CN110437317A - 具有变异衣壳蛋白的腺相关病毒及其用途 - Google Patents
具有变异衣壳蛋白的腺相关病毒及其用途 Download PDFInfo
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Abstract
本发明提供一种具有变异型衣壳蛋白的腺相关病毒及其用途。所述变异型腺相关病毒AAV‑ie，即在野生型AAV‑DJ衣壳蛋白VP1在N589与R590之间插入氨基酸序列DGTLAVPFK。变异型腺相关病毒AAV‑ie能够高效感染毛细胞和支持细胞，相比于其亲本具有较大的改善，为科学研究提供了更好的技术支持。
Description
技术领域
本发明涉及生物技术领域，特别涉及一种具有变异衣壳蛋白的腺相关病毒及其用途。
背景技术
听力损失是最常见的感官残疾之一，影响到世界6.8％以上的人口(约5亿人)。听力损失最普遍的治疗方法是使用听力设备，这种装置可以放大声音、增强声音传递或直接刺激神经元。这种方法目前是治疗听力损失的最佳选择，但不幸的是，在嘈杂的环境中，听力敏感性和对自然声音的感知仍有局限性。因此，需要更好的替代策略来治疗听力损失。近年来，基因治疗已成为治疗遗传性疾病的一种有希望的策略。
感音神经性听力损失的病例有一半是由于毛细胞(Hair cell)和支持细胞(Supporting cell)的基因突变造成的，螺旋神经元中的基因突变也能导致一部分的听力障碍。100多个耳聋基因的突变已被发现。遗传性听力损失是一种典型的单源性疾病，单一的突变会导致听力损失。因此，基因治疗是一种理想的、有希望的潜在治疗策略，可以保持或重建听力功能。对于遗传性听力损失，大多数耳聋基因在毛细胞中表达，一部分基因的突变可能影响螺旋神经元。大量的基因突变直接影响毛细胞的各种功能。但一些关键耳聋基因如GJB2等主要表达在支持细胞中，其突变会影响支持细胞的功能，进而伴随毛细胞的损伤，最终导致听力丧失。据报道，有超过300个GJB2的突变能够导致遗传听力损失。GJB2的突变是人类遗传性听力损失最常见的原因，约占常染色体隐性听力损失的50％，占所有遗传性听力损失的15％-18％。因此，遗传性听力损失的基因治疗需要同时针对毛细胞和支持细胞。
AAV是一种直径为18-26nm的无包膜的二十面体病毒，衣壳结构由三种衣壳蛋白(VP1、VP2和VP3)构成。衣壳内含有一条4.7kb的单链DNA(ssDNA)，携带两个基因rep和cap，DNA两侧是回文倒置末端重复序列(ITRs)。rep和cap都有多个open reading frames(ORFs)，它们表达基因组复制和包装所需的蛋白质。由于ITRs序列可以直接应用于普通质粒中，因此可以直接将外源DNA插入ITRs之间构成重组质粒，将AAV病毒基因克隆进入第二个质粒，在帮助质粒Helper的作用下包装形成新的重组AAV病毒。目前常用的AAV有AAV-1，AAV-2，AAV-5，AAV-8，AAV-9以及AAV-DJ等。
一些病毒和非病毒基因转移策略已在基因治疗中得到测试。在这些方法中，AAV介导的基因治疗在过去十年中进展迅速，已经有170多个人体试验(7.2％)使用了AAV载体。AAV载体具有出色的安全性、低免疫原性和较高的转染效率。美国FDA在2017年批准了首个AAV介导的基因治疗，其用于治疗罕见的遗传性眼病。然而，迄今为止，还没有基于AAV载体的耳蜗基因治疗。这可能是由于耳蜗中不同细胞类型缺乏具有较高传导效率的AAV所导致。几项研究在动物模型中测试了AAV介导的耳蜗基因疗法，并显示出有希望的结果。然而，大多数研究的重点是毛细胞和螺旋神经元，这可能是由于缺乏安全和有效的针对支持细胞的AAV。一些AAV血清型已在耳蜗中进行了评估，所有检测的AAV对支持细胞的感染效率极低，低于13％。因此，设计新型可以有效地转染支持细胞的AAV，成为进一步应用基因治疗遗传性听力损失的里程碑。
发明内容
鉴于以上所述现有技术的缺点，本发明的目的在于提供一种具有变异衣壳蛋白的腺相关病毒及其用途，用于解决现有技术中AAV对毛细胞和支持细胞感染率不高的问题。
基于在野生型AAV-DJ的衣壳蛋白VP1的N589与R590之间，插入氨基酸序列如SEQID NO.1所示的氨基酸片段之后，构建的获得的变体AAV-ie具有更强的毛细胞和支持细胞的感染性，申请人提出本发明创造。
为实现上述目的及其他相关目的，本发明一方面提供一种变异型腺相关病毒AAV-ie衣壳蛋白VP1，所述变异型腺相关病毒AAV-ie衣壳蛋白VP1，相比于野生型AAV-DJ衣壳蛋白VP1，在N589与R590之间插入氨基酸片段，所述氨基酸片段的氨基酸序列如SEQ ID NO.1所示。
DGTLAVPFK(SEQ ID NO.1)
在本发明一些实施方式中，所述野生型AAV-DJ衣壳蛋白VP1的氨基酸序列如SEQID NO.3所示。
MAADGYLPDWLEDTLSEGIRQWWKLKPGPPPPKPAERHKDDSRGLVLPGYKYLGPFNGLDKGEPVNEADAAALEHDKAYDRQLDSGDNPYLKYNHADAEFQERLKEDTSFGGNLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEHSPVEPDSSSGTGKAGQQPARKRLNFGQTGDADSVPDPQPIGEPPAAPSGVGSLTMAAGGGAPMADNNEGADGVGNSSGNWHCDSTWMGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLSFKLFNIQVKEVTQNEGTKTIANNLTSTIQVFTDSEYQLPYVLGSAHQGCLPPFPADVFMIPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFQFTYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTQTTGGTTNTQTLGFSQGGPNTMANQAKNWLPGPCYRQQRVSKTSADNNNSEYSWTGATKYHLNGRDSLVNPGPAMASHKDDEEKFFPQSGVLIFGKQGSEKTNVDIEKVMITDEEEIRTTNPVATEQYGSVSTNLQRGNRQAATADVNTQGVLPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPLMGGFGLKHPPPQILIKNTPVPADPPTTFNQSKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSTSVDFAVNTEGVYSEPRPIGTRYLTRNL(SEQ ID NO.3)
在本发明一些实施方式中，所述变异型腺相关病毒AAV-ie衣壳蛋白VP1的氨基酸序列如SEQ ID NO.4所示。
MAADGYLPDWLEDTLSEGIRQWWKLKPGPPPPKPAERHKDDSRGLVLPGYKYLGPFNGLDKGEPVNEADAAALEHDKAYDRQLDSGDNPYLKYNHADAEFQERLKEDTSFGGNLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEHSPVEPDSSSGTGKAGQQPARKRLNFGQTGDADSVPDPQPIGEPPAAPSGVGSLTMAAGGGAPMADNNEGADGVGNSSGNWHCDSTWMGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLSFKLFNIQVKEVTQNEGTKTIANNLTSTIQVFTDSEYQLPYVLGSAHQGCLPPFPADVFMIPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFQFTYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTQTTGGTTNTQTLGFSQGGPNTMANQAKNWLPGPCYRQQRVSKTSADNNNSEYSWTGATKYHLNGRDSLVNPGPAMASHKDDEEKFFPQSGVLIFGKQGSEKTNVDIEKVMITDEEEIRTTNPVATEQYGSVSTNLQRGNDGTLAVPFKRQAATADVNTQGVLPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPLMGGFGLKHPPPQILIKNTPVPADPPTTFNQSKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSTSVDFAVNTEGVYSEPRPIGTRYLTRNL(SEQ IDNO.4)
本发明的另外一个方面提供了一种分离的核酸，所述核酸含有编码上述变异型腺相关病毒AAV-ie衣壳蛋白VP1的核苷酸序列。
本发明的另外一个方面提供了一种构建体，所述构建体含有上述的分离的核酸。所述构建体通常可以通过将上述分离的核酸插入合适的表达载体中构建获得，本领域技术人员可选择合适的表达载体，例如，上述表达载体可以是Rep2-Capsid等。
本发明的另外一个方面提供了一种宿主细胞，所述宿主细胞包含上述的构建体或基因组中整合有外源的上述的分离的核酸，或所述宿主细胞包含上述变异型腺相关病毒AAV-ie。所述宿主细胞可以是真核细胞和/或原核细胞，更具体可以是小鼠细胞、人细胞等，更具体可以是人胚胎肾细胞等，更具体可以是HEK293FT细胞等。
本发明的另外一个方面提供了一种变异型腺相关病毒AAV-ie，所述变异型腺相关病毒AAV-ie含有如上述变异型腺相关病毒AAV-ie衣壳蛋VP1。即在野生型AAV-DJ衣壳蛋白VP1在N589与R590之间，插入氨基酸序列如SEQ ID NO.1所示的氨基酸片段。
进一步地，所述变异型腺相关病毒AAV-ie还包括编码目的产物的异源核苷酸序列，所述编码目的产物的异源核苷酸序可以是各种衣壳蛋白能包裹携带的。上述编码目的产物的异源核苷酸序列通常可以是构建体，所述构建体通常可以含有编码目的产物的核酸。所述构建体通常可以通过将编码目的产物的核酸插入合适的表达载体中构建获得，本领域技术人员可选择合适的表达载体，例如，上述表达载体可以是包括但不限于pAAV-CAG、pAAV-TRE、pAAV-EF1a、pAAV-GFAP promoter、pAAV-Lgr5 promoter、pAAV-Sox2 promoter表达载体等。
进一步地，所述目的产物为核酸或蛋白质，所述核酸可以是小向导RNA(sgRNA)、干扰RNA(RNAi)等。
所述变异型腺相关病毒AAV-ie可以作为载体材料，将外源的基因导入受试个体的细胞中，相比于亲本野生型AAV-DJ，变异型腺相关病毒AAV-ie对毛细胞和支持细胞的感染率明显提高。
本发明的另外一个方面提供了一种药物组合物，所述药物组合物包含如上所述的变异型腺相关病毒AAV-ie以及药学上可接受的辅料。
本发明的另外一个方面提供了上述变异型腺相关病毒AAV-ie在用于将目的产物递送至个体的毛细胞和/或支持细胞中的用途。所述目的产物递送可以是非诊断治疗目的的，例如，可以是体外的，对离体的毛细胞和/或支持细胞进行目的产物递送。所述毛细胞通常包括外毛细胞和/或内毛细胞。
进一步地，所述目的产物为核酸或蛋白质，所述核酸可以是小向导RNA(sgRNA)、干扰RNA(RNAi)等。
本发明的另外一个方面提供了上述变异型腺相关病毒AAV-ie在制备治疗个体中耳蜗损伤导致的听力障碍疾病的药物中的用途。
进一步地，所述听力障碍疾病为毛细胞和/或支持细胞和/或螺旋神经元细胞相关疾病。
进一步地，所述听力障碍疾病为基因缺陷、环境损伤或衰老的相关疾病，例如，可以是基因突变等所导致的相关疾病，再例如，可以是噪声所致的、药物所导致的相关疾病，再例如，可以是衰老所导致的相关疾病。
进一步地，听力障碍疾病可以为细胞损伤等的相关疾病，具体可以是耳蜗毛细胞损伤、支持细胞损伤等，更具体可以是基因突变导致的耳蜗毛细胞损伤、基因突变导致的支持细胞损伤等、噪声所致的细胞损伤、药物所导致的细胞损伤或者衰老所导致。
进一步地，所述变异型腺相关病毒AAV-ie作为递送目的产物的载体。如上所述，本发明的具有变异衣壳蛋白的腺相关病毒及其用途，具有以下有益效果：
具有更强的毛细胞和支持细胞的感染性，能够高效感染毛细胞和支持细胞，也能高效感染螺旋神经元，相比于其亲本具有较大的改善，为科学研究提供了更好的技术支持。
附图说明
图1a显示为实施例一中野生型AAV-DJ感染HEK 293T细胞后绿色荧光蛋白mNeonGreen的表达图。
图1b显示为实施例一中变异型AAV-ie感染HEK 293T细胞后绿色荧光蛋白mNeonGreen的表达图。
图1c显示为实施例一中野生型AAV-DJ和变异型AAV-ie感染HEK 293T细胞的荧光比例的统计。
图2a显示了实施例二中AAV1、AAV6、AAV9、AAV-DJ、AAV-ie、Anc80L65体外感染体外培养的耳蜗组织后绿色荧光mNeonGreen与支持细胞特异性标记蛋白Sox2的免疫染色照片。
图2b显示了对图2a的数据统计结果。
图3a显示了实施例二中AAV1、AAV6、AAV9、AAV-DJ、AAV-ie、Anc80L65感染体外培养的耳蜗组织后绿色荧光mNeonGreen与毛细胞特异性标记蛋白Myo7a的免疫染色照片。
图3b显示了对图3a的数据统计结果。
图4a显示了实施例三中大视野下，耳蜗三个区域：顶转(apex)、中转(middle)和底转(base)的绿色荧光mNeonGreen的表达和品红色所代表的支持细胞标记蛋白Sox2的染色结果。新生小鼠体内圆窗注射AAV-ie病毒后14天，将耳蜗剖出，进行支持细胞特异性标记蛋白Sox2的免疫染色，然后对绿色荧光mNeonGreen和免疫染色的结果拍照。
图4b显示了实施例三中AAV1、AAV6、AAV8、AAV9、DJ8、Anc80L65、AAV-DJ、AAV-ie感染耳蜗组织后对绿色荧光mNeonGreen和Sox2阳性的支持细胞的荧光照片。
图4c显示了对图4b的数据统计结果。
图5a显示了实施例三中AAV-DJ，AAV-ie绿色荧光mNeonGreen的表达和品红色所代表的毛细胞标记蛋白Myo7a的染色结果。
图5b显示了对图5a的数据统计结果。
图6a显示了耳蜗三个区域：顶转(apex)、中转(middle)和底转(base)的绿色荧光mNeonGreen的表达和品红色所代表的螺旋神经元标记蛋白NeurN的染色结果。
图6b是对图6a的统计结果。
图7显示了实施例四中小鼠耳蜗样品免疫染色结果。
图8a显示了实施例五中小鼠椭圆囊样本免疫染色结果。
图8b显示了实施例五中人的椭圆囊样本免疫染色结果。
图8c为图8b的放大结果。
具体实施方式
以下通过特定的具体实例说明本发明的实施方式，本领域技术人员可由本说明书所揭露的内容轻易地了解本发明的其他优点与功效。本发明还可以通过另外不同的具体实施方式加以实施或应用，本说明书中的各项细节也可以基于不同观点与应用，在没有背离本发明的精神下进行各种修饰或改变。
在进一步描述本发明具体实施方式之前，应理解，本发明的保护范围不局限于下述特定的具体实施方案；还应当理解，本发明实施例中使用的术语是为了描述特定的具体实施方案，而不是为了限制本发明的保护范围；在本发明说明书和权利要求书中，除非文中另外明确指出，单数形式“一个”、“一”和“这个”包括复数形式。
当实施例给出数值范围时，应理解，除非本发明另有说明，每个数值范围的两个端点以及两个端点之间任何一个数值均可选用。除非另外定义，本发明中使用的所有技术和科学术语与本技术领域技术人员通常理解的意义相同。除实施例中使用的具体方法、设备、材料外，根据本技术领域的技术人员对现有技术的掌握及本发明的记载，还可以使用与本发明实施例中所述的方法、设备、材料相似或等同的现有技术的任何方法、设备和材料来实现本发明。
除非另外说明，本发明中所公开的实验方法、检测方法、制备方法均采用本技术领域常规的分子生物学、生物化学、染色质结构和分析、分析化学、细胞培养、重组DNA技术及相关领域的常规技术。这些技术在现有文献中已有完善说明，具体可参见Sambrook等MOLECULAR CLONING：A LABORATORY MANUAL，Second edition，Cold Spring HarborLaboratory Press，1989and Third edition，2001；Ausubel等，CURRENT PROTOCOLS INMOLECULAR BIOLOGY，John Wiley&Sons，New York，1987and periodic updates；theseries METHODS IN ENZYMOLOGY，Academic Press，San Diego；Wolffe，CHROMATINSTRUCTURE AND FUNCTION，Third edition，Academic Press，San Diego，1998；METHODS INENZYMOLOGY，Vol.304，Chromatin(P.M.Wassarman and A.P.Wolffe，eds.)，AcademicPress，San Diego，1999；和METHODS IN MOLECULAR BIOLOGY，Vol.119，ChromatinProtocols(P.B.Becker，ed.)Humana Press，Totowa，1999等。
材料与来源：
幼鼠：上海灵畅生物科技有限公司
各种AAV：南京金斯瑞公司合成构建
驴血清：上海翊圣生物科技有限公司
各个抗体与染色时的稀释比：
一抗：myosin 7A(Myo7a,#25-6790Proteus Biosciences,1:1000),Sox2(Sox-2,#sc-17320,Santa Cruz Biotechnology,1:1000),Flag(Flag,#F3165,Sigma Aldrich,1:1000),NeuN(NeuN,#12943S,Cell Signaling Technology,1:500)。
二抗：二抗标记物anti-rat、mouse、rabbit及goat的各三种不同标记颜色(TRIC,FITC,Cy5)的二抗，来源于invitrogen公司。
细胞和组织培养试剂：DMEM(Hyclone公司)，胎牛血清(Lensa公司)，添加物N2(ThermoFisher公司)，氨苄西林(ThermoFisher公司)，双抗(ThermoFisher公司)。
细胞与组织培养耗材：包括各类培养皿、离心管、移液管，一次性滤器等常用耗材购于Corning公司。
实施例一 构建AAV变异体并感染HEK 293T细胞
AAV变异体(命名为AAV-ie)Rep-Cap质粒的构建。
AAV的包装需要三个质粒：含有目的基因的基因组质粒、Rep-Cap质粒和Helper质粒。
其中Rep-Cap质粒中的Cap蛋白的序列决定了AAV的不同血清型，进而影响了AAV感染细胞的偏好性。因此，改造Cap蛋白能够得到新的AAV型。
(1)载体的构建、酶切和纯化
亲本AAV-DJ的Rep-Cap质粒由南京金斯瑞公司(www.genscript.com.cn)合成。
首先，通过聚合酶链式反应(PCR)诱变将独特NheI内切酶位点引入野生型AAV-DJVP1衣壳蛋白氨基酸589和590之间，引物南京金斯瑞公司(www.genscript.com.cn)合成。然后用限制性内切酶NheI消化，并回收(Axygen:AP-GX-250G)。回收后的片段经过Nanodrop2000检测浓度。
编码DGTLAVPFK的DNA序列(SEQ ID NO.2：gatgggactttggcggtgccttttaag)由南京金斯瑞公司(www.genscript.com.cn)合成。将合成的片段用超纯水溶解到10μM。
(2)载体的连接、转化与质粒提取。
回收的骨架载体和编码DGTLAVPFK的DNA序列片段经过重组连接(Novoprotein:NR001A)，连接体系如下：重组连接buffer 2μL，骨架载体30ng，编码DGTLAVPFK的DNA片段1μL，重组连接酶0.5μL，ddH2O补齐到10μL，50℃反应20分钟。
转化步骤如下：取100μL感受态细胞(TransGen:CD201)在冰上解冻；10μL的连接产物与感受态细胞混合，冰上放置20分钟；42℃热激60秒；冰上放置2分钟，加入400μL复苏LB培养基(MDBio:L001-1kg)，摇床30分钟；取70μL涂氨苄平板(50μg/ml，37℃培养箱，培养14个小时。
挑选单克隆菌，在4ml液体LB培养基中扩大培养，14小时后提取质粒(Axygene:AP-MN-P-250G)。
步骤如下：菌液经过4000转/分钟离心10分钟，倒掉上清培养基；加入350μL的buffer S1，将菌体吹散，转移到2ml离心管中；加入250μL的buffer S2，上下颠倒8次；加入250μL的buffer S3，颠倒混匀6次，产生沉淀；12000转/分钟离心10分钟，取上清过柱；离心1分钟，倒掉废液，加入500μL的W1，离心一分钟，倒掉废液；加入750μL的W2，离心，倒掉上清；加入500μL的W2，离心，倒掉上清；空转1分钟；加入50μL的洗脱液，静置2分钟，离心。获得质粒经过浓度检测，取10μL送测序，阳性质粒保存在-20℃。测序结果表明，获得质粒能够编码变异型衣壳蛋白VP1。
进一步的实验结果表明，制备获得的质粒能够表达变异型衣壳蛋白VP1，其中，AAV-ie capsid VP1的多核苷酸编码序列如SEQ ID NO.5所示，构建获得的Rep-Cap质粒的全序列如SEQ ID NO.6所示。
2.AAV变异体(命名为AAV-ie)病毒的包装与纯化。
将得到的Rep-Cap质粒，表达一个绿色荧光蛋白mNeonGreen(GenBank:LC279210.1)的基因组质粒pAAV-CAG-mNeonGreen(质粒全序列如SEQ ID NO.11所示)，pHelper质粒(质粒全序列如SEQ ID NO.12所示)以合适的量共转于HEK-293T细胞中，采用碘二烷醇梯度超高速离心纯化AAV病毒，测量病毒滴度在1E+12-1E+13GC/mL为合适浓度，放置-80℃备用。
在用DMEM+10％胎牛血清培养的HEK 293T细胞中加入衣壳蛋白变异体和其亲本衣壳蛋白产生的AAV病毒(分别为AAV-ie和AAV-DJ)。病毒加入的MOI值为1000。48小时后，用荧光显微镜观察了绿色荧光蛋白mNeonGreen的表达，如图1a-1c。图1a，1b大图表示的绿色荧光蛋白的信号，右上角的是明场自然光条件下的细胞。
结果显示：AAV变异体与其亲本AAV感染HEK 293T细胞的比例相似。
实施例二 AAV变异体体外感染小鼠耳蜗组织
在冰上迅速取出P3野生型C57小鼠(上海灵畅生物科技有限公司)耳蜗，贴于涂有cell-tak的玻片上，置于98％DMEM+1％N2+1％Amp(5ug/mL)培养基中稳定12h后，加入1％FBS和2×1010GC AAV培养48-60h。然后对培养的样品进行免疫染色鉴定。样品浸入4％中PFA中进行固定处理,然后样品浸入含有10％的驴血清和0.3％Triton X-100PBS中，室温下孵育1小时后,加入Myo7a(肌球蛋白7a)、Sox2蛋白的抗体以及相应的二级抗体。样品采用抗荧光淬火剂安装介质封片，并采用共聚焦进行观察。
在利用共聚焦方法拍摄图像时，选择拍摄变异体感染样品的激光功率设置作为标准。所有可见的绿色荧光蛋白mNeonGreen的信号都是通过相同的激光设置捕获的。
在数据处理方面，通过计算mNeonGreen阳性的支持细胞和毛细胞的数量，在耳蜗上手动量化了mNeonGreen的百分比，Myo7a阳性的是毛细胞，Sox2蛋白阳性的是支持细胞。
其中如图2a-2b，品红色表示的是支持细胞，绿色表示的是AAV导入细胞内表达出来的绿色荧光蛋白mNeonGreen,图2b显示的为图2a的统计数据。结果显示：AAV变异体AAV-ie能高效感染体外培养的小鼠耳蜗组织的支持细胞，与其他AAV及亲本AAV相比，其感染Sox2阳性的细胞比例更高。
其中如图3a-3b，品红色表示的是毛细胞，绿色表示的是AAV导入细胞内表达出来的绿色荧光蛋白mNeonGreen，图3b显示的为图3a的统计数据，其中OHC表示外毛细胞，IHC表示内毛细胞。结果显示：AAV变异体能高效感染体外培养的小鼠耳蜗组织中的毛细胞。与AAV1,AAV6,AAV9,Anc80L65(构建方法与实施例一相同，区别仅在于所使用的Rep-Cap质粒不同，质粒序列详见SEQ ID NO.7～10)和AAV-DJ相比，变异体AAV-ie对Myo7a阳性的毛细胞感染率很高。
上述结果表明：AAV变异体AAV-ie能高效感染Myo7a阳性的毛细胞、Sox2蛋白阳性的支持细胞。
实施例三 AAV变异体体内注射后能高效感染小鼠耳蜗中的多种组织细胞
利用耳蜗圆窗注射技术，将1.5uL AAV变异体病毒(各种病毒的浓度如图4中标注)注射入耳蜗的外淋巴液中。具体步骤如下：新生小鼠的麻醉方法采用低温诱导麻醉的方法。P2-3小鼠置于冰浴中2-3min，取出在冰垫上进行后续手术过程。手术仅在每只小鼠的左耳中进行，右耳为阴性对照。手术时，在左耳耳后切口，根据颞骨和面神经的相对位置关系，暴露圆窗。手术时注意避免损伤面神经。接下来，使用微量进样系统(Nanoliter2000，WPI)将AAV通过毛细玻璃电极(直径10mm)从圆窗注射进入耳蜗中。幼鼠耳蜗可容纳2uL的AAV病毒溶液，注射病毒体积为1-2uL。手术后缝合伤口并涂抹止痛药和消炎药剂。
所用的小鼠品系是C57/B6。注射10天后，将耳蜗剥离出来。对样品进行免疫染色鉴定。将样品浸入4％中PFA中进行固定处理，然后样品浸入含有10％的驴血清和0.3％TritonX-100PBS中，室温下孵育1小时后,加入Myo7a、Sox2、NeurN的抗体以及相应的二级抗体。样品采用抗荧光淬火剂安装介质封片,并采用共聚焦进行观察。
如图4a-4c，品红色表示的是Sox2阳性的支持细胞，绿色表示的是AAV导入细胞内表达出来的绿色荧光蛋白mNeonGreen。4c表示的是对4b的计数统计。结果显示：1.图4a是大视野拍摄的耳蜗组织的三个区域：顶转(apex)、中转(middle)和底转(base)的绿色荧光mNeonGreen的表达和品红色所代表的支持细胞标记蛋白Sox2的染色结果。可以发现AAV-ie能高效的在耳蜗各个细胞中导入荧光蛋白。2.图4b和4c的结果显示AAV变异体能高效感染体内小鼠耳蜗组织中的支持细胞，与AAV1、AAV6、AAV8、AAV9、DJ8、Anc80L65和AAV-DJ相比，变异体AAV-ie对Sox2阳性的支持细胞感染率更高。
如图5a-5b，品红色表示的是Myo7a阳性的毛细胞，绿色表示的是AAV导入细胞内表达出来的绿色荧光蛋白mNeonGreen。IHC表示内毛细胞，OHC表示外毛细胞。5b表示的是对5a的计数统计。结果显示AAV变异体AAV-ie能高效感染体内小鼠耳蜗组织中的毛细胞。与亲本AAV相比，变异体AAV-ie对Myo7a阳性的毛细胞感染率更高。
如图6a-6b,品红色表示的是NeurN阳性螺旋神经元细胞，绿色表示的是AAV导入细胞内表达出来的绿色荧光蛋白mNeonGreen。6b表示的是对6a的计数统计。结果显示在耳蜗三个区域：顶转(apex)、中转(middle)和底转(base)，AAV变异体AAV-ie能高效感染体外培养的小鼠耳蜗组织中的螺旋神经元。
