CN110719954B - 用于iii型糖原贮积病的治疗剂 - Google Patents

用于iii型糖原贮积病的治疗剂 Download PDF

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CN110719954B
CN110719954B CN201880036551.XA CN201880036551A CN110719954B CN 110719954 B CN110719954 B CN 110719954B CN 201880036551 A CN201880036551 A CN 201880036551A CN 110719954 B CN110719954 B CN 110719954B
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K·井川清嶽
C·G·佩雷斯-加西亚
P·奇瓦库拉
H·巴斯卡兰
C·W·科博
S·C·多尔蒂
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Abstract

本发明提供了一系列用于表达人淀粉‑α‑1,6‑葡萄糖苷酶,4‑α‑葡聚糖转移酶(amylo‑alpha‑1,6‑glucosidase,4‑alpha‑glucanotransferase)(AGL)或其具有AGL活性的片段的可翻译多核苷酸和低聚物分子。所述多核苷酸和低聚物分子可表达为提供所述人AGL或其具有AGL活性的片段。所述分子可以用作用于在细胞或受试者中表达活性多肽或蛋白的活性剂。所述药剂可以在用于改善、预防、延缓发作或治疗与受试者的淀粉‑α‑1,6‑葡萄糖苷酶,4‑α‑葡聚糖转移酶(AGL)活性降低相关联的疾病或病状的方法中使用。

Description

用于III型糖原贮积病的治疗剂
相关申请的交叉引用
本申请要求于2017年5月31日提交的美国临时申请序号62/513,350的优先权，所述美国临时申请出于所有目的以全文引用的方式并入本文。
技术领域
本发明涉及分子生物学和遗传学领域以及由可翻译分子产生的生物药物和治疗剂。更具体地说，本发明涉及用于具有翻译成活性多肽或蛋白的能力以在体内并且作为治疗剂使用的分子的方法、结构和组合物。
对以电子方式提交的文本文件的描述
与本申请一起以电子方式提交的文本文件的内容以全文引用的方式并入本文：序列表的计算机可读格式副本(文件名称：ULPI_041_01WO_SeqList_ST25.txt，记录日期：2018年5月30日，文件大小：226千字节)。
背景技术
III型糖原贮积病(也称为GSD III或柯里病(Cori disease))是一种由缺乏糖原脱支酶引起的罕见的(发病率1:100,000)先天性糖原代谢缺陷，所述糖原脱支酶被称为淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(amylo-alpha-1,6-glucosidase,4-alpha-glucanotransferase)(AGL)。这种常染色体隐性代谢紊乱表征为可变的肝脏、心肌和骨骼肌受累。
基于有缺陷的酶AGL在组织表达上的差异，存在四种GSD III亚型。GSD IIIa占所有GSD III的大约85％并且表现为由于肝脏和肌肉两者中的酶缺乏引起的肝脏和肌肉受累。GSD IIIb占大约15％并且通常表现为由于仅肝脏中的酶缺乏引起的仅肝脏受累。同时，GSD IIIc和GSD IIId均极为罕见，其中认为GSD IIIc是由于缺乏葡萄糖苷酶脱支活性引起的并且认为GSD IIId是由于缺乏转移酶脱支活性引起的。
在婴儿期和儿童早期，肝脏受累表现为酮性低血糖症、肝肿大、高脂血症和肝转氨酶升高。在青少年期和成人期，肝脏疾病变得不那么突出。大多数患有GSD IIIa的患者通常在儿童时期期间出现肥厚型心肌病。其临床显著性的范围为大多数的无症状到严重的心功能障碍、充血性心力衰竭和罕见的猝死。表现为无力的骨骼肌病通常在儿童时期并不明显，但缓慢进展，从而通常在成人中变得突出。
与受影响的器官一致，GSD III患者的血清中经常存在增强的丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、碱性磷酸酶(ALP)和/或肌酸磷酸激酶(CPK)活性。
如上所述，GSD III是由缺乏AGL引起的。该缺乏通常归因于AGL基因的一种或多种遗传突变，所述一种或多种遗传突变导致患有GSD III的受试者的AGL酶活性部分或全部消除。在几个种族人群中执行了对GSD III患者的AGL蛋白的分子分析，并且描述了100多种不同的AGL突变。参见Goldstein等人,2010,《医学遗传学(Genet.Med.)》12:424-430。还参见Sentner等人，2013,《遗传性代谢病杂志报道(JIMD Rep.)》7:19-26。
目前还没有对GSD III的有效治疗。已经尝试通过频繁摄入碳水化合物含量高的餐食来控制低血糖症，通常通过使用夜间胃滴注喂食或玉米淀粉补充。同时，通过白天期间的高蛋白饮食加上整夜肠内输注对患有肌病的患者进行治疗。已有少数患者的症状得到暂时改善的记载，但尚无长期数据表明高蛋白饮食可预防或治疗进行性肌病。参见Chen YT、Burchell A,《糖原贮积病(Glycogen storage disease)》于Scriver CR、Beaudet AL、SlyWS、Valle D,《遗传性疾病的代谢和分子基础(The metabolic and molecular basis ofinherited disease)》纽约：麦格劳-希尔公司(McGraw Hill),1995:935-65。进行性肌病和/或心肌病是成人发病的主要原因，并且已经报道了有患者表现为进行性肝硬化和肝癌。因此，迫切需要可以解决该疾病的根本原因(即缺乏AGL酶活性)的疗法。
迄今为止，在细胞溶质中存在有缺陷的酶的疾病中尚未发现酶置换，如GSD III中的AGL，这可能是由于缺乏跨质膜将外源酶递送到细胞质的有效且特异的细胞摄取机制所致。
本发明通过提供具有在细胞质中进行翻译以提供活性AGL的能力的分子、结构和组合物来解决上述需求，所述活性AGL可以改善、预防或治疗与AGL缺乏相关联的疾病或病状，如GSD III。
发明内容
本发明提供了包括具有翻译能力的新型分子的组合物，所述分子可以用于提供一种或多种活性多肽和蛋白或其片段。本发明进一步提供了使用包括新型分子的这些组合物来预防或治疗各种病症的方法。更具体地说，本发明的实施例提供了包括用于提供活性淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)的可翻译分子的组合物和使用所述组合物来治疗GSD III的方法。
本发明的可翻译分子可以具有功能性细胞质活性，以用于产生AGL多肽或蛋白。所述多肽和蛋白对于治疗方式可以具有活性。
本发明的可翻译分子尤其在细胞的细胞质中可以具有长半衰期。可翻译分子可以可表达为提供对于改善、预防或治疗与AGL缺陷相关联的疾病或病状具有活性的产物。
本公开针对可翻译分子提供了一系列结构，以用于产生AGL多肽或蛋白。在一些实施例中，可翻译分子可以具有与原生mRNA相比增加的翻译能力和/或延长的半衰期。
本发明的可翻译分子可以在医药中使用并且用于用于生产和递送活性多肽和蛋白的方法和组合物。本发明的可翻译分子可以用于在体外、离体地或在体内提供多肽或蛋白质。
本公开的实施例提供了一系列用于表达人淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)或其具有AGL活性的片段的新型多核苷酸。所述多核苷酸可以包含天然核苷酸和经过化学修饰的核苷酸。所述多核苷酸可以可表达为提供人AGL或其具有AGL活性的片段。
在另外方面，本发明提供了用于表达人淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)或其具有AGL活性的片段的包括一个或多个解锁核酸(UNA)单体的一系列新型可翻译低聚物。可翻译低聚物可以含有一个或多个UNA单体以及天然核苷酸和经过化学修饰的核苷酸。包括一个或多个UNA单体的可翻译低聚物可以可表达为提供人淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)或其具有AGL活性的片段。
在某些方面，本发明的可翻译分子可以提供对多肽或蛋白质或其片段的高效表达。所述表达可以在体外、离体地或在体内。
在一些实施例中，与对同一多肽或蛋白质进行编码的原生成熟的mRNA相比，本发明的分子可以具有增加的细胞质半衰期。相对于原生成熟的mRNA，本发明的分子和组合物可以提供增加的功能性细胞活性。
在另外方面，与原生成熟的mRNA相比，本发明的可翻译分子作为提供多肽或蛋白产物的药剂可以提供增加的活性。本发明的可翻译分子可以减少有效疗法所需的剂量水平。
本发明的实施例包含以下内容。
附图说明
图1示出了使用本发明的可翻译分子在体外表达人淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL，NM_000028)的结果。图1示出了在相对于参考分子534归一化的AML12和C2C12细胞中的相对AGL表达。纵轴反映了相对于参考的倍数增加，例如，10是与参考相比增加了10倍。包含参考分子在内的分子包括烟草蚀纹病毒(TEV)5'UTR和非洲爪蟾(Xenopus)β-珠蛋白(XBG)3'UTR。分子在转录期间被封端并且用Nl-甲基假尿苷合成，使得100％的尿苷被Nl-甲基假尿苷置换。在两个细胞系(AML12、C2C12)中转染对AGL进行编码的可翻译分子。转染后6小时裂解并收获细胞。通过使用对AGL具有特异性的抗体(ab133720，兔)，执行定量蛋白印迹法来检测AGL。
图2示出了使用本发明的可翻译分子在体外表达人淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL，NM_000028)的结果。图2示出了在相对于参考分子534归一化的AML12和C2C12细胞中的相对AGL表达。纵轴反映了相对于参考的倍数增加，例如，10是与参考相比增加了10倍。包含参考分子在内的分子包括烟草蚀纹病毒(TEV)5'UTR和非洲爪蟾β-珠蛋白(XBG)3'UTR。分子在转录期间被封端并且用Nl-甲基假尿苷合成，使得100％的尿苷被Nl-甲基假尿苷置换。在两个细胞系(AML12、C2C12)中转染对AGL进行编码的可翻译分子。转染后24小时裂解并收获细胞。通过使用对AGL具有特异性的抗体(ab133720，兔)，执行定量蛋白印迹法来检测AGL。
图3示出了使用本发明的可翻译分子在体内表达人淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL，NM_000028)的结果。图3示出了在24小时和48小时的时间点处针对可翻译分子528和534(参考)在WT小鼠体内的相对AGL表达。分子包括烟草蚀纹病毒(TEV)5'UTR和非洲爪蟾β-珠蛋白(XBG)3'UTR。分子在转录期间被封端并且用Nl-甲基假尿苷合成，使得100％的尿苷被Nl-甲基假尿苷置换。在脂质纳米颗粒调配物中各自制备对AGL进行编码的可翻译分子，并且以10mg/kg将其静脉注射到WT小鼠体内。在24小时和48小时时收获小鼠肝脏，并且通过使用对AGL具有特异性的抗体(ab133720，兔)，执行定量蛋白印迹法来检测AGL。
图4示出了使用本发明的可翻译分子在体内表达人淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL，NM_000028)的结果。图4示出了与基线PBS(100)相比，在经过可翻译分子525、527、528、529和546给药后的WT小鼠体内的相对肝脏AGL表达。在脂质纳米颗粒调配物中制备对AGL进行编码的经过合成的可翻译分子525、527、528、529和546，并将其IP注射到WT小鼠体内。注射的剂量为10mpk，并且在6小时时收集肝脏，以供进一步分析。通过使用对AGL具有特异性的抗体(ab133720，兔)，执行定量蛋白印迹法来检测AGL。
图5示出了从以下三个可翻译分子中表达人AGL的结果：546、1783和1784。用密码子优化的mRNA来转染人原代肝细胞，并且在转染后6小时、24小时、48小时和72小时时通过In-Cell WesternTM测量AGL蛋白表达。将mRNA序列的表达与未处理对照物(“unt”)进行比较。
图6示出了在野生型C57BL/6小鼠中人AGL从用脂质纳米颗粒调配的各种mRNA分子的表达。通过多重反应监测测定来确定在肝脏活检样品的匀浆中从mRNA分子表达的外源人AGL的蛋白浓度(ng/mg)。在曲线图中示出为546.1、736.1、738.1、737.1、731.1和1783.1的可翻译分子分别与实例2中描述的546、736、738、737、731和1783相同。同时，可翻译分子546.7的核碱基序列与实例2中描述的546的核碱基序列相同，但用5-甲氧基尿苷代替尿苷而非用于合成可翻译分子546的N1-甲基假尿苷合成。
图7示出了在用脂质纳米颗粒调配的各种mRNA分子治疗的野生型C57BL/6小鼠中的内源性小鼠AGL表达。通过多重反应监测测定来确定表达在肝脏活检样品的匀浆中的内源性小鼠AGL的蛋白浓度(ng/mg)。在曲线图中示出为546.1、736.1、738.1、737.1、731.1和1783.1的可翻译分子分别与实例2中描述的546、736、738、737、731和1783相同。同时，可翻译分子546.7的核碱基序列与实例2中描述的546的核碱基序列相同，但用5-甲氧基尿苷代替尿苷而非用于合成可翻译分子546的N1-甲基假尿苷合成。
图8示出了来自用媒剂(“VEH”)治疗的AGL敲除小鼠的肝脏的组织病理学。在用媒剂治疗的肝细胞内观察到显著到严重肝细胞空泡化和中度到显著糖原累积增加。
图9示出了来自用ATX2脂质纳米颗粒调配的可翻译分子546治疗的AGL敲除小鼠的肝脏的组织病理学。在用ATX2脂质纳米颗粒调配的可翻译分子546治疗的肝细胞内，观察到在肝细胞有仅轻度到中度肝细胞空泡化和仅轻度到中度糖原累积增加。
具体实施方式
本发明提供了一系列用于治疗性应用的新型药剂和组合物。本发明的分子和组合物可以用于改善、预防和治疗GSD III和/或与受试者的淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)的存在或功能的减少相关联的疾病。
在一些实施例中，本发明包含用于表达人淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶的合成的经过纯化的可翻译多核苷酸分子。所述分子可以含有天然核苷酸和经过化学修饰的核苷酸并且对人淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)或其具有AGL活性的片段进行编码。
在某些实施例中，本公开包含用于表达人淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)或其具有AGL活性的片段的包括一个或多个UNA单体的合成的经过纯化的可翻译低聚物分子。可翻译低聚物可以含有一个或多个UNA单体以及天然核苷酸和经过化学修饰的核苷酸。包括一个或多个UNA单体的可翻译低聚物可以可表达为提供人淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)或其具有AGL活性的片段。
如本文所使用的，术语“可翻译”可以与术语“可表达”可互换地使用并且是指多核苷酸或其一部分能够被宿主细胞转化为多肽。如本领域所理解的，翻译是细胞的细胞质中的核糖体产生多肽的过程。在翻译中，由核糖体复合物中的tRNA对信使RNA(mRNA)进行解码以产生特异性氨基酸链或多肽。此外，当在本说明书中关于低聚物使用时，术语“可翻译”意味着低聚物的至少一部分，例如低聚物序列的编码区(也称为编码序列或CDS)能够转化为蛋白质或其片段。
如本文所使用的，术语“单体”是指可以与相同或不同类型的另一种分子连接以形成低聚物的单个单元，例如单个核酸。在一些实施例中，单体可以是解锁核酸，即UNA单体。
同时，术语“低聚物”可以与“多核苷酸”可互换地使用并且是指包括至少两个单体的分子并且包含如DNA和RNA等寡核苷酸。在含有RNA单体和/或解锁核酸(UNA)单体的低聚物的情况下，本发明的低聚物除了编码序列(CDS)之外可以另外含有序列。这些另外的序列可以是非翻译序列，即未被宿主细胞转化为蛋白质的序列。这些非翻译序列可以包含5'端帽、5'非翻译区(5'UTR)、3'非翻译区(3'UTR)和尾区，例如polyA尾区。如本文进一步详细描述的，这些非翻译序列中的任何非翻译序列均可以含有一个或多个UNA单体—这些UNA单体无法被宿主细胞的机器翻译。在本发明的上下文中，“可翻译低聚物”、“可翻译分子”、“可翻译多核苷酸”或“可翻译化合物”是指包括能够转换成蛋白质或其片段(例如人AGL蛋白质或其片段)的区域(例如RNA的编码区)的序列(例如人AGL的编码序列或其密码子优化版本)。
