGAUGCUAGAGAUGCCUCCAAGAAUCCAAAAUCCCCUGCUGUCCGGAAUCAGGUUGGGCCAAUUACCAACUGGCGCUCAUUAUAAAAUUCGGAGUAUAUUACAUGGAAUGGGAAUCCAUUACAGGGACUUCUUGAGUUGUGGAGACGGCUCCGGAGGGAUGACUGCUGCAUUACUACGAGAAAAUGUGCAUAGCAGAGGAAUAUUCAAUAGUCUGUUAGAAUUAUCAGGGUCAGUCAUGCGAGGCGCCUCUCCUGAGCCCCCCAGUGCCCUAGAAACUUUAGGAGGAGAUAAAUCGAGAUGUGUAAAUGGUGAAACAUGUUGGGAAUAUCCAUCUGACUUAUGUGACCCAAGGACUUGGGACUAUUUCCUCCGACUCAAAGCAGGCUUGGGGCUUCAAAUUGAUUUAAUUGUAAUGGAUAUGGAAGUUCGGGAUUCUUCUACUAGCCUGAAAAUUGAGACGAAUGUUAGAAAUUAUGUGCACCGGAUUUUGGAUGAGCAAGGAGUUUUAAUCUACAAGACUUAUGGAACAUAUAUUUGUGAGAGCGAAAAGAAUGCAGUAACAAUCCUUGGUCCCAUGUUCAAGACGGUCGACUUAGUUCAAACAGAAUUUAGUAGUUCUCAAACGUCUGAAGUAUAUAUGGUAUGUAAAGGUUUGAAGAAAUUAAUCGAUGAACCCAAUCCCGAUUGGUCUUCCAUCAAUGAAUCCUGGAAAAACCUGUACGCAUUCCAGUCAUCAGAACAGGAAUUUGCCAGAGCAAAGAAGGUUAGUACAUACUUUACCUUGACAGGUAUUCCCUCCCAAUUCAUUCCUGAUCCUUUUGUAAACAUUGAGACUAUGCUACAAAUAUUCGGAGUACCCACGGGUGUGUCUCAUGCGGCUGCCUUAAAAUCAUCUGAUAGACCUGCAGAUUUAUUGACCAUUAGCCUUUUUUAUAUGGCGAUUAUAUCGUAUUAUAACAUCAAUCAUAUCAGAGUAGGACCGAUACCUCCGAACCCCCCAUCAGAUGGAAUUGCACAAAAUGUGGGGAUCGCUAUAACUGGUAUAAGCUUUUGGCUGAGUUUGAUGGAGAAAGACAUUCCACUAUAUCAACAGUGUUUAGCAGUUAUCCAGCAAUCAUUCCCGAUUAGGUGGGAGGCUGUUUCAGUAAAAGGAGGAUACAAGCAGAAGUGGAGUACUAGAGGUGAUGGGCUCCCAAAAGAUACCCGAACUUCAGACUCCUUGGCCCCAAUCGGGAACUGGAUCAGAUCUCUGGAAUUGGUCCGAAACCAAGUUCGUCUAAAUCCAUUCAAUGAGAUCUUGUUCAAUCAGCUAUGUCGUACAGUGGAUAAUCAUUUGAAAUGGUCAAAUUUGCGAAGAAACACAGGAAUGAUUGAAUGGAUCAAUAGACGAAUUUCAAAAGAAGACCGGUCUAUACUGAUGUUGAAGAGUGACCUACACGAGGAAAACUCUUGGAGAGAUUAA(SEQ ID NO:12)。

AUGGAUUUCGAUCUAAUAGAAGAUUCUGCUAACUGGGAAGAUGAUGAGUCGGACUUUUUUUUGCGAGACAUUCUAUCUCAAGAAGAUCAGAUGUCUUACUUAAACACCGCAGACUAUAAUUUGAAUUCUCCACUCAUUUCCGAUGACAUGGUUUAUAUAAUUAAAAGAAUGAAUCACGAGGAGGUUCCCCCCAUUUGGAGAUCUAAGGAGUGGGAUUCUCCUCUUGAUAUGCUUAGAGGUUGUCAAGCUCAACCCAUGUCUCACCAAGAGAUGCACAAUUGGUUUGGAACUUGGAUUCAAAAUGUUCAGCAUGAUUCCGCCCAACCCUUCACAUUCUUAAAAGAGGUGGACAAAGAGUCAGAAAUGACCUAUGACUUGGUCUCCACUUUUCUGAAGGGGUGGGUGGGGAAAGAUUACCCAUUCAAGUCAAAGAACAAAGAAAUUGAUAGUAUGGCAUUAGUCGGUCCUCUUUGUCAAAAAUUCCUUGAUUUGCAUAAAAUCACUCUGAUAUUAAAUGCAGUUUCUUUGGGGGAAACUAAAGAACUCCUAACCACUUUCAAGGGUAAAUACCGCAUGAGUUGUGAAAACAUUCCUAUUGCUCGCUUAAGACUUCCAAGUCUAGGCCCUGUAUUCAUGUGCAAAGGUUGGACUUAUAUCCACAAAGAAAGAGUGCUAAUGGAUCGAACUUCCUUCUCAUGUGUAAAAGAUGUGAUAAUAGGACGAAUGCAGACAUUUUUGUCUAUGAUCGGACGAUCGGAUAACAAGUUUAGUCCUGAUCAAAUCUAUACUCUGGCAAAUGUAUACAGAAUUGGAGAUAAAAUUUUAGAACAAUGCGGCAAUAAGGCUUAUGAUUUGAUUAAAAUGAUUGAACCCAUCUGCAAUCUGAAAAUGAUGGAAUUAGCUAGAUUGCAUCGUCCCAAGAUUCCGAAGUUCCCUCAUUUUGAAGAGCAUGUCAAAGGCUCAGUGCAAGAAUUGACACAAAGGUCCAAUAGAAUACAAACAUUGUAUGAUCUGAUUAUGUCUAUGAAGGAUGUUGACCUCGUAUUAGUGGUUUACGGAUCUUUCCGUCAUUGGGGUCACCCAUUCAUCGAUUAUUUUCAGGGAUUGGAAAAAUUGCACACUCAAGUGAAUAUGGAAAAGCAUAUUGACAAGGAGUACCCACAGCAAUUGGCCAGUGACUUAGCCAGACUAGUAUUAAAUAAACAAUUCAGCGAAUCAAAAAAAUGGUUUGUAGACCCUUCAAAAAUGUCUCCCAAACACCCUUUUUAUGAACAUGUUAUCAAUAAGACAUGGCCUACAGCAGCUAAAAUUCAAGACUUUGGAGACAAUUGGCACAAGCUCCCAUUGAUUCAAUGUUUUGAAAUACCAGACUUGAUAGAUCCAUCAGUGAUUUAUUCAGACAAAAGUCAUUCCAUGAACAAAAAGGAGGUGAUACAGCAUGUUCGGUCUAAGCCAAAUAUUCCCAUCCCAAGCAACAAAGUUCUGCAAACAAUGCUUACCAAUAGAGCAACAAAUUGGAAAGCCUUUCUGAAAGAUAUUGACGAAAAUGGACUGGAUGAUGACGAUUUAAUAAUCGGAUUAAAGGGAAAAGAAAGAGAAUUGAAAAUAGCUGGGAGAUUCUUCUCAUUAAUGUCAUGGAGAUUAAGGGAAUAUUUUGUGAUCACAGAGUACCUAAUCAAGACAUACUACGUUCCCUUGUUUAAGGGGUUAACUAUGGCAGAUGACCUCACAUCGGUGAUCAAGAAGAUGAUGGACAGUUCAUCAGGUCAAGGGCUUGAUGAUUAUUCAUCUGUUUGUUUAGCCAACCAUAUCGAUUAUGAGAAGUGGAAUAACCAUCAGAGAAAGGAAUCGAAUGGUCCCAUCUUCAGAGUCAUGGGACAGUUUCUAGGUUAUCCAUCACUUAUAGAGAGAACUCAUGAGUUUUUUGAGAAGAGCUUAAUUUACUAUAACGGGCGUCCAGAUUUGAUGACAAUUCGAAAUGGUACAUUGUGUAAUUCCACGAAGCACCGAGUUUGCUGGAAUGGGCAGAAAGGAGGAUUGGAAGGACUAAGACAAAAGGGAUGGAGUAUUGUGAAUCUCUUGGUUAUUCAAAGGGAAGCUAAGAUCCGAAACACAGCAGUGAAGGUCUUAGCACAGGGAGAUAAUCAGGUUAUAUGCACUCAAUACAAGACAAAGAAGACUAGAUCAGAGUUAGAGUUGAGAGCAGUCUUACAUCAGAUGGCCGGGAAUAAUAACAAAAUUAUGGAAGAAAUUAAGAGAGGUACAGAAAAGUUGGGCCUAAUCAUAAAUGAUGAUGAGACUAUGCAAUCAGCAGACUAUUUGAACUAUGGGAAAAUUCCAAUUUUCCGAGGAGUCAUCAGAGGACUAGAAACAAAGAGAUGGUCACGGGUGACAUGCGUCACAAAUGAUCAAAUCCCGACGUGUGCCAAUUUGAUGUCCUCAGUCUCUACUAAUGCCUUGACUGUUGCUCAUUUUGCUGAAAAUCCUAUAAAUGCAAUGAUUCAAUACAAUUACUUUGGAACCUUUGCACGAUUAUUGCUUUUUAUGCAUGACCCCGCAAUCCGGCAAUCCUUGUAUAAGGUCCAAGAAAAAAUACCUGGUUUGCACACCAGAACAUUCAAAUACGCCAUGUUGUACCUAGAUCCUUCAAUCGGAGGAGUAUGUGGUAUGGCAUUAUCUCGUUUUUUAAUUAGAGCAUUUCCAGAUCCAGUAACAGAGAGUCUCUCAUUCUGGAAGUUUAUCUAUGAACAUGCUUCUGAGCCUCAUCUAAAAAAGAUGGAAGUGAUGUUCGAGACCCCCCAAGGUGCCAAAUUCAGAAUAGAACACAUCAACAAAUUGUUAGAGGA

CCCUACCUCUCUAAACAUCUCAAUGGGUAUGAGUCCCGCAAAUUUGUUAAAGAGUGAGGUCAAAAAAUGUUUAAUAGAAUCAAGAUCAUCCAUCAAGAAUGAAAUCAUAAAAGAUGCUACUAUCUACAUGCAUCAAGAAGAAGAGAAGCUUAGAGGAUUUUUGUGGUCCAUCAAACCAUUAUUCCCUCGUUUCUUGAGUGAAUUCAAAGCUGGAACUUUUUUGGGAGUAUCUGAGGGCUUUAUAAAUUUAUUUCAAAACUCACGUACCAUAAGAAACUCAUUUAAAAAGAGAUAUCACAAGGAUCUUGAUGAAUUAAUAAUCAAGAGAGAAAUAUCGUCCUUGAGUCAUCUCGGAUCUAUGCAUUAUCGAUUAGGGGAUAAUCAGAUAUGGUCCUGUUCUGCAUCUCGAGCCGACAUAUUAAGAUACAAGUCAUGGACCAGGAAAGUUGUGGGAACUACAGUGCCUCAUCCUUUGGAAAUGCACGGACCACCCUCAAAGAAGGAAAGACCUUGCCAAUUGUGUAAUUCAUCCGGUCUCACAUAUAUUUCGGUACAUUGUCCAAAAGGAAUUAUUGAUGUAUUCAAUAGAAGGGGACCUUUGCCGGCUUACUUGGGGUCCAAUACAUCUGAGUCUACAUCUAUUUUACAGCCGUGGGAAAAAGAGAGUAAAAUACCAAUAAUUAAAAGAGCAACUAGAUUAAGGGACGCCAUUUCUUGGUUCAUCCCACCCGAGUCUCCUCUGUCUACAUGCAUUUUGAAUAAUAUUCAGGCUUUAACAGGAGAAGAUUGGAGCUCUAAACAACAUGGCUUUAAGAGGACAGGAUCAGCAUUACAUAGAUUUUCCACCUCCCGGAUGAGCAAUGGAGGAUUUGCCUCACAAAGCCCGGCCACCUUAACUCGCAUGAUUGCAACUACAGACACAAUGAGAGACUUUGGUACAAAAAAUUAUGAUUUCAUGUUCCAAGCAUCUUUGUUAUAUGGACAGAUGACAACAAGUAUUUCAAGAUAUGGAACCCCAGGGUCUUGCACGGAUCAUUACCAUAUCAGAUGUAAAGGAUGCAUUAGAGAGAUUGAAGAAGUAGAACUGAACACUAGUCUAGAAUACAAGACGCCCGAUGUUUCUCACAUAUUGGAAAAAUGGAGGAACAAUACUGGAUCUUGGGGUCAUCAAAUCAAACAAUUGAAACCUGCCGAGGGAAACUGGGAAUCAUUGUCUCCUGUAGAGCAAUCAUAUCAAGUUGCAAGAUGUAUUGGAUUUCUUUAUGGUGAACUGACACACAAGAAAUCGAGACAGGCUGAUGAUAGUUCUCUAUUCCCCUUGAGCAUCCAACUAAAAGUGAGAGGGAGAGGUUUCUUGCGAGGCCUUCUAGACGGUUUGAUGAGAUCCAGUUGCUGUCAGGUAAUUCAUAGACGAAGUGUUUCUACCUUAAAGAGACCUGCAAAUGCAGUUUACGGUGGCCUCAUAUACCUCAUUGAUAAACUAAGUGCCUCAAGUCCCUUCUUAUCACUUGUAAGAACCGGACCUAUUCGACAAGAAUUAGAACAGGUGCCACACAAGAUGUCUACAUCGUAUCCAACGAACAUUAGGGACUUGGGCUCCAUUGUGAGAAAUUAUUUUAAAUAUCAGUGUCGACCAGUUGAGAGAGGAAACUAUAAAACUUAUUAUAAUCAAAUAUGGUUAUUUUCUGAUGUUUUGUCCACUGAAUUUAUAGGGCCAAUGCGUAUAUCCAGCUCUCUCCUUAGACUCCUUUAUCGACCUUCUCUGACAAAGAAGGAUAGAGAAGAAUUGAGGGAGUUAGCAGCACUAUCAUCUAAUUUACGAAGUGGAGAAGAUUGGGAUGAUUUACAUAUCAAAUUUUUCUCAAACGAUCUCCUCUUUUGCUCACAAGAGAUAAGACAUGCUUGCAAAUUUGGGAUUAAAAAAGAUAAUGAAGAUAUCACUUUUUACCCGAAUUGGGGAACAGAGUACAUUGGGAAUGUCACUGAUAUUCCUGUGUUUUACCGAGCUCAAAAUGUCAAAAAGGAUAUUAGAGUACCCCCUCGGAUCCAAAAUCCUUUAAUGUCAGGACUCAGGCUUGGACAACUGCCAACUGGGGCUCAUUACAAAAUGAGAGCCAUUGUAUUUCGUCUUAAAAUUCCGUAUCAUGACUUUCUGGAUUGCGGAGAUGGGUCAGGAGGAAUGACUGCAGCAUUACUCCGGUAUAAUAGGACAUCCAGAGGAAUCUUUAACAGUUUGUUGGAUCUGUCAGACACAAUGCUAAGAGGAUCCUCACCAGAACCUCCCAGUGCCCUUGAGACCCUGGGGGGAGAGAGAGUAAGAUGUGUGAAUGGAGACAGUUGCUGGGAACAUCCUUCCGAUCUCAGCGAUGAGAACACAUGGAAGUAUUUUCUCCAUUUAAAAAAGGGAUGUGGAAUGAGUAUCAAUCUUAUUACCAUGGAUAUGGAAGUCCAGGAUUCUGUUAUAUCAUACAAAAUCGAGUCAUUGGUCAGACAGUAUGUUCCGGUUCUAUUGGAAUCCGACGGUUGCCUUAUUUAUAAAACAUAUGGAACAUAUAUUGCCACACAAGAAGACAAUUCCCUGACUCUUAUAGGAUCACUUUUUCACUCUGUCCAACUUGUCCAAACGGAUUUAAGCUCUAGUAAUACAUCCGAAUUGUACUUGGUAUGCAGGAGAUUGAAGGAUUAUGUAGACACUCCCUUUGUUGAUUGGAUAGAAUUAUACGACAACUGGGAGAAACAGUAUGCUUUCAGAAGCUUCAAAGAUGAAUUUCAAAGAGCUCAAUCACUUACCCCAGAGACAACCCUAAUAGGAAUACCUCCUCAAUUUGUACCGGAUCCUGGAGUCAAUCUAGAGACCCUGUUUCAAAUAGCAGGGGUUCCCACCGGAGUUGCGCAUGGAAUCACACAUCACAUAUUGCAGUCUAAGGAUAAAUUAAUAUCAAAUGCCAUCGGAAGCAUGUGUGUUAUCUCUCAUUUCCACAUUAACACAAUACGGACCACAGAUAGUAUGCCUGGACCCCCAUCAGAUGGGGAUGUCAACAAAAUGUGUUCAGCAUUGAUUGGGGCAUGUUUCUGGCUGGACUGGAUGGAAUCUGAUCUUAACUUAUACAAACAUUGUCUAAGAUCAAUAAUGAAGUCUAUGCCUGUGAGAUGGUUUAGAACAUUAAAAAAUGAAAAAUGGUCGCAAAAAUGGGAUUGUAAAGGAGAUGCAAUCCCAAAAGAUUCCAGAUUAGGAGACAGCCUUGCUAAUAUCGGCAAUUGGAUAAGAGCUUGGGAAUUAAUUAGAAAUGGAAAUAAGUCUGAGCCUUUUGAUUCAAUGGUAGCAGAAGCAUUGACAAAAUCUGUGGAUAAAUCACUUAGUUGGAGGAAAAUCUCAAAGUCAACUGGAAUUCCGAGACUUCUAAACAGUGAUAUCGAUUUGGUUGAUCAAUCUAUACUAAAUGUUCAGAUCGACAUCGUAGAAAAUCAAGCUUGGCAAAAUUGA(SEQID NO:18)。