上述结果表明：采用圆窗注射的方法AAV变异体能高效感染Myo7a阳性的毛细胞、Sox2阳性的支持细胞以及NeurN阳性的螺旋神经元。
实施例四 体内注射携带Atoh1基因的AAV-ie进入耳蜗使得大量的毛细胞再生
利用耳蜗圆窗注射技术，将携带小鼠Atoh1基因(NCBI Reference Sequence:NM_007500.5)的1.5uL AAV-ie-Atoh1病毒(病毒浓度为5E+12GC/mL)(构建方法参照实施例一，其中所使用的质粒替换为表达Atoh1基因(NCBI Reference Sequence:NM_007500.5)的质粒)注射入耳蜗的外淋巴液中，具体方法参照实施例三。所用的小鼠品系是C57/B6。所用的小鼠为出生2-3天后的幼鼠。注射10天后，将耳蜗剥离出来。对样品进行免疫染色鉴定。样品将浸入4％中PFA中进行固定处理,然后样品浸入含有10％的驴血清和0.3％Triton X-100PBS中，室温下孵育1小时后,加入Myo7a、Sox2的抗体以及相应的二级抗体。样品采用抗荧光淬火剂安装介质封片,并采用共聚焦进行观察。结果如图7所示,品红色表示的是毛细胞标记基因Myo7a的免疫染色，绿色表示的是支持细胞的标记基因Sox2的免疫染色，白色箭头表示异位再生的毛细胞。
上述结果表明：AAV-ie-Atoh1病毒导入Atoh1基因到支持细胞后，在sensoryregion和GER区域能够显著再生毛细胞，
实施例五 AAV-ie能高效感染小鼠和人的椭圆囊(utricle)细胞
椭圆囊(utricle)是内耳中感知重力和维持平衡的器官。利用耳蜗圆窗注射技术，将1.5uL AAV-ie病毒(病毒浓度为6E+12GC/mL)注射入耳蜗的外淋巴液中，具体方法参照实施例三。所用的小鼠品系是C57/B6。所用的小鼠为出生2-3天后的幼鼠。注射10天后，将椭圆囊剥离出来。对样品进行免疫染色鉴定。样品将浸入4％中PFA中进行固定处理,然后样品浸入含有10％的驴血清和0.3％Triton X-100PBS中，室温下孵育1小时后,加入Myo7a、Sox2的抗体以及相应的二级抗体。样品采用抗荧光淬火剂安装介质封片,并采用共聚焦进行观察，结果如图8a所示，其中品红色分别表示了毛细胞和支持细胞的标记蛋白Myo7a和Sox2，绿色表征了由AAV-ie传递的绿色荧光蛋白mNeonGreen。可以发现AAV-ie能够高效感染小鼠的椭圆囊的毛细胞和支持细胞。
取人的椭圆囊，用5×1010GCs的AAV-ie病毒感染。7天后，4％人样本在4％PFA中进行固定处理,然后样品浸入含有10％的驴血清和0.3％Triton X-100PBS中，室温下孵育1小时后,加入Myo7a、Sox2的抗体以及相应的二级抗体。样品采用抗荧光淬火剂安装介质封片,并采用共聚焦进行观察，结果如图8b和图8c所示，其中绿色表征了由AAV-ie传递的绿色荧光蛋白mNeonGreen。红色表示毛细胞的标记蛋白Myo7a，品红色支持细胞的标记蛋白Sox2。结果显示，AAV-ie也能够高效感染人的椭圆囊的毛细胞和支持细胞。
以上的实施例是为了说明本发明公开的实施方案，并不能理解为对本发明的限制。此外，本文所列出的各种修改以及发明中方法、组合物的变化，在不脱离本发明的范围和精神的前提下对本领域内的技术人员来说是显而易见的。虽然已结合本发明的多种具体优选实施例对本发明进行了具体的描述，但应当理解，本发明不应仅限于这些具体实施例。事实上，各种如上所述的对本领域内的技术人员来说显而易见的修改来获取发明都应包括在本发明的范围内。
序列表
<110> 上海科技大学
<120> 具有变异衣壳蛋白的腺相关病毒及其用途
<150> 2019100930003
<151> 2019-01-30
<160> 12
<170> SIPOSequenceListing 1.0
<210> 1
<211> 9
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
Asp Gly Thr Leu Ala Val Pro Phe Lys
1 5
<210> 2
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
gatgggactt tggcggtgcc ttttaag 27
<210> 3
<211> 737
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 3
Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Thr Leu Ser
1 5 10 15
Glu Gly Ile Arg Gln Trp Trp Lys Leu Lys Pro Gly Pro Pro Pro Pro
20 25 30
Lys Pro Ala Glu Arg His Lys Asp Asp Ser Arg Gly Leu Val Leu Pro
35 40 45
Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro
50 55 60
Val Asn Glu Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp
65 70 75 80
Arg Gln Leu Asp Ser Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala
85 90 95
Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly
100 105 110
Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro
115 120 125
Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg
130 135 140
Pro Val Glu His Ser Pro Val Glu Pro Asp Ser Ser Ser Gly Thr Gly
145 150 155 160
Lys Ala Gly Gln Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln Thr
165 170 175
Gly Asp Ala Asp Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro
180 185 190
Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ala Gly Gly Gly
195 200 205
Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ser
210 215 220
Ser Gly Asn Trp His Cys Asp Ser Thr Trp Met Gly Asp Arg Val Ile
225 230 235 240
Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu
245 250 255
Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn
260 265 270
Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg
275 280 285
Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn
290 295 300
Asn Trp Gly Phe Arg Pro Lys Arg Leu Ser Phe Lys Leu Phe Asn Ile
305 310 315 320
Gln Val Lys Glu Val Thr Gln Asn Glu Gly Thr Lys Thr Ile Ala Asn
325 330 335
Asn Leu Thr Ser Thr Ile Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu
340 345 350
Pro Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro
355 360 365
Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn
370 375 380
Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe
385 390 395 400
Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Thr Tyr Thr
405 410 415
Phe Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu
420 425 430
Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser
435 440 445
Arg Thr Gln Thr Thr Gly Gly Thr Thr Asn Thr Gln Thr Leu Gly Phe
450 455 460
Ser Gln Gly Gly Pro Asn Thr Met Ala Asn Gln Ala Lys Asn Trp Leu
465 470 475 480
Pro Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Ser Ala Asp
485 490 495
Asn Asn Asn Ser Glu Tyr Ser Trp Thr Gly Ala Thr Lys Tyr His Leu
500 505 510
Asn Gly Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Ser His
515 520 525
Lys Asp Asp Glu Glu Lys Phe Phe Pro Gln Ser Gly Val Leu Ile Phe
530 535 540
Gly Lys Gln Gly Ser Glu Lys Thr Asn Val Asp Ile Glu Lys Val Met
545 550 555 560
Ile Thr Asp Glu Glu Glu Ile Arg Thr Thr Asn Pro Val Ala Thr Glu
565 570 575
Gln Tyr Gly Ser Val Ser Thr Asn Leu Gln Arg Gly Asn Arg Gln Ala
580 585 590
Ala Thr Ala Asp Val Asn Thr Gln Gly Val Leu Pro Gly Met Val Trp
595 600 605
Gln Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro
610 615 620
His Thr Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly
625 630 635 640
Leu Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro
645 650 655
Ala Asp Pro Pro Thr Thr Phe Asn Gln Ser Lys Leu Asn Ser Phe Ile
660 665 670
Thr Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu
675 680 685
Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser
690 695 700
Asn Tyr Tyr Lys Ser Thr Ser Val Asp Phe Ala Val Asn Thr Glu Gly
705 710 715 720
Val Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn
725 730 735
Leu
<210> 4
<211> 746
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 4
Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Thr Leu Ser
1 5 10 15
Glu Gly Ile Arg Gln Trp Trp Lys Leu Lys Pro Gly Pro Pro Pro Pro
20 25 30
Lys Pro Ala Glu Arg His Lys Asp Asp Ser Arg Gly Leu Val Leu Pro
35 40 45
Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro
50 55 60
Val Asn Glu Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp
65 70 75 80
Arg Gln Leu Asp Ser Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala
85 90 95
Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly
100 105 110
Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro
115 120 125
Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg
130 135 140
Pro Val Glu His Ser Pro Val Glu Pro Asp Ser Ser Ser Gly Thr Gly
145 150 155 160
Lys Ala Gly Gln Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln Thr
165 170 175
Gly Asp Ala Asp Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro
180 185 190
Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ala Gly Gly Gly
195 200 205
Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ser
210 215 220
Ser Gly Asn Trp His Cys Asp Ser Thr Trp Met Gly Asp Arg Val Ile
225 230 235 240
Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu
245 250 255
Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn
260 265 270
Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg
275 280 285
Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn
290 295 300
Asn Trp Gly Phe Arg Pro Lys Arg Leu Ser Phe Lys Leu Phe Asn Ile
305 310 315 320
Gln Val Lys Glu Val Thr Gln Asn Glu Gly Thr Lys Thr Ile Ala Asn
325 330 335
Asn Leu Thr Ser Thr Ile Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu
340 345 350
Pro Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro
355 360 365
Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn
370 375 380
Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe
385 390 395 400
Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Thr Tyr Thr
405 410 415
Phe Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu
420 425 430
Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser
435 440 445
Arg Thr Gln Thr Thr Gly Gly Thr Thr Asn Thr Gln Thr Leu Gly Phe
450 455 460
Ser Gln Gly Gly Pro Asn Thr Met Ala Asn Gln Ala Lys Asn Trp Leu
465 470 475 480
Pro Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Ser Ala Asp
485 490 495
Asn Asn Asn Ser Glu Tyr Ser Trp Thr Gly Ala Thr Lys Tyr His Leu
500 505 510
Asn Gly Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Ser His
515 520 525
Lys Asp Asp Glu Glu Lys Phe Phe Pro Gln Ser Gly Val Leu Ile Phe
530 535 540
Gly Lys Gln Gly Ser Glu Lys Thr Asn Val Asp Ile Glu Lys Val Met
545 550 555 560
Ile Thr Asp Glu Glu Glu Ile Arg Thr Thr Asn Pro Val Ala Thr Glu
565 570 575
Gln Tyr Gly Ser Val Ser Thr Asn Leu Gln Arg Gly Asn Asp Gly Thr
580 585 590
Leu Ala Val Pro Phe Lys Arg Gln Ala Ala Thr Ala Asp Val Asn Thr
595 600 605
Gln Gly Val Leu Pro Gly Met Val Trp Gln Asp Arg Asp Val Tyr Leu
610 615 620
Gln Gly Pro Ile Trp Ala Lys Ile Pro His Thr Asp Gly His Phe His
625 630 635 640
Pro Ser Pro Leu Met Gly Gly Phe Gly Leu Lys His Pro Pro Pro Gln
645 650 655
Ile Leu Ile Lys Asn Thr Pro Val Pro Ala Asp Pro Pro Thr Thr Phe
660 665 670
Asn Gln Ser Lys Leu Asn Ser Phe Ile Thr Gln Tyr Ser Thr Gly Gln
675 680 685
Val Ser Val Glu Ile Glu Trp Glu Leu Gln Lys Glu Asn Ser Lys Arg
690 695 700
Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn Tyr Tyr Lys Ser Thr Ser
705 710 715 720
Val Asp Phe Ala Val Asn Thr Glu Gly Val Tyr Ser Glu Pro Arg Pro
725 730 735
Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu
740 745
<210> 5
<211> 2241
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
atggctgccg atggttatct tccagattgg ctcgaggaca ctctctctga aggaataaga 60
cagtggtgga agctcaaacc tggcccacca ccaccaaagc ccgcagagcg gcataaggac 120
gacagcaggg gtcttgtgct tcctgggtac aagtacctcg gacccttcaa cggactcgac 180
aagggagagc cggtcaacga ggcagacgcc gcggccctcg agcacgacaa agcctacgac 240
cggcagctcg acagcggaga caacccgtac ctcaagtaca accacgccga cgccgagttc 300
caggagcggc tcaaagaaga tacgtctttt gggggcaacc tcgggcgagc agtcttccag 360
gccaaaaaga ggcttcttga acctcttggt ctggttgagg aagcggctaa gacggctcct 420
ggaaagaaga ggcctgtaga gcactctcct gtggagccag actcctcctc gggaaccgga 480
aaggcgggcc agcagcctgc aagaaaaaga ttgaattttg gtcagactgg agacgcagac 540
tcagtcccag accctcaacc aatcggagaa cctcccgcag ccccctcagg tgtgggatct 600
cttacaatgg ctgcaggcgg tggcgcacca atggcagaca ataacgaggg cgccgacgga 660
gtgggtaatt cctcgggaaa ttggcattgc gattccacat ggatgggcga cagagtcatc 720
accaccagca cccgaacctg ggccctgccc acctacaaca accacctcta caagcaaatc 780
tccaacagca catctggagg atcttcaaat gacaacgcct acttcggcta cagcaccccc 840
tgggggtatt ttgactttaa cagattccac tgccactttt caccacgtga ctggcagcga 900
ctcatcaaca acaactgggg attccggccc aagagactca gcttcaagct cttcaacatc 960
caggtcaagg aggtcacgca gaatgaaggc accaagacca tcgccaataa cctcaccagc 1020
accatccagg tgtttacgga ctcggagtac cagctgccgt acgttctcgg ctctgcccac 1080
cagggctgcc tgcctccgtt cccggcggac gtgttcatga ttccccagta cggctaccta 1140
acactcaaca acggtagtca ggccgtggga cgctcctcct tctactgcct ggaatacttt 1200
ccttcgcaga tgctgagaac cggcaacaac ttccagttta cttacacctt cgaggacgtg 1260
cctttccaca gcagctacgc ccacagccag agcttggacc ggctgatgaa tcctctgatt 1320
gaccagtacc tgtactactt gtctcggact caaacaacag gaggcacgac aaatacgcag 1380
actctgggct tcagccaagg tgggcctaat acaatggcca atcaggcaaa gaactggctg 1440
ccaggaccct gttaccgcca gcagcgagta tcaaagacat ctgcggataa caacaacagt 1500
gaatactcgt ggactggagc taccaagtac cacctcaatg gcagagactc tctggtgaat 1560
ccgggcccgg ccatggcaag ccacaaggac gatgaagaaa agttttttcc tcagagcggg 1620
gttctcatct ttgggaagca aggctcagag aaaacaaatg tggacattga aaaggtcatg 1680
attacagacg aagaggaaat caggacaacc aatcccgtgg ctacggagca gtatggttct 1740
gtatctacca acctccagag aggcaacgat gggactttgg cggtgccttt taagagacaa 1800
gcagctaccg cagatgtcaa cacacaaggc gttcttccag gcatggtctg gcaggacaga 1860
gatgtgtacc ttcaggggcc catctgggca aagattccac acacggacgg acattttcac 1920
ccctctcccc tcatgggtgg attcggactt aaacaccctc cgcctcagat cctgatcaag 1980
aacacgcctg tacctgcgga tcctccgacc accttcaacc agtcaaagct gaactctttc 2040
atcacccagt attctactgg ccaagtcagc gtggagatcg agtgggagct gcagaaggaa 2100
aacagcaagc gctggaaccc cgagatccag tacacctcca actactacaa atctacaagt 2160
gtggactttg ctgttaatac agaaggcgtg tactctgaac cccgccccat tggcacccgt 2220
tacctcaccc gtaatctgta a 2241
<210> 6
<211> 7360
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