如本文所使用的，术语“密码子优化”意指天然编码序列(或天然编码序列的有目的地设计的变体)通过在不改变经过编码的蛋白质氨基酸序列的情况下选择不同的密码子而被重新设计从而提高蛋白质表达水平(Gustafsson等人,《密码子偏倚和异源蛋白表达(Codon bias and heterologous protein expression)》2004,《生物科技趋势(TrendsBiotechnol)》,22:346-53)。已经显示，如高密码子适应指数(CAI)、LowU法、mRNA二级结构、顺式调控序列、GC含量等变量以及许多其它类似的变量在一定程度上与蛋白表达水平相关(Villalobos等人,《基因设计器：用于构建人工DNA区段的合成生物学工具(GeneDesigner:a synthetic biology tool for constructing artificial DNA segments)》2006,《BMC生物信息学(BMC Bioinformatics)》7:285)。高CAI(密码子适应指数)法为整个蛋白质编码序列挑选最常用的同义密码子。从人类基因组的74218个蛋白质编码基因中推导出每个氨基酸的最常用密码子。LowU法针对可以用具有较少U部分的同义密码子置换的仅含U密码子。如果存在几种替换选择，则将选择更常用的密码子。LowU法不会改变序列中其余的密码子。该方法可以与公开的mRNA结合使用，以设计用5-甲氧基尿苷合成的编码序列。
如配备有本公开的技术人员将了解的，本发明的可翻译分子可以用于改善、预防或治疗与受试者的淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)的活性降低(例如由于浓度、存在和/或功能的减少引起的)相关联的任何疾病或病症。在一些实施例中，本发明的可翻译分子可以在用于改善、预防或治疗GSD IIIa、GSD IIIb、GSD IIIc和GSD IIId(在本文中统一地或单独地称为“GSD III”或“III型糖原贮积病”)中的一种或多种的方法中使用。本文要治疗的疾病或病症(例如GSD IIIa、GSD IIIb、GSD IIIc和GSD IIId)可以与以下相关联：低血糖(低血糖症)、肝脏增大(肝肿大)、血液中脂肪过量(高脂血症)、肝酶血液水平升高、慢性肝病(肝硬化)、肝衰竭、生长缓慢、身材矮小、良性肿瘤(腺瘤)、肥厚型心肌病、心功能障碍、充血性心力衰竭、骨骼肌病和/或肌张力低下(张力减退)。在一些实施例中，本发明的可翻译分子可以用于改善、预防或治疗这些上述症状中的任何或全部症状。
如技术人员所理解的，GSD III在本领域可以由许多替代名称来指代，包含但不限于AGL缺乏症、柯里病、柯里氏病(Cori's disease)、脱支酶缺乏症、福布斯病(Forbesdisease)、糖原脱支酶缺乏症、GSD3或极限糊精糖增多症(由于细胞溶质中的极限糊精样结构)。因此，GSD III可以与本说明书、实例、附图和权利要求书中的这些替代名称中的任何替代名称可互换地使用。
本发明的对功能性AGL部分进行编码的可翻译分子可以递送到有需要的患者(例如GSD III患者)的肝脏，尤其是肝细胞，并且可以提升患者的活性AGL水平。可翻译分子可以用于预防、治疗、减轻或逆转患者的GSD III的任何症状。
在另外方面，本发明的可翻译分子还可以用于减少GSD III患者对特定饮食的依赖，以控制疾病。例如，本发明的可翻译分子可以用于减少GSD III患者对频繁摄入碳水化合物含量高的餐食和/或蛋白质异常高的饮食的依赖。
本发明的实施例进一步涵盖用于制备用于表达人淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)的可翻译分子的方法。所述方法包含：在天然核苷三磷酸和经过化学修饰的核苷三磷酸存在的情况下在体外转录AGL DNA模板以形成产物混合物；以及纯化所述产物混合物以分离出可翻译分子。还可以通过本领域已知的方法来制备可翻译分子。
本发明的分子可以是含有RNA和/或UNA单体的可翻译分子。这些可翻译分子尤其在细胞质中可以具有长半衰期。长持续时间可翻译分子可以用于改善、预防或治疗与受试者的淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)的活性降低(例如由于浓度、存在和/或功能的减少引起的)相关联的疾病或病症。
本发明的可翻译分子的性质是根据其分子结构产生的，并且基于那些性质，整个分子的结构作为整体可以提供显著的益处。本发明的实施例可以提供具有有利地提供提高的蛋白浓度或增加的蛋白活性的一种或多种性质的可翻译分子。本发明的分子和组合物可以提供包括用于改善、预防或治疗与受试者的淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)的活性降低(例如由于浓度、存在和/或功能的减少引起的)相关联的任何疾病或病症的治疗剂的调配物。
本发明提供了令人惊讶地可翻译以在体外、离体地以及在体内提供活性多肽或蛋白的一系列可翻译分子。
本发明的可翻译分子可表达为提供一种或多种活性多肽或蛋白或其片段。
可翻译结构和组合物可以具有增加的翻译活性或细胞质半衰期。在这些实施例中，与原生mRNA相比，可翻译结构和组合物可以在哺乳动物细胞的细胞质中提供增加的功能性半衰期。
如本文所使用的，术语“半衰期”是量(如核酸或蛋白浓度或活性)下降到其在时间段开始时测量的值的一半所需的时间。
本文提供了本发明的可翻译分子的一系列结构，所述一系列结构包含含有一个或多个UNA单体的低聚物。含有一个或多个UNA单体的低聚物可以合并专门的连接子基团。连接子基团可以在可翻译分子中的链中附接。还可以将每个连接子基团附接到核碱基。
在一些方面，连接子基团可以是单体。单体可以附接以形成链分子。在本发明的链分子中，连接子基团单体可以附接在链中的任何点处。
在某些方面，连接子基团单体可以在本发明的链分子中附接，使得连接子基团单体驻留在链的末端附近或链中的任何位置处。
在另外方面，链分子的连接子基团可以各自附接到核碱基。链分子中存在核碱基可以在链分子中提供核碱基序列。
在某些实施例中，本发明提供了具有合并连接子基团单体与某些天然核苷酸或非天然核苷酸或经过修饰的核苷酸或经过化学修饰的核苷酸的新型组合的链结构的可翻译低聚物分子。
本发明的低聚物分子可以展示核碱基序列并且可以被设计成在体外、离体地或在体内表达多肽或蛋白质。表达的多肽或蛋白质可以具有各种形式的活性，包含与从天然mRNA、原生mRNA或野生型mRNA表达的蛋白质相对应的活性或与阴性蛋白质或显性阴性蛋白质相对应的活性。
在一些方面，本发明可以提供具有与细胞的原生核酸分子的至少片段相同的碱基序列的活性可翻译低聚物分子。
在一些实施例中，细胞可以是真核细胞、哺乳动物细胞或人细胞。
本发明提供了用于合并连接子基团单体的可翻译低聚剂的结构、方法和组合物。本发明的低聚分子可以在用于治疗的调配物中用作活性剂。
本发明提供了由于其能够表达为受试者的细胞中的多肽或蛋白质而因此可用于提供治疗效果的一系列可翻译分子。
在某些实施例中，可翻译分子可以被构造为由单体构成的低聚物。本发明的低聚结构可以含有一个或多个连接子基团单体连同某些核苷酸。
在某些实施例中，可翻译分子可以含有核碱基序列并且可以被设计成部分地通过与原生多核苷酸序列具有足够的同源性来表达任何同种型的肽或蛋白质。
在一些实施例中，可翻译分子的长度可以为约200到约12,000个单体或更多。在某些实施例中，可翻译分子可以为1,000到9,000个单体长、3,000到7,000个单体长、或4,000到6,000个单体长。在一个示例性实施例中，可翻译分子的长度为4,500到5,500个单体。在另外的示例性实施例中，可翻译分子的长度为约5,000个单体。
在一些实施例中，可翻译分子可以含有1到约800个UNA单体。在某些实施例中，可翻译分子可以含有1到600个UNA单体、或1到100个UNA单体、或1到12个UNA单体。
在一些实施例中，可翻译分子可以含有1到约800个锁核酸(LNA)单体。在某些实施例中，可翻译分子可以含有1到600个LNA单体、或1到100个LNA单体、或1到12个LNA单体。
本发明的可翻译分子可以包括5'端帽、单体的5'非翻译区、单体的编码区、单体的3'非翻译区和单体的尾区。
本发明的可翻译分子可以包括含有一个或多个UNA单体的单体的3'非翻译区。
本发明的可翻译分子可以包括含有一个或多个UNA单体的单体的尾区。
本发明的可翻译分子可以包括序列或结构的区域，所述区域可操作以用于在细胞中翻译或者具有mRNA的包含例如5'端帽、5'非翻译区、编码区、3'非翻译区和polyA尾在内的区域的功能。
本发明进一步设想了用于向细胞递送包括一个或多个可翻译分子的一个或多个载体的方法。在另外的实施例中，本发明还设想了向细胞递送一个或多个可翻译分子。
在一些实施例中，可以将一个或多个可翻译分子在体外、离体地或在体内递送到细胞。可以使用如本领域已知的病毒和非病毒转移方法来将可翻译分子引入哺乳动物细胞中。可翻译分子可以与药学上可接受媒剂或者例如与纳米颗粒或脂质体一起递送。
在一些实施例中，与利用原生组合物相比，本发明的可翻译结构和组合物可以减少在培养物中进行细胞命运操纵所需的转染的数量和频率。
在另外方面，与利用原生mRNA相比，本发明针对作为活性剂的可翻译分子提供了增加的活性。
在一些方面，本发明可以提供与有原生核酸、多肽或蛋白质诱导的分子相比可以减少细胞的固有免疫应答的可翻译分子。
本发明可以提供与原生分子相比难以脱腺苷酸化的合成可翻译分子。
在某些实施例中，本发明可以提供具有与原生分子相比增加的比活性和更长的功能半衰期的合成可翻译分子。本发明的合成可翻译分子可以提供提高的异位蛋白表达水平。当使用载体表达可翻译分子时，细胞递送可以处于提高的水平，并且细胞毒素的固有免疫应答可以是有限的，从而使得可以实现更高的异位蛋白表达水平。与原生mRNA相比，本发明的可翻译分子可以具有增加的比活性和更长的功能半衰期。
在某些方面，可翻译分子可以具有许多相对于原生mRNA的突变。
在另外的实施例中，本发明可以提供具有可裂解递送并且靶向附接在3'端和/或5'端的部分的可翻译分子。
通常，可以将通过转染递送的合成可翻译分子的比活性视为每单位时间内每个递送的转录物表达的蛋白质的分子的数量。
如本文所使用的，翻译效率是指通过在体外或在体内翻译可翻译分子来产生蛋白质或多肽的量度。
本发明提供了可以含有一个或多个UNA单体和许多核酸单体的一系列可翻译低聚物分子，其中所述可翻译分子可以可表达为提供多肽或蛋白质。
在一些实施例中，本发明包含可以在一个或多个非翻译区中含有一个或多个UNA单体以及许多核酸单体的一系列可翻译低聚物分子，其中所述可翻译分子可以可表达为提供多肽或蛋白质。
在一些实施例中，本发明包含在尾区中含有一个或多个UNA单体以及许多核酸单体的一系列可翻译分子，其中所述可翻译分子可以可表达为提供多肽或蛋白质。
在一些实施例中，可翻译分子可以含有经过修饰的5'端帽。
在另外的实施例中，可翻译分子可以含有单体的翻译增强性5'非翻译区。
在另外的实施例中，可翻译分子可以含有单体的翻译增强性3'非翻译区。
在另外的实施例中，可翻译分子在单体的3'非翻译区中可以含有一个或多个UNA单体。
在另外的实施例中，可翻译分子在单体的尾区中可以含有一个或多个UNA单体。
在另外的实施例中，可翻译分子在polyA尾中可以含有一个或多个UNA单体。
在一些实施例中，可翻译分子在单体的3'非翻译区中或在单体的尾区中(例如在polyA尾中)可以含有一个或多个LNA单体。
另一方面，与对同一翻译产物进行编码的原生mRNA相比，本发明的可翻译分子在体内可以表现出提高至少2倍、3倍、5倍或10倍的翻译效率。
在另外方面，与对同一多肽或蛋白质进行编码的原生mRNA相比，可翻译分子在体内可以产生提高至少2倍、3倍、5倍或10倍的多肽或蛋白质水平。
在某些实施例中，与对同一多肽或蛋白质进行编码的原生mRNA相比，可翻译分子在体内可以提供提高的多肽或蛋白质水平。例如，多肽或蛋白质的水平可以提高10％、或20％、或30％、或40％、或50％或更多。
在另外的实施例中，本发明提供了用于通过向受试者施用含有本发明的可翻译分子的组合物来治疗受试者的疾病或病状的方法。
本发明的可翻译分子可以用于改善、预防或治疗疾病或病症，例如与受试者的淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)的活性降低(例如由于浓度、存在和/或功能的减少引起的)相关联的疾病或病症。在这些实施例中，可以施用包括本发明的可翻译分子的组合物，以调控、调节或增加AGL酶在受试者体内的浓度或有效性。在一些方面，所述酶可以是未经过修饰的天然酶，患者具有异常量的未经过修饰的天然酶(例如患者具有部分地或完全地消除AGL活性的AGL突变版本)。在一些方面，所述酶可以是可以用于治疗携带AGL突变版本的患者的未经过修饰的天然AGL酶。在示例性实施例中，本发明的可翻译分子可以用于改善、预防或治疗GSD III。
在一些实施例中，可翻译分子可以递送到细胞或受试者并且被翻译以提高细胞或受试者中的AGL水平。
如本文所使用的，术语“受试者”是指人类或任何非人类动物(例如小鼠、大鼠、兔、狗、猫、牛、猪、绵羊、马或灵长类动物)。人类包含出生前形式和出生后形式。在许多实施例中，受试者是人类。受试者可以是患者，所述患者是指呈现给医疗提供者以进行疾病的诊断或治疗的人类。术语“受试者”在本文中与“个体”或“患者”可互换地使用。受试者可以患有或易于患上疾病或病症，但是可以或可以不展示出疾病或病症的症状。
在示例性实施例中，本发明的受试者是淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)的活性降低(例如由于浓度、存在和/或功能的减少引起的)的受试者。在另外的示例性实施例中，受试者是人类。
在一些实施例中，施用包括本发明的可翻译分子的组合物可以使经过治疗的受试者的肝脏AGL蛋白水平提高。在一些实施例中，施用包括本发明的可翻译分子的组合物使肝脏AGL蛋白水平相对于在治疗之前受试者的基线AGL蛋白水平提高5％、10％、20％、30％、40％、50％、60％、70％、80％、90％或95％。在一个示例性实施例中，施用包括本发明的可翻译分子的组合物使肝脏AGL水平相对于治疗之前的受试者体内的基线AGL水平提高。在一些实施例中，肝脏AGL水平可以提高至少约5％、10％、20％、30％、40％、50％、100％、200％或更多。
在一些实施例中，从本发明的可翻译分子表达的AGL蛋白可在肝脏、血清、血浆、肾脏、心脏、肌肉、大脑、脑脊液或淋巴结中检测。在示例性实施例中，AGL蛋白表达在肝细胞，例如经过治疗的受试者的肝细胞中。
在一些实施例中，施用包括本发明的可翻译分子的组合物使天然的未突变的人AGL(即与异常或突变AGL相反的正常或野生型AGL)蛋白表达水平处于或高于经过治疗的受试者的肝脏中总蛋白的约10ng/mg、约20ng/mg、约50ng/mg、约100ng/mg、约150ng/mg、约200ng/mg、约250ng/mg、约300ng/mg、约350ng/mg、约400ng/mg、约450ng/mg、约500ng/mg、约600ng/mg、约700ng/mg、约800ng/mg、约900ng/mg、约1000ng/mg、约1200ng/mg或约1500ng/mg。
如本文使用的，如应用于一个或多个感兴趣的值的术语“约”或“大约”是指与规定的参考值类似的值。在某些实施例中，除非另有规定或另外从上下文显而易见(除了此些数字将会超过可能值的100％的情况)，否则术语“大约”或“约”是指在规定的参考值的在任一方向(大于或小于)的10％、9％、8％、7％、6％、5％、4％、3％、2％、1％或更小的值的范围。
在一些实施例中，可在施用包括本发明的可翻译分子的组合物之后6小时、12小时、18小时、24小时、30小时、36小时、48小时、60小时和/或72小时检测天然的未突变的人AGL蛋白的表达。在一些实施例中，可在施用包括本发明的可翻译分子的组合物之后1天、2天、3天、4天、5天、6天和/或7天检测天然的未突变的人AGL蛋白的表达。在一些实施例中，可在施用后1周、2周、3周和/或4周检测天然的未突变的人AGL蛋白的表达。在一些实施例中，可在施用包括本发明的可翻译分子的组合物之后检测天然的未突变人AGL蛋白的表达。在一些实施例中，可在施用包括本发明的可翻译分子的组合物之后在肝脏例如肝细胞中检测天然的未突变的人AGL蛋白的表达。
用于制备可翻译分子的变体模板
在本文描述的各个实施例中，可翻译低聚物可以包括mRNA编码的AGL，其中所述mRNA编码AGL是密码子优化的。在一些实施例中，AGL是人AGL(即hAGL)。在一些实施例中，人AGL包括氨基酸序列SEQ ID NO:2。在一些实施例中，人AGL由氨基酸序列SEQ ID NO:2组成。