AUGGAUCUCAACCCGGUCGAUGAUGCUGCAGAGUUAUCAGAGGAGAACUUCUUCUCUGGGAAAUUGUCCAAGGAAUGCCGAAUUAGAGGAUUGAAUUCAGUUGAUUACAAUCUGAACUCCCCGUUGGUGUCCGACGAUUUGACAUACUUGCUGAACAAAUUUAAGGGAAAACCUGUGCCUAUCCGAUGGAAGAUGAAGAAAUGGGACUCUAUUCUAGACCAACUGCGUAAACAUGAUCUGGAGUAUCUGAGACCCUCGGAUCUACACCAAUGGUUUGCAGAAUGGAUGUUGUACUCAAAGCAUGGCAGUAAGCAGGGAGAAGACUUCUUAAAGACAGUAGACGAAGAGGCUAGUGAUACGUUUGAGGUUGUAAGGUCGUUCAUAAGGGGAUGGACUGGUGGAGAAAUCAACUUUGUCAGGAAGAGCGGAAAGCAUAUGGGGUAUUGUGCGGAGUUGUGCCAGAAAUUCUUGGAUUUGCAUAAAUUGACCCUCCUUGGAAAUGCCGCCACCGAUAAUGAGUUGUUACAACUGAGCAAAACUUUCGGGGAUGACAAGAUUUACAAGAAACGGCUAAUCAAACUUCCAAGUCUUGGUCGCGUCAUCUUUGAUUCUGGAUUCUUCAUUGUCCUGGAUCAGCGAGUUCUAAUGGAUCGUAAUUUUAUGUUGAUGAUGAAAGACGUAAUAAUUGGAAGAAUGCAAACAGUUCUGUCCAUGAUUAGCAGAUGUGAUGAUAAAUUCAGUUCCAAGGAUAUUGAUUUUCUAUUAAAGGUCUACUCAACAGGCGAUAAGAUCAUAAGAAAGUUGGGUAACGAUGGGUAUGAGUUGAUCAAGACCGUAGAACCUAUGUGCAAUCUUCGUCUAUCAGACCUGGCCAGAAGGUUCCGUCCACUCGUACCUCCUUUCCCUCAUUUCAGGAGACAUAUUGAAUCGACUGUAGAUGAACUGAGUGCGAAGACUCCAUUGAUCAGGGAAUUGUUCUCAUUGAUUGAUACUUCUCCUAAUGUAGAUUCAACGUUGGUCGUUUACGGUUCAUUUCGUCAUUGGGGUCAUCCUUUUAUUAAUUAUUUUGAAGGCCUGGAGAAAUUGCAUAAGCAAGUGACAAUGGAAAAAGAAAUUGAUACCAAUUAUUCGGAAGCGUUGGCCAGUGAUCUGGCUAGGAUAGUCUUGACUAAAGAGUUCAAUGAGAAAAAACAAUGGGCUGUAGAUUACCACAGAGUGCCUACAAGCCAUCCCUUUAAAAACCACAUCCGAGACAACACAUGGCCAACAGCUGCAGUGAUCCAAGAUUUUGGAGAUCACUGGCAUGAGCUCCCAUUAAUUCAAUGCUUCGACAUACCUGAUCUGAUUGAUCCAUCCAUUAUCUACUCAGACAAGAGCCAUUCCAUGAAUCGAUCAGAGGUGUUAAAUCAUGUAAGAACCAAGCCUCAUACACCGAUUCCAAGCAAGAAAGUUUUAGAGAGUAUGAUCGACAAACCAGCGACAAAUUGGCUUGAGUUUCUUGAAGAGAUCGACAAAAAUGGAUUAUCUGAUGAGGAUUUGGUGAUCGGAUUAAAAGGAAAAGAAAGAGAACUGAAGAUUGCAGGUAGAUUUUUCUCAUUGAUGUCUUGGAAGCUUAGGGAAUACUUUGUUGUCACAGAGUAUUUGAUCAAGACUCACUUUGUGCCAUUGUUCCAUGGGCUGACCAUGGCUGAUGAUAUGACAGCAGUGAUCAAAAAGAUGCUGGAAAGCUCGUCUGGUCAAGGGCUAACAAACUACGACAGCGUUUGUAUUGCAAAUCAUAUUGACUAUGAAAAAUGGAAUAAUCAUCAACGAAAAUUGUCCAAUGGACCUGUGUUUAAAGUCAUGGGACAAUUCUUGGGUUAUCCAAAUUUGAUAUACCGGACACAUGAGUUUUUUGAGAAAAGUUUAAUUUACUAUAAUGAAAGACCGGAUCUCAUGAAAGUCAGAAAUGGGAUCUUAGAAAAUAGCACACAUCAGAGGGUUUGCUGGAAUGGACAGGCAGGUGGUUUAGAAGGUUUGAGACAGAAAGGAUGGAGUAUUCUUAAUUUGCUGGUCAUACAGAGAGAAGCGAAAAUCCGGAAUACCGCAGUUAAAGUGCUAGCACAAGGCGACAAUCAAGUCAUAUGUACACAGUACAAGACCAAACAGUAUCGAAACGAUAUCGAGUUGAGACAAGCCUUAAACCAAAUGGCAGCAAACAAUGAUGUCAUCAUGAAGGCCAUAGAGUCAGGAACCAAUAAGUUGGGUUUGUUGAUCAAUCAGGAUGAGACGAUGCAAUCUGCUGAUUAUCUGAACUAUGGGAAGGUACCUAUCUUUAGAGGAGUAAUCAGAGGGCUGGAAACAAAGCGAUGGUCCCGAGUCACUUGUGUGACAAAUGACCAAUUACCAACGUGUGCAAAUCUCAUGUCAUCUGUAUCUACAAAUGCUUUGACAGUGGCACAUUUCGAUGUACACCCAUUAAAUGCAAUGAUCCAAUUUAACUUCUUCGGGAAUUUUGCUCGCCUUCUCUUGAUUAUGCACGAUCCGGCGAUUCGACAAUCAUUAAAUCAGUUGAAAGGACCAAACAUCAAUGUCCAUUCAUACGGGUUCAAAGUUGCGAUGUUGUACUUGGACCCUUCAAUAGGAGGAGUGUGUGGUACAGCCCUUUCUCGAUUUUUGAUUAGGAGUUUCCCUGACCCUGUGACGGAGAGUCUCUCAUUUUGGAAGCUUAUCCAUCAUAGCACCAGUGAUAUCAGAUUAAAAAAUCUCUCAGAACAGUUUGGCAAUCCUAAGAUAGCAGUUUUUAGAGAAUCUCACAUAGAAAAGUUGUUGGAAGAUCCUACUAGUUUGAAUAUAUCGAUGGGAAUGAGUGCUGCAAAUUUGUUGAAAACAGAGAUAAAGAAAAAUCUCCUACAAAAGAAAUCAAGCAUAGGGAAUCAGAUAGUCAAAGACGCCGUAUACUACAUACACUCGGAAGAUGAGAAAUUGAGAACUUUUUUGUGGUCCAUUACUCCAUUGUUUCCUCGGUUUCUGAGCGAAUUUAAAGCAGGGACGUUCAUGGGAGUAGCGAGCAGCAUUGUAUCUUUGUUCCAAAACUCUCGAACCAUUCGAAAUGUUUUCAGAGACUAUAUGAGUCAAACAAUUGAUGACUUGAUUGUCAAGAGUGAACUAACAUCUCUGGAGCAUUUGUCUAACUAUACUGACAGGAAAGGGAGUGGUGGGAUCUGGAGUUGCUCUGCGGAACAGGCUGACAAACUUAGAAGAAUGUCCUGGAAGCGUCCAGUGCUAGGGACAACAGUCCCUCACCCGUUGGAAAUGCACGGAAGGGGAACACUGAAAUCCCCGCUAUCCAAGUGUUGCAAGGAGUCCCGAAUGGAUUAUAUUUCUGUCCAUAUUCCUGAAGGAUUGAACAAAGUUUUGGACGGCCGAGGAAGCCUACCAGCAUAUUUGGGUUCAAAAACAUCAGAGUCUACCUCAAUUCUUCAGCCUUGGGAGAAGGAAAGCAAAAUUCCCAUCAUAAGAAGAGCUACUAGAUUACGAGAUGCAAUUCAUUGGUUUGUGGAUCCGGACAGUAACUUAGCCCGAAGCAUAUUGAACAAUAUUGAAUCCUUAACUGGAGAAAAGUGGGAAGGGGCUUUAAAAGGAUAUAAGAGAACGGGUUCUGCCCUCCAUAGAUUUUCCACCUCGCGGGUAAGCCAUGGUGGUUUUUCUUCACAAAGUCCUGCAUGUCUCACCAGAAUGAUGGCAACCACAGACACCAUGCGCGAUUAUGCUCAGCUCAAUUAUGACUUCAUGUUCCAAGCAAGUCUUCUCUACUCUCAAAUGACCAGUUCCGUCAUUUUAAUGGGAACCACUGUCUCAAACACUAUUCAUUUCCAUGUUACCUGUCGAAAAUGUAUCCGAGAAAUAACAGAACCGAUGUUAGAAAGUCCUAGGGAGUACAGAGGGAAGGACGUUCAUUUAGUCCUGGCAAAAUGGAAGAAUUCUUCAAAUGGUUGGGGAGAAACCCUCCAACUACUAAAGCCAGUUGAGGGAGAUUGGGACACUAUCCCUCCUGUUGAAAAAUCUUAUCAUGUGGGUAGGAUCUUGGGUUUUCUUUAUGGUGACCUAAAGAGUCAAAAUUCCAGUCGUGCAGACGACAGUUCAAUCUUCCCCCUCAGCAUUCAAAUGAGAUUAAGAGGUCGAGGCUUUUUACGAGGAAUCCUUGACGGAUUAGUCCGAGCUAGUGCUUGUCAGGUUAUCCACAGGAGAAGUGUUGCUUUGCUUUCAAAACCAGCAAAUGCAAUUUAUGGAGGAUUGAUUUACUUGAUAGACAAGAUAAGUGCAUCCACCUCGUUUACGACACUGUGUAGGGACGGACCCAUUAGAGAAGAACUAUCAUCUAUACCUCACAAAAUUCCUACUUCGUAUCCAACUUCAAAUUCAGAUAUGGGACUCCACAUUAGAAAUUAUUUGAAGUUUCAGUGCAAGACAGUCGAAUUGGGAAAAUAUCAAUCUGAUAUAAAGGAUUUAUGGUUGUUCUCGGAUGUGAUGACGUCGAAUAUUGCAGGACCCUUUGCAUUGUCUACGAAAAUUUUGAAGUGCUUAUACAAGCCUGCUUUGUCCCAGAAAGAUCGAAACAAUAUCAGGAAGAUCAGCAAUUUCUCCAAGAUGAUGCGAUCACAAUUGAGCUGGGACCCCACAAGCUCUGAAUUUAUUACAUCUCAAAUCCUAGUAUGCAAUGAGGAAAUUAGGCAUGCCUGCAAGUUUGGAAUUCCAAAAUUAUCACUUAAGUUUGAUGAUCCAGUCUGGGGACCCGAGGAUUAUGGAUUGAUCUGGUCCAUUCCAGUUGAUUAUUCCAGUCAAUCAGUUCCAAAGAACUUGAAGCCCUGUCCUAGAAUUCAAAACCCUAGCAUAUCUGGUUUUAGGUUAGGACAACUUCCAACAGGAGCUCACUACAAGCUGAGAUCAAUCCUAAGGAAGAAAAAUAUACAUUACAGGGAUGCACUGUGUGGGGGUGAUGGGUCAGGAGGUAUGACAGCGGCAGUCCUACGUUAUAAUCUAAAGGCCAGGGCCAUAUUUAAUAGCAUCCUUGAUUUUGAUGGAUCAACAAUGAAGGGAGCUUCACCAGAUCCACCUAGUGCAUUGGAAACAGUGGUGAAUGGCCGGACUCGGUGCGUCAACGCGGAGAGUUGUUGGGAGAACCCGUCAGAUCUGAGUGAACAGAGAACAUGGGAUUAUUUCAAAUUCUUGAAAACCCAUCACGGAUUAAAGAUAGACCUGAUUGUAUUGGACAUGGAAGUGAGAGACUUUGCAAUCUCGGCAUCCAUAGAGAAGUGUGUCCGAAACAAUGUAUCGUCAAUAUUGGAAGAAGACGGAGUCCUAAUAUACAAGACCUAUGGAUCAACAAUUGCAGCAGAAAGUAGCAAUGCAGUUGUAAAUAUUGGAGUAUUGUUUGAGAGCGUAGAGUUGAUUCAGACGGAGUAUAGCAGUACAUCCACUUCAGAAGUGUACAUGUAUUGCCGAAAGAUUAAGAAGUUUGUGGACGCUCCUCACCCAGAUUGGAUAUCACUUGAUUAUUAUUGGAGCAAAUUAUUUUGCUUUAGGAGUUACAAAGAAGAAUUCUUCCGAUCCUACGAAGUGUCUCGUAAAGAAUCCCUAAAAGGAAUACCCAAUUCCUUCAUCCCUGACCCAUUGGUCAAUCUAGAGACAUUACUUCAGAUAGCAGGAGUUCCUUCUGGGAUAUCACACCAGUUGGCAAUUGAUAUUAAGGAAUCACAGCUGACUCAGAUCACCGCUGCAAUGGUGCUUUGCGGAAUGAUCGCAAACUACACUUUGGAUGUGACAAAAAAGAGAGAUUCAUACAAUCCUCCUUCCGACGGAAGAUUAAUUCGAAUGAGUGCUGCUUUAGUCGGAAUAAGUUUCUGGAUUUCCGUCAAGUAUUAUGACAAAGAGUUGAAUUUCGAAUUGGAACAGAUAUUAUCCAGAUCAUUUCCCAUAAGGUGGAUGCUUUCACGCAACUAUUUAUUCUGGACAACAAAAAAGGGUUUUAGGAAUGCAAAAGACGUAAGACUAUCUGGGAAUAUGGCUAAUAUAGGGAAUUGGAUUCGAUGCAUGGAGCUCCUUCAUCUCCCCCCGGGAUCCCUGUCCAAGGAUGAGGUCACGACCACUUGUGGGAAAUACAUUAGAAGCUUGAAGUAUUCUGUCAUACUCCAACAGACCGGGAUCAUUGAUUUGUGGAAAUCAAGGGUCGCAAGUGAUGAUCGAUCAUUGAUGGAGGUUAAGACCGAAUUCAUUGAAUCUGAGCAUUGGGUAGAUUAA(SEQ ID NO:24)。