gcgcgccgat atcgttaacg ccccgcgccg gccgctctag aactagtgga tcccccggaa 60
gatcagaagt tcctattccg aagttcctat tctctagaaa gtataggaac ttctgatctg 120
cgcagccgcc atgccggggt tttacgagat tgtgattaag gtccccagcg accttgacga 180
gcatctgccc ggcatttctg acagctttgt gaactgggtg gccgagaagg aatgggagtt 240
gccgccagat tctgacatgg atctgaatct gattgagcag gcacccctga ccgtggccga 300
gaagctgcag cgcgactttc tgacggaatg gcgccgtgtg agtaaggccc cggaggccct 360
tttctttgtg caatttgaga agggagagag ctacttccac atgcacgtgc tcgtggaaac 420
caccggggtg aaatccatgg ttttgggacg tttcctgagt cagattcgcg aaaaactgat 480
tcagagaatt taccgcggga tcgagccgac tttgccaaac tggttcgcgg tcacaaagac 540
cagaaatggc gccggaggcg ggaacaaggt ggtggatgag tgctacatcc ccaattactt 600
gctccccaaa acccagcctg agctccagtg ggcgtggact aatatggaac agtatttaag 660
cgcctgtttg aatctcacgg agcgtaaacg gttggtggcg cagcatctga cgcacgtgtc 720
gcagacgcag gagcagaaca aagagaatca gaatcccaat tctgatgcgc cggtgatcag 780
atcaaaaact tcagccaggt acatggagct ggtcgggtgg ctcgtggaca aggggattac 840
ctcggagaag cagtggatcc aggaggacca ggcctcatac atctccttca atgcggcctc 900
caactcgcgg tcccaaatca aggctgcctt ggacaatgcg ggaaagatta tgagcctgac 960
taaaaccgcc cccgactacc tggtgggcca gcagcccgtg gaggacattt ccagcaatcg 1020
gatttataaa attttggaac taaacgggta cgatccccaa tatgcggctt ccgtctttct 1080
gggatgggcc acgaaaaagt tcggcaagag gaacaccatc tggctgtttg ggcctgcaac 1140
taccgggaag accaacatcg cggaggccat agcccacact gtgcccttct acgggtgcgt 1200
aaactggacc aatgagaact ttcccttcaa cgactgtgtc gacaagatgg tgatctggtg 1260
ggaggagggg aagatgaccg ccaaggtcgt ggagtcggcc aaagccattc tcggaggaag 1320
caaggtgcgc gtggaccaga aatgcaagtc ctcggcccag atagacccga ctcccgtgat 1380
cgtcacctcc aacaccaaca tgtgcgccgt gattgacggg aactcaacga ccttcgaaca 1440
ccagcagccg ttgcaagacc ggatgttcaa atttgaactc acccgccgtc tggatcatga 1500
ctttgggaag gtcaccaagc aggaagtcaa agactttttc cggtgggcaa aggatcacgt 1560
ggttgaggtg gagcatgaat tctacgtcaa aaagggtgga gccaagaaaa gacccgcccc 1620
cagtgacgca gatataagtg agcccaaacg ggtgcgcgag tcagttgcgc agccatcgac 1680
gtcagacgcg gaagcttcga tcaactacgc agacaggtac caaaacaaat gttctcgtca 1740
cgtgggcatg aatctgatgc tgtttccctg cagacaatgc gagagaatga atcagaattc 1800
aaatatctgc ttcactcacg gacagaaaga ctgtttagag tgctttcccg tgtcagaatc 1860
tcaacccgtt tctgtcgtca aaaaggcgta tcagaaactg tgctacattc atcatatcat 1920
gggaaaggtg ccagacgctt gcactgcctg cgatctggtc aatgtggatt tggatgactg 1980
catctttgaa caataaatga tttaaatcag gtatggctgc cgatggttat cttccagatt 2040
ggctcgagga cactctctct gaaggaataa gacagtggtg gaagctcaaa cctggcccac 2100
caccaccaaa gcccgcagag cggcataagg acgacagcag gggtcttgtg cttcctgggt 2160
acaagtacct cggacccttc aacggactcg acaagggaga gccggtcaac gaggcagacg 2220
ccgcggccct cgagcacgac aaagcctacg accggcagct cgacagcgga gacaacccgt 2280
acctcaagta caaccacgcc gacgccgagt tccaggagcg gctcaaagaa gatacgtctt 2340
ttgggggcaa cctcgggcga gcagtcttcc aggccaaaaa gaggcttctt gaacctcttg 2400
gtctggttga ggaagcggct aagacggctc ctggaaagaa gaggcctgta gagcactctc 2460
ctgtggagcc agactcctcc tcgggaaccg gaaaggcggg ccagcagcct gcaagaaaaa 2520
gattgaattt tggtcagact ggagacgcag actcagtccc agaccctcaa ccaatcggag 2580
aacctcccgc agccccctca ggtgtgggat ctcttacaat ggctgcaggc ggtggcgcac 2640
caatggcaga caataacgag ggcgccgacg gagtgggtaa ttcctcggga aattggcatt 2700
gcgattccac atggatgggc gacagagtca tcaccaccag cacccgaacc tgggccctgc 2760
ccacctacaa caaccacctc tacaagcaaa tctccaacag cacatctgga ggatcttcaa 2820
atgacaacgc ctacttcggc tacagcaccc cctgggggta ttttgacttt aacagattcc 2880
actgccactt ttcaccacgt gactggcagc gactcatcaa caacaactgg ggattccggc 2940
ccaagagact cagcttcaag ctcttcaaca tccaggtcaa ggaggtcacg cagaatgaag 3000
gcaccaagac catcgccaat aacctcacca gcaccatcca ggtgtttacg gactcggagt 3060
accagctgcc gtacgttctc ggctctgccc accagggctg cctgcctccg ttcccggcgg 3120
acgtgttcat gattccccag tacggctacc taacactcaa caacggtagt caggccgtgg 3180
gacgctcctc cttctactgc ctggaatact ttccttcgca gatgctgaga accggcaaca 3240
acttccagtt tacttacacc ttcgaggacg tgcctttcca cagcagctac gcccacagcc 3300
agagcttgga ccggctgatg aatcctctga ttgaccagta cctgtactac ttgtctcgga 3360
ctcaaacaac aggaggcacg acaaatacgc agactctggg cttcagccaa ggtgggccta 3420
atacaatggc caatcaggca aagaactggc tgccaggacc ctgttaccgc cagcagcgag 3480
tatcaaagac atctgcggat aacaacaaca gtgaatactc gtggactgga gctaccaagt 3540
accacctcaa tggcagagac tctctggtga atccgggccc ggccatggca agccacaagg 3600
acgatgaaga aaagtttttt cctcagagcg gggttctcat ctttgggaag caaggctcag 3660
agaaaacaaa tgtggacatt gaaaaggtca tgattacaga cgaagaggaa atcaggacaa 3720
ccaatcccgt ggctacggag cagtatggtt ctgtatctac caacctccag agaggcaacg 3780
atgggacttt ggcggtgcct tttaagagac aagcagctac cgcagatgtc aacacacaag 3840
gcgttcttcc aggcatggtc tggcaggaca gagatgtgta ccttcagggg cccatctggg 3900
caaagattcc acacacggac ggacattttc acccctctcc cctcatgggt ggattcggac 3960
ttaaacaccc tccgcctcag atcctgatca agaacacgcc tgtacctgcg gatcctccga 4020
ccaccttcaa ccagtcaaag ctgaactctt tcatcaccca gtattctact ggccaagtca 4080
gcgtggagat cgagtgggag ctgcagaagg aaaacagcaa gcgctggaac cccgagatcc 4140
agtacacctc caactactac aaatctacaa gtgtggactt tgctgttaat acagaaggcg 4200
tgtactctga accccgcccc attggcaccc gttacctcac ccgtaatctg taattgcttg 4260
ttaatcaata aaccgtttaa ttcgtttcag ttgaactttg gtctctgcgt atttctttct 4320
tatctagttt ccatggctac gtagataagt agcatggcgg gttaatcatt aactacagcc 4380
cgggcgttta aacagcgggc ggaggggtgg agtcgtgacg tgaattacgt catagggtta 4440
gggaggtcct gtattagagg tcacgtgagt gttttgcgac attttgcgac accatgtggt 4500
ctcgctgggg gggggggccc gagtgagcac gcagggtctc cattttgaag cgggaggttt 4560
gaacgagcgc tggcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct 4620
ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc 4680
gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggaaa 4740
ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt 4800
tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata 4860
gggttgagtg ttgttccagt ttggaacaag agtccactat taagaacgtg gactccaacg 4920
tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat 4980
caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc 5040
gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga 5100
aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac 5160
ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc 5220
gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac 5280
aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt 5340
tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag 5400
aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg 5460
aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa 5520
tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc 5580
aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag 5640
tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa 5700
ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc 5760
taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg 5820
agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa 5880
caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa 5940
tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg 6000
gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag 6060
cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg 6120
caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt 6180
ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt 6240
aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac 6300
gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 6360
atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 6420
tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 6480
gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga 6540
actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 6600
gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 6660
agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 6720
ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 6780
aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 6840
cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 6900
gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 6960
cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat 7020
cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca 7080
gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca 7140
aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg 7200
actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac 7260
cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac 7320
aatttcacac aggaaacagc tatgaccatg attacgccaa 7360
<210> 7
<211> 7330
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
gcgcgccgat atcgttaacg ccccgcgccg gccgctctag aactagtgga tcccccggaa 60
gatcagaagt tcctattccg aagttcctat tctctagaaa gtataggaac ttctgatctg 120
cgcagccgcc atgccggggt tttacgagat tgtgattaag gtccccagcg accttgacga 180
gcatctgccc ggcatttctg acagctttgt gaactgggtg gccgagaagg aatgggagtt 240
gccgccagat tctgacatgg atctgaatct gattgagcag gcacccctga ccgtggccga 300
gaagctgcag cgcgactttc tgacggaatg gcgccgtgtg agtaaggccc cggaggccct 360
tttctttgtg caatttgaga agggagagag ctacttccac atgcacgtgc tcgtggaaac 420
caccggggtg aaatccatgg ttttgggacg tttcctgagt cagattcgcg aaaaactgat 480
tcagagaatt taccgcggga tcgagccgac tttgccaaac tggttcgcgg tcacaaagac 540
cagaaatggc gccggaggcg ggaacaaggt ggtggatgag tgctacatcc ccaattactt 600
gctccccaaa acccagcctg agctccagtg ggcgtggact aatatggaac agtatttaag 660
cgcctgtttg aatctcacgg agcgtaaacg gttggtggcg cagcatctga cgcacgtgtc 720
gcagacgcag gagcagaaca aagagaatca gaatcccaat tctgatgcgc cggtgatcag 780
atcaaaaact tcagccaggt acatggagct ggtcgggtgg ctcgtggaca aggggattac 840
ctcggagaag cagtggatcc aggaggacca ggcctcatac atctccttca atgcggcctc 900
caactcgcgg tcccaaatca aggctgcctt ggacaatgcg ggaaagatta tgagcctgac 960
taaaaccgcc cccgactacc tggtgggcca gcagcccgtg gaggacattt ccagcaatcg 1020
gatttataaa attttggaac taaacgggta cgatccccaa tatgcggctt ccgtctttct 1080
gggatgggcc acgaaaaagt tcggcaagag gaacaccatc tggctgtttg ggcctgcaac 1140
taccgggaag accaacatcg cggaggccat agcccacact gtgcccttct acgggtgcgt 1200
aaactggacc aatgagaact ttcccttcaa cgactgtgtc gacaagatgg tgatctggtg 1260
ggaggagggg aagatgaccg ccaaggtcgt ggagtcggcc aaagccattc tcggaggaag 1320
caaggtgcgc gtggaccaga aatgcaagtc ctcggcccag atagacccga ctcccgtgat 1380
cgtcacctcc aacaccaaca tgtgcgccgt gattgacggg aactcaacga ccttcgaaca 1440
ccagcagccg ttgcaagacc ggatgttcaa atttgaactc acccgccgtc tggatcatga 1500
ctttgggaag gtcaccaagc aggaagtcaa agactttttc cggtgggcaa aggatcacgt 1560
ggttgaggtg gagcatgaat tctacgtcaa aaagggtgga gccaagaaaa gacccgcccc 1620
cagtgacgca gatataagtg agcccaaacg ggtgcgcgag tcagttgcgc agccatcgac 1680
gtcagacgcg gaagcttcga tcaactacgc agacaggtac caaaacaaat gttctcgtca 1740
cgtgggcatg aatctgatgc tgtttccctg cagacaatgc gagagaatga atcagaattc 1800
aaatatctgc ttcactcacg gacagaaaga ctgtttagag tgctttcccg tgtcagaatc 1860
tcaacccgtt tctgtcgtca aaaaggcgta tcagaaactg tgctacattc atcatatcat 1920
gggaaaggtg ccagacgctt gcactgcctg cgatctggtc aatgtggatt tggatgactg 1980
catctttgaa caataaatga tttaaatcag gtatggctgc cgatggttat cttccagatt 2040
ggctcgagga caacctctct gagggcattc gcgagtggtg ggacttgaaa cctggagccc 2100
cgaagcccaa agccaaccag caaaagcagg acgacggccg gggtctggtg cttcctggct 2160
acaagtacct cggacccttc aacggactcg acaaggggga gcccgtcaac gcggcggacg 2220
cagcggccct cgagcacgac aaggcctacg accagcagct caaagcgggt gacaatccgt 2280
acctgcggta taaccacgcc gacgccgagt ttcaggagcg tctgcaagaa gatacgtctt 2340
ttgggggcaa cctcgggcga gcagtcttcc aggccaagaa gcgggttctc gaacctctcg 2400
gtctggttga ggaaggcgct aagacggctc ctggaaagaa acgtccggta gagcagtcgc 2460
cacaagagcc agactcctcc tcgggcatcg gcaagacagg ccagcagccc gctaaaaaga 2520
gactcaattt tggtcagact ggcgactcag agtcagtccc cgatccacaa cctctcggag 2580
aacctccagc aacccccgct gctgtgggac ctactacaat ggcttcaggc ggtggcgcac 2640
caatggcaga caataacgaa ggcgccgacg gagtgggtaa tgcctcagga aattggcatt 2700
gcgattccac atggctgggc gacagagtca tcaccaccag cacccgcacc tgggccttgc 2760
ccacctacaa taaccacctc tacaagcaaa tctccagtgc ttcaacgggg gccagcaacg 2820
acaaccacta cttcggctac agcaccccct gggggtattt tgatttcaac agattccact 2880