在一些实施例中，可以利用变体DNA模板来制备能够对AGL进行编码的可翻译分子。本公开的变体DNA模板在用于制备可翻译分子的方法和可翻译分子的效率方面可以表现出优势。可以利用改变模板来增强经过修饰的核苷酸或单体在本发明的可翻译分子中的合并。在某些方面，可以利用改变模板来增强可翻译分子的结构特征。可翻译分子的增强的结构特征可以提供意想不到的有利性质，包含用于提供多肽或蛋白产物的翻译效率。
在本发明的一些方面，改变模板可以包含减少某些核苷酸在模板链中的出现或出现频率。减少某种核苷酸的出现可以改变本公开的结构和方法以提供非原生形式，这令人惊奇地实现了改善对AGL进行编码的可翻译RNA产物的性质。
本发明的各方面在用于制备可翻译分子的方法中可能需要变体DNA模板。DNA分子可以具有可以转录以提供对AGL进行编码的靶可翻译分子的核苷酸的非编码模板链。
靶可翻译分子可以是任何RNA，无论是原生的或经过修饰的、合成的或源自天然来源。
在一些实施例中，可以使用变体DNA模板，所述变体DNA模板的模板链的开放阅读框被转化成替代形式，同时维持了密码子分配。
在某些实施例中，可以使用DNA模板，所述DNA模板的替代性核苷酸是基于替代性密码子优化和/或序列简并使用的。
在另外的实施例中，DNA模板可以具有在维持密码子分配的同时用替代性核苷酸置换的某些核苷酸。
本发明的实施例有利地利用了本发明的DNA模板中的替代性密码子以在用于制备对AGL进行编码的可翻译分子的方法中使用。与在许多方法中可能需要优选密码子的细胞和生物体相比，可以在本发明的DNA模板中实现的改变的范围要大得多。在本发明中，可以在DNA模板中使用广泛范围的替代性密码子和位置以用于转录可翻译分子。
在本发明的另外方面，改变模板可以包含减少某些核苷酸在模板链中的出现或出现频率。例如，可以将核苷酸在模板中的出现减少到低于模板中核苷酸的25％的水平。在另外的实例中，可以将核苷酸在模板中的出现减少到低于模板中核苷酸的20％的水平。在一些实例中，可以将核苷酸在模板中的出现减少到低于模板中核苷酸的16％的水平。在某些实例中，可以将核苷酸在模板中的出现减少到低于模板中核苷酸的12％的水平。
人AGL
人AGL基因对分子质量为大约174.8kDa的1532个氨基酸蛋白质进行编码。AGL是在糖原降解时充当1,4-α-D-葡聚糖:1,4-α-D-葡聚糖-4-α-D-糖基转移酶和淀粉-1,6-葡萄糖苷酶的多功能酶。如上所述，正常AGL活性的遗传缺陷引起糖原贮积病III。
共有人AGL编码序列的RNA序列具有SEQ ID NO:1中示出的4,599个核碱基。
共有人AGL编码序列—见于NCBI登录号NP_000019.2—翻译成SEQ ID NO:2。
在一些实施例中，可以制备可翻译分子并将其用于表达人淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(hAGL)，所述可翻译分子与hAGL的原生mRNA相比具有有利地增加的翻译效率。表达hAGL的可翻译分子可以表现出适合于用于改善、预防或治疗疾病的方法的活性。在一些实施例中，可翻译分子可以包括一个或多个UNA单体。
在一些实施例中，可翻译分子可以包含5'端帽、5'UTR、翻译起始序列(例如Kozak序列)、人AGL CDS、3'UTR和/或尾区。在示例性实施例中，可翻译分子可以包含5'端帽(m7GpppGm)、烟草蚀纹病毒(TEV)的5'UTR、Kozak序列、人AGL CDS、非洲爪蟾β-珠蛋白的3'UTR和尾区。在另外的示例性实施例中，人AGL CDS可以包括下文进一步详细描述的SEQ IDNO:7-32或SEQ ID NO:41-45中的密码子优化序列。在本文所描述的这些和其它实施例中的任何实施例中，可翻译分子可以包括一个或多个UNA单体。在本文所描述的这些和其它实施例中的任何实施例中，可翻译分子可以包括一个或多个LNA单体。
与AGL的原生mRNA相比，分子的翻译效率可以提高。具体地说，在48小时后，与AGL的原生mRNA相比，分子的翻译效率可以提高了多于一倍。
在一些实施例中，本发明的适合的mRNA序列包括对人AGL蛋白进行编码的mRNA序列。SEQ ID NO:2中示出了天然存在的人AGL蛋白的序列。
在一些实施例中，适合的mRNA序列可以是对人AGL的同系物或变体进行编码的mRNA序列。如本文所使用的，人AGL蛋白的同系物或变体可以是经过修饰的人AGL蛋白，所述经过修饰的人AGL蛋白与野生型人AGL蛋白或天然存在的人AGL蛋白相比，在保留基本的AGL蛋白活性的同时含有一个或多个氨基酸取代、缺失和/或插入。在一些实施例中，适合于本发明的mRNA对与人AGL蛋白基本上相同的蛋白质进行编码。在一些实施例中，适合于本发明的mRNA对与SEQ ID NO:2的一致性为至少80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更大的氨基酸序列进行编码。在一些实施例中，适合于本发明的mRNA对人AGL蛋白的片段或一部分进行编码。
在一些实施例中，适合于本发明的mRNA对人AGL蛋白的片段或一部分进行编码，其中所述蛋白质的所述片段或所述部分仍维持与野生型蛋白的活性类似的AGL活性。
在一些实施例中，适合于本发明的mRNA包括与SEQ ID NO:7-32或SEQ ID NO:41-45的一致性为至少80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更大的序列。
在一些实施例中，本发明的可翻译低聚分子包括与SEQ ID NO:7-32或SEQ ID NO:41-45的一致性为至少80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更大的编码序列。在一些实施例中，包括与SEQ ID NO:7-32或SEQ ID NO:41-45的一致性为至少80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更大的编码序列的可翻译低聚分子进一步包括选自5'端帽、5'UTR、翻译起始序列、3'UTR和尾区的一个或多个序列。
在一些实施例中，本发明的可翻译低聚分子包括在全长人AGL编码序列SEQ IDNO:1上与野生型人AGL编码序列的一致性小于80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％的编码序列并且表达功能性人AGL蛋白。在一个示例性实施例中，本发明的可翻译低聚分子包括在全长人AGL编码序列SEQ ID NO:1上与野生型人AGL编码序列的一致性小于80％的编码序列并且表达功能性人AGL蛋白质。在另一个示例性实施例中，本发明的可翻译低聚分子包括在全长人AGL编码序列SEQ ID NO:1上与野生型人AGL编码序列的一致性小于80％的编码序列并且表达功能性人AGL蛋白，其中所述编码序列与选自SEQ ID NO:7-32或SEQ ID NO:41-45的序列的一致性为至少80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更大。在又一个示例性实施例中，本发明的可翻译低聚分子包括在全长人AGL编码序列SEQ ID NO:1上与野生型人AGL编码序列的一致性小于80％的编码序列并且表达功能性人AGL蛋白，其中所述编码序列与选自SEQ ID NO:7-32或SEQ ID NO:41-45的序列的一致性为至少95％。在又一个示例性实施例中，本发明的可翻译低聚分子包括在全长人AGL编码序列SEQ ID NO:1上与野生型人AGL编码序列的一致性小于80％的编码序列并且表达功能性人AGL蛋白，其中所述编码序列与选自SEQ ID NO:19、SEQ ID NO:31或SEQ ID NO:45的序列的一致性为至少95％或更大。因此，在一些实施例中，本申请提供了包括在全长人AGL编码序列SEQ ID NO:1上与野生型人AGL编码序列的一致性小于80％的核碱基序列或由其组成的多核苷酸，并且其中所述人AGL编码序列与选自SEQ ID NO:7-32或SEQ ID NO:41-45的序列的一致性为至少80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更大。在示例性实施例中，本申请提供了包括在全长人AGL编码序列SEQ ID NO:1上与野生型人AGL编码序列的一致性小于80％的核碱基序列或由其组成的多核苷酸，并且其中所述人AGL编码序列与选自SEQ ID NO:19、SEQ ID NO:31或SEQ ID NO:45的序列的一致性为至少95％。在一个具体实施例中，本申请提供了包括在全长人AGL编码序列SEQ ID NO:1上与野生型人AGL编码序列的一致性小于80％的核碱基序列的多核苷酸，并且其中所述人AGL编码序列与SEQ ID NO:19的一致性为至少95％。在另一个具体实施例中，本申请提供了包括在全长人AGL编码序列SEQ ID NO:1上与野生型人AGL编码序列的一致性小于80％的核碱基序列的多核苷酸，并且其中所述人AGL编码序列与SEQ ID NO:31的一致性为至少95％。在另一个具体实施例中，本申请提供了包括在全长人AGL编码序列SEQ ID NO:1上与野生型人AGL编码序列的一致性小于80％的核碱基序列的多核苷酸，并且其中所述人AGL编码序列与SEQ ID NO:45的一致性为至少95％。
在一些实施例中，本发明的可翻译低聚分子对融合蛋白进行编码，所述融合蛋白包括与另一个序列融合(例如N或C末端融合)的AGL蛋白的全长、片段或一部分。在一些实施例中，N或C末端序列是信号序列或细胞靶向序列。
UNA单体和低聚物
在一些实施例中，连接子基团单体可以是解锁核单体(UNA单体)，所述UNA单体是基于如下所示的丙烷-1,2,3-三-基-三氧基结构的小有机分子：
其中R1和R2是H，并且R1和R2可以是磷酸二酯键，碱基可以是核碱基，并且R3是下文描述的官能团。
在另一个视图中，UNA单体的主要原子可以以IUPAC表示法绘制如下：
其中低聚物链行进的方向是从丙烷残基的1-端到3-端。
核碱基的实例包含尿嘧啶、胸腺嘧啶、胞嘧啶、5-甲基胞嘧啶、腺嘌呤、鸟嘌呤、肌苷以及天然和非天然核碱基类似物。
核碱基的实例包含假尿嘧啶、1-甲基假尿嘧啶(m1Ψ)，即N1-甲基假尿嘧啶和5-甲氧基尿嘧啶。
不是核苷酸的UNA单体通常可以是低聚物中的内部连接子单体。低聚物中的内部UNA单体两侧的侧翼是其它单体。
例如，当低聚物形成复合物或双链体时，UNA单体可以参与碱基配对，并且所述复合物或双链体中存在具有核碱基的其它单体。
以下示出了作为内部单体侧翼在丙烷-1-基位置和丙烷-3-基位置两者处的UNA单体的实例，其中R3是-OH。
UNA单体可以是低聚物的末端单体，其中所述UNA单体附接到丙烷1-基位置或丙烷3-基位置处的仅一个单体。因为UNA单体是柔性有机结构，与核苷酸不同，所以末端UNA单体可以是低聚物的柔性终止子。
下文示出作为附接在丙烷-3-基位置处的末端单体的UNA单体的实例。
因为UNA单体可以是柔性分子，所以作为末端单体的UNA单体可以设想大为不同的构象。下文示出连接在丙烷-3-基位置处作为末端单体的能量最小化UNA单体构象的实例。
UNA-A末端形式：虚线键示出了丙烷-3-基附接
除其它事项外，UNA单体的结构允许其附接到天然存在的核苷酸。
UNA低聚物可以是由UNA单体以及可以基于天然存在的核苷酸的各种核苷酸够构成的链。
在一些实施例中，UNA单体的官能团R3可以是-OR4、-SR4、-NR4 2、-NH(C＝O)R4、吗啉基、吗啉-1-基、哌嗪-1-基或4-烷酰基-哌嗪-1-基，其中R4每次出现时是相同或不同的并且可以是H、烷基、胆固醇、脂质分子、多胺、氨基酸或多肽。
UNA单体是有机分子。UNA单体不是核酸单体或核苷酸，也不是天然存在的核苷或经过修饰的天然存在的核苷。
本发明的UNA低聚物是合成链分子。
在一些实施例中，如上所示，UNA单体可以是UNA-A(指定为)、UNA-U(指定为/>)、UNA-C(指定为/>)和UNA-G(指定为/>)。
本文可以使用的代号包含指代2'-O-甲基修饰的核糖核苷酸的mA、mG、mC和mU。
本文可以使用的代号包含指代2'-脱氧T核苷酸的dT。
如本文所使用的，在低聚物序列的上下文中，符号N可以表示任何天然核苷酸单体或任何经过修饰的核苷酸单体。
如本文所使用的，在低聚物序列的上下文中，符号Q表示非天然的、经过修饰的或经过化学修饰的核苷酸单体。
如本文所使用的，在低聚物序列的上下文中，符号X可以用于表示UNA单体。
经过修饰的核苷酸和经过化学修饰的核苷酸
在本文的经过修饰的或经过化学修饰的核苷酸的实例中，烷基、环烷基或苯基取代基可以是未取代的或者进一步由一个或多个烷基、卤素、卤代烷基、氨基或硝基取代基取代。
核酸单体的实例包含非天然的、经过修饰的和经过化学修饰的核苷酸，所述核苷酸包含本领域已知的任何此类核苷酸。
经过修饰的或经过化学修饰的核苷酸的实例包含5-羟基胞苷、5-烷基胞苷、5-羟烷基胞苷、5-羧基胞苷、5-甲酰基胞苷、5-烷氧基胞苷、5-炔基胞苷、5-卤代胞苷、2-硫代胞苷、N4-烷基胞苷，N4-氨基胞苷、N4-乙酰基胞苷和N4,N4-二烷基胞苷。
经过修饰的或经过化学修饰的核苷酸的实例包含5-羟基胞苷、5-甲基胞苷、5-羟甲基胞苷、5-羧基胞苷、5-甲酰基胞苷、5-甲氧基胞苷、5-丙炔基胞苷、5-溴代胞苷、5-碘代胞苷、2-硫代胞苷、N4-甲基胞苷、N4-氨基胞苷、N4-乙酰基胞苷和N4,N4-二甲苯胞苷。
经过修饰的或经过化学修饰的核苷酸的实例包含5-羟基尿苷、5-烷基尿苷、5-羟烷基尿苷、5-羧基尿苷、5-羧烷基酯尿苷、5-甲酰基尿苷、5-烷氧基尿苷、5-炔基尿苷、5-卤代尿苷、2-硫代尿苷和6-烷基尿苷。
经过修饰的或经过化学修饰的核苷酸的实例包含5-羟基尿苷、5-甲基尿苷、5-羟甲基尿苷、5-羧基尿苷、5-羧甲基酯尿苷、5-甲酰基尿苷、5-甲氧基尿苷、5-丙炔基尿苷、5-溴代尿苷、5-氟代尿苷、5-碘代尿苷、2-硫代尿苷和6-甲基尿苷。
经过修饰的或经过化学修饰的核苷酸的实例包含5-甲氧基羰基甲基-2-硫代尿苷、5-甲基氨甲基-2-硫代尿苷，5-氨基甲酰基甲基尿苷、5-氨基甲酰基甲基-2'-O-甲基尿苷、1-甲基-3-(3-氨基-3-羧基丙基)假尿苷、5-甲基氨甲基-2-硒基尿苷、5-羧甲基尿苷、5-甲基二氢尿苷、5-牛磺甲基尿苷(5-taurinomethyluridine)、5-牛磺甲基-2-硫代尿苷、5-(异戊烯基氨甲基)尿苷、2'-O-甲基假尿苷、2-硫代-2'O-甲基尿苷和3,2'-O-二甲基尿苷。
经过修饰的或经过化学修饰的核苷酸的实例包含N6-甲基腺苷、2-氨基腺苷、3-甲基腺苷、8-氮腺苷、7-脱氮腺苷、8-氧代腺苷、8-溴代腺苷、2-甲硫基-N6-甲基腺苷、N6-异戊烯基腺苷、2-甲硫基-N6-异戊烯基腺苷、N6-(顺式-羟基异戊烯基)腺苷、2-甲硫基-N6-(顺式-羟基异戊烯基)腺苷、N6-甘氨酰基氨基甲酰基腺苷、N6-苏氨酰基氨基甲酰基-腺苷、N6-甲基-N6-苏氨酰基氨基甲酰基-腺苷、2-甲硫基-N6-苏氨酰基氨基甲酰基-腺苷、N6,N6-二甲基腺苷、N6-羟基正缬氨酰基氨基甲酰基腺苷、2-甲硫基-N6-羟基正缬氨酰基氨基甲酰基-腺苷、N6-乙酰基-腺苷、7-甲基-腺嘌呤、2-甲硫基-腺嘌呤、2-甲氧基-腺嘌呤、α-硫代-腺苷、2'-O-甲基-腺苷、N6,2'-O-二甲基-腺苷、N6,N6,2'-O-三甲基-腺苷、1,2'-O-二甲基-腺苷、2'-O-腺嘌呤核糖苷、2-氨基-N6-甲基-嘌呤、1-硫代-腺苷、2'-F-阿糖-腺苷、2'-F-腺苷、2'-OH-阿糖-腺苷和N6-(19-氨基-五氧杂十九基)-腺苷。
经过修饰的或经过化学修饰的核苷酸的实例包含Nl-烷基鸟苷、N2-烷基鸟苷、噻吩并鸟苷、7-脱氮鸟苷、8-氧代鸟苷、8-溴代鸟苷、O6-烷基鸟苷、黄嘌呤核苷、肌苷和Nl-烷基肌苷。
经过修饰的或经过化学修饰的核苷酸的实例包含Nl-甲基鸟苷、N2-甲基鸟苷、噻吩并鸟苷、7-脱氮鸟苷、8-氧代鸟苷、8-溴代鸟苷、O6-甲基鸟苷、黄嘌呤核苷、肌苷和Nl-甲基肌苷。
经过修饰的或经过化学修饰的核苷酸的实例包含假尿苷。假尿苷的实例包含Nl-烷基假尿苷、Nl-环烷基假尿苷、N1-羟基假尿苷、N1-羟烷基假尿苷、Nl-苯基假尿苷、Nl-苯基烷基假尿苷、Nl-氨烷基假尿苷、N3-烷基假尿苷、N6-烷基假尿苷、N6-烷氧基假尿苷、N6-羟基假尿苷、N6-羟烷基假尿苷、N6-吗啉基假尿苷、N6-苯基假尿苷和N6-卤代假尿苷。假尿苷的实例包含Nl-烷基-N6-烷基假尿苷、Nl-烷基-N6-烷氧基假尿苷、Nl-烷基-N6-羟基假尿苷、Nl-烷基-N6-羟烷基假尿苷、Nl-烷基-N6-吗啉基假尿苷、Nl-烷基-N6-苯基假尿苷和Nl-烷基-N6-卤代假尿苷。在这些实例中，烷基、环烷基和苯基取代基可以是未取代的或者进一步由烷基、卤素、卤代烷基、氨基或硝基取代基取代。
假尿苷的实例包含Nl-甲基假尿苷、Nl-乙基假尿苷、Nl-丙基假尿苷、Nl-环丙基假尿苷、Nl-苯基假尿苷、Nl-氨甲基假尿苷、N3-甲基假尿苷、N1-羟基假尿苷和N1-羟甲基假尿苷。