AUGCUCGAUCCUGGAGAGGUCUAUGAUGACCCUAUUGACCCAAUCGAGUUAGAGGCUGAACCCAGAGGAACCCCCAUUGUCCCCAACAUCUUGAGGAACUCUGACUACAAUCUCAACUCUCCUUUGAUAGAAGAUCCUGCUAGACUAAUGUUAGAAUGGUUAAAAACAGGGAAUAGACCUUAUCGGAUGACUCUAACAGACAAUUGCUCCAGGUCUUUCA

GAGUUUUGAAAGAUUAUUUCAAGAAGGUAGAUUUGGGUUCUCUCAAGGUGGGCGGAAUGGCUGCACAGUCAAUGAUUUCUCUCUGGUUAUAUGGUGCCCACUCUGAAUCCAACAGGAGCCGGAGAUGUAUAACAGACUUGGCCCAUUUCUAUUCCAAGUCGUCCCCCAUAGAGAAGCUGUUGAAUCUCACGCUAGGAAAUAGAGGGCUGAGAAUCCCCCCAGAGGGAGUGUUAAGUUGCCUUGAGAGGGUUGAUUAUGAUAAUGCAUUUGGAAGGUAUCUUGCCAACACGUAUUCCUCUUACUUGUUCUUCCAUGUAAUCACCUUAUACAUGAACGCCCUAGACUGGGAUGAGGAAAAGACCAUCCUAGCAUUAUGGAAAGAUUUAACCUCAGUGGACAUCGGGAAGGACUUGGUAAAGUUCAAAGACCAAAUAUGGGGACUGCUGAUCGUGACAAAGGACUUUGUUUACUCCCAAAGUUCCAAUUGUCUUUUUGACAGAAACUACACACUUAUGCUAAAAGAUCUUUUCUUGUCUCGCUUCAACUCCUUAAUGGUCUUGCUCUCUCCCCCAGAGCCCCGAUACUCAGAUGACUUGAUAUCUCAACUAUGCCAGCUGUACAUUGCUGGGGAUCAAGUCUUGUCUAUGUGUGGAAACUCCGGCUAUGAAGUCAUCAAAAUAUUGGAGCCAUAUGUCGUGAAUAGUUUAGUCCAGAGAGCAGAAAAGUUUAGGCCUCUCAUUCAUUCCUUGGGAGACUUUCCUGUAUUUAUAAAAGACAAGGUAAGUCAACUUGAAGAGACGUUCGGUCCCUGUGCAAGAAGGUUCUUUAGGGCUCUGGAUCAAUUCGACAACAUACAUGACUUGGUUUUUGUGUUUGGCUGUUACAGGCAUUGGGGGCACCCAUAUAUAGAUUAUCGAAAGGGUCUGUCAAAACUAUAUGAUCAGGUUCACCUUAAAAAAGUGAUAGAUAAGUCCUACCAGGAGUGCUUAGCAAGCGACCUAGCCAGGAGGAUCCUUAGAUGGGGUUUUGAUAAGUACUCCAAGUGGUAUCUGGAUUCAAGAUUCCUAGCCCGAGACCACCCCUUGACUCCUUAUAUCAAAACCCAAACAUGGCCACCCAAACAUAUUGUAGACUUGGUGGGGGAUACAUGGCACAAGCUCCCGAUCACGCAGAUCUUUGAGAUUCCUGAAUCAAUGGAUCCGUCAGAAAUAUUGGAUGACAAAUCACAUUCUUUCACCAGAACGAGACUAGCUUCUUGGCUGUCAGAAAACCGAGGGGGGCCUGUUCCUAGCGAAAAAGUUAUUAUCACGGCCCUGUCUAAGCCGCCUGUCAAUCCCCGAGAGUUUCUGAGGUCUAUAGACCUCGGAGGAUUGCCAGAUGAAGACUUGAUAAUUGGCCUCAAGCCAAAGGAACGGGAAUUGAAGAUUGAAGGUCGAUUCUUUGCUCUAAUGUCAUGGAAUCUAAGAUUGUAUUUUGUCAUCACUGAAAAACUCUUGGCCAACUACAUCUUGCCACUUUUUGACGCGCUGACUAUGACAGACAACCUGAACAAGGUGUUUAAAAAGCUGAUCGACAGGGUCACCGGGCAAGGGCUUUUGGACUAUUCAAGGGUCACAUAUGCAUUUCACCUGGACUAUGAAAAGUGGAACAACCAUCAAAGAUUAGAGUCAACAGAGGAUGUAUUUUCUGUCCUAGAUCAAGUGUUUGGAUUGAAGAGAGUGUUUUCUAGAACACACGAGUUUUUUCAAAAGGCCUGGAUCUAUUAUUCAGACAGAUCAGACCUCAUCGGGUUACGGGAGGAUCAAAUAUACUGCUUAGAUGCGUCCAACGGCCCAACCUGUUGGAAUGGCCAGGAUGGCGGGCUAGAAGGCUUACGGCAGAAGGGCUGGAGUCUAGUCAGCUUAUUGAUGAUAGAUAGAGAAUCUCAAAUCAGGAACACAAGAACCAAAAUACUAGCUCAAGGAGACAACCAGGUUUUAUGUCCGACAUACAUGUUGUCGCCAGGGCUAUCUCAAGAGGGGCUCCUCUAUGAAUUGGAGAGAAUAUCAAGGAAUGCACUUUCGAUAUACAGAGCCGUCGAGGAAGGGGCAUCUAAGCUAGGGCUGAUCAUCAAGAAAGAAGAGACCAUGUGUAGUUAUGACUUCCUCAUCUAUGGAAAAACCCCUUUGUUUAGAGGUAACAUAUUGGUGCCUGAGUCCAAAAGAUGGGCCAGAGUCUCUUGCGUCUCUAAUGACCAAAUAGUCAACCUCGCCAAUAUAAUGUCGACAGUGUCCACCAAUGCGCUAACAGUGGCACAACACUCUCAAUCUUUGAUCAAACCGAUGAGGGAUUUUCUGCUCAUGUCAGUACAGGCAGUCUUUCACUACCUGCUAUUUAGCCCAAUCUUAAAGGGAAGAGUUUACAAGAUUCUGAGCGCUGAAGGGGAGAGCUUUCUCCUAGCCAUGUCAAGGAUAAUCUAUCUAGAUCCUUCUUUGGGAGGGAUAUCUGGAAUGUCCCUCGGAAGAUUCCAUAUACGACAGUUCUCAGACCCUGUCUCUGAAGGGUUAUCCUUCUGGAGAGAGAUCUGGUUAAGCUCCCAAGAGUCCUGGAUUCACGCGUUGUGUCAAGAGGCUGGAAACCCAGAUCUUGGAGAGAGAACACUCGAGAGCUUCACUCGCCUUCUAGAAGAUCCGACCACCUUAAAUAUCAGAGGAGGGGCCAGUCCUACCAUUCUACUCAAGGAUGCAAUCAGAAAGGCUUUAUAUGACGAGGUGGACAAGGUGGAAAAUUCAGAGUUUCGAGAGGCAAUCCUGUUGUCCAAGACCCAUAGAGAUAAUUUUAUACUCUUCUUAAUAUCUGUUGAGCCUCUGUUUCCUCGAUUUCUCAGUGAGCUAUUCAGUUCGUCUUUUUUGGGAAUCCCCGAGUCAAUCAUUGGAUUGAUACAAAACUCCCGAACGAUAAGAAGGCAGUUUAGAAAGAGUCUCUCAAAAACUUUAGAAGAAUCCUUCUACAACUCAGAGAUCCACGGGAUUAGUCGGAUGACCCAGACACCUCAGAGGGUUGGGGGGGUGUGGCCUUGCUCUUCAGAGAGGGCAGAUCAACUUAGGGAGAUCUCUUGGGGAAGAAAAGUGGUAGGCACGACAGUUCCUCACCCUUCUGAGAUGUUGGGAUUACUUCCCAAGUCCUCUAUUUCUUGCACUUGUGGAGCAACAGGAGGAGGCAAUCCUAGAGUUUCUGUAUCAGUACUCCCGUCCUUUGAUCAGUCAUUUUUUUCACGAGGCCCCCUAAAGGGAUACUUGGGCUCGUCCACCUCUAUGUCGACCCAGCUAUUCCAUGCAUGGGAAAAAGUCACUAAUGUUCAUGUGGUGAAGAGAGCUCUAUCGUUAAAAGAAUCUAUAAACUGGUUCAUUACUAGAGAUUCCAACUUGGCUCAAGCUCUAAUUAGGAACAUUAUGUCUCUGACAGGCCCUGAUUUCCCUCUAGAGGAGGCCCCUGUCUUCAAAAGGACGGGGUCAGCCUUGCAUAGGUUCAAGUCUGCCAGAUACAGCGAAGGAGGGUAUUCUUCUGUCUGCCCGAACCUCCUCUCUCAUAUUUCUGUUAGUACAGACACCAUGUCUGAUUUGACCCAAGACGGGAAGAACUACGAUUUCAUGUUCCAGCCAUUGAUGCUUUAUGCACAGACAUGGACAUCAGAGCUGGUACAGAGAGACACAAGGCUAAGAGACUCUACGUUUCAUUGGCACCUCCGAUGCAACAGGUGUGUGAGACCCAUUGACGACGUGACCCUGGAGACCUCUCAGAUCUUCGAGUUUCCGGAUGUGUCGAAAAGAAUAUCCAGAAUGGUUUCUGGGGCUGUGCCUCACUUCCAGAGGCUUCCCGAUAUCCGUCUGAGACCAGGAGAUUUUGAAUCUCUAAGCGGUAGAGAAAAGUCUCACCAUAUCGGAUCAGCUCAGGGGCUCUUAUACUCAAUCUUAGUGGCAAUUCACGACUCAGGAUACAAUGAUGGAACCAUCUUCCCUGUCAACAUAUACGGCAAGGUUUCCCCUAGAGACUAUUUGAGAGGGCUCGCAAGGGGAGUAUUGAUAGGAUCCUCGAUUUGCUUCUUGACAAGAAUGACAAAUAUCAAUAUUAAUAGACCUCUUGAAUUGGUCUCAGGGGUAAUCUCAUAUAUUCUCCUGAGGCUAGAUAACCAUCCCUCCUUGUACAUAAUGCUCAGAGAACCGUCUCUUAGAGGAGAGAUAUUUUCUAUCCCUCAGAAAAUCCCCGCCGCUUAUCCAACCACUAUGAAAGAAGGCAACAGAUCAAUCUUGUGUUAUCUCCAACAUGUGCUACGCUAUGAGCGAGAGAUAAUCACGGCGUCUCCAGAGAAUGACUGGCUAUGGAUCUUUUCAGACUUUAGAAGUGCCAAAAUGACGUACCUAUCCCUCAUUACUUACCAGUCUCAUCUUCUACUCCAGAGGGUUGAGAGAAACCUAUCUAAGAGUAUGAGAGAUAACCUGCGACAAUUGAGUUCUUUGAUGAGGCAGGUGCUGGGCGGGCACGGAGAAGAUACCUUAGAGUCAGACGACAACAUUCAACGACUGCUAAAAGACUCUUUACGAAGGACAAGAUGGGUGGAUCAAGAGGUGCGCCAUGCAGCUAGAACCAUGACUGGAGAUUACAGCCCCAACAAGAAGGUGUCCCGUAAGGUAGGAUGUUCAGAAUGGGUCUGCUCUGCUCAACAGGUUGCAGUCUCUACCUCAGCAAACCCGGCCCCUGUCUCGGAGCUUGACAUAAGGGCCCUCUCUAAGAGGUUCCAGAACCCUUUGAUCUCGGGCUUGAGAGUGGUUCAGUGGGCAACCGGUGCUCAUUAUAAGCUUAAGCCUAUUCUAGAUGAUCUCAAUGUUUUCCCAUCUCUCUGCCUUGUAGUUGGGGACGGGUCAGGGGGGAUAUCAAGGGCAGUCCUCAACAUGUUUCCAGAUGCCAAGCUUGUGUUCAACAGUCUUUUAGAGGUGAAUGACCUGAUGGCUUCCGGAACACAUCCACUGCCUCCUUCAGCAAUCAUGAGGGGAGGAAAUGAUAUCGUCUCCAGAGUGAUAGAUCUUGACUCAAUCUGGGAAAAACCGUCCGACUUGAGAAACUUGGCAACCUGGAAAUACUUCCAGUCAGUCCAAAAGCAGGUCAACAUGUCCUAUGACCUCAUUAUUUGCGAUGCAGAAGUUACUGACAUUGCAUCUAUCAACCGGAUCACCCUGUUAAUGUCCGAUUUUGCAUUGUCUAUAGAUGGACCACUCUAUUUGGUCUUCAAAACUUAUGGGACUAUGCUAGUAAAUCCAAACUACAAGGCUAUUCAACACCUGUCAAGAGCGUUCCCCUCGGUCACAGGGUUUAUCACCCAAGUAACUUCGUCUUUUUCAUCUGAGCUCUACCUCCGAUUCUCCAAACGAGGGAAGUUUUUCAGAGAUGCUGAGUACUUGACCUCUUCCACCCUUCGAGAAAUGAGCCUUGUGUUAUUCAAUUGUAGCAGCCCCAAGAGUGAGAUGCAGAGAGCUCGUUCCUUGAACUAUCAGGAUCUUGUGAGAGGAUUUCCUGAAGAAAUCAUAUCAAAUCCUUACAAUGAGAUGAUCAUAACUCUGAUUGACAGUGAUGUAGAAUCUUUUCUAGUCCACAAGAUGGUUGAUGAUCUUGAGUUACAGAGGGGAACUCUGUCUAAAGUGGCUAUCAUUAUAGCCAUCAUGAUAGUUUUCUCCAACAGAGUCUUCAACGUUUCCAAACCCCUAACUGACCCCUCGUUCUAUCCACCGUCUGAUCCCAAAAUCCUGAGGCACUUCAACAUAUGUUGCAGUACUAUGAUGUAUCUAUCUACUGCUUUAGGUGACGUCCCUAGCUUCGCAAGACUUCACGACCUGUAUAACAGACCUAUAACUUAUUACUUCAGAAAGCAAGUCAUUCGAGGGAACGUUUAUCUAUCUUGGAGUUGGUCCAACGACACCUCAGUGUUCAAAAGGGUAGCCUGUAAUUCUAGCCUGAGUCUGUCAUCUCACUGGAUCAGGUUGAUUUACAAGAUAGUGAAGACUACCAGACUCGUUGGCAGCAUCAAGGAUCUAUCCAGAGAAGUGGAAAGACACCUUCAUAGGUACAACAGGUGGAUCACCCUAGAGGAUAUCAGAUCUAGAUCAUCCCUACUAGACUACAGUUGCCUGUGA(SEQ ID NO:30)。