gccacttttc accacgtgac tggcagcgac tcatcaacaa caattgggga ttccggccca 2940
agagactcaa cttcaaactc ttcaacatcc aagtcaagga ggtcacgacg aatgatggcg 3000
tcacaaccat cgctaataac cttaccagca cggttcaagt cttctcggac tcggagtacc 3060
agcttccgta cgtcctcggc tctgcgcacc agggctgcct ccctccgttc ccggcggacg 3120
tgttcatgat tccgcaatac ggctacctga cgctcaacaa tggcagccaa gccgtgggac 3180
gttcatcctt ttactgcctg gaatatttcc cttctcagat gctgagaacg ggcaacaact 3240
ttaccttcag ctacaccttt gaggaagtgc ctttccacag cagctacgcg cacagccaga 3300
gcctggaccg gctgatgaat cctctcatcg accaatacct gtattacctg aacagaactc 3360
aaaatcagtc cggaagtgcc caaaacaagg acttgctgtt tagccgtggg tctccagctg 3420
gcatgtctgt tcagcccaaa aactggctac ctggaccctg ttatcggcag cagcgcgttt 3480
ctaaaacaaa aacagacaac aacaacagca attttacctg gactggtgct tcaaaatata 3540
acctcaatgg gcgtgaatcc atcatcaacc ctggcactgc tatggcctca cacaaagacg 3600
acgaagacaa gttctttccc atgagcggtg tcatgatttt tggaaaagag agcgccggag 3660
cttcaaacac tgcattggac aatgtcatga ttacagacga agaggaaatt aaagccacta 3720
accctgtggc caccgaaaga tttgggaccg tggcagtcaa tttccagagc agcagcacag 3780
accctgcgac cggagatgtg catgctatgg gagcattacc tggcatggtg tggcaagata 3840
gagacgtgta cctgcagggt cccatttggg ccaaaattcc tcacacagat ggacactttc 3900
acccgtctcc tcttatgggc ggctttggac tcaagaaccc gcctcctcag atcctcatca 3960
aaaacacgcc tgttcctgcg aatcctccgg cggagttttc agctacaaag tttgcttcat 4020
tcatcaccca atactccaca ggacaagtga gtgtggaaat tgaatgggag ctgcagaaag 4080
aaaacagcaa gcgctggaat cccgaagtgc agtacacatc caattatgca aaatctgcca 4140
acgttgattt tactgtggac aacaatggac tttatactga gcctcgcccc attggcaccc 4200
gttaccttac ccgtcccctg taattgcttg ttaatcaata aaccgtttaa ttcgtttcag 4260
ttgaactttg gtctctgcgt atttctttct tatctagttt ccatggctac gtagataagt 4320
agcatggcgg gttaatcatt aactacagcc cgggcgttta aacagcgggc ggaggggtgg 4380
agtcgtgacg tgaattacgt catagggtta gggaggtcct gtattagagg tcacgtgagt 4440
gttttgcgac attttgcgac accatgtggt ctcgctgggg gggggggccc gagtgagcac 4500
gcagggtctc cattttgaag cgggaggttt gaacgagcgc tggcgcgctc actggccgtc 4560
gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca 4620
catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa 4680
cagttgcgca gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg ttaaaattcg 4740
cgttaaattt ttgttaaatc agctcatttt tttaaccaat aggccgaaat cggcaaaatc 4800
ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag 4860
agtccactat taagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg 4920
atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag 4980
cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga 5040
acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg 5100
tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg 5160
cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 5220
tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 5280
gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 5340
cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 5400
atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 5460
agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 5520
gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt 5580
ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 5640
cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 5700
ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 5760
atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 5820
gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 5880
tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 5940
gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 6000
gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 6060
tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 6120
ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 6180
tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 6240
ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 6300
ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 6360
tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 6420
ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 6480
tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 6540
tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 6600
actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 6660
cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 6720
gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 6780
tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 6840
ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 6900
ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 6960
cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 7020
cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga 7080
gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc 7140
attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa 7200
ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc 7260
gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg 7320
attacgccaa 7330
<210> 8
<211> 7330
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
gcgcgccgat atcgttaacg ccccgcgccg gccgctctag aactagtgga tcccccggaa 60
gatcagaagt tcctattccg aagttcctat tctctagaaa gtataggaac ttctgatctg 120
cgcagccgcc atgccggggt tttacgagat tgtgattaag gtccccagcg accttgacga 180
gcatctgccc ggcatttctg acagctttgt gaactgggtg gccgagaagg aatgggagtt 240
gccgccagat tctgacatgg atctgaatct gattgagcag gcacccctga ccgtggccga 300
gaagctgcag cgcgactttc tgacggaatg gcgccgtgtg agtaaggccc cggaggccct 360
tttctttgtg caatttgaga agggagagag ctacttccac atgcacgtgc tcgtggaaac 420
caccggggtg aaatccatgg ttttgggacg tttcctgagt cagattcgcg aaaaactgat 480
tcagagaatt taccgcggga tcgagccgac tttgccaaac tggttcgcgg tcacaaagac 540
cagaaatggc gccggaggcg ggaacaaggt ggtggatgag tgctacatcc ccaattactt 600
gctccccaaa acccagcctg agctccagtg ggcgtggact aatatggaac agtatttaag 660
cgcctgtttg aatctcacgg agcgtaaacg gttggtggcg cagcatctga cgcacgtgtc 720
gcagacgcag gagcagaaca aagagaatca gaatcccaat tctgatgcgc cggtgatcag 780
atcaaaaact tcagccaggt acatggagct ggtcgggtgg ctcgtggaca aggggattac 840
ctcggagaag cagtggatcc aggaggacca ggcctcatac atctccttca atgcggcctc 900
caactcgcgg tcccaaatca aggctgcctt ggacaatgcg ggaaagatta tgagcctgac 960
taaaaccgcc cccgactacc tggtgggcca gcagcccgtg gaggacattt ccagcaatcg 1020
gatttataaa attttggaac taaacgggta cgatccccaa tatgcggctt ccgtctttct 1080
gggatgggcc acgaaaaagt tcggcaagag gaacaccatc tggctgtttg ggcctgcaac 1140
taccgggaag accaacatcg cggaggccat agcccacact gtgcccttct acgggtgcgt 1200
aaactggacc aatgagaact ttcccttcaa cgactgtgtc gacaagatgg tgatctggtg 1260
ggaggagggg aagatgaccg ccaaggtcgt ggagtcggcc aaagccattc tcggaggaag 1320
caaggtgcgc gtggaccaga aatgcaagtc ctcggcccag atagacccga ctcccgtgat 1380
cgtcacctcc aacaccaaca tgtgcgccgt gattgacggg aactcaacga ccttcgaaca 1440
ccagcagccg ttgcaagacc ggatgttcaa atttgaactc acccgccgtc tggatcatga 1500
ctttgggaag gtcaccaagc aggaagtcaa agactttttc cggtgggcaa aggatcacgt 1560
ggttgaggtg gagcatgaat tctacgtcaa aaagggtgga gccaagaaaa gacccgcccc 1620
cagtgacgca gatataagtg agcccaaacg ggtgcgcgag tcagttgcgc agccatcgac 1680
gtcagacgcg gaagcttcga tcaactacgc agacaggtac caaaacaaat gttctcgtca 1740
cgtgggcatg aatctgatgc tgtttccctg cagacaatgc gagagaatga atcagaattc 1800
aaatatctgc ttcactcacg gacagaaaga ctgtttagag tgctttcccg tgtcagaatc 1860
tcaacccgtt tctgtcgtca aaaaggcgta tcagaaactg tgctacattc atcatatcat 1920
gggaaaggtg ccagacgctt gcactgcctg cgatctggtc aatgtggatt tggatgactg 1980
catctttgaa caataaatga tttaaatcag gtatggctgc cgatggttat cttccagatt 2040
ggctcgagga caacctctct gagggcattc gcgagtggtg ggacttgaaa cctggagccc 2100
cgaaacccaa agccaaccag caaaagcagg acgacggccg gggtctggtg cttcctggct 2160
acaagtacct cggacccttc aacggactcg acaaggggga gcccgtcaac gcggcggatg 2220
cagcggccct cgagcacgac aaggcctacg accagcagct caaagcgggt gacaatccgt 2280
acctgcggta taaccacgcc gacgccgagt ttcaggagcg tctgcaagaa gatacgtctt 2340
ttgggggcaa cctcgggcga gcagtcttcc aggccaagaa gagggttctc gaaccttttg 2400
gtctggttga ggaaggtgct aagacggctc ctggaaagaa acgtccggta gagcagtcgc 2460
cacaagagcc agactcctcc tcgggcattg gcaagacagg ccagcagccc gctaaaaaga 2520
gactcaattt tggtcagact ggcgactcag agtcagtccc cgacccacaa cctctcggag 2580
aacctccagc aacccccgct gctgtgggac ctactacaat ggcttcaggc ggtggcgcac 2640
caatggcaga caataacgaa ggcgccgacg gagtgggtaa tgcctcagga aattggcatt 2700
gcgattccac atggctgggc gacagagtca tcaccaccag cacccgaaca tgggccttgc 2760
ccacctataa caaccacctc tacaagcaaa tctccagtgc ttcaacgggg gccagcaacg 2820
acaaccacta cttcggctac agcaccccct gggggtattt tgatttcaac agattccact 2880
gccatttctc accacgtgac tggcagcgac tcatcaacaa caattgggga ttccggccca 2940
agagactcaa cttcaagctc ttcaacatcc aagtcaagga ggtcacgacg aatgatggcg 3000
tcacgaccat cgctaataac cttaccagca cggttcaagt cttctcggac tcggagtacc 3060
agttgccgta cgtcctcggc tctgcgcacc agggctgcct ccctccgttc ccggcggacg 3120
tgttcatgat tccgcagtac ggctacctaa cgctcaacaa tggcagccag gcagtgggac 3180
ggtcatcctt ttactgcctg gaatatttcc catcgcagat gctgagaacg ggcaataact 3240
ttaccttcag ctacaccttc gaggacgtgc ctttccacag cagctacgcg cacagccaga 3300
gcctggaccg gctgatgaat cctctcatcg accagtacct gtattacctg aacagaactc 3360
agaatcagtc cggaagtgcc caaaacaagg acttgctgtt tagccggggg tctccagctg 3420
gcatgtctgt tcagcccaaa aactggctac ctggaccctg ttaccggcag cagcgcgttt 3480
ctaaaacaaa aacagacaac aacaacagca actttacctg gactggtgct tcaaaatata 3540
accttaatgg gcgtgaatct ataatcaacc ctggcactgc tatggcctca cacaaagacg 3600
acaaagacaa gttctttccc atgagcggtg tcatgatttt tggaaaggag agcgccggag 3660
cttcaaacac tgcattggac aatgtcatga tcacagacga agaggaaatc aaagccacta 3720
accccgtggc caccgaaaga tttgggactg tggcagtcaa tctccagagc agcagcacag 3780
accctgcgac cggagatgtg catgttatgg gagccttacc tggaatggtg tggcaagaca 3840
gagacgtata cctgcagggt cctatttggg ccaaaattcc tcacacggat ggacactttc 3900
acccgtctcc tctcatgggc ggctttggac ttaagcaccc gcctcctcag atcctcatca 3960
aaaacacgcc tgttcctgcg aatcctccgg cagagttttc ggctacaaag tttgcttcat 4020
tcatcaccca gtattccaca ggacaagtga gcgtggagat tgaatgggag ctgcagaaag 4080
aaaacagcaa acgctggaat cccgaagtgc agtatacatc taactatgca aaatctgcca 4140
acgttgattt cactgtggac aacaatggac tttatactga gcctcgcccc attggcaccc 4200
gttacctcac ccgtcccctg taattgcttg ttaatcaata aaccgtttaa ttcgtttcag 4260
ttgaactttg gtctctgcgt atttctttct tatctagttt ccatggctac gtagataagt 4320
agcatggcgg gttaatcatt aactacagcc cgggcgttta aacagcgggc ggaggggtgg 4380
agtcgtgacg tgaattacgt catagggtta gggaggtcct gtattagagg tcacgtgagt 4440
gttttgcgac attttgcgac accatgtggt ctcgctgggg gggggggccc gagtgagcac 4500
gcagggtctc cattttgaag cgggaggttt gaacgagcgc tggcgcgctc actggccgtc 4560
gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca 4620
catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa 4680
cagttgcgca gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg ttaaaattcg 4740
cgttaaattt ttgttaaatc agctcatttt tttaaccaat aggccgaaat cggcaaaatc 4800
ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag 4860
agtccactat taagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg 4920
atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag 4980
cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga 5040
acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg 5100
tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg 5160
cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 5220
tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 5280
gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 5340
cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 5400
atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 5460
agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 5520
gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt 5580
ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 5640
cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 5700
ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 5760
atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 5820
gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 5880
tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 5940
gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 6000
gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 6060
tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 6120
ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 6180
tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 6240
ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 6300
ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 6360
tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 6420
ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 6480
tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 6540
tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 6600
actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 6660
cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 6720
gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 6780
tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 6840
ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 6900
ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 6960
cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 7020
cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga 7080
gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc 7140
attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa 7200
ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc 7260
gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg 7320
attacgccaa 7330
<210> 9
<211> 7330
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
gcgcgccgat atcgttaacg ccccgcgccg gccgctctag aactagtgga tcccccggaa 60
gatcagaagt tcctattccg aagttcctat tctctagaaa gtataggaac ttctgatctg 120
cgcagccgcc atgccggggt tttacgagat tgtgattaag gtccccagcg accttgacga 180
gcatctgccc ggcatttctg acagctttgt gaactgggtg gccgagaagg aatgggagtt 240
gccgccagat tctgacatgg atctgaatct gattgagcag gcacccctga ccgtggccga 300
gaagctgcag cgcgactttc tgacggaatg gcgccgtgtg agtaaggccc cggaggccct 360
tttctttgtg caatttgaga agggagagag ctacttccac atgcacgtgc tcgtggaaac 420
caccggggtg aaatccatgg ttttgggacg tttcctgagt cagattcgcg aaaaactgat 480
tcagagaatt taccgcggga tcgagccgac tttgccaaac tggttcgcgg tcacaaagac 540
cagaaatggc gccggaggcg ggaacaaggt ggtggatgag tgctacatcc ccaattactt 600
gctccccaaa acccagcctg agctccagtg ggcgtggact aatatggaac agtatttaag 660
cgcctgtttg aatctcacgg agcgtaaacg gttggtggcg cagcatctga cgcacgtgtc 720
gcagacgcag gagcagaaca aagagaatca gaatcccaat tctgatgcgc cggtgatcag 780
atcaaaaact tcagccaggt acatggagct ggtcgggtgg ctcgtggaca aggggattac 840
ctcggagaag cagtggatcc aggaggacca ggcctcatac atctccttca atgcggcctc 900
caactcgcgg tcccaaatca aggctgcctt ggacaatgcg ggaaagatta tgagcctgac 960
taaaaccgcc cccgactacc tggtgggcca gcagcccgtg gaggacattt ccagcaatcg 1020
gatttataaa attttggaac taaacgggta cgatccccaa tatgcggctt ccgtctttct 1080
gggatgggcc acgaaaaagt tcggcaagag gaacaccatc tggctgtttg ggcctgcaac 1140
taccgggaag accaacatcg cggaggccat agcccacact gtgcccttct acgggtgcgt 1200
aaactggacc aatgagaact ttcccttcaa cgactgtgtc gacaagatgg tgatctggtg 1260
ggaggagggg aagatgaccg ccaaggtcgt ggagtcggcc aaagccattc tcggaggaag 1320
caaggtgcgc gtggaccaga aatgcaagtc ctcggcccag atagacccga ctcccgtgat 1380
cgtcacctcc aacaccaaca tgtgcgccgt gattgacggg aactcaacga ccttcgaaca 1440
ccagcagccg ttgcaagacc ggatgttcaa atttgaactc acccgccgtc tggatcatga 1500
ctttgggaag gtcaccaagc aggaagtcaa agactttttc cggtgggcaa aggatcacgt 1560
ggttgaggtg gagcatgaat tctacgtcaa aaagggtgga gccaagaaaa gacccgcccc 1620
cagtgacgca gatataagtg agcccaaacg ggtgcgcgag tcagttgcgc agccatcgac 1680
gtcagacgcg gaagcttcga tcaactacgc agacaggtac caaaacaaat gttctcgtca 1740
cgtgggcatg aatctgatgc tgtttccctg cagacaatgc gagagaatga atcagaattc 1800
aaatatctgc ttcactcacg gacagaaaga ctgtttagag tgctttcccg tgtcagaatc 1860
tcaacccgtt tctgtcgtca aaaaggcgta tcagaaactg tgctacattc atcatatcat 1920
gggaaaggtg ccagacgctt gcactgcctg cgatctggtc aatgtggatt tggatgactg 1980
catctttgaa caataaatga tttaaatcag gtatggctgc cgatggttat cttccagatt 2040
ggctcgagga caaccttagt gaaggaattc gcgagtggtg ggctttgaaa cctggagccc 2100
ctcaacccaa ggcaaatcaa caacatcaag acaacgctcg aggtcttgtg cttccgggtt 2160
acaaatacct tggacccggc aacggactcg acaaggggga gccggtcaac gcagcagacg 2220
cggcggccct cgagcacgac aaggcctacg accagcagct caaggccgga gacaacccgt 2280
acctcaagta caaccacgcc gacgccgagt tccaggagcg gctcaaagaa gatacgtctt 2340
ttgggggcaa cctcgggcga gcagtcttcc aggccaaaaa gaggcttctt gaacctcttg 2400
gtctggttga ggaagcggct aagacggctc ctggaaagaa gaggcctgta gagcagtctc 2460
ctcaggaacc ggactcctcc gcgggtattg gcaaatcggg tgcacagccc gctaaaaaga 2520
gactcaattt cggtcagact ggcgacacag agtcagtccc agaccctcaa ccaatcggag 2580
aacctcccgc agccccctca ggtgtgggat ctcttacaat ggcttcaggt ggtggcgcac 2640
cagtggcaga caataacgaa ggtgccgatg gagtgggtag ttcctcggga aattggcatt 2700
gcgattccca atggctgggg gacagagtca tcaccaccag cacccgaacc tgggccctgc 2760
ccacctacaa caatcacctc tacaagcaaa tctccaacag cacatctgga ggatcttcaa 2820
atgacaacgc ctacttcggc tacagcaccc cctgggggta ttttgacttc aacagattcc 2880
actgccactt ctcaccacgt gactggcagc gactcatcaa caacaactgg ggattccggc 2940
ctaagcgact caacttcaag ctcttcaaca ttcaggtcaa agaggttacg gacaacaatg 3000
gagtcaagac catcgccaat aaccttacca gcacggtcca ggtcttcacg gactcagact 3060
atcagctccc gtacgtgctc gggtcggctc acgagggctg cctcccgccg ttcccagcgg 3120
acgttttcat gattcctcag tacgggtatc tgacgcttaa tgatggaagc caggccgtgg 3180
gtcgttcgtc cttttactgc ctggaatatt tcccgtcgca aatgctaaga acgggtaaca 3240
acttccagtt cagctacgag tttgagaacg tacctttcca tagcagctac gctcacagcc 3300
aaagcctgga ccgactaatg aatccactca tcgaccaata cttgtactat ctctcaaaga 3360
ctattaacgg ttctggacag aatcaacaaa cgctaaaatt cagtgtggcc ggacccagca 3420
acatggctgt ccagggaaga aactacatac ctggacccag ctaccgacaa caacgtgtct 3480
caaccactgt gactcaaaac aacaacagcg aatttgcttg gcctggagct tcttcttggg 3540
ctctcaatgg acgtaatagc ttgatgaatc ctggacctgc tatggccagc cacaaagaag 3600
gagaggaccg tttctttcct ttgtctggat ctttaatttt tggcaaacaa ggaactggaa 3660
gagacaacgt ggatgcggac aaagtcatga taaccaacga agaagaaatt aaaactacta 3720
acccggtagc aacggagtcc tatggacaag tggccacaaa ccaccagagt gcccaagcac 3780
aggcgcagac cggctgggtt caaaaccaag gaatacttcc gggtatggtt tggcaggaca 3840
gagatgtgta cctgcaagga cccatttggg ccaaaattcc tcacacggac ggcaactttc 3900
acccttctcc gctgatggga gggtttggaa tgaagcaccc gcctcctcag atcctcatca 3960
aaaacacacc tgtacctgcg gatcctccaa cggccttcaa caaggacaag ctgaactctt 4020
tcatcaccca gtattctact ggccaagtca gcgtggagat cgagtgggag ctgcagaagg 4080
aaaacagcaa gcgctggaac ccggagatcc agtacacttc caactattac aagtctaata 4140
atgttgaatt tgctgttaat actgaaggtg tatatagtga accccgcccc attggcacca 4200
gatacctgac tcgtaatctg taattgcttg ttaatcaata aaccgtttaa ttcgtttcag 4260
ttgaactttg gtctctgcgt atttctttct tatctagttt ccatggctac gtagataagt 4320
agcatggcgg gttaatcatt aactacagcc cgggcgttta aacagcgggc ggaggggtgg 4380
agtcgtgacg tgaattacgt catagggtta gggaggtcct gtattagagg tcacgtgagt 4440
gttttgcgac attttgcgac accatgtggt ctcgctgggg gggggggccc gagtgagcac 4500
gcagggtctc cattttgaag cgggaggttt gaacgagcgc tggcgcgctc actggccgtc 4560
gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca 4620
catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa 4680
cagttgcgca gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg ttaaaattcg 4740
cgttaaattt ttgttaaatc agctcatttt tttaaccaat aggccgaaat cggcaaaatc 4800
ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag 4860
agtccactat taagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg 4920
atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag 4980
cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga 5040
acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg 5100
tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg 5160
cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 5220
tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 5280
gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 5340
cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 5400
atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 5460
agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 5520
gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt 5580
ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 5640
cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 5700
ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 5760
atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 5820
gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 5880
tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 5940
gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 6000
gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 6060
tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 6120
ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 6180
tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 6240
ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 6300
ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 6360
tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 6420
ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 6480
tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 6540
tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 6600
actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 6660
cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 6720
gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 6780
tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 6840
ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 6900
ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 6960
cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 7020
cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga 7080
gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc 7140
attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa 7200
ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc 7260
gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg 7320
attacgccaa 7330
<210> 10
<211> 7330
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
gcgcgccgat atcgttaacg ccccgcgccg gccgctctag aactagtgga tcccccggaa 60
gatcagaagt tcctattccg aagttcctat tctctagaaa gtataggaac ttctgatctg 120
cgcagccgcc atgccggggt tttacgagat tgtgattaag gtccccagcg accttgacga 180
gcatctgccc ggcatttctg acagctttgt gaactgggtg gccgagaagg aatgggagtt 240
gccgccagat tctgacatgg atctgaatct gattgagcag gcacccctga ccgtggccga 300
gaagctgcag cgcgactttc tgacggaatg gcgccgtgtg agtaaggccc cggaggccct 360
tttctttgtg caatttgaga agggagagag ctacttccac atgcacgtgc tcgtggaaac 420
caccggggtg aaatccatgg ttttgggacg tttcctgagt cagattcgcg aaaaactgat 480
tcagagaatt taccgcggga tcgagccgac tttgccaaac tggttcgcgg tcacaaagac 540
cagaaatggc gccggaggcg ggaacaaggt ggtggatgag tgctacatcc ccaattactt 600
gctccccaaa acccagcctg agctccagtg ggcgtggact aatatggaac agtatttaag 660
cgcctgtttg aatctcacgg agcgtaaacg gttggtggcg cagcatctga cgcacgtgtc 720
gcagacgcag gagcagaaca aagagaatca gaatcccaat tctgatgcgc cggtgatcag 780
atcaaaaact tcagccaggt acatggagct ggtcgggtgg ctcgtggaca aggggattac 840
ctcggagaag cagtggatcc aggaggacca ggcctcatac atctccttca atgcggcctc 900
caactcgcgg tcccaaatca aggctgcctt ggacaatgcg ggaaagatta tgagcctgac 960
taaaaccgcc cccgactacc tggtgggcca gcagcccgtg gaggacattt ccagcaatcg 1020
gatttataaa attttggaac taaacgggta cgatccccaa tatgcggctt ccgtctttct 1080
gggatgggcc acgaaaaagt tcggcaagag gaacaccatc tggctgtttg ggcctgcaac 1140
taccgggaag accaacatcg cggaggccat agcccacact gtgcccttct acgggtgcgt 1200
aaactggacc aatgagaact ttcccttcaa cgactgtgtc gacaagatgg tgatctggtg 1260
ggaggagggg aagatgaccg ccaaggtcgt ggagtcggcc aaagccattc tcggaggaag 1320
caaggtgcgc gtggaccaga aatgcaagtc ctcggcccag atagacccga ctcccgtgat 1380
cgtcacctcc aacaccaaca tgtgcgccgt gattgacggg aactcaacga ccttcgaaca 1440
ccagcagccg ttgcaagacc ggatgttcaa atttgaactc acccgccgtc tggatcatga 1500
ctttgggaag gtcaccaagc aggaagtcaa agactttttc cggtgggcaa aggatcacgt 1560
ggttgaggtg gagcatgaat tctacgtcaa aaagggtgga gccaagaaaa gacccgcccc 1620
cagtgacgca gatataagtg agcccaaacg ggtgcgcgag tcagttgcgc agccatcgac 1680
gtcagacgcg gaagcttcga tcaactacgc agacaggtac caaaacaaat gttctcgtca 1740