核酸单体的实例包含经过修饰的和经过化学修饰的核苷酸，所述核苷酸包含本领域已知的任何此类核苷酸。
经过修饰的和经过化学修饰的核苷酸单体的实例包含本领域已知的任何此类核苷酸，例如，2'-O-甲基核糖核苷酸、2'-O-甲基嘌呤核苷酸、2'-脱氧-2'-氟核糖核苷酸、2'-脱氧-2'-氟嘧啶核苷酸、2'-脱氧核糖核苷酸、2'-脱氧嘌呤核苷酸、通用碱基核苷酸、5-C-甲基-核苷酸和反向脱氧碱基单体残基。
经过修饰的和经过化学修饰的核苷酸单体的实例包含3'端稳定的核苷酸、3'-甘油基核苷酸、3'-反向碱基核苷酸和3'-反向胸苷。
经过修饰的和经过化学修饰的核苷酸单体的实例包含锁核酸核苷酸(LNA)、2'-O,4'-C-亚甲基-(D-呋喃核糖基)核苷酸、2'-甲氧基乙氧基(MOE)核苷酸、2'-甲基-硫代-乙基、2'-脱氧-2'-氟核苷酸和2'-O-甲基核苷酸。在示例性实施例中，经过修饰的单体是锁核酸核苷酸(LNA)。
经过修饰的和经过化学修饰的核苷酸单体的实例包含2',4'-约束的2'-O-甲氧乙基(cMOE)和2'-O-乙基(cEt)修饰的DNA。
经过修饰的和经过化学修饰的核苷酸单体的实例包含2'-氨基核苷酸、2'-O-氨基核苷酸、2'-C-烯丙基核苷酸和2'-O-烯丙基核苷酸。
经过修饰的和经过化学修饰的核苷酸单体的实例包含N6-甲基腺苷核苷酸。
经过修饰的和经过化学修饰的核苷酸单体的实例包含具有经过修饰的碱基的核苷酸单体5-(3-氨基)丙基尿苷、5-(2-巯基)乙基尿苷、5-溴代尿苷、8-溴代鸟苷或7-脱氮腺苷。
经过修饰的和经过化学修饰的核苷酸单体的实例包含2'-O-氨丙基取代的核苷酸。
经过修饰的和经过化学修饰的核苷酸单体的实例包含用2'-R、2'-OR、2'-卤素、2'-SR或2'-氨基置换核苷酸的2'-OH基团，其中R可以是H、烷基、烯基或炔基。
Saenger,《核酸结构原理(Principles of Nucleic Acid Structure)》,施普林格出版社(Springer-Verlag),1984中给出了经过修饰的核苷酸的一些实例。
可以将上文描述的碱基修饰的实例与核苷或核苷酸结构的另外的修饰组合，所述另外的修饰包含糖修饰和键修饰。
在自然界中可以发现某些经过修饰的或经过化学修饰的核苷酸单体。
含有一个或多个UNA单体的可翻译分子
本发明的各方面提供了是含有一个或多个UNA单体的低聚化合物的可翻译分子的结构和组合物。可翻译低聚物可以是药物组合物的活性剂。在一些实施例中，可翻译低聚物对人AGL或其变体进行编码。
本发明的低聚可翻译分子可以含有一个或多个UNA单体。本发明的低聚分子可以用作用于供应肽和蛋白质治疗剂的调配物的活性剂。在一些实施例中，可翻译低聚物对人AGL或其变体进行编码。
在一些实施例中，本发明提供了具有并入UNA单体与某些天然核苷酸、非天然核苷酸、经过修饰的核苷酸或经过化学修饰的核苷酸的新型组合的结构的低聚可翻译化合物。
本发明的可翻译低聚化合物的长度可以为约200到约12,000个碱基。本发明的可翻译低聚化合物的长度可以为约1000个、2000个、3000个、4000个、5000个、6000个、7000个、8000个或约9000个碱基。在一些实施例中，本发明的可翻译低聚化合物的长度可以为约4000个、4100个、4200个、4300个、4400个、4500个、4600个、4700个、4800个、4900个、5000个、5100个、5200个、5300个、5400个或约5500个碱基。在示例性实施例中，本发明的可翻译低聚化合物的长度为约5000个碱基。
在另外方面，本发明的包括一个或多个UNA单体的低聚可翻译化合物可以是药理学活性分子。可翻译低聚分子可以用作用于在体外、在体内或离体地产生肽或蛋白质活性剂的活性药物成分。在示例性实施例中，本发明的可翻译低聚化合物对人AGL或其变体进行编码。
本发明的可翻译低聚分子可以具有式I的结构：
其中L1是键，n为200到12,000，并且每次出现时，L2是具有式-C1-C2-C3-的UNA连接子基团，其中R附接到C2并且具有式-OCH(CH2R3)R5，其中R3是-OR4、-SR4、-NR4 2、-NH(C＝O)R4、吗啉基、吗啉-1-基、哌嗪-1-基或4-烷酰基哌嗪-1-基，其中R4在每次出现时是相同或不同的并且是H、烷基、胆固醇、脂质分子、多胺、氨基酸或多肽，并且其中R5是核碱基，或者L2(R)是如核糖等糖并且R是核碱基，或者L2是如经过修饰的核糖等经过修饰的糖并且R是核碱基。在某些实施例中，核碱基可以是经过修饰的核碱基。L1可以是磷酸二酯键。
可翻译低聚分子的碱基序列可以是任何核碱基序列。
在一些方面，本发明的可翻译低聚分子在任何单体间(intermonomer)位置处可以具有任何数量的硫代磷酸酯单体间键。
在一些实施例中，可翻译低聚分子的单体间键中的任何一个或多个单体间键可以是磷酸二酯、包括二硫酸酯的硫代磷酸酯、手性硫代磷酸酯和其它经过化学修饰的形式。
当可翻译低聚分子在UNA单体中终止时，根据上文所示的位置编号，末端位置具有1-端，或者末端位置具有3-端。
增强的翻译
本发明的可翻译分子可以并入增强分子的翻译效率的区域。
通常，可以将本领域已知的翻译增强子区并入可翻译分子的结构中，以增加肽或蛋白质产率。
含有翻译增强子区的可翻译分子可以增加肽或蛋白质的产生。
在一些实施例中，翻译增强子区可以包括或位于可翻译分子的5'非翻译区或3'非翻译区。
翻译增强子区的实例包含来自TEV 5'UTR和非洲爪蟾β-珠蛋白3'UTR的天然存在的增强子区。
分子结构和序列
可翻译分子可以被设计成表达靶肽或蛋白。在一些实施例中，靶肽或蛋白可以与受试者的病状或疾病相关联。
在一些方面，可翻译分子的碱基序列可以包含与mRNA的碱基序列的至少有效部分或结构域相同的部分，其中有效部分足以向可翻译分子的翻译产物赋予治疗活性。
在一些方面，本发明提供了具有与细胞的原生核酸分子的至少片段相同的碱基序列的活性可翻译分子。
在某些实施例中，可翻译分子的碱基序列可以包含除了一个或多个碱基突变外与mRNA的碱基序列相同的部分。可翻译分子的突变数量应当不超过将会产生可翻译分子的具有基本上低于mRNA的活性的翻译产物的量。
本发明的低聚可翻译UNA分子可以展示核碱基序列并且可以被设计成在体外、离体地或在体内表达肽或蛋白质。所表达的肽或蛋白质可以具有各种形式的活性，所述活性包含与从原生或天然mRNA表达的蛋白质相对应的活性。
在一些实施例中，本发明的可翻译分子的链长可以为约400到15,000个单体，其中不是UNA单体的任何单体可以是N单体或Q单体。
分子端帽结构
本发明的可翻译分子可以具有5'端，所述5'端被本领域已知的各种基团及其类似物封端。在示例性实施例中，所述5'端帽可以是m7GpppGm端帽。在另外的实施例中，所述5'端帽可以选自m7GpppA、m7GpppC；非甲基化的端帽类似物(例如，GpppG)；二甲基化的端帽类似物(例如，m2,7GpppG)、三甲基化的端帽类似物(例如，m2,2,7GpppG)、二甲基化的对称端帽类似物(例如，m7Gpppm7G)或抗-反向端帽类似物(例如，ARCA；m7、2'OmeGpppG、m72'dGpppG、m7,3'OmeGpppG、m7,3'dGpppG及其四核糖基衍生物)(参见，例如，Jemelity,J.等人,《RNA》9:1108-1122(2003))。在其它实施例中，所述5'端帽可以是ARCA端帽(3'-OMe-m7G(5')pppG)。所述5'端帽可以是mCAP(m7G(5')ppp(5')G、N7-甲基-鸟苷-5'-三磷酸-5'-鸟苷)。所述5'端帽可以抗水解。
WO2015/051169A2、WO/2015/061491以及美国专利第8,093,367号和第8,304,529号中给出了5'端帽结构的一些实例。
尾区
在一些实施例中，对AGL进行编码的可翻译低聚物包括尾区，所述尾区能够用于保护mRNA免受外切核酸酶降解。在一些实施例中，所述尾区可以是polyA尾。
可以使用本领域已知的各种方法添加polyA尾，例如，使用polyA聚合酶将尾添加到合成的或体外转录的RNA。其它方法包含使用转录载体对polyA尾进行编码或使用连接酶(例如，通过使用T4RNA连接酶和/或T4DNA连接酶的配体)，其中polyA可以连接到有义RNA的3'端。在一些实施例中，使用上述方法中任一种方法的组合。
在一些实施例中，可翻译低聚物包括3'polyA尾结构。在一些实施例中，所述polyA尾的长度可以为至少约5个、10个、15个、20个、25个、30个、35个、40个、45个、50个、100个、200个或300个核苷酸。在一些实施例中，3'polyA尾含有约5到300个腺苷核苷酸(例如，约30到250个腺苷核苷酸、约60到220个腺苷核苷酸、约80到200个腺苷核苷酸、约90到约150个腺苷核苷酸或约100到约120个腺苷核苷酸)。在示例性实施例中，所述3'polyA尾的长度为约100个核苷酸。在另一个示例性实施例中，所述3'polyA尾的长度为约115个核苷酸。在另一个示例性实施例中，所述3'polyA尾的长度为约250个核苷酸。
在一些实施例中，所述3'polyA尾包括一个或多个UNA单体。在一些实施例中，所述3'polyA尾含有2个、3个、4个、5个、10个、15个、20个或更多个UNA单体。在示例性实施例中，所述3'polyA尾含有2个UNA单体。在另一个示例性实施例中，所述3'polyA尾含有连续发现的2个UNA单体，即，二者在所述3'polyA尾中彼此相邻。
在示例性实施例中，所述3'polyA尾包括SEQ ID NO:6中所示的序列或由其组成。在另一个示例性实施例中，所述3'polyA尾包括SEQ ID NO:38中所示的序列或由其组成。在又一个示例性实施例中，所述3'polyA尾包括SEQ ID NO:39中所示的序列或由其组成。
在一些实施例中，所述可翻译低聚物包括3'polyC尾结构。在一些实施例中，所述polyC尾的长度可以为至少约5个、10个、15个、20个、25个、30个、35个、40个、45个、50个、100个、200个或300个核苷酸。在一些实施例中，3'polyC尾含有约5到300个胞嘧啶核苷酸(例如，约30到250个胞嘧啶核苷酸、约60到220个胞嘧啶核苷酸、约80到200个胞嘧啶核苷酸、约90到约150个胞嘧啶核苷酸或约100到约120个胞嘧啶核苷酸)。在示例性实施例中，所述3'polyC尾的长度为约100个核苷酸。在另一个示例性实施例中，所述3'polyC尾的长度为约115个核苷酸。所述polyC尾可以添加至所述polyA尾，或者可以取代所述polyA尾。所述polyC尾可以添加至所述polyA尾的5'端或所述polyA尾的3'端。
在一些实施例中，调节所述polyA尾和/或所述polyC尾的长度以控制本发明的经过修饰的可翻译低聚分子的稳定性，从而控制蛋白质的转录。例如，由于polyA尾的长度可能影响可翻译分子的半衰期，因此可以调整所述polyA尾的长度以改变mRNA对核酸酶的抗性水平，从而控制靶细胞中多核苷酸表达和/或多肽生成的时间进程。
5'和3'非翻译区(UTR)
在一些实施例中，对AGL进行编码的可翻译低聚物可以包括5'非翻译区和/或3'非翻译区。如本领域所理解的，5'UTR和/或3'UTR可能影响mRNA的稳定性或翻译的效率。在示例性实施例中，所述可翻译低聚物包括5'UTR和3'UTR。
在一些实施例中，所述可翻译低聚物可以包括至少有约25个、50个、75个、100个、125个、150个、175个、200个、300个、400个或500个核苷酸的5'UTR。在一些实施例中，5'UTR含有约50到300个核苷酸(例如，约75到250个核苷酸、约100到200个核苷酸、约120到约150个核苷酸或约135个核苷酸)。在示例性实施例中，所述5'UTR的长度为约135个核苷酸。
在一些实施例中，所述5'UTR源自本领域已知相对稳定的mRNA分子(例如，组蛋白、微管蛋白、珠蛋白、GAPDH、肌动蛋白或柠檬酸循环酶)，以提高所述可翻译低聚物的稳定性。在其它实施例中，5'UTR序列可以包含CMV即刻早期1(IE1)基因的部分序列。5'UTR序列的实例可以在美国专利第9,149,506号中找到。在一些实施例中，所述5'UTR包括选自以下的序列：人IL-6、丙氨酸氨基转移酶1、人载脂蛋白E、人纤维蛋白原α链、人转甲状腺素蛋白、人触珠蛋白、人α-1-抗糜蛋白酶、人抗凝血酶、人α-1-抗胰蛋白酶、人白蛋白、人β珠蛋白、人补体C3、人补体C5、Synk、AT1G58420、小鼠β珠蛋白、小鼠白蛋白和烟草蚀纹病毒的5'UTR或前述任何一种的片段。在示例性实施例中，所述5'UTR源自烟草蚀纹病毒(TEV)。在另一个示例性实施例中，所述5'UTR包括SEQ ID NO:3中阐述的序列或由其组成。在又一个示例性实施例中，所述5'UTR是SEQ ID NO:3中阐述的序列的片段，如SEQ ID NO:3中的至少10个、20个、30个、40个、50个、60个、70个、80个、90个、100个、110个、120个或125个相邻核苷酸的片段。
在一些实施例中，所述可翻译低聚分子包括内部核糖体进入位点(IRES)。如本领域所理解的，IRES是允许以不依赖末端的方式启动翻译的RNA元件。在示例性实施例中，所述IRES位于所述5'UTR中。在其它实施例中，所述IRES可以位于所述5'UTR之外。
在一些实施例中,所述可翻译低聚物可以包括至少有约25个、50个、75个、100个、125个、150个、175个、200个、300个、400个或500个核苷酸的3'UTR。在一些实施例中，3'UTR含有约50到300个核苷酸(例如，约75到250个核苷酸、约100到200个核苷酸、约140到约175个核苷酸或约160个核苷酸)。在示例性实施例中，所述3'UTR的长度为约160个核苷酸。
在一些实施例中，所述3'UTR包括一个或多个UNA单体。在一些实施例中，所述3'UTR含有2个、3个、4个、5个、10个、15个、20个或更多个UNA单体。
3'UTR序列的实例可以在美国专利第9,149,506号中找到。在一些实施例中，所述3'UTR包括选自以下的序列：丙氨酸氨基转移酶1、人载脂蛋白E、人纤维蛋白原α链、人触珠蛋白、人抗凝血酶、人α珠蛋白、人β珠蛋白、人补体C3、人生长因子、人七肽、MALAT-1、小鼠β珠蛋白、小鼠白蛋白和非洲爪蟾β珠蛋白的3'UTR或前述任何一种的片段。在示例性实施例中，所述3'UTR源自非洲爪蟾β珠蛋白。在另一个示例性实施例中，所述3'UTR源自非洲爪蟾β珠蛋白并含有一个或多个UNA单体。在另一个示例性实施例中，所述3'UTR包括SEQ ID NO:5和33-37中阐述的序列或由其组成。在又一个示例性实施例中，所述3'UTR是SEQ ID NO:5和33-37中阐述的序列的片段，如SEQ ID NO:5和33-37中的至少10个、20个、30个、40个、50个、60个、70个、80个、90个、100个、110个、120个、130个、140个或150个相邻核苷酸的片段。
在某些示例性实施例中，对AGL进行编码的可翻译低聚物包括SEQ ID NO:3的5'UTR序列和选自SEQ ID NO:5和33-37的3'UTR序列。在一些实施例中，对AGL进行编码的可翻译低聚物进一步包括SEQ ID NO:6、SEQ ID NO:38或SEQ ID NO:39中所示的polyA尾。在一些实施例中，AGL的mRNA编码序列包括选自SEQ ID NO:7-32或SEQ ID NO:41-45的序列。
三联终止密码子
在一些实施例中，对AGL进行编码的可翻译低聚物可以包括紧邻CDS下游的序列，所述序列产生三联终止密码子。可以引入所述三联终止密码子以提高翻译效率。在一些实施例中，所述可翻译低聚物可以包括紧邻本文所述的PAH CDS下游的序列AUAAGUGAA(SEQID NO:40)，如SEQ ID NO:7-32或SEQ ID NO:41-45所示。
翻译起始位点
在一些实施例中，对AGL进行编码的可翻译低聚物可以包括翻译起始位点。此类序列在本领域中是已知的，并且包含Kozak序列。参见，例如Kozak,Marilyn(1988)《分子和细胞生物学(Mol.and Cell Biol.)》,8:2737-2744；Kozak,Marilyn(1991)《生物化学杂志(J.Biol.Chem.)》,266:19867-19870；Kozak,Marilyn(1990)《美国国家科学院院刊(ProcNatl.Acad.Sci.USA)》,87:8301-8305；和Kozak,Marilyn(1989)《细胞生物学杂志(J.CellBiol.)》,108:229–241；以及其中引用的参考文献。如本领域所理解的，Kozak序列是围绕真核mRNA的翻译起始位点为中心的短共有序列，所述短共有序列允许有效地启动mRNA的翻译。核糖体翻译机器在Kozak序列的背景下识别AUG起始密码子。
在一些实施例中，翻译起始位点，例如Kozak序列，被插入到AGL的编码序列的上游。在一些实施例中，翻译起始位点被插入到5'UTR的下游。在某些示例性实施例中，翻译起始位点被插入到AGL的编码序列的上游和5'UTR的下游。