根据本发明的实施例，所述第四RNA序列具有SEQ ID NO：35～38所示碱基序列的至少之一。

AUGGAGAAGCCAGUAGUAAACGUAGACGUAGACCCCCAGAGUCCGUUUGUCGUGCAACUGCAAAAAAGCUUCCCGCAAUUUGAGGUAGUAGCACAGCAGGUCACUCCAAAUGACCAUGCUAAUGCCAGAGCAUUUUCGCAUCUGGCCAGUAAACUAAUCGAGCUGGAGGUUCCUACCACAGCGACGAUCUUGGACAUAGGCAGCGCACCGGCUCGUAGAAUGUUUUCCGAGCACCAGUAUCAUUGUGUCUGCCCCAUGCGUAGUCCAGAAGACCCGGACCGCAUGAUGAAAUACGCCAGUAAACUGGCGGAAAAAGCGUGCAAGAUUACAAACAAGAACUUGCAUGAGAAGAUUAAGGAUCUCCGGACCGUACUUGAUACGCCGGAUGCUGAAACACCAUCGCUCUGCUUUCACAACGAUGUUACCUGCAACAUGCGUGCCGAAUAUUCCGUCAUGCAGGACGUGUAUAUCAACGCUCCCGGAACUAUCUAUCAUCAGGCUAUGAAAGGCGUGCGGACCCUGUACUGGAUUGGCUUCGACACCACCCAGUUCAUGUUCUCGGCUAUGGCAGGUUCGUACCCUGCGUACAACACCAACUGGGCCGACGAGAAAGUCCUUGAAGCGCGUAACAUCGGACUUUGCAGCACAAAGCUGAGUGAAGGUAGGACAGGAAAAUUGUCGAUAAUGAGGAAGAAGGAGUUGAAGCCCGGGUCGCGGGUUUAUUUCUCCGUAGGAUCGACACUUUAUCCAGAACACAGAGCCAGCUUGCAGAGCUGGCAUCUUCCAUCGGUGUUCCACUUGAAUGGAAAGCAGUCGUACACUUGCCGCUGUGAUACAGUGGUGAGUUGCGAAGGCUACGUAGUGAAGAAAAUCACCAUCAGUCCCGGGAUCACGGGAGAAACCGUGGGAUACGCGGUUACACACAAUAGCGAGGGCUUCUUGCUAUGCAAAGUUACUGACACAGUAAAAGGAGAACGGGUAUCGUUCCCUGUGUGCACGUACAUCCCGGCCACCAUAUGCGAUCAGAUGACUGGUAUAAUGGCCACGGAUAUAUCACCUGACGAUGCACAAAAACUUCUGGUUGGGCUCAACCAGCGAAUUGUCAUUAACGGUAGGACUAACAGGAACACCAACACCAUGCAAAAUUACCUUCUGCCGAUCAUAGCACAAGGGUUCAGCAAAUGGGCUAAGGAGCGCAAGGAUGAUCUUGAUAACGAGAAAAUGCUGGGUACUAGAGAACGCAAGCUUACGUAUGGCUGCUUGUGGGCGUUUCGCACUAAGAAAGUACAUUCGUUUUAUCGCCCACCUGGAACGCAGACCUGCGUAAAAGUCCCAGCCUCUUUUAGCGCUUUUCCCAUGUCGUCCGUAUGGACGACCUCUUUGCCCAUGUCGCUGAGGCAGAAAUUGAAACUGGCAUUGCAACCAAAGAAGGAGGAAAAACUGCUGCAGGUCUCGGAGGAAUUAGUCAUGGAGGCCAAGGCUGCUUUUGAGGAUGCUCAGGAGGAAGCCAGAGCGGAGAAGCUCCGAGAAGCACUUCCACCAUUAGUGGCAGACAAAGGCAUCGAGGCAGCCGCAGAAGUUGUCUGCGAAGUGGAGGGGCUCCAGGCGGACAUCGGAGCA(SEQ ID NO:35)。

GCAUUAGUUGAAACCCCGCGCGGUCACGUAAGGAUAAUACCUCAAGCAAAUGACCGUAUGAUCGGACAGUAUAUCGUUGUCUCGCCAAACUCUGUGCUGAAGAAUGCCAAACUCGCACCAGCGCACCCGCUAGCAGAUCAGGUUAAGAUCAUAACACACUCCGGAAGAUCAGGAAGGUACGCGGUCGAACCAUACGACGCUAAAGUACUGAUGCCAGCAGGAGGUGCCGUACCAUGGCCAGAAUUCCUAGCACUGAGUGAGAGCGCCACGUUAGUGUACAACGAAAGAGAGUUUGUGAACCGCAAACUAUACCACAUUGCCAUGCAUGGCCCCGCCAAGAAUACAGAAGAGGAGCAGUACAAGGUUACAAAGGCAGAGCUUGCAGAAACAGAGUACGUGUUUGACGUGGACAAGAAGCGUUGCGUUAAGAAGGAAGAAGCCUCAGGUCUGGUCCUCUCGGGAGAACUGACCAACCCUCCCUAUCAUGAGCUAGCUCUGGAGGGACUGAAGACCCGACCUGCGGUCCCGUACAAGGUCGAAACAAUAGGAGUGAUAGGCACACCGGGGUCGGGCAAGUCAGCUAUUAUCAAGUCAACUGUCACGGCACGAGAUCUUGUUACCAGCGGAAAGAAAGAAAAUUGUCGCGAAAUUGAGGCCGACGUGCUAAGACUGAGGGGUAUGCAGAUUACGUCGAAGACAGUAGAUUCGGUUAUGCUCAACGGAUGCCACAAAGCCGUAGAAGUGCUGUACGUUGACGAAGCGUUCGCGUGCCACGCAGGAGCACUACUUGCCUUGAUUGCUAUCGUCAGGCCCCGCAAGAAGGUAGUACUAUGCGGAGACCCCAUGCAAUGCGGAUUCUUCAACAUGAUGCAACUAAAGGUACAUUUCAAUCACCCUGAAAAAGACAUAUGCACCAAGACAUUCUACAAGUAUAUCUCCCGGCGUUGCACACAGCCAGUUACAGCUAUUGUAUCGACACUGCAUUACGAUGGAAAGAUGAAAACCACGAACCCGUGCAAGAAGAACAUUGAAAUCGAUAUUACAGGGGCCACAAAGCCGAAGCCAGGGGAUAUCAUCCUGACAUGUUUCCGCGGGUGGGUUAAGCAAUUGCAAAUCGACUAUCCCGGACAUGAAGUAAUGACAGCCGCGGCCUCACAAGGGCUAACCAGAAAAGGAGUGUAUGCCGUCCGGCAAAAAGUCAAUGAAAACCCACUGUACGCGAUCACAUCAGAGCAUGUGAACGUGUUGCUCACCCGCACUGAGGACAGGCUAGUGUGGAAAACCUUGCAGGGCGACCCAUGGAUUAAGCAGCCCACUAACAUACCUAAAGGAAACUUUCAGGCUACUAUAGAGGACUGGGAAGCUGAACACAAGGGAAUAAUUGCUGCAAUAAACAGCCCCACUCCCCGUGCCAAUCCGUUCAGCUGCAAGACCAACGUUUGCUGGGCGAAAGCAUUGGAACCGAUACUAGCCACGGCCGGUAUCGUACUUACCGGUUGCCAGUGGAGCGAACUGUUCCCACAGUUUGCGGAUGACAAACCACAUUCGGCCAUUUACGCCUUAGACGUAAUUUGCAUUAAGUUUUUCGGCAUGGACUUGACAAGCGGACUGUUUUCUAAACAGAGCAUCCCACUAACGUACCAUCCCGCCGAUUCAGCGAGGCCGGUAGCUCAUUGGGACAACAGCCCAGGAACCCGCAAGUAUGGGUACGAUCACGCCAUUGCCGCCGAACUCUCCCGUAGAUUUCCGGUGUUCCAGCUAGCUGGGAAGGGCACACAACUUGAUUUGCAGACGGGGAGAACCAGAGUUAUCUCUGCACAGCAUAACCUGGUCCCGGUGAACCGCAAUCUUCCUCACGCCUUAGUCCCCGAGUACAAGGAGAAGCAACCCGGCCCGGUCAAAAAAUUCUUGAACCAGUUCAAACACCACUCAGUACUUGUGGUAUCAGAGGAAAAAAUUGAAGCUCCCCGUAAGAGAAUCGAAUGGAUCGCCCCGAUUGGCAUAGCCGGUGCAGAUAAGAACUACAACCUGGCUUUCGGGUUUCCGCCGCAGGCACGGUACGACCUGGUGUUCAUCAACAUUGGAACUAAAUACAGAAACCACCACUUUCAGCAGUGCGAAGACCAUGCGGCGACCUUAAAAACCCUUUCGCGUUCGGCCCUGAAUUGCCUUAACCCAGGAGGCACCCUCGUGGUGAAGUCCUAUGGCUACGCCGACCGCAACAGUGAGGACGUAGUCACCGCUCUUGCCAGAAAGUUUGUCAGGGUGUCUGCAGCGAGACCAGAUUGUGUCUCAAGCAAUACAGAAAUGUACCUGAUUUUCCGACAACUAGACAACAGCCGUACACGGCAAUUCACCCCGCACCAUCUGAAUUGCGUGAUUUCGUCCGUGUAUGAGGGUACAAGAGAUGGAGUUGGAGCC(SEQID NO:36)。