cgtgggcatg aatctgatgc tgtttccctg cagacaatgc gagagaatga atcagaattc 1800
aaatatctgc ttcactcacg gacagaaaga ctgtttagag tgctttcccg tgtcagaatc 1860
tcaacccgtt tctgtcgtca aaaaggcgta tcagaaactg tgctacattc atcatatcat 1920
gggaaaggtg ccagacgctt gcactgcctg cgatctggtc aatgtggatt tggatgactg 1980
catctttgaa caataaatga tttaaatcag gtatggctgc cgatggttat cttccagatt 2040
ggctcgagga caacctctct gagggcattc gcgagtggtg ggacttgaaa cctggagccc 2100
cgaaacccaa agccaaccag caaaagcagg acgacggccg gggtctggtg cttcctggct 2160
acaagtacct cggacccttc aacggactcg acaaggggga gcccgtcaac gcggcggacg 2220
cagcggccct cgagcacgac aaggcctacg accagcagct caaagcgggt gacaatccgt 2280
acctgcggta taaccacgcc gacgccgagt ttcaggagcg tctgcaagaa gatacgtctt 2340
ttgggggcaa cctcgggcga gcagtcttcc aggccaagaa gcgggttctc gaacctctcg 2400
gtctggttga ggaaggcgct aagacggctc ctggaaagaa gagaccggta gagcaatcac 2460
cccaggaacc agactcctct tcgggcatcg gcaagaaagg ccagcagccc gcgagaaaga 2520
gactcaactt tgggcagact ggcgactcag agtcagtgcc cgaccctcaa ccactcggag 2580
aaccccccgc agccccctct ggtgtgggat ctaatacaat ggctgcaggc ggtggcgctc 2640
caatggcaga caataacgaa ggcgccgacg gagtgggtaa cgcctcagga aattggcatt 2700
gcgattccac atggctgggc gacagagtca tcaccaccag cacccgaacc tgggccctcc 2760
ccacctacaa caaccacctc tacaagcaaa tctccagcca atcgggaggc agcaccaacg 2820
acaacaccta cttcggctac agcaccccct gggggtattt tgactttaac agattccact 2880
gccacttctc accacgtgac tggcagcgac tcatcaacaa caactgggga ttccggccca 2940
agaagctcaa cttcaagctc ttcaacatcc aggtcaagga ggtcacgacg aatgatggca 3000
ccacgaccat cgccaataac cttaccagca cggttcaggt ctttacggac tcggaatacc 3060
agctcccgta cgtcctcggc tctgcgcacc agggctgcct gcctccgttc ccggcggacg 3120
tcttcatgat tcctcagtac gggtacctga ctctgaacaa tggcagtcag gccgtgggcc 3180
gttcctcctt ctactgcctg gaatactttc cttctcaaat gctgagaacg ggcaacaact 3240
ttcagttcag ctacacgttt gaggacgtgc cttttcacag cagctacgcg cacagccaaa 3300
gcctggaccg gctgatgaac cccctcatcg accagtacct gtactacctg tctcggactc 3360
agaccacgag tggtaccgca ggaaatcgga cgttgcaatt ttctcaggcc gggcctagta 3420
gcatggcgaa tcaggccaaa aactggctac ccgggccctg ctaccggcag caacgcgtct 3480
ccaagacaac caatcaaaat aacaacagca actttgcctg gaccggtgcc accaagtatc 3540
atctgaatgg cagagactct ctggtaaatc ccggtcccgc tatggcaacc cacaaggacg 3600
acgaagacaa attttttccg atgagcggag tcttaatatt tgggaaacag ggagctggaa 3660
atagcaacgt ggaccttgac aacgttatga taaccaacga ggaagaaatt aaaaccacca 3720
acccagtggc cacagaagag tacggcacgg tggccactaa cctgcaatcg gccaacaccg 3780
ctcctgctac agggaccgtc aacagtcaag gagccttacc tggcatggtc tggcaggacc 3840
gggacgtgta cctgcagggt cctatctggg ccaagattcc tcacacggac ggacactttc 3900
atccctcgcc gctgatggga ggctttggac tgaaacaccc gcctcctcag atcctgatta 3960
agaatacacc tgttcccgcg aatcctccaa ctaccttcag tccagctaag tttgcgtcgt 4020
tcatcacgca gtacagcacc ggacaggtca gcgtggaaat tgaatgggag ctgcagaaag 4080
aaaacagcaa acgctggaac ccagagattc aatacacttc caactacaac aaatctacaa 4140
atgtggactt tgctgttgac acaaatggcg tttattctga gcctcgcccc atcggcaccc 4200
gttacctcac ccgtaatctg taattgcttg ttaatcaata aaccgtttaa ttcgtttcag 4260
ttgaactttg gtctctgcgt atttctttct tatctagttt ccatggctac gtagataagt 4320
agcatggcgg gttaatcatt aactacagcc cgggcgttta aacagcgggc ggaggggtgg 4380
agtcgtgacg tgaattacgt catagggtta gggaggtcct gtattagagg tcacgtgagt 4440
gttttgcgac attttgcgac accatgtggt ctcgctgggg gggggggccc gagtgagcac 4500
gcagggtctc cattttgaag cgggaggttt gaacgagcgc tggcgcgctc actggccgtc 4560
gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca 4620
catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa 4680
cagttgcgca gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg ttaaaattcg 4740
cgttaaattt ttgttaaatc agctcatttt tttaaccaat aggccgaaat cggcaaaatc 4800
ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag 4860
agtccactat taagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg 4920
atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag 4980
cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga 5040
acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg 5100
tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg 5160
cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 5220
tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 5280
gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 5340
cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 5400
atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 5460
agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 5520
gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt 5580
ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 5640
cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 5700
ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 5760
atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 5820
gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 5880
tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 5940
gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 6000
gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 6060
tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 6120
ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 6180
tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 6240
ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 6300
ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 6360
tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 6420
ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 6480
tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 6540
tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 6600
actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 6660
cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 6720
gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 6780
tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 6840
ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 6900
ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 6960
cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 7020
cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga 7080
gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc 7140
attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa 7200
ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc 7260
gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg 7320
attacgccaa 7330
<210> 11
<211> 5704
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60
gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120
actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt 180
ttaaacagag aggaatctct cgagccattg acgtcaataa tgacgtatgt tcccatagta 240
acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac 300
ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt 360
aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag 420
tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc 480
actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta 540
ttttgtgcag cgatgggggc gggggggggg gggggggggc ggggcgaggc ggagaggtgc 600
ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg 660
gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg 720
ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt 780
actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt 840
ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg 900
ccctttgtgc ggggggagcg gctcggggct gtccgcgggg ggacggctgc cttcgggggg 960
gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta 1020
accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc 1080
tgtctcatca ttttggcaaa gaattggatc gaattcgcca ccatgggctc cggagccacc 1140
aacttttctc tgctgaagca ggctggcgac gtggaggaga acccaggacc tatggtgtct 1200
aagggcgagg aggataacat ggccagcctg cctgctacac acgagctgca catcttcgga 1260
agcatcaacg gcgtggactt tgatatggtg ggccagggaa ccggcaaccc aaacgacgga 1320
tacgaggagc tgaacctgaa gagcacaaag ggcgatctgc agttctcccc ctggattctg 1380
gtgcctcaca tcggatacgg ctttcaccag tacctgccat acccagacgg aatgagccca 1440
ttccaggctg ctatggtgga tggcagcggc taccaggtgc acaggaccat gcagtttgag 1500
gacggcgcct ctctgaccgt gaactacaga tacacatacg agggaagcca catcaagggc 1560
gaggcccagg tgaagggaac cggcttccct gctgacggac cagtgatgac caactccctg 1620
acagccgctg actggtgccg gtctaagaaa acatacccta acgataagac catcatctct 1680
acattcaagt ggagctacac cacaggaaac ggcaagaggt acagatccac cgcccggacc 1740
acatacacat ttgctaagcc aatggccgct aactacctga agaaccagcc catgtacgtg 1800
ttccgcaaga ccgagctgaa gcactccaag acagagctga acttcaagga gtggcagaag 1860
gcctttaccg acgtgatggg aatggatgag ctgtacaagg gatccgacta caaggaccac 1920
gatggcgact acaaggatca cgacatcgat tacaaggacg atgacgataa gaagcggaca 1980
gctgatggca gcgagttcga gtcccccaag aagaagagga aggtggagtg agctagcgat 2040
atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt 2100
cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat 2160
gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct 2220
ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct 2280
gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc 2340
gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg 2400
acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc 2460
tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac 2520
gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg 2580
cctcttccgc gacttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc 2640
ccgcagatct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 2700
aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 2760
caatgtatct tatcatgtct ggctagacac gtggccgcta ccccgaccac atgaagcagc 2820
acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca 2880
aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga 2940
accgcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc 3000
tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt 3060
tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg 3120
gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag 3180
tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 3240
gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 3300
acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 3360
agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg 3420
ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 3480
ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta 3540
taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 3600
aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc 3660
tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc 3720
tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc 3780
tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg 3840
atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc 3900
acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat 3960
atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag 4020
agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt 4080
cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt 4140
gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc 4200
cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta 4260
tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac 4320
ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa 4380
ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg 4440
atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc 4500
cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg 4560
atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta 4620
gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg 4680
cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg 4740
tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc 4800
tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt 4860
gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt 4920
gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc 4980
atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag 5040
atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 5100
aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 5160
aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag 5220
ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 5280
ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 5340
tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 5400
ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 5460
acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 5520
gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 5580
cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 5640
aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac 5700
atgt 5704
<210> 12
<211> 11635
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
ggtacccaac tccatgctta acagtcccca ggtacagccc accctgcgtc gcaaccagga 60
acagctctac agcttcctgg agcgccactc gccctacttc cgcagccaca gtgcgcagat 120
taggagcgcc acttcttttt gtcacttgaa aaacatgtaa aaataatgta ctaggagaca 180
ctttcaataa aggcaaatgt ttttatttgt acactctcgg gtgattattt accccccacc 240
cttgccgtct gcgccgttta aaaatcaaag gggttctgcc gcgcatcgct atgcgccact 300
ggcagggaca cgttgcgata ctggtgttta gtgctccact taaactcagg cacaaccatc 360
cgcggcagct cggtgaagtt ttcactccac aggctgcgca ccatcaccaa cgcgtttagc 420
aggtcgggcg ccgatatctt gaagtcgcag ttggggcctc cgccctgcgc gcgcgagttg 480
cgatacacag ggttgcagca ctggaacact atcagcgccg ggtggtgcac gctggccagc 540
acgctcttgt cggagatcag atccgcgtcc aggtcctccg cgttgctcag ggcgaacgga 600
gtcaactttg gtagctgcct tcccaaaaag ggtgcatgcc caggctttga gttgcactcg 660
caccgtagtg gcatcagaag gtgaccgtgc ccggtctggg cgttaggata cagcgcctgc 720
atgaaagcct tgatctgctt aaaagccacc tgagcctttg cgccttcaga gaagaacatg 780
ccgcaagact tgccggaaaa ctgattggcc ggacaggccg cgtcatgcac gcagcacctt 840
gcgtcggtgt tggagatctg caccacattt cggccccacc ggttcttcac gatcttggcc 900
ttgctagact gctccttcag cgcgcgctgc ccgttttcgc tcgtcacatc catttcaatc 960
acgtgctcct tatttatcat aatgctcccg tgtagacact taagctcgcc ttcgatctca 1020
gcgcagcggt gcagccacaa cgcgcagccc gtgggctcgt ggtgcttgta ggttacctct 1080
gcaaacgact gcaggtacgc ctgcaggaat cgccccatca tcgtcacaaa ggtcttgttg 1140
ctggtgaagg tcagctgcaa cccgcggtgc tcctcgttta gccaggtctt gcatacggcc 1200
gccagagctt ccacttggtc aggcagtagc ttgaagtttg cctttagatc gttatccacg 1260
tggtacttgt ccatcaacgc gcgcgcagcc tccatgccct tctcccacgc agacacgatc 1320
ggcaggctca gcgggtttat caccgtgctt tcactttccg cttcactgga ctcttccttt 1380
tcctcttgcg tccgcatacc ccgcgccact gggtcgtctt cattcagccg ccgcaccgtg 1440
cgcttacctc ccttgccgtg cttgattagc accggtgggt tgctgaaacc caccatttgt 1500
agcgccacat cttctctttc ttcctcgctg tccacgatca cctctgggga tggcgggcgc 1560
tcgggcttgg gagaggggcg cttctttttc tttttggacg caatggccaa atccgccgtc 1620
gaggtcgatg gccgcgggct gggtgtgcgc ggcaccagcg catcttgtga cgagtcttct 1680
tcgtcctcgg actcgagacg ccgcctcagc cgcttttttg ggggcgcgcg gggaggcggc 1740
ggcgacggcg acggggacga cacgtcctcc atggttggtg gacgtcgcgc cgcaccgcgt 1800
ccgcgctcgg gggtggtttc gcgctgctcc tcttcccgac tggccatttc cttctcctat 1860
aggcagaaaa agatcatgga gtcagtcgag aaggaggaca gcctaaccgc cccctttgag 1920
ttcgccacca ccgcctccac cgatgccgcc aacgcgccta ccaccttccc cgtcgaggca 1980
cccccgcttg aggaggagga agtgattatc gagcaggacc caggttttgt aagcgaagac 2040
gacgaggatc gctcagtacc aacagaggat aaaaagcaag accaggacga cgcagaggca 2100
aacgaggaac aagtcgggcg gggggaccaa aggcatggcg actacctaga tgtgggagac 2160
gacgtgctgt tgaagcatct gcagcgccag tgcgccatta tctgcgacgc gttgcaagag 2220
cgcagcgatg tgcccctcgc catagcggat gtcagccttg cctacgaacg ccacctgttc 2280
tcaccgcgcg taccccccaa acgccaagaa aacggcacat gcgagcccaa cccgcgcctc 2340
aacttctacc ccgtatttgc cgtgccagag gtgcttgcca cctatcacat ctttttccaa 2400
aactgcaaga tacccctatc ctgccgtgcc aaccgcagcc gagcggacaa gcagctggcc 2460
ttgcggcagg gcgctgtcat acctgatatc gcctcgctcg acgaagtgcc aaaaatcttt 2520
gagggtcttg gacgcgacga gaaacgcgcg gcaaacgctc tgcaacaaga aaacagcgaa 2580
aatgaaagtc actgtggagt gctggtggaa cttgagggtg acaacgcgcg cctagccgtg 2640
ctgaaacgca gcatcgaggt cacccacttt gcctacccgg cacttaacct accccccaag 2700
gttatgagca cagtcatgag cgagctgatc gtgcgccgtg cacgacccct ggagagggat 2760
gcaaacttgc aagaacaaac cgaggagggc ctacccgcag ttggcgatga gcagctggcg 2820
cgctggcttg agacgcgcga gcctgccgac ttggaggagc gacgcaagct aatgatggcc 2880
gcagtgcttg ttaccgtgga gcttgagtgc atgcagcggt tctttgctga cccggagatg 2940
cagcgcaagc tagaggaaac gttgcactac acctttcgcc agggctacgt gcgccaggcc 3000
tgcaaaattt ccaacgtgga gctctgcaac ctggtctcct accttggaat tttgcacgaa 3060
aaccgcctcg ggcaaaacgt gcttcattcc acgctcaagg gcgaggcgcg ccgcgactac 3120
gtccgcgact gcgtttactt atttctgtgc tacacctggc aaacggccat gggcgtgtgg 3180
cagcaatgcc tggaggagcg caacctaaag gagctgcaga agctgctaaa gcaaaacttg 3240
aaggacctat ggacggcctt caacgagcgc tccgtggccg cgcacctggc ggacattatc 3300
ttccccgaac gcctgcttaa aaccctgcaa cagggtctgc cagacttcac cagtcaaagc 3360
atgttgcaaa actttaggaa ctttatccta gagcgttcag gaattctgcc cgccacctgc 3420
tgtgcgcttc ctagcgactt tgtgcccatt aagtaccgtg aatgccctcc gccgctttgg 3480
ggtcactgct accttctgca gctagccaac taccttgcct accactccga catcatggaa 3540
gacgtgagcg gtgacggcct actggagtgt cactgtcgct gcaacctatg caccccgcac 3600
cgctccctgg tctgcaattc gcaactgctt agcgaaagtc aaattatcgg tacctttgag 3660
ctgcagggtc cctcgcctga cgaaaagtcc gcggctccgg ggttgaaact cactccgggg 3720
ctgtggacgt cggcttacct tcgcaaattt gtacctgagg actaccacgc ccacgagatt 3780
aggttctacg aagaccaatc ccgcccgcca aatgcggagc ttaccgcctg cgtcattacc 3840
cagggccaca tccttggcca attgcaagcc atcaacaaag cccgccaaga gtttctgcta 3900
cgaaagggac ggggggttta cctggacccc cagtccggcg aggagctcaa cccaatcccc 3960
ccgccgccgc agccctatca gcagccgcgg gcccttgctt cccaggatgg cacccaaaaa 4020
gaagctgcag ctgccgccgc cgccacccac ggacgaggag gaatactggg acagtcaggc 4080
agaggaggtt ttggacgagg aggaggagat gatggaagac tgggacagcc tagacgaagc 4140
ttccgaggcc gaagaggtgt cagacgaaac accgtcaccc tcggtcgcat tcccctcgcc 4200
ggcgccccag aaattggcaa ccgttcccag catcgctaca acctccgctc ctcaggcgcc 4260
gccggcactg cctgttcgcc gacccaaccg tagatgggac accactggaa ccagggccgg 4320
taagtctaag cagccgccgc cgttagccca agagcaacaa cagcgccaag gctaccgctc 4380
gtggcgcggg cacaagaacg ccatagttgc ttgcttgcaa gactgtgggg gcaacatctc 4440
cttcgcccgc cgctttcttc tctaccatca cggcgtggcc ttcccccgta acatcctgca 4500
ttactaccgt catctctaca gcccctactg caccggcggc agcggcagcg gcagcaacag 4560
cagcggtcac acagaagcaa aggcgaccgg atagcaagac tctgacaaag cccaagaaat 4620
ccacagcggc ggcagcagca ggaggaggag cgctgcgtct ggcgcccaac gaacccgtat 4680
cgacccgcga gcttagaaat aggatttttc ccactctgta tgctatattt caacaaagca 4740
ggggccaaga acaagagctg aaaataaaaa acaggtctct gcgctccctc acccgcagct 4800
gcctgtatca caaaagcgaa gatcagcttc ggcgcacgct ggaagacgcg gaggctctct 4860
tcagcaaata ctgcgcgctg actcttaagg actagtttcg cgccctttct caaatttaag 4920
cgcgaaaact acgtcatctc cagcggccac acccggcgcc agcacctgtc gtcagcgcca 4980
ttatgagcaa ggaaattccc acgccctaca tgtggagtta ccagccacaa atgggacttg 5040
cggctggagc tgcccaagac tactcaaccc gaataaacta catgagcgcg ggaccccaca 5100
tgatatcccg ggtcaacgga atccgcgccc accgaaaccg aattctcctc gaacaggcgg 5160
ctattaccac cacacctcgt aataacctta atccccgtag ttggcccgct gccctggtgt 5220
accaggaaag tcccgctccc accactgtgg tacttcccag agacgcccag gccgaagttc 5280
agatgactaa ctcaggggcg cagcttgcgg gcggctttcg tcacagggtg cggtcgcccg 5340
ggcgttttag ggcggagtaa cttgcatgta ttgggaattg tagttttttt aaaatgggaa 5400
gtgacgtatc gtgggaaaac ggaagtgaag atttgaggaa gttgtgggtt ttttggcttt 5460
cgtttctggg cgtaggttcg cgtgcggttt tctgggtgtt ttttgtggac tttaaccgtt 5520
acgtcatttt ttagtcctat atatactcgc tctgtacttg gcccttttta cactgtgact 5580
gattgagctg gtgccgtgtc gagtggtgtt ttttaatagg tttttttact ggtaaggctg 5640
actgttatgg ctgccgctgt ggaagcgctg tatgttgttc tggagcggga gggtgctatt 5700
ttgcctaggc aggagggttt ttcaggtgtt tatgtgtttt tctctcctat taattttgtt 5760
atacctccta tgggggctgt aatgttgtct ctacgcctgc gggtatgtat tcccccgggc 5820
tatttcggtc gctttttagc actgaccgat gttaaccaac ctgatgtgtt taccgagtct 5880
tacattatga ctccggacat gaccgaggaa ctgtcggtgg tgctttttaa tcacggtgac 5940
cagttttttt acggtcacgc cggcatggcc gtagtccgtc ttatgcttat aagggttgtt 6000
tttcctgttg taagacaggc ttctaatgtt taaatgtttt tttttttgtt attttatttt 6060
gtgtttaatg caggaacccg cagacatgtt tgagagaaaa atggtgtctt tttctgtggt 6120
ggttccggaa cttacctgcc tttatctgca tgagcatgac tacgatgtgc ttgctttttt 6180
gcgcgaggct ttgcctgatt ttttgagcag caccttgcat tttatatcgc cgcccatgca 6240
acaagcttac ataggggcta cgctggttag catagctccg agtatgcgtg tcataatcag 6300
tgtgggttct tttgtcatgg ttcctggcgg ggaagtggcc gcgctggtcc gtgcagacct 6360
gcacgattat gttcagctgg ccctgcgaag ggacctacgg gatcgcggta tttttgttaa 6420
tgttccgctt ttgaatctta tacaggtctg tgaggaacct gaatttttgc aatcatgatt 6480
cgctgcttga ggctgaaggt ggagggcgct ctggagcaga tttttacaat ggccggactt 6540
aatattcggg atttgcttag agacatattg ataaggtggc gagatgaaaa ttatttgggc 6600
atggttgaag gtgctggaat gtttatagag gagattcacc ctgaagggtt tagcctttac 6660
gtccacttgg acgtgagggc agtttgcctt ttggaagcca ttgtgcaaca tcttacaaat 6720
gccattatct gttctttggc tgtagagttt gaccacgcca ccggagggga gcgcgttcac 6780
ttaatagatc ttcattttga ggttttggat aatcttttgg aataaaaaaa aaaaaacatg 6840
gttcttccag ctcttcccgc tcctcccgtg tgtgactcgc agaacgaatg tgtaggttgg 6900
ctgggtgtgg cttattctgc ggtggtggat gttatcaggg cagcggcgca tgaaggagtt 6960
tacatagaac ccgaagccag ggggcgcctg gatgctttga gagagtggat atactacaac 7020
tactacacag agcgagctaa gcgacgagac cggagacgca gatctgtttg tcacgcccgc 7080
acctggtttt gcttcaggaa atatgactac gtccggcgtt ccatttggca tgacactacg 7140
accaacacga tctcggttgt ctcggcgcac tccgtacagt agggatcgcc tacctccttt 7200
tgagacagag acccgcgcta ccatactgga ggatcatccg ctgctgcccg aatgtaacac 7260
tttgacaatg cacaacgtga gttacgtgcg aggtcttccc tgcagtgtgg gatttacgct 7320
gattcaggaa tgggttgttc cctgggatat ggttctgacg cgggaggagc ttgtaatcct 7380
gaggaagtgt atgcacgtgt gcctgtgttg tgccaacatt gatatcatga cgagcatgat 7440
gatccatggt tacgagtcct gggctctcca ctgtcattgt tccagtcccg gttccctgca 7500
gtgcatagcc ggcgggcagg ttttggccag ctggtttagg atggtggtgg atggcgccat 7560
gtttaatcag aggtttatat ggtaccggga ggtggtgaat tacaacatgc caaaagaggt 7620
aatgtttatg tccagcgtgt ttatgagggg tcgccactta atctacctgc gcttgtggta 7680
tgatggccac gtgggttctg tggtccccgc catgagcttt ggatacagcg ccttgcactg 7740
tgggattttg aacaatattg tggtgctgtg ctgcagttac tgtgctgatt taagtgagat 7800
cagggtgcgc tgctgtgccc ggaggacaag gcgtctcatg ctgcgggcgg tgcgaatcat 7860
cgctgaggag accactgcca tgttgtattc ctgcaggacg gagcggcggc ggcagcagtt 7920
tattcgcgcg ctgctgcagc accaccgccc tatcctgatg cacgattatg actctacccc 7980
catgtaggcg tggacttccc cttcgccgcc cgttgagcaa ccgcaagttg gacagcagcc 8040
tgtggctcag cagctggaca gcgacatgaa cttaagcgag ctgcccgggg agtttattaa 8100
tatcactgat gagcgtttgg ctcgacagga aaccgtgtgg aatataacac ctaagaatat 8160
gtctgttacc catgatatga tgctttttaa ggccagccgg ggagaaagga ctgtgtactc 8220
tgtgtgttgg gagggaggtg gcaggttgaa tactagggtt ctgtgagttt gattaaggta 8280
cggtgatcaa tataagctat gtggtggtgg ggctatacta ctgaatgaaa aatgacttga 8340
aattttctgc aattgaaaaa taaacacgtt gaaacataac atgcaacagg ttcacgattc 8400
tttattcctg ggcaatgtag gagaaggtgt aagagttggt agcaaaagtt tcagtggtgt 8460
attttccact ttcccaggac catgtaaaag acatagagta agtgcttacc tcgctagttt 8520
ctgtggattc actagaatcg atgtaggatg ttgcccctcc tgacgcggta ggagaagggg 8580
agggtgccct gcatgtctgc cgctgctctt gctcttgccg ctgctgagga ggggggcgca 8640
tctgccgcag caccggatgc atctgggaaa agcaaaaaag gggctcgtcc ctgtttccgg 8700
aggaatttgc aagcggggtc ttgcatgacg gggaggcaaa cccccgttcg ccgcagtccg 8760
gccggcccga gactcgaacc gggggtcctg cgactcaacc cttggaaaat aaccctccgg 8820
ctacagggag cgagccactt aatgctttcg ctttccagcc taaccgctta cgccgcgcgc 8880
ggccagtggc caaaaaagct agcgcagcag ccgccgcgcc tggaaggaag ccaaaaggag 8940
cgctcccccg ttgtctgacg tcgcacacct gggttcgaca cgcgggcggt aaccgcatgg 9000
atcacggcgg acggccggat ccggggttcg aaccccggtc gtccgccatg atacccttgc 9060
gaatttatcc accagaccac ggaagagtgc ccgcttacag gctctccttt tgcacggtct 9120
agagcgtcaa cgactgcgca cgcctcaccg gccagagcgt cccgaccatg gagcactttt 9180
tgccgctgcg caacatctgg aaccgcgtcc gcgactttcc gcgcgcctcc accaccgccg 9240
ccggcatcac ctggatgtcc aggtacatct acggattacg tcgacgttta aaccatatga 9300
tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 9360
aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 9420
tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 9480
tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 9540
cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 9600
agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 9660
tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 9720
aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 9780
ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 9840
cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 9900
accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 9960
ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 10020
ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 10080
gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 10140
aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 10200
gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 10260
gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 10320
cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 10380
gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 10440
gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 10500
ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 10560
tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 10620
ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 10680
cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 10740
accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 10800
cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 10860
tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 10920
cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 10980
acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 11040
atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 11100
tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 11160
aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt 11220
aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag 11280
aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga 11340
acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg 11400
aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc 11460
ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg 11520
aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc 11580
gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgatg gatcc 11635
Claims (14)
1.一种变异型腺相关病毒AAV-ie衣壳蛋白VP1，所述变异型腺相关病毒AAV-ie衣壳蛋白VP1，相比于野生型AAV-DJ衣壳蛋白VP1，在N589与R590之间插入氨基酸片段，所述氨基酸片段的氨基酸序列如SEQ ID NO.1所示。
2.如权利要求1所述的一种变异型腺相关病毒AAV-ie衣壳蛋白VP1，其特征在于，所述变异型腺相关病毒AAV-ie衣壳蛋白VP1的氨基酸序列如SEQ ID NO.4所示。
3.一种分离的核酸，其特征在于，所述核酸含有编码如权利要求1中所述的变异型腺相关病毒AAV-ie衣壳蛋白VP1的核苷酸序列。
4.一种构建体，所述构建体含有如权利要求3所述的分离的核酸。
5.一种宿主细胞，其特征在于，所述宿主细胞包含如权利要求4所述的构建体或基因组中整合有外源的如权利要求3所述的分离的核酸。
6.一种变异型腺相关病毒AAV-ie，其特征在于，所述变异型腺相关病毒AAV-ie含有如权利要求1-2任一权利要求所述的变异型腺相关病毒AAV-ie衣壳蛋白VP1。
7.如权利要求6所述的变异型腺相关病毒AAV-ie，其特征在于，所述变异型腺相关病毒AAV-ie还包括编码目的产物的异源核苷酸序列。
8.如权利要求6所述的变异型腺相关病毒AAV-ie，其特征在于，所述目的产物为核酸或者蛋白质，所述核酸优选选自小向导RNA、干扰RNA。
9.一种药物组合物，其特征在于，所述药物组合物包含如权利要求6～8任意一项所述的变异型腺相关病毒AAV-ie以及药学上可接受的辅料。
10.如权利要求6～8任意一项所述的变异型腺相关病毒AAV-ie在用于将目的产物递送至个体的毛细胞和/或支持细胞中的用途。
11.根据权利要求10所述的用途，其特征在于，所述目的产物为核酸或蛋白质，所述核酸优选为小向导RNA、干扰RNA。
12.如权利要求6～8任意一项所述的变异型腺相关病毒AAV-ie在制备治疗个体中耳蜗损伤导致的听力障碍疾病的药物中的用途。
13.如权利要求12所述的用途，其特征在于，所述听力障碍疾病为基因缺陷、环境损伤或衰老的相关疾病。
14.根据权利要求12所述的用途，其特征在于，所述听力障碍疾病为细胞损伤的相关疾病。
Priority Applications (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US17/284,673 US20210355171A1 (en) | 2019-01-30 | 2019-09-24 | Adeno-associated virus having a variant capsid protein, and use thereof |
| EP19912419.9A EP3943503A4 (en) | 2019-01-30 | 2019-09-24 | ADENO-ASSOCIATED VIRUS WITH VARIANT CAPSID PROTEIN AND USE THEREOF |
| PCT/CN2019/107474 WO2020155655A1 (zh) | 2019-01-30 | 2019-09-24 | 具有变异衣壳蛋白的腺相关病毒及其用途 |
| CA3140506A CA3140506A1 (en) | 2019-01-30 | 2019-09-24 | Adeno-associated virus having a variant capsid protein, and use thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910093000 | 2019-01-30 | ||
| CN2019100930003 | 2019-01-30 |
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| CN110437317A true CN110437317A (zh) | 2019-11-12 |
| CN110437317B CN110437317B (zh) | 2023-05-02 |
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| CN201910807643.XA Active CN110437317B (zh) | 2019-01-30 | 2019-08-29 | 具有变异衣壳蛋白的腺相关病毒及其用途 |
Country Status (5)
| Country | Link |
|---|---|
| US (1) | US20210355171A1 (zh) |
| EP (1) | EP3943503A4 (zh) |
| CN (1) | CN110437317B (zh) |
| CA (1) | CA3140506A1 (zh) |
| WO (1) | WO2020155655A1 (zh) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110950934A (zh) * | 2019-12-31 | 2020-04-03 | 复旦大学 | 一种腺相关病毒衣壳蛋白、载体及其构建方法与应用 |
| CN111825772A (zh) * | 2020-07-30 | 2020-10-27 | 中国科学院精密测量科学与技术创新研究院 | 具有变异衣壳蛋白的腺相关病毒及其应用 |
| CN112011571A (zh) * | 2020-04-26 | 2020-12-01 | 辉大(上海)生物科技有限公司 | 一种用于治疗脊髓性肌萎缩的基因治疗药物 |
| CN112813037A (zh) * | 2021-01-08 | 2021-05-18 | 中国科学院动物研究所 | 一种高效感染原代小胶质细胞的重组突变腺相关病毒及其相关生物材料 |
| CN115340998A (zh) * | 2021-05-14 | 2022-11-15 | 上海科技大学 | 一种重组病毒载体、包含其的重组病毒、新冠病毒疫苗及其应用 |
| WO2023143366A1 (zh) * | 2022-01-30 | 2023-08-03 | 上海玮美基因科技有限责任公司 | 变异型腺相关病毒及其在疾病治疗中的应用 |
| CN116925192A (zh) * | 2022-04-12 | 2023-10-24 | 合肥星眸生物科技有限公司 | 一种融合型腺相关病毒及其应用 |
| CN117065035A (zh) * | 2023-10-12 | 2023-11-17 | 首都医科大学附属北京友谊医院 | 联合调控细胞周期和Atoh1表达在治疗前庭功能障碍中的应用 |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113874512A (zh) * | 2018-11-07 | 2021-12-31 | 阿库斯股份有限公司 | 诱导毛细胞分化的组合物和方法 |
| AU2021225035A1 (en) | 2020-02-21 | 2022-10-13 | Akouos, Inc. | Compositions and methods for treating non-age-associated hearing impairment in a human subject |
| WO2024186275A1 (en) * | 2023-03-09 | 2024-09-12 | Agency For Science, Technology And Research | Novel compositions of adeno-associated viruses engineered for enhanced tissue transduction and specificity |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6156303A (en) * | 1997-06-11 | 2000-12-05 | University Of Washington | Adeno-associated virus (AAV) isolates and AAV vectors derived therefrom |
| CN101124328A (zh) * | 2004-12-15 | 2008-02-13 | 北卡罗来纳查佩尔山大学 | 嵌合载体 |
| US20100129405A1 (en) * | 2005-04-29 | 2010-05-27 | Michael Schmidt | Isolation, cloning and characterization of new adeno-associated virus (aav) serotypes |
| US20120253018A1 (en) * | 2003-12-04 | 2012-10-04 | Human Services | Bovine Adeno-Associated Viral (BAAV) Vector and Uses Thereof |
| WO2013102492A1 (en) * | 2012-01-04 | 2013-07-11 | Instytut Biochemii I Biofizyki Pan | Synthetic genes encoding peptide fragments of natural myelin proteins for induction of oral tolerance, dna fragment comprising these genes, means of obtaining these peptides in a microbial (bacterial) system and their medical application |
| CN105473723A (zh) * | 2012-05-18 | 2016-04-06 | 宾夕法尼亚大学托管会 | 亚家族e猿腺病毒a1302、a1320、a1331和a1337及其用途 |
| CN107012171A (zh) * | 2011-04-22 | 2017-08-04 | 加利福尼亚大学董事会 | 具有变异衣壳的腺相关病毒病毒体及其使用方法 |
| CN108603174A (zh) * | 2015-12-01 | 2018-09-28 | 星火治疗有限公司 | 在适用于临床应用的无血清悬浮细胞培养体系中产生重组腺相关病毒(aav)载体的可扩展方法 |
Family Cites Families (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7588772B2 (en) * | 2006-03-30 | 2009-09-15 | Board Of Trustees Of The Leland Stamford Junior University | AAV capsid library and AAV capsid proteins |
| EP3561062A1 (en) * | 2013-09-13 | 2019-10-30 | California Institute of Technology | Selective recovery |
| WO2017100671A1 (en) * | 2015-12-11 | 2017-06-15 | California Institute Of Technology | TARGETING PEPTIDES FOR DIRECTING ADENO-ASSOCIATED VIRUSES (AAVs) |
| WO2018204803A1 (en) * | 2017-05-05 | 2018-11-08 | Voyager Therapeutics, Inc. | Compositions and methods of treating huntington's disease |
| JOP20190269A1 (ar) * | 2017-06-15 | 2019-11-20 | Voyager Therapeutics Inc | بولي نوكليوتيدات aadc لعلاج مرض باركنسون |
| RU2770922C2 (ru) * | 2017-09-20 | 2022-04-25 | 4Д Молекьюлар Терапьютикс Инк. | Капсиды вариантов аденоассоциированных вирусов и методы их применения |
| US11905533B2 (en) * | 2017-12-19 | 2024-02-20 | The University Of North Carolina At Chapel Hill | Methods and compositions for delivery of viral vectors across the blood-brain barrier |
| SG11202006056RA (en) * | 2018-01-17 | 2020-07-29 | Meiragtx Uk Ii Ltd | A modified raav capsid protein for gene therapy |
| WO2020072683A1 (en) * | 2018-10-02 | 2020-04-09 | Voyager Therapeutics, Inc. | Redirection of tropism of aav capsids |
-
2019
- 2019-08-29 CN CN201910807643.XA patent/CN110437317B/zh active Active
- 2019-09-24 US US17/284,673 patent/US20210355171A1/en active Pending
- 2019-09-24 CA CA3140506A patent/CA3140506A1/en active Pending
- 2019-09-24 WO PCT/CN2019/107474 patent/WO2020155655A1/zh unknown
- 2019-09-24 EP EP19912419.9A patent/EP3943503A4/en not_active Withdrawn
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6156303A (en) * | 1997-06-11 | 2000-12-05 | University Of Washington | Adeno-associated virus (AAV) isolates and AAV vectors derived therefrom |
| US20120253018A1 (en) * | 2003-12-04 | 2012-10-04 | Human Services | Bovine Adeno-Associated Viral (BAAV) Vector and Uses Thereof |
| CN101124328A (zh) * | 2004-12-15 | 2008-02-13 | 北卡罗来纳查佩尔山大学 | 嵌合载体 |
| US20100129405A1 (en) * | 2005-04-29 | 2010-05-27 | Michael Schmidt | Isolation, cloning and characterization of new adeno-associated virus (aav) serotypes |
| US20140024013A1 (en) * | 2005-04-29 | 2014-01-23 | Human Services | Isolation, Cloning and Characterization of New Adeno-Associated Virus (AAV) Serotypes |
| CN107012171A (zh) * | 2011-04-22 | 2017-08-04 | 加利福尼亚大学董事会 | 具有变异衣壳的腺相关病毒病毒体及其使用方法 |
| WO2013102492A1 (en) * | 2012-01-04 | 2013-07-11 | Instytut Biochemii I Biofizyki Pan | Synthetic genes encoding peptide fragments of natural myelin proteins for induction of oral tolerance, dna fragment comprising these genes, means of obtaining these peptides in a microbial (bacterial) system and their medical application |
| CN105473723A (zh) * | 2012-05-18 | 2016-04-06 | 宾夕法尼亚大学托管会 | 亚家族e猿腺病毒a1302、a1320、a1331和a1337及其用途 |
| CN108603174A (zh) * | 2015-12-01 | 2018-09-28 | 星火治疗有限公司 | 在适用于临床应用的无血清悬浮细胞培养体系中产生重组腺相关病毒(aav)载体的可扩展方法 |
Non-Patent Citations (4)
| Title |
|---|
| FANGZHI TAN ET AL.: "AAV-ie enables safe and efficient gene transfer to inner ear cells", 《NATURE COMMUNICATIONS》 * |
| KEN Y CHAN ET AL.: "Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems", 《NAT NEUROSCI.》 * |
| LERCH,T.F.ET AL.: "Chain A, Adeno-associated Virus Dj，PDB: 3J1Q_A", 《GENBANK》 * |
| 邓怡晨等: "AAV载体靶向修饰策略在基因治疗中的研究进展", 《中国比较医学杂志》 * |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110950934B (zh) * | 2019-12-31 | 2022-11-04 | 复旦大学 | 一种腺相关病毒衣壳蛋白、载体及其构建方法与应用 |
| CN110950934A (zh) * | 2019-12-31 | 2020-04-03 | 复旦大学 | 一种腺相关病毒衣壳蛋白、载体及其构建方法与应用 |
| CN112011571A (zh) * | 2020-04-26 | 2020-12-01 | 辉大(上海)生物科技有限公司 | 一种用于治疗脊髓性肌萎缩的基因治疗药物 |
| CN111825772B (zh) * | 2020-07-30 | 2023-10-20 | 中国科学院精密测量科学与技术创新研究院 | 具有变异衣壳蛋白的腺相关病毒及其应用 |
| CN111825772A (zh) * | 2020-07-30 | 2020-10-27 | 中国科学院精密测量科学与技术创新研究院 | 具有变异衣壳蛋白的腺相关病毒及其应用 |
| CN112813037A (zh) * | 2021-01-08 | 2021-05-18 | 中国科学院动物研究所 | 一种高效感染原代小胶质细胞的重组突变腺相关病毒及其相关生物材料 |
| CN112813037B (zh) * | 2021-01-08 | 2022-05-10 | 中国科学院动物研究所 | 一种高效感染原代小胶质细胞的重组突变腺相关病毒及其相关生物材料 |
| CN115340998A (zh) * | 2021-05-14 | 2022-11-15 | 上海科技大学 | 一种重组病毒载体、包含其的重组病毒、新冠病毒疫苗及其应用 |
| CN115340998B (zh) * | 2021-05-14 | 2023-12-01 | 上海科技大学 | 一种重组病毒载体、包含其的重组病毒、新冠病毒疫苗及其应用 |
| WO2023143366A1 (zh) * | 2022-01-30 | 2023-08-03 | 上海玮美基因科技有限责任公司 | 变异型腺相关病毒及其在疾病治疗中的应用 |
| CN116925192A (zh) * | 2022-04-12 | 2023-10-24 | 合肥星眸生物科技有限公司 | 一种融合型腺相关病毒及其应用 |
| CN117065035A (zh) * | 2023-10-12 | 2023-11-17 | 首都医科大学附属北京友谊医院 | 联合调控细胞周期和Atoh1表达在治疗前庭功能障碍中的应用 |
| CN117065035B (zh) * | 2023-10-12 | 2024-02-06 | 首都医科大学附属北京友谊医院 | 联合调控细胞周期和Atoh1表达在治疗前庭功能障碍中的应用 |
Also Published As
| Publication number | Publication date |
|---|---|
| EP3943503A4 (en) | 2023-10-18 |
| WO2020155655A1 (zh) | 2020-08-06 |
| US20210355171A1 (en) | 2021-11-18 |
| CA3140506A1 (en) | 2020-08-06 |
| CN110437317B (zh) | 2023-05-02 |
| EP3943503A1 (en) | 2022-01-26 |
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