如本领域所理解的，Kozak序列的长度可以变化。通常，增加前导序列的长度会增强翻译。
在一些实施例中，对AGL进行编码的可翻译低聚物包括具有SEQ ID NO:4的序列的Kozak序列。在某些示例性实施例中，对AGL进行编码的可翻译低聚物包括具有SEQ ID NO:4的序列的Kozak序列,其中所述Kozak序列紧邻5'UTR的下游且紧邻AGL的编码序列的上游。
合成方法
在各个方面，本发明提供了用于合成可翻译信使分子的方法。
可以使用本文公开的方法以及本领域已知的任何相关技术来合成和分离本发明的可翻译分子。
一些用于制备核酸的方法在下列文献中给出：例如，Merino,《核苷酸类似物的化学合成(Chemical Synthesis of Nucleoside Analogues)》,(2013)；Gait,《寡核苷酸合成：一种实用的方法(Oligonucleotide synthesis:a practical approach)》(1984)；Herdewijn,《寡核苷酸合成，分子生物学方法(Oligonucleotide Synthesis,Methods inMolecular Biology)》,第288卷(2005)。
在一些实施例中，可翻译分子可以通过体外转录(IVT)反应生成。可以使用T7试剂聚合核苷三磷酸酯(NTP)的混合物以便，例如，从DNA模板产生RNA。可以用无RNase的DNase降解DNA模板，并且对RNA进行柱分离。
在一些实施例中，可以使用连接酶将合成的低聚物连接到RNA分子或RNA转录物的3'端以形成可翻译分子。连接到3'端的合成的低聚物可以提供polyA尾的功能，并有利地提供了抵抗其被3'-外切核糖核酸酶去除的能力。连接产物可翻译分子可以具有提高的比活性，并提供增加水平的异位蛋白表达。
在某些实施例中，本发明的可翻译分子的连接产物可以通过具有原生特异性的RNA转录物生成。连接产物可以是合成分子，所述合成分子保留了在5'端的RNA转录物的结构以确保与原生特异性相容。
在另外的实施例中，本发明的可翻译分子的连接产物可以通过外源性RNA转录物或者非天然RNA生成。连接产物可以是保留了RNA的结构的合成分子。
在不希望受到理论约束的情况下，细胞中规范的mRNA降解途径包含以下步骤：(i)通过3'外切核酸酶将polyA尾逐渐切回到末端，从而终止了有效翻译所需的环相互作用，并使端帽易受到攻击；(ii)使复合物去端化(decapping)以去除5'端帽；(iii)通过5'和3'外切核酸酶活性降解转录物的未受保护且翻译能力低的残基。
本发明的实施例涉及新的可翻译结构，其可以具有比原生转录物的翻译活性更高的翻译活性。除其它外，本文的可翻译分子可以防止外切核酸酶在去腺苷酸化过程中修剪polyA尾。
本发明的实施例提供了可翻译分子的结构、组合物和方法。本发明的实施例可以提供含有一个或多个UNA单体且功能半衰期增长的可翻译分子。
已经发现，通过将mRNA转录物高度转化为连接产物，可以出人意料地完成合成的低聚物与mRNA转录物的3'端的连接。
如本文所用，术语polyA尾和polyA低聚物是指单体的低聚物，其中单体可以包含基于腺嘌呤的核苷酸、UNA单体、天然存在的核苷酸、经过修饰的核苷酸或核苷酸类似物。
用于连接至RNA的3'端的低聚物的长度可以为2到120个单体、或长度为3到120个单体、或长度为4到120个单体、或长度为5到120个单体或更长。在示例性实施例中，用于连接的低聚物的长度为约30个单体。
脂基制剂
脂基制剂由于其生物相容性和易于大规模生产而越来越多地被视为是最有希望的RNA递送系统之一。阳离子脂质作为递送RNA的合成材料已经得到广泛的研究。混合在一起后，核酸被阳离子脂质凝聚而形成被称为阳离子脂质体的脂质/核酸复合物。这些脂质复合物能够保护遗传物质免受核酸酶的作用，并通过与带负电荷的细胞膜相互作用而将所述遗传物质递送到细胞中。通过直接将生理pH值下的带正电荷的脂质与带负电荷的核酸混合，可以制备出阳离子脂质体。
常规的脂质体由脂质双层组成，所述脂质双层可以由阳离子、阴离子或中性(磷)脂质和胆固醇构成并且包含水核。脂质双层和水空间均可以分别掺入疏水性或亲水性化合物。可以通过在脂质体表面添加亲水性聚合物涂层，例如聚乙二醇(PEG)来改变脂质体特性和体内行为，从而获得立体稳定。此外，通过将配体(例如，抗体、肽和碳水化合物)附接到其表面或附接的PEG链的末端，脂质体可以用于特异性靶向(《前沿药理学(FrontPharmacol.)》2015年12月1日；6:286)。
脂质体是胶体脂质基和基于表面活性剂的递送系统，其由围绕水性隔室的磷脂双层构成。脂质体可以呈现为球形小泡，并且大小范围为20纳米到几微米。基于阳离子脂质的脂质体能够通过静电相互作用与带负电荷的核酸结合，从而形成具有生物相容性、低毒性和能够大规模生产以用于体内临床应用的复合物。脂质体可以与质膜融合，从而被摄取；一旦进入细胞内，脂质体就会通过内吞途径被处理，然后遗传物质从内体/载剂释放到细胞质中。因为脂质体基本上是生物膜的类似物且可以由天然和合成的磷脂制备，所以脂质体因其优越的生物相容性而长期以来被视为药物递送媒剂。(《国家纳米医学杂志(Int JNanomedicine.)》2014；9:1833-1843)。
阳离子脂质体传统上是寡核苷酸最常用的非病毒递送系统，包含质粒DNA、反义寡核苷酸和siRNA/小发夹R A-shRNA)。阳离子脂质，如DOTAP(1,2-二烯酰-3-三甲基丙烷)和DOTMA(N-[l-(2,3-二烯酰氧基)丙基]-N,N,N-三甲基硫酸铵)可以与带负电荷的核酸形成复合物或阳离子脂质体，以通过静电相互作用形成纳米颗粒，从而提供较高的体外转染效率。此外，还开发了用于RNA递送的中性脂基纳米脂质体，例如中性l,2-二油酰基-sn-甘油-3-磷脂酰胆碱(DOPC)基纳米脂质体。(《高级药物递送综述(Adv Drug Deliv Rev.)》2014年2月；66:110-116)
根据一些实施例，本文描述的可表达的多核苷酸和异源mRNA结构是脂质配制的。脂质制剂优选地选自但不限于脂质体、阳离子脂质体、共聚物(如PLGA)和脂质纳米颗粒。在一个优选的实施例中，脂质纳米颗粒(LNP)包括：
(a)核酸；
(b)阳离子脂质；
(c)聚合还原剂(如聚乙二醇(PEG)脂质或经过PEG修饰的脂质)；
(d)任选地，非阳离子脂质(如中性脂质)；以及
(e)任选地，固醇。
在一个实施例中，脂质纳米颗粒制剂由以下组成：(i)至少一个阳离子脂质；(ii)中性脂质；(iii)固醇，例如，胆固醇；以及(iv)PEG-脂质，摩尔比约为：20-60％阳离子脂质：5-25％中性脂质：25-55％固醇；0.5-15％PEG-脂质。
含硫代氨基甲酸酯和氨基甲酸酯的脂质制剂
WO/2015/074085和USSN 15/387,067中给出了用于递送本发明的活性分子的脂质和脂质组合物的一些实例，所述文献中的每一个均通过引用整体并入本文。在某些实施例中，脂质是下式的化合物：
其中
R1和R2均包括由1至14个碳组成的直链烷基或由2至14个碳组成的烯基或炔基；
L1和L2均包括由5至18个碳组成的或与N形成杂环的直链亚烷基或亚烯基；
X是S；
L3包括键或由1至6个碳组成的或与N形成杂环的直链亚烷基；
R3包括由1至6个碳组成的直链或支链亚烷基；并且
R4和R5相同或不同，各自包括氢或由1至6个碳组成的直链或支链烷基；
或其药学上可接受的盐。
脂质制剂可以含有一个或多个选自以下的可电离阳离子脂质：
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阳离子脂质
脂质纳米颗粒优选地包含适于形成脂质纳米颗粒的阳离子脂质。优选地，所述阳离子脂质在接近生理pH值下携带净正电荷。
例如，所述阳离子脂质可以为N,N-二油基-N,N-二甲基氯化铵(DODAC)、N,N-二硬脂基-N,N-二甲基溴化铵(DDAB)、1,2-二油基三甲基氯化铵(DOTAP)(也被称为N-(2,3-二油基氧基)丙基)-N,N,N-三甲基氯化铵和1,2-二油酰氧基-3-三甲基氨基丙烷氯盐)、N-(1-(2,3-二油酰氧基)丙基)-N,N,N-三甲基氯化铵(DOTMA)、N,N-二甲基-2,3-二油酰氧基)丙基胺(DODMA)、1,2-二亚油酰氧基-N,N-二甲基氨基丙烷(DLinDMA)、1,2-二氢亚油酰氧基-N,N-二甲基氨基丙烷(DLenDMA)、1,2-二氢亚油酰氧基-N,N-二甲基氨基丙烷(γ-DLenDMA)、1,2-二烷基氨基甲酰氨基甲酰氧基-3-二甲基氨基丙烷(DLin-C-DAP)、1,2-二烷基氨基甲酰氧基-3-(二甲基氨基)乙酰氧基丙烷(DLin-DAC)、1,2-二烷基氨基甲酰氧基-3-吗啉丙烷(DLin-MA)、1,2-二氢油酰基-3-二甲基氨基丙烷(DLinDAP)、1,2-二氢油酰基硫代-3-二甲基氨基丙烷(DLin-S-DMA)、1-亚油酰基-2-亚油酰基-3-二甲基氨基丙烷(DLin-2-DMAP)、1,2-二氢油酰氧基-3-三甲氨基丙烷氯盐(DLin-TMA.CI)、1,2-二氢油酰-3-三甲氨基丙烷氯盐(DLin-TAP.CI)、1,2-二氢油酰氧基-3-(N-甲基哌嗪)丙烷(DLin-MPZ)、或3-(N,N-二油基氨基)-1,2-丙二醇(DLinAP)、3-(N,N-二油基氨基)-1,2-丙二醇(DOAP)、1,2-二油基氧代-3-(2-N,N-二甲基氨基)乙氧基丙烷(DLin-EG-DM A)、2,2-二苯基-4-二甲基氨基甲基-[1,3]-二氧戊烷(DLin-K-DMA)或其类似物、(3aR,5s,6aS)-N,N-二甲基-2,2-二((9Z,12Z)-十八烷基-9,12-二烯基)四氢-3aH-环戊烯[d][1,3]二噁醇-5-胺、(6Z,9Z,28Z,31Z)-三十七碳-6,9,28,31-四烯-19-基4-(二甲基氨基)丁酸甲酯(MC3)、1,1'-(2-(4-(2-((2-(双(2-羟基十二烷基)氨基)乙基)(2-羟基十二烷基)氨基)乙基)哌嗪-1-基)乙基氮杂二基)二十二烷-2-醇(C12-200)、2,2-二氢吲哚基-4-(2-二甲基氨基乙基)-[1,3]-二氧戊环(DLin-K-C2-DMA)、2,2-二氢吲哚基-4-二甲基氨基甲基-[1,3]-二氧戊环(DLin-K-DMA)、(6Z,9Z,28Z,31Z)-三十七碳6,9,28,31-四烯-19-基4-(二甲基氨基)丁酸(DLin-M-C3-DMA)、3-((6Z,9Z,28Z,31Z)-三十七碳-6,9,28,31-四烯-19-氧基)-N,N-二甲基丙-1-胺(MC3醚)、4-((6Z,9Z,28Z,31Z)-三十七碳-6,9,28,31-四烯-19-氧基)-N,N-二甲基丁-1-胺(MC4醚)或前述任何一种的任何组合。其它阳离子脂质包含但不限于N,N-二硬脂基-N,N-二甲基溴化铵(DDAB)、3P-(N-(N',N'-二甲基氨基乙烷)-氨基甲酰基)胆固醇(DC-Choi)、N-(l-(2,3-二油酰氧基)丙基)-N-2-(精胺)乙基)-N,N-二甲基三氟乙酸铵(DOSPA)、二十八烷基酰胺基糖基羧精胺(DOGS)、1,2-二亮氨酰-sn-3-磷酸乙醇胺(DOPE)、1,2-二油酰-3-二甲基氨丙烷(DODAP)、N-(1,2-二异丙基-3-基)-N,N-二甲基-N-羟乙基溴化铵(DMRIE)和2,2-二苯基-4-二甲基氨基乙基-[1,3]-二氧戊环(XTC)。此外，还可以使用阳离子脂质的商业制剂，例如脂质体(LIPOFECTIN)(包含DOTMA和DOPE，可从GIBCO/BRL获得)和脂质体(Lipofectamine)(包括DOSPA和DOPE，可从GIBCO/BRL获得)。
其它合适的阳离子脂质公开于以下文献中：国际出版物第WO 09/086558、WO 09/127060、WO 10/048536、WO 10/054406、WO 10/088537、WO 10/129709和WO 2011/153493号；美国专利出版物第2011/0256175、2012/0128760和2012/0027803号；美国专利第8,158,601号；以及Love等人,《美国科学院院报(PNAS)》,107(5),1864-69,2010。其它合适的氨基脂质包含具有替代脂肪酸基团和其它二烷基氨基的脂质，包含烷基取代基不同的脂质(例如，N-乙基-N-甲基氨基和N-丙基-N-乙基氨基)。通常，具有较少饱和酰基链的氨基脂质更易于调整大小，尤其是当出于过滤灭菌的目的，复合物的大小必须小于约0.3微米时。可以使用含有碳链长度在C14到C22范围内的不饱和脂肪酸的氨基脂质。也可以使用其它支架来分离氨基脂质的氨基基团和脂肪酸或脂肪烷基部分。
在另外的优选实施例中，根据专利申请PCT/EP2017/064066，LNP包括具有式(III)的阳离子脂质。在此上下文中，PCT/EP2017/064066的公开内容也通过引用并入本文。
在某些实施例中，本发明的氨基或阳离子脂质具有至少一个可质子化或可去质子化基团，使得脂质在等于或低于生理pH值(例如pH 7.4)的pH值下带正电荷，且在第二pH值下为中性，所述第二pH值优选地等于或高于生理pH值。当然，应当理解的是，作为pH的函数的质子的添加或去除是平衡过程，并且对带电或中性脂质的提及是指占优势物种的性质，而不要求所有的脂质都以带电或中性形式存在。不排除在本发明中使用具有多个可质子化或可去质子化基团的脂质或两性离子脂质。在某些实施例中，可质子化脂质的可质子化基团的pKa在约4到约11的范围内，例如，pKa为约5到约7。
阳离子脂质可以包括存在于颗粒中的总脂质的约20mol％到约70或75mol％、或约45到约65mol％、或约20、25、30、35、40、45、50、55、60、65或约70mol％。在另一个实施例中，脂质纳米颗粒包含摩尔占比为约25％到约75％的阳离子脂质，例如，摩尔占比为约20％到约70％、约35％到约65％、约45％到约65％、约60％、约57.5％、约57.1％、约50％或约40％(基于脂质纳米颗粒中脂质的100％摩尔百分比)。在一个实施例中，阳离子脂质与核酸的比率为约3到约15，例如约5到约13或约7到约11。
药物组合物
在一些方面，本发明提供了含有可翻译化合物和药学上可接受的载剂的药物组合物。
药物组合物能够进行局部或全身施用。在一些方面，药物组合物可以具有任何施用方式。在某些方面，可以通过任何途径进行施用，包含静脉内、皮下、肺脏、肌内、腹膜内、皮肤、口服、吸入或鼻腔施用。
本发明的实施例包含含有脂质制剂中的可翻译化合物的药物组合物。
在一些实施例中，药物组合物可以包括选自阳离子脂质、阴离子脂质、固醇、聚乙二醇脂质和前述任何组合的一种或多种脂质。在一些实施例中，含有可翻译化合物的药物组合物包括阳离子脂质、磷脂、胆固醇和聚乙二醇脂质。
在某些实施例中，药物组合物可以基本上不含脂质体。
在另外的实施例中，药物组合物可以包含纳米颗粒。
WO/2015/074085中给出了用于递送本发明的活性分子的脂质和脂质组合物的一些实例，所述文献通过引用整体并入本文。在某些实施例中，脂质是阳离子。在一些实施例中，阳离子脂质包括式II的化合物：
其中R1和R2相同或不同，各自为直链或支链烷基、烯基或炔基，L1和L2相同或不同，各自为具有至少五个碳原子的直链烷基，或与N形成杂环，X1为键或为--CO--O--，通过其形成L2--CO--O--R2，X2为S或O，L3为键或低级烷基，R3为低级烷基，R4和R5相同或不同，各自为低级烷基。本文还描述了式II的化合物，其中不存在L3，R1和R2各由至少7个碳原子组成，R3为乙烯或正丙烯，R4和R5为甲基或乙基，并且L1和L2各由具有至少5个碳原子的直链烷基组成。本文还描述了式II的化合物，其中不存在L3，R1和R2各由至少7个碳原子组成，R3为乙烯或正丙烯，R4和R5为甲基或乙基，并且L1和L2各由具有至少5个碳原子的直链烷基组成。本文还描述了式II的化合物，其中不存在L3，R1和R2各由具有至少9个碳原子的烯基组成，R3为乙烯或正丙烯，R4和R5为甲基或乙基，并且L1和L2各由具有至少5个碳原子的直链烷基组成。本文还描述了式II的化合物，其中是L3为亚甲基，R1和R2各由至少7个碳原子组成，R3为乙烯或正丙烯，R4和R5为甲基或乙基，并且L1和L2各由具有至少5个碳原子的直链烷基组成。本文还描述了式II的化合物，其中是L3为亚甲基，R1和R2各由至少9个碳原子组成，R3为乙烯或正丙烯，R4和R5各为甲基，L1和L2各由具有至少7个碳原子的直链烷基组成。本文还描述了式II的化合物，其中是L3为亚甲基，R1由具有至少9个碳原子的烯基组成且R2由具有至少7个碳原子的烯基组成，R3为正丙烯，R4和R5各为甲基，L1和L2各由具有至少7个碳原子的直链烷基组成。本文还描述了式II的化合物，其中是L3为亚甲基，R1和R2各由具有至少9个碳原子的烯基组成，R3为乙烯，R4和R5各为甲基，L1和L2各由具有至少7个碳原子的直链烷基组成。