GCGCCGUCAUACCGCACCAAAAGGGAGAAUAUUGCUGACUGUCAAGAGGAAGCAGUUGUCAACGCAGCCAAUCCGCUGGGUAGACCAGGCGAAGGAGUCUGCCGUGCCAUCUAUAAACGUUGGCCGACCAGUUUUACCGAUUCAGCCACGGAGACAGGCACCGCAAGAAUGACUGUGUGCCUAGGAAAGAAAGUGAUCCACGCGGUCGGCCCUGAUUUCCGGAAGCACCCAGAAGCAGAAGCCUUGAAAUUGCUACAAAACGCCUACCAUGCAGUGGCAGACUUAGUAAAUGAACAUAACAUCAAGUCUGUCGCCAUUCCACUGCUAUCUACAGGCAUUUACGCAGCCGGAAAAGACCGCCUUGAAGUAUCACUUAACUGCUUGACAACCGCGCUAGACAGAACUGACGCGGACGUAACCAUCUAUUGCCUGGAUAAGAAGUGGAAGGAAAGAAUCGACGCGGCACUCCAACUUAAGGAGUCUGUAACAGAGCUGAAGGAUGAAGAUAUGGAGAUCGACGAUGAGUUAGUAUGGAUCCAUCCAGACAGUUGCUUGAAGGGAAGAAAGGGAUUCAGUACUACAAAAGGAAAAUUGUAUUCGUACUUCGAAGGCACCAAAUUCCAUCAAGCAGCAAAAGACAUGGCGGAGAUAAAGGUCCUGUUCCCUAAUGACCAGGAAAGUAAUGAACAACUGUGUGCCUACAUAUUGGGUGAGACCAUGGAAGCAAUCCGCGAAAAGUGCCCGGUCGACCAUAACCCGUCGUCUAGCCCGCCCAAAACGUUGCCGUGCCUUUGCAUGUAUGCCAUGACGCCAGAAAGGGUCCACAGACUUAGAAGCAAUAACGUCAAAGAAGUUACAGUAUGCUCCUCCACCCCCCUUCCUAAGCACAAAAUUAAGAAUGUUCAGAAGGUUCAGUGCACGAAAGUAGUCCUGUUUAAUCCGCACACUCCCGCAUUCGUUCCCGCCCGUAAGUACAUAGAAGUGCCAGAACAGCCUACCGCUCCUCCUGCACAGGCCGAGGAGGCCCCCGAAGUUGUAGCGACACCGUCACCAUCUACAGCUGAUAACACCUCGCUUGAUGUCACAGACAUCUCACUGGAUAUGGAUGACAGUAGCGAAGGCUCACUUUUUUCGAGCUUUAGCGGAUCGGACAACUCUAUUACUAGUAUGGACAGUUGGUCGUCAGGACCUAGUUCACUAGAGAUAGUAGACCGAAGGCAGGUGGUGGUGGCUGACGUUCAUGCCGUCCAAGAGCCUGCCCCUAUUCCACCGCCAAGGCUAAAGAAGAUGGCCCGCCUGGCAGCGGCAAGAAAAGAGCCCACUCCACCGGCAAGCAAUAGCUCUGAGUCCCUCCACCUCUCUUUUGGUGGGGUAUCCAUGUCCCUCGGAUCAAUUUUCGACGGAGAGACGGCCCGCCAGGCAGCGGUACAACCCCUGGCAACAGGCCCCACGGAUGUGCCUAUGUCUUUCGGAUCGUUUUCCGACGGAGAGAUUGAUGAGCUGAGCCGCAGAGUAACUGAGUCCGAACCCGUCCUGUUUGGAUCAUUUGAACCGGGCGAAGUGAACUCAAUUAUAUCGUCCCGAUCAGCCGUAUCUUUUCCACUACGCAAGCAGAGACGUAGACGCAGGAGCAGGAGGACUGAAUAC(SEQ ID NO:37)。

AUGAUGCCCACCGAAGCCAACAAAAGUAGGUACCAGUCUCGUAAAGUAGAAAAUCAGAAAGCCAUAACCACUGAGCGACUACUGUCAGGACUACGACUGUAUAACUCUGCCACAGAUCAGCCAGAAUGCUAUAAGAUCACCUAUCCGAAACCAUUGUACUCCAGUAGCGUACCGGCGAACUACUCCGAUCCACAGUUCGCUGUAGCUGUCUGUAACAACUAUCUGCAUGAGAACUAUCCGACAGUAGCAUCUUAUCAGAUUACUGACGAGUACGAUGCUUACUUGGAUAUGGUAGACGGGACAGUCGCCUGCCUGGAUACUGCAACCUUCUGCCCCGCUAAGCUUAGAAGUUACCCGAAAAAACAUGAGUAUAGAGCCCCGAAUAUCCGCAGUGCGGUUCCAUCAGCGAUGCAGAACACGCUACAAAAUGUGCUCAUUGCCGCAACUAAAAGAAAUUGCAACGUCACGCAGAUGCGUGAACUGCCAACACUGGACUCAGCGACAUUCAAUGUCGAAUGCUUUCGAAAAUAUGCAUGUAAUGACGAGUAUUGGGAGGAGUUCGCUCGGAAGCCAAUUAGGAUUACCACUGAGUUUGUCACCGCAUAUGUAGCUAGACUGAAAGGCCCUAAGGCCGCCGCACUAUUUGCAAAGACGUAUAAUUUGGUCCCAUUGCAAGAAGUGCCUAUGGAUAGAUUCGUCAUGGACAUGAAAAGAGACGUGAAAGUUACACCAGGCACGAAACACACAGAAGAAAGACCGAAAGUACAAGUGAUACAAGCCGCAGAACCCCUGGCGACUGCUUACUUAUGCGGGAUUCACCGGGAAUUAGUGCGUAGGCUUACGGCCGUCUUGCUUCCAAACAUUCACACGCUUUUUGACAUGUCGGCGGAGGAUUUUGAUGCAAUCAUAGCAGAACACUUCAAGCAAGGCGACCCGGUACUGGAGACGGAUAUCGCAUCAUUCGACAAAAGCCAAGACGACGCUAUGGCGUUAACCGGUCUGAUGAUCUUGGAGGACCUGGGUGUGGAUCAACCACUACUCGACUUGAUCGAGUGCGCCUUUGGAGAAAUAUCAUCCACCCAUCUACCUACGGGUACUCGUUUUAAAUUCGGGGCGAUGAUGAAAUCCGGAAUGUUCCUCACACUUUUUGUCAACACAGUUUUGAAUGUCGUUAUCGCCAGCAGAGUACUAGAAGAGCGGCUUAAAACGUCCAGAUGUGCAGCGUUCAUUGGCGACGACAACAUCAUACAUGGAGUAGUAUCUGACAAAGAAAUGGCUGAGAGGUGCGCCACCUGGCUCAACAUGGAGGUUAAGAUCAUCGACGCAGUCAUCGGUGAGAGACCACCUUACUUCUGCGGCGGAUUUAUCUUGCAAGAUUCGGUUACUUCCACAGCGUGCCGCGUGGCGGAUCCCCUGAAAAGGCUGUUUAAGUUGGGUAAACCGCUCCCAGCCGACGACGAGCAAGACGAAGACAGAAGACGCGCUCUGCUAGAUGAAACAAAGGCGUGGUUUAGAGUAGGUAUAACAGGCACUUUAGCAGUGGCCGUGACGACCCGGUAUGAGGUAGACAAUAUUACACCUGUCCUACUGGCAUUGAGAACUUUUGCCCAGAGCAAAAGAGCAUUCCAAGCCAUCAGAGGGGAAAUAAAGCAUCUCUACGGUGGUCCUAAAUAG(SEQ ID NO:38)。

根据本发明的一些具体实施例，所述自复制RNA分子包括以下6种碱基序列组合中的一种：

序列组合1：

所述第一RNA序列具有SEQ ID NO：4所示碱基序列，所述第二RNA序列具有SEQ IDNO：5所示碱基序列，所述第三RNA序列具有SEQ ID NO：6所示碱基序列。

序列组合2：

所述第一RNA序列具有SEQ ID NO：10所示碱基序列，所述第二RNA序列具有SEQ IDNO：11所示碱基序列，所述第三RNA序列具有SEQ ID NO：12所示碱基序列。

序列组合3：

所述第一RNA序列具有SEQ ID NO：16所示碱基序列，所述第二RNA序列具有SEQ IDNO：17所示碱基序列，所述第三RNA序列具有SEQ ID NO：18所示碱基序列。

序列组合4：

所述第一RNA序列具有SEQ ID NO：22所示碱基序列，所述第二RNA序列具有SEQ IDNO：23所示碱基序列，所述第三RNA序列具有SEQ ID NO：24所示碱基序列。

序列组合5：

所述第一RNA序列具有SEQ ID NO：28所示碱基序列，所述第二RNA序列具有SEQ IDNO：29所示碱基序列，所述第三RNA序列具有SEQ ID NO：30所示碱基序列。

序列组合6：

所述第四RNA序列具有SEQ ID NO：35～38所示碱基序列。该序列组合是正链RNA病毒来源的自复制RNA序列，其核心复合物为NSP1、NSP2、NSP3和NSP4。

根据本发明的实施例，进一步包括：靶分子编码区，所述靶分子区编码至少一个靶分子。

根据本发明的实施例，所述靶分子适于作为核酸类药物、蛋白类药物、病原体疫苗、肿瘤疫苗和罕见病治疗制剂的至少之一。由此，利用reRNA^TM(在本文中，自复制RNA分子和自扩增RNA分子与reRNA^TM是可以互换使用的)的自我扩增的特性，可以使用极低剂量的reRNA^TM来实现蛋白类药物的递送，自我扩增的reRNA^TM可以极大地提高蛋白的合成量，并且可以实现蛋白的长效表达。基于此特点，reRNA^TM技术可以应用于生命科学及医学的各个方面，如核酸类药物递送、蛋白类药物递送、病原体疫苗、肿瘤疫苗、罕见病治疗等。

根据本申请的实施例，所述靶分子编码区可以编码荧光蛋白、粒细胞-巨噬细胞集落刺激因子和卵清蛋白OVA的至少之一，可选的，所述荧光蛋白包括绿色荧光蛋白、红色荧光蛋白以及黄色荧光蛋白的至少之一。例如，通过将卵清蛋白在肿瘤细胞中表达，可以激活动物的免疫体系清除肿瘤细胞，从而达到治疗癌症的目的。

本发明所述自复制RNA分子还可含有一种或多种修饰核苷酸，并因此具有改善的稳定性，在体内耐降解和清除，以及其它优点。这些修饰核苷酸的类型包括但不限于m5C(5-甲基胞苷)、m5U(5-甲基尿苷)、m6A(N6-甲基腺苷)、s2U(2-硫代尿苷)、Um(2'-O-甲基尿苷)、m1A(l-甲基腺苷)；m2A(2-甲基腺苷)；Am(2-1-O-甲基腺苷)；ms2m6A(2-甲硫基-N6-甲基腺苷)；i6A(N6-异戊烯基腺苷)；ms2i6A(2-甲硫基-N6异戊烯基腺苷)；io6A(N6-(顺-羟基异戊烯基)腺苷)；ms2io6A(2-甲硫基-N6-(顺-羟基异戊烯基)腺苷)；g6A(N6-甘氨酰氨甲酰基腺苷)；t6A(N6-苏氨酰氨甲酰基腺苷)；ms2t6A(2-甲硫基-N6-苏氨酰氨甲酰基腺苷)；m6t6A(N6-甲基-N6-苏氨酰氨甲酰基腺苷)；hn6A(N6-羟基正缬氨酰(norvalyl)氨甲酰基腺苷)；ms2hn6A(2-甲硫基-N6-羟基正缬氨酰氨甲酰基腺苷)；Ar(p)(2'-O-核糖基腺苷(磷酸盐))；I(肌苷)；m1I(1-甲基肌苷)；m'Im(1,2'-O-二甲基肌苷)；m3C(3-甲基胞苷)；Cm(2T-O-甲基胞苷)；s2C(2-硫胞苷)；ac4C(N4-乙酰胞苷)；f5C(5-甲酰基(fonnyl)胞苷)；m5Cm(5,2-O-二甲基胞苷)；ac4Cm(N4乙酰基2TO甲基胞苷)；k2C(赖西丁)；m1G(1-甲基鸟苷)；m2G(N2-甲基鸟苷)；m7G(7-甲基鸟苷)；Gm(2'-O-甲基鸟苷)；m22G(N2,N2-二甲基鸟苷)；m2Gm(N2,2'-O-二甲基鸟苷)；m22Gm(N2,N2,2'-O-三甲基鸟苷)；Gr(p)(2'-O-核糖基鸟苷(磷酸盐))；yW(怀丁苷)；o2yW(过氧化怀丁苷)；OHyW(羟基怀丁苷)；OHyW*(改性不足的羟基怀丁苷)；imG(怀俄苷)；mimG(甲基鸟苷)；Q(辫苷)；oQ(环氧辫苷)；galQ(半乳糖基-辫苷)；manQ(甘露糖-辫苷)；preQo(7-氰基-7-脱氮(deaza)鸟苷)；preQi(7-氨甲基-7-脱氮鸟苷)；G*(古嘌苷)；D(二氢尿苷)；m5Um(5,2'-O-二甲基尿苷)；s4U(4-硫代尿苷)；m5s2U(5-甲基-2-硫代尿苷)；s2Um(2-硫-2'-O-甲基尿苷)；acp3U(3-(3-氨基-3-羧基丙基)尿苷)；ho5U(5-羟基尿苷)；mo5U(5-甲氧基尿苷)；cmo5U(尿苷5-氧乙酸)；mcmo5U(尿苷5-氧乙酸甲酯)；chm5U(5-(羧基羟基甲基)尿苷)；mchm5U(5-(羧基羟基甲基)尿苷甲酯)；mcm5U(5-甲氧基羰基甲基尿苷)；mcm5Um(S-甲氧基羰基甲基-2-O-甲基尿苷)；mcm5s2U(5-甲氧基羰基甲基-2-硫代尿苷)；nm5s2U(5-氨基甲基-2-硫代尿苷)；mnm5U(5-甲基氨基甲基尿苷)；mnm5s2U(5-甲基氨甲基-2-硫代尿苷)；mnm5se2U(5-甲基氨甲基-2硒基尿苷)；ncm5U(5-氨甲酰基甲基尿苷)；ncm5Um(5-氨甲酰基甲基-2'-O-甲基尿苷)；cmnm5U(5-羧基甲基氨甲基尿苷)；cnmm5Um(5-羧甲基氨甲基-2-L-O甲基尿苷)；cmnm5s2U(5-羧基甲基氨甲基-2-硫代尿苷)；m62A(N6,N6-二甲基腺苷)；Tm(2'-O-甲基肌苷)；m4C(N4-甲基胞苷)；m4Cm(N4,2-O-二甲基胞苷)；hm5C(5-羟基甲基胞苷)；m3U(3-甲基尿苷)；cm5U(5-羧基甲基尿苷)；m6Am(N6,T-O-二甲基腺苷)；rn62Am(N6,N6,O-2-三甲基腺苷)；m2'7G(N2,7-二甲基鸟苷)；m2'2'7G(N2,N2,7-三甲基鸟苷)；m3Um(3,2T-O-二甲基尿苷)；m5D(5-甲基二氢尿苷)；f5Cm(5-甲酰基-2'-O-甲基胞苷)；m1Gm(1,2'-O-二甲基鸟苷)；m'Am(1,2-O-二甲基腺苷)虹膜甲基尿苷(irinomethyluridine))；tm5s2U(S-牛磺酸(taurino)甲基-2-硫代尿苷))；imG-14(4-去甲基鸟苷)；imG2(异鸟苷)；ac6A(N6-乙酰腺苷)、次黄嘌呤、肌苷、8-氧代-腺嘌呤、其7-取代的衍生物、二氢尿嘧啶、假尿嘧啶、2-硫尿嘧啶、4-硫尿嘧啶、5-氨基尿嘧啶、5-(C1-C6)-烷基尿嘧啶、5-甲基尿嘧啶、5-(C2-C6)-烯基尿嘧啶、5-(C2-C6)-炔基尿嘧啶、5-(羟基甲基)尿嘧啶、5-氯尿嘧啶、5-氟尿嘧啶、5-溴尿嘧啶、5-羟基胞嘧啶、5-(C1-C6)-烷基胞嘧啶、5-甲基胞嘧啶、5-(C2-C6)-烯基胞嘧啶、5-(C2-C6)-炔基胞嘧啶、5-氯胞嘧啶、5-氟胞嘧啶、5-溴胞嘧啶、N2-二甲基鸟嘌呤、7-脱氮鸟嘌呤、8-氮杂鸟嘌呤、7-脱氮-7-取代鸟嘌呤、7-脱氮-7-(C2-C6)炔基鸟嘌呤、7-脱氮-8-取代鸟嘌呤、8-羟基鸟嘌呤、6-硫代鸟嘌呤、8-氧代鸟嘌呤、2-氨基嘌呤、2-氨基-6-氯嘌呤、2,4-二氨基嘌呤、2,6-二氨基嘌呤、8-氮杂嘌呤、取代的7-脱氮嘌呤、7-氮杂-7-取代嘌呤、7-氮杂-8-取代嘌呤、氢(脱碱基残基)、m5C、m5U、m6A、s2U、W或2'-O-甲基-U。

制备自复制RNA分子的手段

在本发明的第二方面，本发明提出了一种DNA分子，其编码前面任一项所述的自复制RNA分子。本领域技术人员能够理解的是，可以以DNA的形式为细胞提供编码自复制RNA分子的核酸，从而在细胞内合成相应的自复制RNA分子，进一步，自复制RNA分子可以在细胞内复制和翻译成相应的蛋白以及相应的靶分子。

需要说明的是，对于本发明说明书和权利要求书中所提及的DNA分子，本领域技术人员应当理解，实际包括互补双链的任意一条，或者两条。为了方便，在本说明书和权利要求书中，虽然多数情况下只给出了一条链，但实际上也公开了与之互补的另一条链。另外，本申请中的DNA分子的具体序列可以依据RNA分子的序列以及碱基互补配对原则获得，公开其中一种，意味着另一种也被公开。

在本发明的第三方面，本发明提出了一种表达载体，其携带前面所述的DNA分子。这里的表达载体的类型并不受特别限制，只要能够在宿主细胞中复制表达相应的RNA分子就可以。如前所述，可以以DNA的形式为细胞提供编码自复制RNA分子的核酸，从而在细胞内合成相应的自复制RNA分子，进一步，自复制RNA分子可以在细胞内复制和翻译成相应的蛋白以及相应的靶分子。

在将前面所述DNA分子连接到所述表达载体上时，可以将所述DNA分子与表达载体上的控制元件直接或者间接相连，只要这些控制元件能够控制所述DNA分子的翻译和表达等即可，即所述DNA分子与控制元件进行可操作地连接。当然，这些控制元件可以直接来自于载体本身，也可以是外源性的，即并非来自于载体本身。

本文中“可操作地连接”是指将外源基因连接到载体上，使得载体内的控制元件，例如转录控制序列和翻译控制序列等等，能够发挥其预期的调节外源基因的转录和翻译的功能。常用的载体例如可以为质粒、噬菌体等等。

在本发明的第四方面，本发明提出了一种制备前面所述自复制RNA分子的方法，其包括：使前面所述的DNA分子或者表达载体在宿主细胞中表达所述自复制RNA分子；和收集所述自复制RNA分子。

根据本申请的实施例，通过采用这些DNA分子和表达载体，在宿主细胞例如动物细胞中所表达的自复制RNA分子通常是以复合物的形式存在，例如参考图2，自复制RNA分子与N蛋白、P蛋白和L蛋白形成复合物，从而在输入至动物细胞后方便启动自复制RNA分子的自编码效率。

在本发明的第五方面，参考图2，本发明提出了一种RNA-蛋白质复合物，其包括：前面所述自复制RNA分子和蛋白质，所述蛋白质包括N蛋白或其功能片段、P蛋白或其功能片段和L蛋白或其功能片段。由此，输入至动物细胞后方便启动自复制RNA分子的自编码效率。

在本发明的第六方面，本发明提出了一种药物组合物，其特征在于，包括前面所述的自复制RNA分子、DNA分子、表达载体或者RNA-蛋白质复合物。

在本发明的第七方面，本发明提出了一种在个体中表达靶分子的方法，其包括：为个体给予前面所述的自复制RNA分子、DNA分子、表达载体、RNA-蛋白质复合物或者药物组合物。

本申请的附加方面和优点将在下面的描述中部分给出，部分将从下面的描述中变得明显，或通过本申请的实践了解到。其中，实施例中未注明具体技术或条件的，按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者，均为可以通过市购获得的常规产品。

实施例1NPL形成reRNA^TM核心区域最小单位，实现reRNA^TM负载基因的胞内表达

在本实施例中，按照GeneBank数据库的信息，获取编码水泡性口炎病毒(VSV)印第安那株(IND)N蛋白的DNA序列(SEQ ID NO：39所示碱基序列)、P蛋白的DNA序列(SEQ ID NO：40所示碱基序列)、L蛋白的DNA序列(SEQ ID NO：41所示碱基序列)以及编码绿色荧光蛋白(GFP)的DNA序列，其中，表达翻译后获得的水泡性口炎病毒(VSV)印第安那株(IND)N蛋白的氨基酸序列如SEQ ID NO：1所示，P蛋白的氨基酸序列如SEQ ID NO：2所示，L蛋白的氨基酸序列如SEQ ID NO：3所示。具体地，分别构建携带下列序列的reRNA^TMDNA序列：

IND-N-GFP；

IND-P-GFP；

IND-L-GFP；

IND-P+L-GFP；

IND-N+L-GFP；

IND-N+P-GFP；和

IND-N+P+L-GFP。

需要说明的是，在本文中，如无特别说明，N表示N蛋白的编码序列，P表示P蛋白的编码序列，L表示L蛋白的编码序列，IND表示后面所连N、P或L的来源为水泡性口炎病毒(VSV)印第安那株，NJ表示后面所连N、P或L的来源为水泡性口炎病毒(VSV)新泽西株，GFP表示绿色荧光蛋白的编码序列。

进一步，利用上述所构建的DNA序列，分别对293T细胞进行体外转染，在转染后培养293T细胞24小时，通过观察绿色荧光蛋白的荧光信号判断GFP编码序列是否得到正确翻译和表达。结果显示在图3中(图3中为了描述方便，省去了GFP的标注)，结果表明N蛋白、P蛋白和L蛋白中的任何单个蛋白及两个蛋白的组合均不能启动reRNA^TM负载基因(GFP)的表达，只有在N蛋白、P蛋白和L蛋白共同存在的情况下(在本文中也会简称为“NPL”)，才能启动负载基因GFP的表达。

ATGTCTGTTACAGTCAAGAGAATCATTGACAACACAGTCGTAGTTCCAAAACTTCCTGCAAATGAGGATCCAGTGGAATACCCGGCAGATTACTTCAGAAAATCAAAGGAGATTCCTCTTTACATCAATACTACAAAAAGTTTGTCAGATCTAAGAGGATATGTCTACCAAGGCCTCAAATCCGGAAATGTATCAATCATACATGTCAACAGCTACTTGTATGGAGCATTAAAGGACATCCGGGGTAAGTTGGATAAAGATTGGTCAAGTTTCGGAATAAACATCGGGAAAGCAGGGGATACAATCGGAATATTTGACCTTGTATCCTTGAAAGCCCTGGACGGCGTACTTCCAGATGGAGTATCGGATGCTTCCAGAACCAGCGCAGATGACAAATGGTTGCCTTTGTATCTACTTGGCTTATACAGAGTGGGCAGAACACAAATGCCTGAATACAGAAAAAAGCTCATGGATGGGCTGACAAATCAATGCAAAATGATCAATGAACAGTTTGAACCTCTTGTGCCAGAAGGTCGTGACATTTTTGATGTGTGGGGAAATGACAGTAATTACACAAAAATTGTCGCTGCAGTGGACATGTTCTTCCACATGTTCAAAAAACATGAATGTGCCTCGTTCAGATACGGAACTATTGTTTCCAGATTCAAAGATTGTGCTGCATTGGCAACATTTGGACACCTCTGCAAAATAACCGGAATGTCTACAGAAGATGTAACGACCTGGATCTTGAACCGAGAAGTTGCAGATGAAATGGTCCAAATGATGCTTCCAGGCCAAGAAATTGACAAGGCCGATTCATACATGCCTTATTTGATCGACTTTGGATTGTCTTCTAAGTCTCCATATTCTTCCGTCAAAAACCCTGCCTTCCACTTCTGGGGGCAATTGACAGCTCTTCTGCTCAGATCCACCAGAGCAAGGAATGCCCGACAGCCTGATGACATTGAGTATACATCTCTTACTACAGCAGGTTTGTTGTACGCTTATGCAGTAGGATCCTCTGCCGACTTGGCACAACAGTTTTGTGTTGGAGATAACAAATACACTCCAGATGATAGTACCGGAGGATTGACGACTAATGCACCGCCACAAGGCAGAGATGTGGTCGAATGGCTCGGATGGTTTGAAGATCAAAACAGAAAACCGACTCCTGATATGATGCAGTATGCGAAAAGAGCAGTCATGTCACTGCAAGGCCTAAGAGAGAAGACAATTGGCAAGTATGCTAAGTCAGAATTTGACAAATGA(SEQ ID NO：39)。

ATGGATAATCTCACAAAAGTTCGTGAGTATCTCAAGTCCTATTCTCGTCTGGATCAGGCGGTAGGAGAGATAGATGAGATCGAAGCACAACGAGCTGAAAAGTCCAATTATGAGTTGTTCCAAGAGGATGGAGTGGAAGAGCATACTAAGCCCTCTTATTTTCAGGCAGCAGATGATTCTGACACAGAATCTGAACCAGAAATTGAAGACAATCAAGGCTTGTATGCACCAGATCCAGAAGCTGAGCAAGTTGAAGGCTTTATACAGGGGCCTTTAGATGACTATGCAGATGAGGAAGTGGATGTTGTATTTACTTCGGACTGGAAACAGCCTGAGCTTGAATCTGACGAGCATGGAAAGACCTTACGGTTGACATCGCCAGAGGGTTTAAGTGGAGAGCAGAAATCCCAGTGGCTTTCGACGATTAAAGCAGTCGTGCAAAGTGCCAAATACTGGAATCTGGCAGAGTGCACATTTGAAGCATCGGGAGAAGGGGTCATTATGAAGGAGCGCCAGATAACTCCGGATGTATATAAGGTCACTCCAGTGATGAACACACATCCGTCCCAATCAGAAGCAGTATCAGATGTTTGGTCTCTCTCAAAGACATCCATGACTTTCCAACCCAAGAAAGCAAGTCTTCAGCCTCTCACCATATCCTTGGATGAATTGTTCTCATCTAGAGGAGAGTTCATCTCTGTCGGAGGTGACGGACGAATGTCTCATAAAGAGGCCATCCTGCTCGGCCTGAGATACAAAAAGTTGTACAATCAGGCGAGAGTCAAATATTCTCTGTAG(SEQ ID NO：40)。