在示例性实施例中，所述阳离子脂质包括选自由以下组成的组的化合物：ATX-001、ATX-002、ATX-003、ATX-004、ATX-005、ATX-006、ATX-007、ATX-008、ATX-009、ATX-010、ATX-011、ATX-012、ATX-013、ATX-014、ATX-015、ATX-016、ATX-017、ATX-018、ATX-019、ATX-020、ATX-021、ATX-022、ATX-023、ATX-024、ATX-025、ATX-026、ATX-027、ATX-028、ATX-029、ATX-030、ATX-031、ATX-032、ATX-081、ATX-095和ATX-126或其药学上可接受的盐。
在某些示例性实施例中，所述阳离子脂质包括ATX-002、ATX-081、ATX-095或ATX-126。
在一些实施例中，所述阳离子脂质或其药学上可接受的盐可以呈现为包括纳米颗粒或脂质分子双层的脂质组合物。优选地，所述脂质双层进一步包括中性脂质或聚合物。优选地，所述脂质组合物包括液体介质。优选地，所述组合物进一步包含本发明的可翻译化合物。优选地，所述脂质组合物进一步包括本发明的可翻译化合物和中性脂质或聚合物。优选地，所述脂质组合物包含该可翻译化合物。
在另外的实施例中，所述阳离子脂质包括式III的化合物：
其中R1和R2相同或不同，各自为由1至9个碳组成的直链或支链烷基、由2至11个碳组成的烯基或炔基或胆甾醇基，L1和L2相同或不同，各自为由5至18个碳组成的直链亚烷基或烯基，X1为--CO--O--，通过其形成-L2-CO--O--R2，X2为S或O，X3为--CO--O--，通过其形成-L1-CO--O--R1，L3为键，R3为由1至6个碳组成的直链或支链亚烷基，并且R4和R5相同或不同，各自为氢或由1至6个碳组成的直链或支链烷基；或其药学上可接受的盐。在一个实施例中，X2为S。在另一个实施例中，R3选自乙烯、正丙烯或异丁烯。在又一个实施例中，R4和R5分别为甲基、乙基或异丙基。在又一个实施例中，L1和L2是相同的。在又一个实施例中，L1和L2是不同的。在又一个实施例中，L1或L2由具有7个碳的直链亚烷基组成。在又一个实施例中，L1或L2由具有9个碳的直链亚烷基组成。在又一个实施例中，R1和R2是相同的。在又一个实施例中，R1和R2是不同的。在又一个实施例中，R1和R2各自由烯基组成。在又一个实施例中，R1和R2各自由烷基组成。在又一个实施例中，烯基由单个双键组成。在又一个实施例中，R1或R2由9个碳组成。在又一个实施例中，R1或R2由11个碳组成。在又一个实施例中，R1或R2由7个碳组成。在又一个实施例中，L3为键，R3为乙烯，X2为S，并且R4和R5各自为甲基。在又一个实施例中，L3为键，R3为正丙烯，X2为S，R4和R5各自为甲基。在又一个实施例中，L3为键，R3为乙烯，X2为S，并且R4和R5各自为乙基。
如本领域技术人员所了解的，式II和式III的化合物形成的盐也属于本公开范围内。除非另外说明，否则本文中对式II和式III的化合物的提及应理解为包含对其盐的提及。如本文所用，术语“盐”是指由无机和/或有机酸形成的酸性盐，以及由无机和/或有机碱形成的碱性盐。此外，当式II或式III的化合物同时包含有碱性部分(如但不限于吡啶或咪唑)和酸性部分(如但不限于羧酸)时，两性离子(“内盐”)可以形成并包含在本文所用的术语“盐”中。这些盐可以是药学上可接受的(即，无毒的、生理学上可接受的)盐，尽管其它盐也是有用的。式II或式III的化合物的盐可以通过以下形成：例如，使式Ⅱ或式Ⅲ化合物与一定量(如当量)的酸或碱在盐沉淀的介质中或在水性介质中反应，然后冻干。
示例性的酸加成盐包含乙酸盐、己二酸盐、藻酸盐、抗坏血酸盐、天冬氨酸盐、苯甲酸盐、苯磺酸盐、硫酸氢盐、硼酸盐、丁酸盐、柠檬酸盐、樟脑酸盐、樟脑磺酸盐、环戊烷丙酸盐、双葡萄糖酸盐、十二烷基硫酸盐、乙磺酸盐、富马酸盐、葡萄糖庚烷酸盐、甘油磷酸酯、半硫酸盐、庚烷酸盐、己酸盐、盐酸盐、氢溴酸盐、氢碘酸盐、2-羟乙烷磺酸盐、乳酸盐、马来酸盐、甲烷磺酸盐、2-萘磺酸盐、烟碱酸盐、硝酸盐、草酸盐、果胶酸盐、过硫酸盐、3-苯基丙酸盐、磷酸盐、苦味酸盐、新戊酸盐、丙酸盐、水杨酸盐、琥珀酸盐、硫酸盐、磺酸盐(如本文所述)、酒石酸盐、硫氰酸盐、甲苯磺酸盐(也被称为甲苯磺酸盐)十一烷酸盐等。此外，下列文献中讨论了通常被认为适于从碱性药物化合物中形成药学上有用的盐的酸：例如，S.Berge等人,《药学期刊(J.Pharmaceutical Sciences)》(1977)66(1)1-19；P.Gould,《国际药学期刊(International J.Pharmaceutics)》(1986)33 201-217；Anderson等人,《医学化学实践(The Practice of Medicinal Chemistry)》(1996),学术出版社(Academic Press),纽约；以及《橙皮书(The Orange Book)》(华盛顿特区食品和药物管理局(Food&DrugAdministration)网站)。这些公开内容通过引用并入本文。
示例性的碱性盐包含铵盐、碱金属盐(如钠盐、锂盐和钾盐)、碱土金属盐(如钙盐和镁盐)、含有机碱(例如，有机胺)的盐(如苄星青霉素、二环己基胺、肼类(由N,N-双(脱氢异己基)乙二胺形成)、N-甲基-D-葡糖胺、N-甲基-D-葡糖酰胺、叔丁基胺)和含氨基酸(如精氨酸、赖氨酸)的盐等。可以用如低级烷基卤化物(例如，甲基、乙基、丙基和丁基氯化物、溴化物和碘化物)、硫酸二烷基酯(例如，二甲基、二乙基、二丁基和二烷基硫酸盐)、长链卤化物(例如，癸基、月桂基、肉豆蔻基和硬脂基氯化物、溴化物和碘化物)、芳基烷基卤化物(例如，苄基和苯乙基溴化物)等试剂对碱性含氮基团进行四分之一化。
在本公开的范围内，所有此类酸盐和碱盐都旨在是药学上可接受的盐，并且出于本公开的目的，所有酸盐和碱盐均被视为等同于对应化合物的自由形式。式II或式III的化合物可以以非溶剂化和溶剂化形式存在，包含水合形式。通常，出于本公开的目的，具有药学上可接受的溶剂(如水、乙醇等)的溶剂化形式等同于非溶剂化形式。式II或III的化合物及其盐和溶剂可能以其互变异构形式存在(例如，作为酰胺或亚胺醚)。本文中所有此类互变异构形式都被视为本公开的一部分。
本文所述的阳离子脂质化合物可以与本发明的可翻译化合物组合以形成微粒、纳米颗粒、脂质体或胶束。由颗粒、脂质体或胶束递送的本发明的可翻译化合物可以是气体、液体或固体的形式。所述阳离子脂质化合物和可翻译化合物可以与其它阳离子脂质化合物、聚合物(合成的或天然的)、表面活性剂、胆固醇、碳水化合物、蛋白质、脂质等组合形成颗粒。然后，可以将这些颗粒任选地与药物赋形剂组合以形成药物组合物。
在某些实施例中，所述阳离子脂质化合物相对无细胞毒性。所述阳离子脂质化合物可以具有生物相容性和生物降解性。阳离子脂质的pKa可以在约5.5到约7.5范围内，更优选地在约6.0到约7.0范围内。可以将其设计为具有在约3.0到约9.0之间或在约5.0到约8.0之间的期望pKa。
含有阳离子脂质化合物的组合物可以为30-70％阳离子脂质化合物、0-60％胆固醇、0-30％磷脂和1-10％聚乙二醇(PEG)。优选地，所述组合物为30-40％阳离子脂质化合物、40-50％胆固醇和10-20％PEG。在其它优选实施例中，所述组合物为50-75％阳离子脂质化合物、20-40％胆固醇、5-10％磷脂和1-10％PEG。所述组合物可以含有60-70％阳离子脂质化合物、25-35％胆固醇和5-10％PEG。所述组合物可以含有多达90％阳离子脂质化合物和2-15％辅助脂。制剂可以是脂质颗粒制剂，例如含有8-30％化合物、5-30％辅助脂和0-20％胆固醇；4-25％阳离子脂质、4-25％辅助脂、2-25％胆固醇、10-35％胆固醇-PEG和5％胆固醇-胺；或2-30％阳离子脂质、2-30％辅助脂、1-15％胆固醇、2-35％胆固醇-PEG和1-20％胆固醇-胺；或多达90％阳离子脂质和2-10％辅助脂或甚至100％阳离子脂质。
在一些实施例中，所述一个或多个基于胆固醇的脂质选自胆固醇、聚乙二醇化的胆固醇和DC-Chol(N,N-二甲基-N-乙基羧基酰胺胆固醇)和1,4-双(3-N-油酰氨基-丙基)哌嗪。在示例性实施例中，所述基于胆固醇的脂质是胆固醇。
在一些实施例中，所述一个或多个聚乙二醇化的脂质，即，经过PEG修饰的脂质。在一些实施例中，所述一个或多个经过PEG修饰的脂质包括长度多达5kDa的聚(乙烯)乙二醇链，所述聚(乙烯)乙二醇链共价附接到具有一个或多个长度为C6-C20的烷基链的脂质上。在一些实施例中，经过PEG修饰的脂质是衍生的神经酰胺，如N-辛酰基鞘氨醇-1-[琥珀酰(甲氧基聚乙二醇)-2000]。在一些实施例中，经过PEG修饰的或聚乙二醇化的脂质是聚乙二醇化的胆固醇或二肉豆蔻酰甘油(DMG)-PEG-2K。在示例性实施例中，经过PEG修饰的脂质是聚乙二醇化的胆固醇。
在另外的实施例中，药物组合物可以含有病毒或细菌载体内的低聚化合物。
本公开的药物组合物可以包含本领域已知的载剂、稀释剂或赋形剂。药物组合物和方法的实例描述于以下文献中：例如，《雷明顿药物科学(Remington's PharmaceuticalSciences)》,麦克出版公司(Mack Publishing Co.)(编辑：A.R.Gennaro 1985)和《雷明顿药学技术与实践(Remington,The Science and Practice of Pharmacy)》,第21版(2005)。
药物组合物的赋形剂的实例包含抗氧化剂、悬浮剂、分散剂、防腐剂、缓冲剂、强力剂和表面活性剂。
本发明的试剂或药物制剂的有效剂量可以是足以在细胞中引起可翻译分子的翻译的量。
治疗有效剂量可以是足以达到治疗效果的试剂或制剂的量。治疗有效剂量可以一次或多次分开施用并通过不同的途径进行施用。如本领域所理解的，治疗有效剂量或治疗有效量在很大程度上取决于本发明的药物组合物中所含的治疗剂的总量。通常，治疗有效量足以对受试者产生有意义的益处(例如，治疗、调节、固化、预防和/或改善GSD III)。例如，治疗有效量可以是足以达到期望的治疗和/或预防效果的量。通常，施用给需要治疗的受试者的治疗剂(例如，对AGL进行编码的可翻译低聚物)的量将取决于所述受试者的特征。这些特征包含所述受试者的病状、疾病严重程度、总体健康、年龄、性别和体重。本领域的普通技术人员将能够容易地根据这些和其它相关因素确定合适的剂量。此外，可以任选地使用客观和主观分析来鉴定最佳剂量范围。。
本文提供的方法设想单次和多次施用治疗有效量的本文所述的可翻译化合物(例如，对AGL进行编码的可翻译低聚物)。可以定期施用包括对AGL进行编码的可翻译化合物的药物组合物，着具体取决于受试者病状的性质、严重性和程度(例如，受试者的GSD III疾病状态的严重性和GSD III的相关症状和/或受试者的AGL活性水平)。在一些实施例中，可以每隔一段时间(例如，一年一次、六个月一次、四个月一次、三个月一次、两个月一次、每月一次)、每两周一次、每周一次、每天一次、一天两次、一天三次、一天四次、一天五次、一天六次或者不间断地定期施用治疗有效量的本发明的可翻译化合物(例如，对AGL进行编码的可翻译低聚物)。
在一些实施例中，本发明的药物组合物的配方使其适用于其中所包含的对AGL进行编码的可翻译化合物的缓释。可以以延长的给药间隔将此类缓释组合物方便地施用于受试者。例如，在一个实施例中，每天两次、每天一次或每隔一天向受试者施用本发明的药物组合物。在一些实施例中，每周两次、一周一次、每10天一次、每两周一次、每28天一次、每月一次、每六周一次、每八周一次、每隔一月、每三个月一次、每四个月一次、每六个月一次、每九个月一次或一年一次向受试者施用本发明的药物组合物。本文还设想了被配制用于补给施用(例如，皮下、肌肉注射)以在延长的时间段内递送或释放对AGL进行编码的可翻译化合物的药物组合物。优选地，所使用的缓释手段与对AGL进行编码的可翻译化合物的修饰相结合，以增强稳定性。
在一些实施例中，施用后，治疗有效剂量可能导致AGL的血清或血浆水平为1-1000pg/ml、或1-1000ng/ml，或1-1000μg/ml或更多。
在一些实施例中，施用治疗有效剂量的包括本发明的可翻译分子的组合物可能导致经过治疗的受试者的肝脏AGL蛋白质水平提高。在一些实施例中，施用包括本发明的可翻译分子的组合物导致肝脏AGL蛋白质水平相对于治疗之前的受试者体内的基线AGL蛋白质水平提高5％、10％、20％、30％、40％、50％、60％、70％、80％、90％或95％。在某些实施例中，施用治疗有效剂量的包括本发明的可翻译分子的组合物会导致肝脏AGL水平相对于治疗之前的受试者体内的基线肝脏AGL水平提高。在一些实施例中，肝脏AGL水平相对于基线肝脏AGL水平增加至少约5％、10％、20％、30％、40％、50％、100％、200％或更多。
在一些实施例中，与治疗前的基线水平相比，定期施用治疗有效剂量导致肝脏中AGL的表达增加。在一些实施例中，施用治疗有效剂量的包括本发明的可翻译分子的组合物导致AGL蛋白质水平的表达处于或高于经过治疗的受试者的肝脏中总蛋白的约10ng/mg、约20ng/mg、约50ng/mg、约100ng/mg、约150ng/mg、约200ng/mg、约250ng/mg、约300ng/mg、约350ng/mg、约400ng/mg、约450ng/mg、约500ng/mg、约600ng/mg、约700ng/mg、约800ng/mg、约900ng/mg、约1000ng/mg、约1200ng/mg或约1500ng/mg。
在一些实施例中，施用治疗有效剂量的包括对AGL进行编码的可翻译低聚物的组合物会导致选自丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、碱性磷酸酶(ALP)、肌酸磷酸激酶(CPK)、糖原和极限糊精(即糖原水解产生的低分子碳水化合物)的一个或多个标志物的水平降低。
在一些实施例中，定期施用治疗有效剂量导致生物样品中ALT、AST、ALP和/或CPK水平降低。在一些实施例中，施用治疗有效剂量的包括本发明的可翻译分子的组合物导致生物样品(例如，血浆或血清样品)中的ALT、AST、ALP和/或CPK水平相对于治疗前的基线ALT、AST、ALP和/或CPK水平降低至少约5％、至少约10％、至少约15％、至少约20％、至少约25％、至少约30％、至少约35％、至少约40％、至少约45％、至少约50％、至少约55％、至少约60％、至少约65％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％。在一些实施例中，生物样品选自血浆、血清、全血、尿液或脑脊液。
在某些示例性实施例中，定期施用治疗有效剂量导致血清或血浆样品中的ALT水平降低，例如，以ALT活性/升(U/l)为单位测量。在一些实施例中，施用治疗有效剂量的包括本发明的可翻译分子的组合物导致生物样品(例如，血浆或血清样品)中的ALT水平相对于治疗前的基线ALT水平降低至少约5％、至少约10％、至少约15％、至少约20％、至少约25％、至少约30％、至少约35％、至少约40％、至少约45％、至少约50％、至少约55％、至少约60％、至少约65％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％。在示例性实施例中，施用治疗有效剂量的包括本发明的可翻译分子的组合物导致生物样品(例如，血浆或血清样品)中的ALT水平相对于治疗前的基线ALT水平降低至少约50％。在另一个示例性实施例中，在空腹后测量ALT水平，例如在空腹6小时、8小时、10小时、12小时、18或24小时后测量。
在其它示例性实施例中，定期施用治疗有效剂量导致血清或血浆样品中的AST水平降低，例如，以AST活性/升(U/l)为单位测量。在一些实施例中，施用治疗有效剂量的包括本发明的可翻译分子的组合物导致生物样品(例如，血浆或血清样品)中的AST水平相对于治疗前的基线AST水平降低至少约5％、至少约10％、至少约15％、至少约20％、至少约25％、至少约30％、至少约35％、至少约40％、至少约45％、至少约50％、至少约55％、至少约60％、至少约65％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％。在示例性实施例中，施用治疗有效剂量的包括本发明的可翻译分子的组合物导致生物样品(例如，血浆或血清样品)中的AST水平相对于治疗前的基线AST水平降低至少约50％。在另一个示例性实施例中，在空腹后测量AST水平，例如在空腹6小时、8小时、10小时、12小时、18小时或24小时后测量。
可以使用本领域已知的任何方法进行ALT、AST、ALP和/或CPK水平的测量，例如，使用Liu等人,2014,《分子遗传与代谢(Mol Genet and Metabolism)》111:467-76中所描述的富士Dri-Chem临床化学分析仪FDC 3500。