ATGGAAGTCCACGATTTTGAGACCGACGAGTTCAATGATTTCAATGAAGATGACTATGCCACAAGAGAATTCCTGAATCCCGATGAGCGCATGACGTACTTGAATCATGCTGATTACAACCTGAATTCTCCTCTAATTAGTGATGATATTGACAATTTAATCAGGAAATTCAATTCTCTTCCAATTCCCTCGATGTGGGATAGTAAGAACTGGGATGGAGTTCTTGAGATGTTAACGTCATGTCAAGCCAATCCCATCCCAACATCTCAGATGCATAAATGGATGGGAAGTTGGTTAATGTCTGATAATCATGATGCCAGTCAAGGGTATAGTTTTTTACATGAAGTGGACAAAGAGGCAGAAATAACATTTGACGTGGTGGAGACCTTCATCCGCGGCTGGGGCAACAAACCAATTGAATACATCAAAAAGGAAAGATGGACTGACTCATTCAAAATTCTCGCTTATTTGTGTCAAAAGTTTTTGGACTTACACAAGTTGACATTAATCTTAAATGCTGTCTCTGAGGTGGAATTGCTCAACTTGGCGAGGACTTTCAAAGGCAAAGTCAGAAGAAGTTCTCATGGAACGAACATATGCAGGATTAGGGTTCCCAGCTTGGGTCCTACTTTTATTTCAGAAGGATGGGCTTACTTCAAGAAACTTGATATTCTAATGGACCGAAACTTTCTGTTAATGGTCAAAGATGTGATTATAGGGAGGATGCAAACGGTGCTATCCATGGTATGTAGAATAGACAACCTGTTCTCAGAGCAAGACATCTTCTCCCTTCTAAATATCTACAGAATTGGAGATAAAATTGTGGAGAGGCAGGGAAATTTTTCTTATGACTTGATTAAAATGGTGGAACCGATATGCAACTTGAAGCTGATGAAATTAGCAAGAGAATCAAGGCCTTTAGTCCCACAATTCCCTCATTTTGAAAATCATATCAAGACTTCTGTTGATGAAGGGGCAAAAATTGACCGAGGTATAAGATTCCTCCATGATCAGATAATGAGTGTGAAAACAGTGGATCTCACACTGGTGATTTATGGATCGTTCAGACATTGGGGTCATCCTTTTATAGATTATTACACTGGACTAGAAAAATTACATTCCCAAGTAACCATGAAGAAAGATATTGATGTGTCATATGCAAAAGCACTTGCAAGTGATTTAGCTCGGATTGTTCTATTTCAACAGTTCAATGATCATAAAAAGTGGTTCGTGAATGGAGACTTGCTCCCTCATGATCATCCCTTTAAAAGTCATGTTAAAGAAAATACATGGCCCACAGCTGCTCAAGTTCAAGATTTTGGAGATAAATGGCATGAACTTCCGCTGATTAAATGTTTTGAAATACCCGACTTACTAGACCCATCGATAATATACTCTGACAAAAGTCATTCAATGAATAGGTCAGAGGTGTTGAAACATGTCCGAATGAATCCGAACACTCCTATCCCTAGTAAAAAGGTGTTGCAGACTATGTTGGACACAAAGGCTACCAATTGGAAAGAATTTCTTAAAGAGATTGATGAGAAGGGCTTAGATGATGATGATCTAATTATTGGTCTTAAAGGAAAGGAGAGGGAACTGAAGTTGGCAGGTAGATTTTTCTCCCTAATGTCTTGGAAATTGCGAGAATACTTTGTAATTACCGAATATTTGATAAAGACTCATTTCGTCCCTATGTTTAAAGGCCTGACAATGGCGGACGATCTAACTGCAGTCATTAAAAAGATGTTAGATTCCTCATCCGGCCAAGGATTGAAGTCATATGAGGCAATTTGCATAGCCAATCACATTGATTACGAAAAATGGAATAACCACCAAAGGAAGTTATCAAACGGCCCAGTGTTCCGAGTTATGGGCCAGTTCTTAGGTTATCCATCCTTAATCGAGAGAACTCATGAATTTTTTGAGAAAAGTCTTATATACTACAATGGAAGACCAGACTTGATGCGTGTTCACAACAACACACTGATCAATTCAACCTCCCAACGAGTTTGTTGGCAAGGACAAGAGGGTGGACTGGAAGGTCTACGGCAAAAAGGATGGAGTATCCTCAATCTACTGGTTATTCAAAGAGAGGCTAAAATCAGAAACACTGCTGTCAAAGTCTTGGCACAAGGTGATAATCAAGTTATTTGCACACAGTATAAAACGAAGAAATCGAGAAACGTTGTAGAATTACAGGGTGCTCTCAATCAAATGGTTTCTAATAATGAGAAAATTATGACTGCAATCAAAATAGGGACAGGGAAGTTAGGACTTTTGATAAATGACGATGAGACTATGCAATCTGCAGATTACTTGAATTATGGAAAAATACCGATTTTCCGTGGAGTGATTAGAGGGTTAGAGACCAAGAGATGGTCACGAGTGACTTGTGTCACCAATGACCAAATACCCACTTGTGCTAATATAATGAGCTCAGTTTCCACAAATGCTCTCACCGTAGCTCATTTTGCTGAGAACCCAATCAATGCCATGATACAGTACAATTATTTTGGGACATTTGCTAGACTCTTGTTGATGATGCATGATCCTGCTCTTCGTCAATCATTGTATGAAGTTCAAGATAAGATACCGGGCTTGCACAGTTCTACTTTCAAATACGCCATGTTGTATTTGGACCCTTCCATTGGAGGAGTGTCGGGCATGTCTTTGTCCAGGTTTTTGATTAGAGCCTTCCCAGATCCCGTAACAGAAAGTCTCTCAT

TCTGGAGATTCATCCATGTACATGCTCGAAGTGAGCATCTGAAGGAGATGAGTGCAGTATTTGGAAACCCCGAGATAGCCAAGTTTCGAATAACTCACATAGACAAGCTAGTAGAAGATCCAACCTCTCTGAACATCGCTATGGGAATGAGTCCAGCGAACTTGTTAAAGACTGAGGTTAAAAAATGCTTAATCGAATCAAGACAAACCATCAGGAACCAGGTGATTAAGGATGCAACCATATATTTGTATCATGAAGAGGATCGGCTCAGAAGTTTCTTATGGTCAATAAATCCTCTGTTCCCTAGATTTTTAAGTGAATTCAAATCAGGCACTTTTTTGGGAGTCGCAGACGGGCTCATCAGTCTATTTCAAAATTCTCGTACTATTCGGAACTCCTTTAAGAAAAAGTATCATAGGGAATTGGATGATTTGATTGTGAGGAGTGAGGTATCCTCTTTGACACATTTAGGGAAACTTCATTTGAGAAGGGGATCATGTAAAATGTGGACATGTTCAGCTACTCATGCTGACACATTAAGATACAAATCCTGGGGCCGTACAGTTATTGGGACAACTGTACCCCATCCATTAGAAATGTTGGGTCCACAACATCGAAAAGAGACTCCTTGTGCACCATGTAACACATCAGGGTTCAATTATGTTTCTGTGCATTGTCCAGACGGGATCCATGACGTCTTTAGTTCACGGGGACCATTGCCTGCTTATCTAGGGTCTAAAACATCTGAATCTACATCTATTTTGCAGCCTTGGGAAAGGGAAAGCAAAGTCCCACTGATTAAAAGAGCTACACGTCTTAGAGATGCTATCTCTTGGTTTGTTGAACCCGACTCTAAACTAGCAATGACTATACTTTCTAACATCCACTCTTTAACAGGCGAAGAATGGACCAAAAGGCAGCATGGGTTCAAAAGAACAGGGTCTGCCCTTCATAGGTTTTCGACATCTCGGATGAGCCATGGTGGGTTCGCATCTCAGAGCACTGCAGCATTGACCAGGTTGATGGCAACTACAGACACCATGAGGGATCTGGGAGATCAGAATTTCGACTTTTTATTCCAAGCAACGTTGCTCTATGCTCAAATTACCACCACTGTTGCAAGAGACGGATGGATCACCAGTTGTACAGATCATTATCATATTGCCTGTAAGTCCTGTTTGAGACCCATAGAAGAGATCACCCTGGACTCAAGTATGGACTACACGCCCCCAGATGTATCCCATGTGCTGAAGACATGGAGGAATGGGGAAGGTTCGTGGGGACAAGAGATAAAACAGATCTATCCTTTAGAAGGGAATTGGAAGAATTTAGCACCTGCTGAGCAATCCTATCAAGTCGGCAGATGTATAGGTTTTCTATATGGAGACTTGGCGTATAGAAAATCTACTCATGCCGAGGACAGTTCTCTATTTCCTCTATCTATACAAGGTCGTATTAGAGGTCGAGGTTTCTTAAAAGGGTTGCTAGACGGATTAATGAGAGCAAGTTGCTGCCAAGTAATACACCGGAGAAGTCTGGCTCATTTGAAGAGGCCGGCCAACGCAGTGTACGGAGGTTTGATTTACTTGATTGATAAATTGAGTGTATCACCTCCATTCCTTTCTCTTACTAGATCAGGACCTATTAGAGACGAATTAGAAACGATTCCCCACAAGATCCCAACCTCCTATCCGACAAGCAACCGTGATATGGGGGTGATTGTCAGAAATTACTTCAAATACCAATGCCGTCTAATTGAAAAGGGAAAATACAGATCACATTATTCACAATTATGGTTATTCTCAGATGTCTTATCCATAGACTTCATTGGACCATTCTCTATTTCCACCACCCTCTTGCAAATCCTATACAAGCCATTTTTATCTGGGAAAGATAAGAATGAGTTGAGAGAGCTGGCAAATCTTTCTTCATTGCTAAGATCAGGAGAGGGGTGGGAAGACATACATGTGAAATTCTTCACCAAGGACATATTATTGTGTCCAGAGGAAATCAGACATGCTTGCAAGTTCGGGATTGCTAAGGATAATAATAAAGACATGAGCTATCCCCCTTGGGGAAGGGAATCCAGAGGGACAATTACAACAATCCCTGTTTATTATACGACCACCCCTTACCCAAAGATGCTAGAGATGCCTCCAAGAATCCAAAATCCCCTGCTGTCCGGAATCAGGTTGGGCCAATTACCAACTGGCGCTCATTATAAAATTCGGAGTATATTACATGGAATGGGAATCCATTACAGGGACTTCTTGAGTTGTGGAGACGGCTCCGGAGGGATGACTGCTGCATTACTACGAGAAAATGTGCATAGCAGAGGAATATTCAATAGTCTGTTAGAATTATCAGGGTCAGTCATGCGAGGCGCCTCTCCTGAGCCCCCCAGTGCCCTAGAAACTTTAGGAGGAGATAAATCGAGATGTGTAAATGGTGAAACATGTTGGGAATATCCATCTGACTTATGTGACCCAAGGACTTGGGACTATTTCCTCCGACTCAAAGCAGGCTTGGGGCTTCAAATTGATTTAATTGTAATGGATATGGAAGTTCGGGATTCTTCTACTAGCCTGAAAATTGAGACGAATGTTAGAAATTATGTGCACCGGATTTTGGATGAGCAAGGAGTTTTAATCTACAAGACTTATGGAACATATATTTGTGAGAGCGAAAAGAATGCAGTAACAATCCTTGGTCCCATGTTCAAGACGGTCGACTTAGTTCAAACAGAATTTAGTAGTTCTCAAACGTCTGAAGTATATATGGTATGTAAAGGTTTGAAGAAATTAATCGATGAACCCAATCCCGATTGGTCTTCCATCAATGAATCCTGGAAAAACCTGTACGCATTCCAGTCATCAGAACAGGAATTTGCCAGAGCAAAGAAGGTTAGTACATACTTTACCTTGACAGGTATTCCCTCCCAATTCATTCCTGATCCTTTTGTAAACATTGAGACTATGCTACAAATATTCGGAGTACCCACGGGTGTGTCTCATGCGGCTGCCTTAAAATCATCTGATAGACCTGCAGATTTATTGACCATTAGCCTTTTTTATATGGCGATTATATCGTATTATAACATCAATCATATCAGAGTAGGACCGATACCTCCGAACCCCCCATCAGATGGAATTGCACAAAATGTGGGGATCGCTATAACTGGTATAAGCTTTTGGCTGAGTTTGATGGAGAAAGACATTCCACTATATCAACAGTGTTTAGCAGTTATCCAGCAATCATTCCCGATTAGGTGGGAGGCTGTTTCAGTAAAAGGAGGATACAAGCAGAAGTGGAGTACTAGAGGTGATGGGCTCCCAAAAGATACCCGAATTTCAGACTCCTTGGCCCCAATCGGGAACTGGATCAGATCTCTGGAATTGGTCCGAAACCAAGTTCGTCTAAATCCATTCAATGAGATCTTGTTCAATCAGCTATGTCGTACAGTGGATAATCATTTGAAATGGTCAAATTTGCGAAGAAACACAGGAATGATTGAATGGATCAATAGACGAATTTCAAAAGAAGACCGGTCTATACTGATGTTGAAGAGTGACCTACACGAGGAAAACTCTTGGAGAGATTAA(SEQ ID NO：41)。

实施例2NPL复合体可以实现reRNA^TM在细胞内的自我扩增

按照和实施例1基本相同的方式，对293T细胞进行基于reRNA^TM的体外递送，区别在于，在本实施例中，构建携带编码所述水泡性口炎病毒(VSV)印第安那株(IND)N蛋白的RNA序列(SEQ ID NO:4所示碱基序列)、P蛋白的RNA序列(SEQ ID NO:5所示碱基序列)和L蛋白的RNA序列(SEQ ID NO:6所示碱基序列)中的至少之一的reRNA^TMRNA序列，具体reRNA^TMRNA序列如下所示：

IND-N-GFP；

IND-P-GFP；

IND-L-GFP；

IND-P+L-GFP；

IND-N+L-GFP；

IND-N+P-GFP；和

IND-N+P+L-GFP。

培养293T细胞24小时，观察绿色荧光蛋白的荧光信号。结果如图4所示，其中，显示N蛋白、P蛋白和L蛋白中的任何单个蛋白及两个蛋白的组合均不能启动reRNA^TM负载基因(GFP)的自我扩增及表达，只有NPL共同存在的情况下，才能启动reRNA^TM负载基因(GFP)的自我扩增及mRNA合成。

实施例3水泡性口炎病毒不同株之间的NPL蛋白可以自由组合，可以实现reRNA^TM负载基因在细胞内的表达

按照GeneBank数据库的信息，进一步获取编码水泡性口炎病毒(VSV)新泽西株(NJ)N蛋白的RNA序列(SEQ ID NO：16所示碱基序列)、P蛋白的RNA序列(SEQ ID NO：17所示碱基序列)、L蛋白的RNA序列(SEQ ID NO：18所示碱基序列)，其中，上述编码序列表达获得的水泡性口炎病毒(VSV)新泽西株(NJ)N蛋白的氨基酸序列如SEQ ID NO：13所示，P蛋白的氨基酸序列如SEQ ID NO：14所示，L蛋白的氨基酸序列如SEQ ID NO：15所示。按照和实施例2基本相同的方式，对293T细胞进行基于reRNA^TM的体外递送，其中，本实施例制备含有负载GFP的reRNA^TMRNA分子，核心区域N、P、L蛋白的编码基因分别来自实施例2所述的水泡性口炎病毒印第安那株(IND)和上述水泡性口炎病毒新泽西株(NJ)的不同组合，具体的，构建携带下列序列的reRNA^TMRNA序列(下列RNA序列中均包含了GFP编码序列，为了描述方便，并未写出)：