在其它示例性实施例中，定期施用治疗有效剂量导致生物样品中的糖原水平降低。在一些实施例中，施用治疗有效剂量的包括本发明的可翻译分子的组合物导致生物样品(例如，肝脏样品)中的糖原累积相对于治疗前的基线糖原水平降低至少约5％、至少约10％、至少约15％、至少约20％、至少约25％、至少约30％、至少约35％、至少约40％、至少约45％、至少约50％、至少约55％、至少约60％、至少约65％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％。在一些实施例中，生物样品是选自肝脏、心脏、膈肌、四头肌和腓肠肌的器官的一部分。在示例性实施例中，生物样品是肝脏切片，例如肝细胞切片。
在其它示例性实施例中，定期施用治疗有效剂量导致生物样品中的极限糊精水平降低。在一些实施例中，施用治疗有效剂量的包括本发明的可翻译分子的组合物导致生物样品(例如，肝脏样品)中的极限糊精累积相对于治疗前的基线糊精水平降低至少约5％、至少约10％、至少约15％、至少约20％、至少约25％、至少约30％、至少约35％、至少约40％、至少约45％、至少约50％、至少约55％、至少约60％、至少约65％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％。在一些实施例中，生物样品是选自肝脏、心脏、膈肌、四头肌和腓肠肌的器官的一部分。在示例性实施例中，生物样品是肝脏切片，例如肝细胞切片。在另一个示例性实施例中，定期施用治疗有效剂量导致肝脏样品中的极限糊精水平相对于治疗前的基线极限糊精水平降低至少50％、60％、70％或80％。
在另外的实施例中，定期施用治疗有效剂量会延迟经过治疗的受试者体内的肝纤维化的发作。在一些实施例中，定期施用治疗有效剂量会减缓肝纤维化的发展或减少患GSDIII的受试者体内的肝纤维化的数量。
体内活性剂(例如，对AGL进行编码的可翻译低聚物)的治疗有效剂量可以是约0.001到约500mg/kg体重的剂量。例如，治疗有效剂量可以为0.001-0.01mg/kg体重、或0.01-0.1mg/kg、或0.1-1mg/kg、或1-10mg/kg或10-100mg/kg。在一些实施例中，以约0.1到约10mg/kg体重(例如，约0.5到约5mg/kg、约1至到4.5mg/kg或约2到约4mg/kg)的剂量提供对AGL进行编码的可翻译低聚物。
体内活性剂(例如，对AGL进行编码的可翻译低聚物)的治疗有效剂量可以是至少约0.001mg/kg体重、或至少约0.01mg/kg、或至少约0.1mg/kg、或至少约1mg/kg、或至少约2mg/kg、或至少约3mg/kg、或至少约4mg/kg、或至少约5mg/kg、至少约10mg/kg、至少约20mg/kg、至少约50mg/kg或更多的剂量。在一些实施例中，以约0.1mg/kg、约0.5mg/kg、约1mg/kg、约1.5mg/kg、约2mg/kg、约2.5mg/kg、约3mg/kg、约3.5mg/kg、约4mg/kg、约5mg/kg或约6mg/kg、7mg/kg、8mg/kg、9mg/kg、10mg/kg、15mg/kg、20mg/kg、25mg/kg、50mg/kg、75mg/kg或100mg/kg的剂量提供对AGL进行编码的可翻译低聚物。
除非另外说明，否则本文所示的核酸酶序列是从左到右，5'到3'。
转染
在一些实验中，将可翻译信使分子转染到96孔板中的Hepa1-6或AML12细胞中。所有转染均按照制造说明使用MessengerMAX转染试剂(赛默飞世尔科技(Thermo FisherScientific))。其它合适的细胞系包含HEK293和Hep3B细胞。
体外转染方案实例如下：
在转染前至少8小时，将肝细胞Hepa1-6细胞以每孔5000个细胞平板接种到96孔板中。
置换含有10％FBS和非必需氨基酸的90μL的DMEM培养基，并在96孔板的每一个孔中分装90μL，然后立即开始转染实验。
根据生产商的指示制备MessengerMAX转染试剂(赛默飞世尔科技)可翻译分子复合物。
将10μL所述复合物转移到96孔板中含有所述细胞的孔中。
在期望的时间点后收集培养基，并在每个孔中加入100μL新鲜培养基。在采用标准生产商方案进行AGL的ELISA分析之前，培养基将保持在-80℃。
体内转染方案的实例如下：
用纳米颗粒配制可翻译分子。
通过侧尾静脉中的标准静脉注射将纳米颗粒配制的可翻译分子(1mg/kg)注入BL57BL/c小鼠(4～6周龄)的体内。
在注射后的适当时间，在肝素包被的微离心管中收集约50μL血液。
在4℃下以3,000X g离心10分钟。
将上清液(血浆)转移到新鲜的微离心管中。在采用标准生产商方案进行AGL的ELISA分析之前，血浆将保持在-80℃。
纳米颗粒制剂
可以使用含有mRNA的乙醇/缓冲液中的适当体积的脂质来制备含有mRNA的脂质纳米颗粒。为此，可以使用Nanossemblr微流体装置，然后进行下游处理。例如，为了制备纳米颗粒，可以将期望量的目标mRNA溶解在5mM柠檬酸缓冲液(pH 3.5)中。脂质可以以适当的摩尔比溶解在乙醇中。组成脂质的摩尔百分比可以是，例如，50％离子化脂质、7％DSPC(1,2-二硬脂基-sn-甘油-3-磷酸胆碱；Avanti极性脂质)、40％胆固醇(Avanti极性脂质)和3％DMG-PEG(1,2-二肉豆蔻酰基-sn-甘油，甲氧基聚乙二醇，PEG链分子量：2000；NOF美国公司)。接着，脂质和mRNA溶液可以在微流体装置(Precision NanoSystems)中以1:3(乙醇：水相)的流量比混合。总混流速度可达12毫升/分钟。可以形成脂质纳米颗粒，然后通过在透析装置(Float-a-lyzer，仕必纯(Spectrum Labs))中使用磷酸盐缓冲液进行过夜透析将所述脂质纳米颗粒纯化，然后使用Amicon Ultra-15离心过滤器(默克密里博(MerckMillipore))对其进行浓缩。可以通过动态光散射(ZEN3600，马尔文仪器公司(MalvernInstruments))来确定颗粒大小。可以通过确定将RiboGreen(分子探针(MolecularProbes))添加到LNP浆液(Fi)时通过荧光测量的未封装的mRNA含量来计算“封装”效率；然后，将该值与通过1％Triton X-100(Ft)裂解LNP所获得的总mRNA含量进行比较，其中“封装”百分比＝(Ft-Fi)/Ft×100。包封可以指在纳米颗粒中包含mRNA，无论其形式如何。
In-Cell Western
使用96孔胶原板以适当的密度将细胞接种在DMEM/FBS培养基中。在最佳交汇处，用在转染试剂混合液(MessengerMax和Opti-MEM)中稀释的目标mRNAs转染细胞。将细胞置于CO2培养箱中，并使其生长。在所需时间点，去除培养基，并将细胞在4％新鲜的PFA上固定20分钟。然后去除固定剂，并将细胞在TBST中多次渗透5分钟。当渗透洗涤完成后，将细胞与封闭缓冲液一起孵育45分钟。然后加入一抗，并在室温下孵育1小时。之后，将细胞在TBST中洗涤数次，然后与稀释于封闭缓冲液中且含有CellTag 700染色的二抗一起孵育1小时。最后，将细胞在TBST中洗涤数次，然后在TBS中洗涤最后一次。然后，使用Licor检测系统对板进行成像，并将数据相对于CellTag 700标记的细胞总数归一化。
生成尾PCR产物
根据生产商的说明，含有每个mRNA表达载体的质粒DNA(10ng)可以用于在50μl的PCR反应中与2X KAPA HiFi PCR混合物(KR0370)一起生成poly A尾120PCR产物。然后，可以在来自赛默飞世尔科技的2％凝胶上检查该产物，并根据低分子量阶梯的强度(赛默飞世尔科技，10068-013)进行近似定量，然后用凯杰(Qiagen)PCR纯化试剂盒清洗产物并将其重新悬浮于50ul的水中。
体外合成转录(IVT)
以下方案针对使用NEB HiScribe T7RNA聚合酶试剂进行200μl IVT反应，所述试剂应该产生约1mg RNA。通过解冻单个100mM NTP储备液(ATP、GTP、CTP和UTP核苷酸或经过化学修饰的相应物)并将其合并在一起来根据需要制备2.5X NTP混合物。对于IVT反应，200μl的反应使用约2-4μg的模板。通过移液将10X IVT反应缓冲液、2.5X dNTP混合液、模板DNA和T7RNA聚合酶充分混合，并在37℃下孵育4小时。为了降解DNA模板，用700ul不含核酸酶的水稀释IVT反应，然后在IVT混合液中加入10X DNaseⅠ缓冲液和20ul不含RNase的DNaseⅠ，并在在37℃下孵育15分钟。然后，根据生产商的说明，使用凯杰RNeasy Maxi色谱柱将稀释后(至1ml)且经过DNase处理的反应纯化，最后在不含RNase水中洗脱。然后，通过UV吸光度对纯化的RNA进行定量，其中A260/A280应该约为1.8-2.2，这取决于所用的重悬缓冲液。
IVT RNA的酶促封端
对于酶促封端，可以使用50倍放大版本的NEB的一步封端和2'O-甲基化反应，其适用于处理多达1mg的IVT转录物。根据转录物长度短至100nt的假设，建议在20μl反应中使用10μg RNA。然而，对于转录物来说，较高的底物-反应体积是可以接受的，其长度通常可以更长(约300-600nt)。在开始封端反应之前，将RNA在65℃下变性5分钟，然后快速冷却以消除任何二级构象。对于总共1ml的封端反应，将700μl不含核酸酶的水中的1mg变性RNA与100μl(10X)封端缓冲液一起使用，将50μl(10mM)GTP、50μl(4mM)SAM、50μl(10U/μl)牛痘封端酶和50μl(50U/μl)mRNA端帽2'-O-甲基转移酶混合，并在37℃下孵育1小时。使用不含RNase的水洗脱所得的封端的mRNA，在RNeasy柱上再次纯化，通过分光光度计进行定量。在变性以及快速冷却以去除二级结构后，通过在变性凝胶中每通道运行500ng纯化产物，还可以在该凝胶上显示mRNA。
实例
实例1：参考可翻译分子534。
在本实例中，制备了参考可翻译分子534，并将其用于表达人WT淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)。可翻译分子包括5'端帽(7mGpppG)、TEV的5'UTR、Kozak序列、WT AGL CDS(SEQ ID NO:1)、非洲爪蟾β-珠蛋白的3'UTR和由114As组成的Poly(A)尾区(即，“Poly(A)114尾区”)。参考可翻译分子进一步包括紧邻AGL CDS下游的SEQ ID NO:40的序列。这个参考可翻译分子是用N1-甲基假尿苷代替尿苷合成的。
这个参考可翻译分子的结构细节如下：SEQ ID NO:3的烟草蚀纹病毒(TEV)5'UTR、SEQ ID NO:4的Kozak序列、SEQ ID NO:5的非洲爪蟾β-珠蛋白(XBG)3'UTR和SEQ ID NO:6的Poly(A)114尾。
可以以5'端帽作为m7GpppGm端帽来合成以下实例中的可翻译分子。以下实例中的可翻译分子可以含有5'-UTR(例如，TEV的5'UTR(SEQ ID NO:3))、翻译起始序列(例如，SEQID NO:4的Kozak序列)、SEQ ID NO:40的序列、3'UTR(例如，非洲爪蟾β-珠蛋白的3'UTR(SEQID NO:5))和poly(A)尾(例如，SEQ ID NO:6、SEQ ID NO:38或SEQ ID NO:39的polyA尾)。
实例2：对AGL进行编码的可翻译分子。
在本实例中，制备了翻译效率提高的可翻译分子522-533、546、730-740和1783-1784，并将其用于表达人淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)。这些表达人淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)的可翻译分子表现出适合在用于改善或治疗GSD III的方法中使用的活性。这些可翻译分子包括5'端帽(7mGpppG)、TEV的5'UTR、Kozak序列、AGL CDS和非洲爪蟾β-珠蛋白的3'UTR。可翻译分子522-533、546和1783-1784进一步包括Poly(A)114尾区。可翻译分子730进一步包括Poly(A)100尾区，而可翻译分子731-740进一步包括Poly(A)110尾区。可翻译分子522-533、546、730-740和1783-1784进一步包括紧邻AGL CDS下游的SEQ ID NO:40的序列。开发了分别与546和1783相同的另外两个可翻译分子2258和2259，不同之处在于其包含Poly(A)100尾区而不是Poly(A)114尾区。可以任选地修改1783和2259的编码序列以包含12个核苷酸差异，如SEQ ID NO:45所示。本实例中描述的可翻译分子是用N1-甲基假尿苷代替尿苷合成的。
每个可翻译分子中的AGL CDS由以下序列构成：
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将本实例中的可翻译分子在AML12和C2C12细胞中翻译以产生人AGL。
实例3：基于非洲爪蟾β-珠蛋白3'UTR的翻译增强子。
在本实例中，示出了用于增强可翻译分子的翻译效率的3'UTR序列的结构。
SEQ ID NO:33-37所示的碱基序列是可翻译分子的部分，其在功能上可以与非洲爪蟾β-珠蛋白的3'-UTR相对应。完整的可翻译分子包括以下序列上游的5'端帽(m7GpppGm)、5'-UTR、和编码区(CDS)以及以下序列下游的polyA尾，其中每一个都对应于原生人mRNA的结构。如上所示，可以任选地使用Kozak序列。因此，掺入以下片段的可翻译分子可以具有增强的翻译效率。非洲爪蟾β-珠蛋白基因序列显示在登录号NM_001096347.1中。
实例4：人原代肝细胞中的AGL表达
在本实例中，用密码子优化的mRNA分子546、1783和1784转染人原代肝细胞。转染后6小时、24小时、48小时和72小时，通过In-Cell WesternTM检测AGL蛋白表达。图5示出了与未处理对照(“unt”)比较的mRNA序列的表达。
实例5：野生型小鼠AGL蛋白表达的体内分析
在本实例中，向野生型C57BL/6小鼠注射了用脂质纳米颗粒配制的人AGL mRNA。注射后6小时处死小鼠。取小鼠肝脏活检样品，并分析肝匀浆中人和小鼠AGL蛋白的表达。图6示出了来自各个mRNA分子的人AGL蛋白的异位表达。图7示出了小鼠AGL蛋白水平，表明其与经过处理的小鼠中的小鼠AGL蛋白的内源性表达水平相似。可翻译分子546.7—如图6和7所示—具有与可翻译分子546相同的核碱基序列，但其是用5-甲氧基尿苷代替尿苷合成的，而不是用N1-甲基假尿苷代替尿苷合成的。
实例6：mRNA处理降低了GSD3小鼠体内的糖原累积
在本实例中，用使用ATX2脂质纳米颗粒配制的媒剂或可翻译分子546治疗AGL基因敲除小鼠。经媒剂(“VEH”)处理的基因敲除小鼠的肝脏显示出显著至重度的肝细胞空泡化以及中度至显著的肝细胞内糖原累积增加(图8)。相反，经使用ATX2脂质纳米颗粒配制的可翻译分子546处理的基因敲除小鼠的肝脏仅有轻度至中度的肝细胞空泡化以及仅轻度至中度的肝细胞内糖原累积增加(图9)。根据本实例所示的组织病理学结果，与用媒剂处理的KO小鼠相比，用mRNA处理的基因敲除小鼠的肝脏中的肝细胞空泡化和糖原累积的严重程度似乎有所降低。
实例7：表达AGL的其它可翻译分子
设计了具有经过另外修饰的密码子优化的人AGL编码序列的另外四个可翻译分子1970、1987、SD1和SD2，所述编码序列分别示于SEQ ID NO:41、42、43和44中。可翻译分子1970、1987、SD1和SD2进一步包括5'端帽(7mGpppG)、TEV的5'UTR、Kozak序列、紧邻编码序列下游的SEQ ID NO:40的序列、非洲爪蟾β-珠蛋白的3'UTR和Poly(A)尾区(例如，Poly(A)100、110或114尾区)。这些可翻译分子可以任选地用N1-甲基假尿苷或5-甲氧基尿苷代替尿苷而合成。
本文具体提到的所有出版物、专利和文献均通过引用并入本文用于所有目的。
应理解的是，本发明不限于所描述的特定方法、方案、材料和试剂，因为所述特定方法、方案、材料和试剂可以变化。还应理解的是，本文所使用的术语仅出于描述特定实施例的目的，并非旨在限制将由所附权利要求书涵盖的本发明的范围。
必须注意的是，除非上下文另外明确指示，否则如本文和所附权利要求中所使用，单数形式“一个/种(a/an)”和“所述”包含复数引用。同样，术语“一个(a或an)”、“一个或多个”和“至少一个”在本文中可以互换使用。还应注意的是，术语“包括(comprises/comprising)”、“含有”、“包含”和“具有”可以互换使用。
在不作进一步阐述的情况下，相信本领域的技术人员能够基于上述描述，最大限度地利用本发明。因此，以下具体实施例应被理解为仅仅是说明性的，而不以任何方式限制本公开的其余部分。
在本说明书中公开的所有特征可以以任何组合来组合。本说明书中公开的每个特征都可以由具有相同、等同或相似目的的替代特征代替。
序列表
<110> Ultragenyx Pharmaceutical Inc.