NJ-N+IND-P+IND-L；

NJ-N+IND-P+NJ-L；

NJ-N+NJ-P+IND-L；

NJ-N+NJ-P+NJ-L；

IND-N+NJ-P+NJ-L；

IND-N+IND-P+NJ-L；

IND-N+NJ-P+IND-L。

分别将相同剂量的上述reRNA^TM导入293T细胞，转染后对细胞进24小时培养，通过观察荧光信号，确认负载基因GFP的表达情况，做qPCR，确认负载基因GFP的自复制情况。

实验结果显示在图5中，结果显示来自两株水泡性口炎病毒的不同NPL组合，都可以启动负载基因(GFP)的表达。

实施例4水泡性口炎病毒3株之间的NPL蛋白的组合也可以实现reRNA^TM在细胞内的表达

本实施例中，发明人发现对上述3株水泡性口炎病毒的N、P、L蛋白进行任意组合均可以有效启动负载基因(GFP)的表达。具体地，按照GeneBank数据库的信息，进一步获取水泡性口炎病毒(VSV)可卡尔株(COC)P蛋白的编码基因(SEQ ID NO：23所示碱基序列)，获得的水泡性口炎病毒(VSV)可卡尔株(COC)P蛋白的氨基酸序列如SEQ ID NO：20所示，按照和实施例2基本相同的方式，对293T细胞进行基于reRNA^TM的体外递送，此处以制备含有负载GFP以及N蛋白来自实施例3所述的水泡性口炎病毒新泽西株(NJ)，P蛋白来自可卡尔株(COC)，L蛋白来自实施例1和2所述的印第安那株(IND)的reRNA^TMRNA分子为例进行实验。

将上述reRNA^TM导入293T细胞，转染后对细胞进行24小时培养，通过观察荧光信号，确认负载基因GFP的表达情况。

实验结果显示在图6中，结果显示来自三株水泡性口炎病毒的NPL组合，可以有效启动负载基因(GFP)的表达。

实施例5水泡性口炎病毒的NPL蛋白可以与其他弹状病毒(狂犬病毒)的NPL进行组合，实现reRNA^TM的胞内表达

按照和实施例2基本相同的方式，对293T细胞进行基于reRNA^TM的体外递送，在本实施例中，制备含有负载GFP的reRNA^TMRNA分子，核心区域NPL基因分别来自水泡性口炎病毒和狂犬病毒的任意不同组合，其中，狂犬病毒N蛋白、P蛋白、L蛋白的氨基酸序列分别如SEQ IDNO：25、26、27所示，编码狂犬病毒N蛋白、P蛋白、L蛋白的RNA序列分别如SEQ ID NO：28、29、30所示。

分别将相同剂量的不同reRNA^TM导入293T细胞，转染后对细胞进行24小时培养，通过观察荧光信号，确认负载基因GFP的表达情况。结果显示来自不同弹状病毒的不同NPL组合，都可以启动负载基因(GFP)的表达。

实施例6将与NPL蛋白具有类似功能的蛋白组合，可以实现reRNA^TM的胞内表达

按照和实施例2基本相同的方式，对293T细胞进行基于reRNA^TM的体外递送，在本实施例中，制备含有负载GFP的reRNA^TMRNA分子，核心区域NPL分别携带不同的保守突变编码N、P、L蛋白的RNA序列分别如SEQ ID NO：10、11、12所示，获得的N、P、L蛋白的氨基酸序列分别如SEQ ID NO：7、8、9所示。分别将相同剂量的不同reRNA^TM转染293T细胞，转染后对细胞进行24小时培养，通过观察荧光信号，确认负载基因GFP的表达情况。结果显示携带保守突变的不同NPL组合，都可以启动负载基因(GFP)的表达。

实施例7reRNA^TM可以实现mGM-CSF分泌蛋白表达

按照和实施例2基本相同的方式，对小鼠MC38细胞进行基于reRNA^TM的体外递送，区别在于，制备含有负载mGM-CSF(鼠粒细胞-巨噬细胞集落刺激因子)的reRNA^TM，其中的核心区域为NPL(来自实施例1和实施例2所述的水泡性口炎病毒印第安那株(IND))，将负载mGM-CSF的reRNA^TM导入小鼠MC38细胞中，不同时间收集细胞培养上清，检测mGM-CSF分泌量。结果显示，可以有效地启动mGM-CSF的表达，并能有效地使其分泌到细胞外。

实施例8reRNA^TM可以实现OVA抗原肽在抗原呈递细胞(APCs)内的递呈

制备含有负载编码OVA抗原的RNA序列的reRNA^TM(reRNA^TMOVA组)和reRNA^TM(Blank组)，其中reRNA^TM的核心区域为NPL(来自实施例1和实施例2所述的水泡性口炎病毒印第安那株(IND))，表达获得的OVA抗原的氨基酸序列如SEQ ID NO:42所示。具体地，将reRNA^TM导入到C57小鼠脾脏细胞中，培养24小时后回收小鼠脾脏细胞。利用流式细胞检测技术对抗原呈递细胞进行OVA抗原呈递检测。

MGSIGAASMEFCFDVFKELKVHHANENIFYCPIAIMSALAMVYLGAKDSTRTQINKVVRFDKLPGFGDSIEAQCGTSVNVHSSLRDILNQITKPNDVYSFSLASRLYAEERYPILPEYLQCVKELYRGGLEPINFQTAADQARELINSWVESQTNGIIRNVLQPSSVDSQTAMVLVNAIVFKGLWEKAFRDEDTQAMPFRVTERESKPVQMMYQTGLFRVASMASEKMKILELPFASGTMSMLVLLPDEVSGLEQLESIINFEKLTEWTSSNVMEERKIKVYLPRMKMEEKYNLTSVLMAMGITDVFSSSANLSGISSAESLKISQAVHAAHAEINEAGREVVGSAEAGVDAASVSEEFRADHPFLFCIKHIATNAVLFFGRCVSP(SEQ ID NO:42)。

实验结果显示在图7中，结果显示抗原呈递细胞能够呈递OVA抗原。这也表明，reRNA^TM所携带的OVA抗原不仅能够在APC细胞内表达，还能够被APC细胞加工呈递至细胞表面。

在本说明书的描述中，参考术语“一个实施例”、“一些实施例”、“示例”、“具体示例”、或“一些示例”等的描述意指结合该实施例或示例描述的具体特征、结构、材料或者特点包含于本申请的至少一个实施例或示例中。在本说明书中，对上述术语的示意性表述不必针对的是相同的实施例或示例。而且，描述的具体特征、结构、材料或者特点可以在任一个或多个实施例或示例中以合适的方式结合。此外，在不相互矛盾的情况下，本领域的技术人员可以将本说明书中描述的不同实施例或示例以及不同实施例或示例的特征进行结合和组合。

尽管上面已经示出和描述了本申请的实施例，可以理解的是，上述实施例是示例性的，不能理解为对本申请的限制，本领域的普通技术人员在本申请的范围内可以对上述实施例进行变化、修改、替换和变型。

Claims (17)

1.一种自复制RNA分子，其特征在于，包括：

第一RNA序列，所述第一RNA序列编码N蛋白或其功能片段；

第二RNA序列，所述第二RNA序列编码P蛋白或其功能片段；和

第三RNA序列，所述第三RNA序列编码L蛋白或其功能片段，

其中，所述自复制RNA分子在动物细胞内适于产生所述N蛋白、所述P蛋白、所述L蛋白或其功能片段。

2.根据权利要求1所述的自复制RNA分子，其特征在于，所述N蛋白、所述P蛋白和所述L蛋白的至少之一分别独立地来自弹状病毒科病毒。

3.根据权利要求2所述的自复制RNA分子，其特征在于，所述弹状病毒科病毒包括选自水泡性病毒属病毒、狂犬病病毒属病毒和短暂热病毒属病毒中的至少之一。

4.根据权利要求2所述的自复制RNA分子，其特征在于，所述弹状病毒科病毒包括选自水疱性口炎新泽西病毒、马拉巴病毒、水疱性口炎阿拉戈斯病毒印第安纳3号、科卡尔病毒印第安纳2号、伊斯法罕病毒、钱迪普拉病毒、鲤鱼春季病毒血症病毒、美国蝙蝠水疱病毒、卡拉哈斯水疱病毒、朱罗纳水疱病毒、马尔帕斯春季水疱病毒、莫雷顿水疱病毒、佩里特水疱病毒、皮里水泡病毒、拉迪水泡病毒、尤格·博格达诺瓦克水泡病毒、西高加索蝙蝠病毒、Aravan狂犬病毒、Khujand狂犬病毒、狂犬病病毒、澳大利亚蝙蝠狂犬病毒、欧洲蝙蝠狂犬病毒1、欧洲蝙蝠狂犬病毒2、伊尔库特病毒、希莫尼蝙蝠病毒、拉各斯蝙蝠病毒、莫科拉病毒、莱伊达蝙蝠狂犬病毒、伊科马狂犬病毒、甘诺鲁瓦蝙蝠狂犬病毒、博克洛蝙蝠狂犬病毒、杜文哈格狂犬病毒、热蜉蝣病毒、阿德莱德河蜉蝣病毒、伯里玛蜉蝣病毒和金伯利蜉蝣病毒的至少之一。

5.根据权利要求1所述的自复制RNA分子，其特征在于，

所述N蛋白具有SEQ ID NO：1、7、13、19或25所示氨基酸序列；

所述P蛋白具有SEQ ID NO：2、8、14、20或26所示氨基酸序列；和

所述L蛋白具有SEQ ID NO：3、9、15、21或27所示氨基酸序列。

6.根据权利要求5所述的自复制RNA分子，其特征在于，

所述N蛋白具有SEQ ID NO：1所示氨基酸序列，所述P蛋白具有SEQ ID NO：2所示氨基酸序列，所述L蛋白具有SEQ ID NO：3所示氨基酸序列；或

所述N蛋白具有SEQ ID NO：7所示氨基酸序列，所述P蛋白具有SEQ ID NO：8所示氨基酸序列，所述L蛋白具有SEQ ID NO：9所示氨基酸序列；或

所述N蛋白具有SEQ ID NO：13所示氨基酸序列，所述P蛋白具有SEQ ID NO：14所示氨基酸序列的，所述L蛋白具有SEQ ID NO：15所示氨基酸序列；或

所述N蛋白具有SEQ ID NO：19所示氨基酸序列，所述P蛋白具有SEQ ID NO：20所示氨基酸序列，所述L蛋白具有SEQ ID NO：21所示氨基酸序列；或

所述N蛋白具有SEQ ID NO：25所示氨基酸序列，所述P蛋白具有SEQ ID NO：26所示氨基酸序列，所述L蛋白具有SEQ ID NO：27所示氨基酸序列。

7.根据权利要求1所述的自复制RNA分子，其特征在于，所述第一RNA序列具有SEQ IDNO：4、10、16、22或28所示碱基序列；

所述第二RNA序列具有SEQ ID NO：5、11、17、23或29所示碱基序列；和

所述第三RNA序列具有SEQ ID NO：6、12、18、24或30所示碱基序列。

8.根据权利要求7所述的自复制RNA分子，其特征在于，所述第一RNA序列具有SEQ IDNO：4所示碱基序列，所述第二RNA序列具有SEQ ID NO：5所示碱基序列，所述第三RNA序列具有SEQ ID NO：6所示碱基序列；或

所述第一RNA序列具有SEQ ID NO：10所示碱基序列，所述第二RNA序列具有SEQ ID NO：11所示碱基序列，所述第三RNA序列具有SEQ ID NO：12所示碱基序列；或

所述第一RNA序列具有SEQ ID NO：16所示碱基序列，所述第二RNA序列具有SEQ ID NO：17所示碱基序列，所述第三RNA序列具有SEQ ID NO：18所示碱基序列；或

所述第一RNA序列具有SEQ ID NO：22所示碱基序列，所述第二RNA序列具有SEQ ID NO：23所示碱基序列，所述第三RNA序列具有SEQ ID NO：24所示碱基序列；或

所述第一RNA序列具有SEQ ID NO：28所示碱基序列，所述第二RNA序列具有SEQ ID NO：29所示碱基序列，所述第三RNA序列具有SEQ ID NO：30所示碱基序列。

9.根据权利要求1所述的自复制RNA分子，其特征在于，进一步包括：

靶分子编码区，所述靶分子编码区编码至少一个靶分子。

10.根据权利要求9所述的自复制RNA分子，其特征在于，所述靶分子适于作为核酸类药物、蛋白类药物、病原体疫苗、肿瘤疫苗和罕见病治疗制剂的至少之一。

11.根据权利要求9所述的自复制RNA分子，其特征在于，所述靶分子编码区编码荧光蛋白、粒细胞-巨噬细胞集落刺激因子和OVA的至少之一，可选的，所述荧光蛋白包括绿色荧光蛋白、红色荧光蛋白以及黄色荧光蛋白的至少之一。

12.一种DNA分子，其编码权利要求1～11任一项所述的自复制RNA分子。

13.一种表达载体，其携带权利要求12所述的DNA分子。

14.一种制备权利要求1～11任一项所述自复制RNA分子的方法，其特征在于，包括：

使权利要求12所述的DNA分子或者权利要求13所述的表达载体在宿主细胞中表达所述自复制RNA分子；和

收集所述自复制RNA分子。

15.一种RNA-蛋白质复合物，其特征在于，包括：

权利要求1～11任一项所述自复制RNA分子；和

蛋白质，所述蛋白质包括N蛋白或其功能片段、P蛋白或其功能片段和L蛋白或其功能片段。

16.一种药物组合物，其特征在于，包括权利要求1～11任一项所述的自复制RNA分子、权利要求12所述的DNA分子、权利要求13所述的表达载体或者权利要求14所述的RNA-蛋白质复合物。

17.一种在个体中表达靶分子的方法，其特征在于，包括：

为个体给予权利要求1～11任一项所述的自复制RNA分子、权利要求12所述的DNA分子、权利要求13所述的表达载体或者权利要求15所述的RNA-蛋白质复合物或者权利要求16所述的药物组合物。

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