Tachikawa, Kiyoshi
Perez-Garcia, Carlos Gustavo
Chivukula, Padmanabh
Bhaskaran, Hari Prakash
Cobaugh, Christian W.
Daugherty, Sean Christopher
<120> 用于III型糖原贮积病的治疗剂
<130> ULPI-041/01WO 315613-2474
<150> US 62/513,350
<151> 2017-05-31
<160> 45
<170> PatentIn version 3.5
<210> 1
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 共有人AGL编码序列
<400> 1
augggacaca guaaacagau ucgaauuuua cuucugaacg aaauggagaa acuggaaaag 60
acccucuuca gacuugaaca aggguaugag cuacaguucc gauuaggccc aacuuuacag 120
ggaaaagcag uuaccgugua uacaaauuac ccauuuccug gagaaacauu uaauagagaa 180
aaauuccguu cucuggauug ggaaaaucca acagaaagag aagaugauuc ugauaaauac 240
uguaaacuua aucugcaaca aucugguuca uuucaguauu auuuccuuca aggaaaugag 300
aaaaguggug gagguuacau aguuguggac cccauuuuac guguuggugc ugauaaucau 360
gugcuacccu uggacugugu uacucuucag acauuuuuag cuaaguguuu gggaccuuuu 420
gaugaauggg aaagcagacu uaggguugca aaagaaucag gcuacaacau gauucauuuu 480
accccauugc agacucuugg acuaucuagg ucaugcuacu cccuugccaa ucaguuagaa 540
uuaaauccug acuuuucaag accuaauaga aaguauaccu ggaaugaugu uggacagcua 600
guggaaaaau uaaaaaagga auggaauguu auuuguauua cugauguugu cuacaaucau 660
acugcugcua auaguaaaug gauccaggaa cauccagaau gugccuauaa ucuugugaau 720
ucuccacacu uaaaaccugc cugggucuua gacagagcac uuuggcguuu cuccugugau 780
guugcagaag ggaaauacaa agaaaaggga auaccugcuu ugauugaaaa ugaucaccau 840
augaauucca uccgaaaaau aauuugggag gauauuuuuc caaagcuuaa acucugggaa 900
uuuuuccaag uagaugucaa caaagcgguu gagcaauuua gaagacuucu uacacaagaa 960
aauaggcgag uaaccaaguc ugauccaaac caacaccuua cgauuauuca agauccugaa 1020
uacagacggu uuggcuguac uguagauaug aacauugcac uaacgacuuu cauaccacau 1080
gacaaggggc cagcagcaau ugaagaaugc uguaauuggu uucauaaaag aauggaggaa 1140
uuaaauucag agaagcaucg acucauuaac uaucaucagg aacaggcagu uaauugccuu 1200
uugggaaaug uguuuuauga acgacuggcu ggccaugguc caaaacuagg accugucacu 1260
agaaagcauc cuuuaguuac cagguauuuu acuuucccau uugaagagau agacuucucc 1320
auggaagaau cuaugauuca ucugccaaau aaagcuuguu uucugauggc acacaaugga 1380
uggguaaugg gagaugaucc ucuucgaaac uuugcugaac cggguucaga aguuuaccua 1440
aggagagaac uuauuugcug gggagacagu guuaaauuac gcuaugggaa uaaaccagag 1500
gacuguccuu aucucugggc acacaugaaa aaauacacug aaauaacugc aacuuauuuc 1560
cagggaguac gucuugauaa cugccacuca acaccucuuc acguagcuga guacauguug 1620
gaugcugcua ggaauuugca acccaauuua uauguaguag cugaacuguu cacaggaagu 1680
gaagaucugg acaaugucuu uguuacuaga cugggcauua guuccuuaau aagagaggca 1740
augagugcau auaauaguca ugaagagggc agauuaguuu accgauaugg aggagaaccu 1800
guuggauccu uuguucagcc cuguuugagg ccuuuaaugc cagcuauugc acaugcccug 1860
uuuauggaua uuacgcauga uaaugagugu ccuauugugc auagaucagc guaugaugcu 1920
cuaccaagua cuacaauugu uucuauggca uguugugcua guggaaguac aagaggcuau 1980
gaugaauuag ugccucauca gauuucagug guuucugaag aacgguuuua cacuaagugg 2040
aauccugaag cauugccuuc aaacacaggu gaaguuaauu uccaaagcgg cauuauugca 2100
gccaggugug cuaucaguaa acuucaucag gagcuuggag ccaaggguuu uauucaggug 2160
uauguggauc aaguugauga agacauagug gcaguaacaa gacacucacc uagcauccau 2220
cagucuguug uggcuguauc uagaacugcu uucaggaauc ccaagacuuc auuuuacagc 2280
aaggaagugc cucaaaugug caucccuggc aaaauugaag aaguaguucu ugaagcuaga 2340
acuauugaga gaaacacgaa accuuauagg aaggaugaga auucaaucaa uggaacacca 2400
gauaucacag uagaaauuag agaacauauu cagcuuaaug aaaguaaaau uguuaaacaa 2460
gcuggaguug ccacaaaagg gcccaaugaa uauauucaag aaauagaauu ugaaaacuug 2520
ucuccaggaa guguuauuau auucagaguu agucuugauc cacaugcaca agucgcuguu 2580
ggaauucuuc gaaaucaucu gacacaauuc aguccucacu uuaaaucugg cagccuagcu 2640
guugacaaug cagauccuau auuaaaaauu ccuuuugcuu cucuugccuc cagauuaacu 2700
uuggcugagc uaaaucagau ccuuuaccga ugugaaucag aagaaaagga agauggugga 2760
gggugcuaug acauaccaaa cuggucagcc cuuaaauaug caggucuuca agguuuaaug 2820
ucuguauugg cagaaauaag accaaagaau gacuuggggc auccuuuuug uaauaauuug 2880
agaucuggag auuggaugau ugacuauguc aguaaccggc uuauuucacg aucaggaacu 2940
auugcugaag uugguaaaug guugcaggcu auguucuucu accugaagca gaucccacgu 3000
uaccuuaucc cauguuacuu ugaugcuaua uuaauuggug cauauaccac ucuucuggau 3060
acagcaugga agcagauguc aagcuuuguu cagaaugguu caaccuuugu gaaacaccuu 3120
ucauuggguu caguucaacu guguggagua ggaaaauucc cuucccugcc aauucuuuca 3180
ccugcccuaa uggauguacc uuauagguua aaugagauca caaaagaaaa ggagcaaugu 3240
uguguuucuc uagcugcagg cuuaccucau uuuucuucug guauuuuccg cugcugggga 3300
agggauacuu uuauugcacu uagagguaua cugcugauua cuggacgcua uguagaagcc 3360
aggaauauua uuuuagcauu ugcggguacc cugaggcaug gacucauucc uaaucuacug 3420
ggugaaggaa uuuaugccag auacaauugu cgggaugcug ugugguggug gcugcagugu 3480
auccaggauu acuguaaaau gguuccaaau ggucuagaca uucucaagug cccaguuucc 3540
agaauguauc cuacagauga uucugcuccu uugccugcug gcacacugga ucagccauug 3600
uuugaaguca uacaggaagc aaugcaaaaa cacaugcagg gcauacaguu ccgagaaagg 3660
aaugcugguc cccagauaga ucgaaacaug aaggacgaag guuuuaauau aacugcagga 3720
guugaugaag aaacaggauu uguuuaugga ggaaaucguu ucaauugugg cacauggaug 3780
gauaaaaugg gagaaaguga cagagcuaga aacagaggaa ucccagccac accaagagau 3840
gggucugcug uggaaauugu gggccugagu aaaucugcug uucgcugguu gcuggaauua 3900
uccaaaaaaa auauuuuccc uuaucaugaa gucacaguaa aaagacaugg aaaggcuaua 3960
aagguaucau augaugagug gaacagaaaa auacaagaca acuuugaaaa gcuauuucau 4020
guuuccgaag acccuucaga uuuaaaugaa aagcauccaa aucugguuca caaacguggc 4080
auauacaaag auaguuaugg agcuucaagu ccuuggugug acuaucagcu caggccuaau 4140
uuuaccauag caaugguugu ggccccugag cucuuuacua cagaaaaagc auggaaagcu 4200
uuggagauug cagaaaaaaa auugcuuggu ccccuuggca ugaaaacuuu agauccagau 4260
gauaugguuu acuguggaau uuaugacaau gcauuagaca augacaacua caaucuugcu 4320
aaagguuuca auuaucacca aggaccugag uggcuguggc cuauugggua uuuucuucgu 4380
gcaaaauuau auuuuuccag auugaugggc ccggagacua cugcaaagac uauaguuuug 4440
guuaaaaaug uucuuucccg acauuauguu caucuugaga gauccccuug gaaaggacuu 4500
ccagaacuga ccaaugagaa ugcccaguac uguccuuuca gcugugaaac acaagccugg 4560
ucaauugcua cuauucuuga gacacuuuau gauuuauag 4599
<210> 2
<211> 1532
<212> PRT
<213> PatentIn版本3.5
<220>
<223> 共有人AGL氨基酸序列
<400> 2
Met Gly His Ser Lys Gln Ile Arg Ile Leu Leu Leu Asn Glu Met Glu
1 5 10 15
Lys Leu Glu Lys Thr Leu Phe Arg Leu Glu Gln Gly Tyr Glu Leu Gln
20 25 30
Phe Arg Leu Gly Pro Thr Leu Gln Gly Lys Ala Val Thr Val Tyr Thr
35 40 45
Asn Tyr Pro Phe Pro Gly Glu Thr Phe Asn Arg Glu Lys Phe Arg Ser
50 55 60
Leu Asp Trp Glu Asn Pro Thr Glu Arg Glu Asp Asp Ser Asp Lys Tyr
65 70 75 80
Cys Lys Leu Asn Leu Gln Gln Ser Gly Ser Phe Gln Tyr Tyr Phe Leu
85 90 95
Gln Gly Asn Glu Lys Ser Gly Gly Gly Tyr Ile Val Val Asp Pro Ile
100 105 110
Leu Arg Val Gly Ala Asp Asn His Val Leu Pro Leu Asp Cys Val Thr
115 120 125
Leu Gln Thr Phe Leu Ala Lys Cys Leu Gly Pro Phe Asp Glu Trp Glu
130 135 140
Ser Arg Leu Arg Val Ala Lys Glu Ser Gly Tyr Asn Met Ile His Phe
145 150 155 160
Thr Pro Leu Gln Thr Leu Gly Leu Ser Arg Ser Cys Tyr Ser Leu Ala
165 170 175
Asn Gln Leu Glu Leu Asn Pro Asp Phe Ser Arg Pro Asn Arg Lys Tyr
180 185 190
Thr Trp Asn Asp Val Gly Gln Leu Val Glu Lys Leu Lys Lys Glu Trp
195 200 205
Asn Val Ile Cys Ile Thr Asp Val Val Tyr Asn His Thr Ala Ala Asn
210 215 220
Ser Lys Trp Ile Gln Glu His Pro Glu Cys Ala Tyr Asn Leu Val Asn
225 230 235 240
Ser Pro His Leu Lys Pro Ala Trp Val Leu Asp Arg Ala Leu Trp Arg
245 250 255
Phe Ser Cys Asp Val Ala Glu Gly Lys Tyr Lys Glu Lys Gly Ile Pro
260 265 270
Ala Leu Ile Glu Asn Asp His His Met Asn Ser Ile Arg Lys Ile Ile
275 280 285
Trp Glu Asp Ile Phe Pro Lys Leu Lys Leu Trp Glu Phe Phe Gln Val
290 295 300
Asp Val Asn Lys Ala Val Glu Gln Phe Arg Arg Leu Leu Thr Gln Glu
305 310 315 320
Asn Arg Arg Val Thr Lys Ser Asp Pro Asn Gln His Leu Thr Ile Ile
325 330 335
Gln Asp Pro Glu Tyr Arg Arg Phe Gly Cys Thr Val Asp Met Asn Ile
340 345 350
Ala Leu Thr Thr Phe Ile Pro His Asp Lys Gly Pro Ala Ala Ile Glu
355 360 365
Glu Cys Cys Asn Trp Phe His Lys Arg Met Glu Glu Leu Asn Ser Glu
370 375 380
Lys His Arg Leu Ile Asn Tyr His Gln Glu Gln Ala Val Asn Cys Leu
385 390 395 400
Leu Gly Asn Val Phe Tyr Glu Arg Leu Ala Gly His Gly Pro Lys Leu
405 410 415
Gly Pro Val Thr Arg Lys His Pro Leu Val Thr Arg Tyr Phe Thr Phe
420 425 430
Pro Phe Glu Glu Ile Asp Phe Ser Met Glu Glu Ser Met Ile His Leu
435 440 445
Pro Asn Lys Ala Cys Phe Leu Met Ala His Asn Gly Trp Val Met Gly
450 455 460
Asp Asp Pro Leu Arg Asn Phe Ala Glu Pro Gly Ser Glu Val Tyr Leu
465 470 475 480
Arg Arg Glu Leu Ile Cys Trp Gly Asp Ser Val Lys Leu Arg Tyr Gly
485 490 495
Asn Lys Pro Glu Asp Cys Pro Tyr Leu Trp Ala His Met Lys Lys Tyr
500 505 510
Thr Glu Ile Thr Ala Thr Tyr Phe Gln Gly Val Arg Leu Asp Asn Cys
515 520 525
His Ser Thr Pro Leu His Val Ala Glu Tyr Met Leu Asp Ala Ala Arg
530 535 540
Asn Leu Gln Pro Asn Leu Tyr Val Val Ala Glu Leu Phe Thr Gly Ser
545 550 555 560
Glu Asp Leu Asp Asn Val Phe Val Thr Arg Leu Gly Ile Ser Ser Leu
565 570 575
Ile Arg Glu Ala Met Ser Ala Tyr Asn Ser His Glu Glu Gly Arg Leu
580 585 590
Val Tyr Arg Tyr Gly Gly Glu Pro Val Gly Ser Phe Val Gln Pro Cys
595 600 605
Leu Arg Pro Leu Met Pro Ala Ile Ala His Ala Leu Phe Met Asp Ile
610 615 620
Thr His Asp Asn Glu Cys Pro Ile Val His Arg Ser Ala Tyr Asp Ala
625 630 635 640
Leu Pro Ser Thr Thr Ile Val Ser Met Ala Cys Cys Ala Ser Gly Ser
645 650 655
Thr Arg Gly Tyr Asp Glu Leu Val Pro His Gln Ile Ser Val Val Ser
660 665 670
Glu Glu Arg Phe Tyr Thr Lys Trp Asn Pro Glu Ala Leu Pro Ser Asn
675 680 685
Thr Gly Glu Val Asn Phe Gln Ser Gly Ile Ile Ala Ala Arg Cys Ala
690 695 700
Ile Ser Lys Leu His Gln Glu Leu Gly Ala Lys Gly Phe Ile Gln Val
705 710 715 720
Tyr Val Asp Gln Val Asp Glu Asp Ile Val Ala Val Thr Arg His Ser
725 730 735
Pro Ser Ile His Gln Ser Val Val Ala Val Ser Arg Thr Ala Phe Arg
740 745 750
Asn Pro Lys Thr Ser Phe Tyr Ser Lys Glu Val Pro Gln Met Cys Ile
755 760 765
Pro Gly Lys Ile Glu Glu Val Val Leu Glu Ala Arg Thr Ile Glu Arg
770 775 780
Asn Thr Lys Pro Tyr Arg Lys Asp Glu Asn Ser Ile Asn Gly Thr Pro
785 790 795 800
Asp Ile Thr Val Glu Ile Arg Glu His Ile Gln Leu Asn Glu Ser Lys
805 810 815
Ile Val Lys Gln Ala Gly Val Ala Thr Lys Gly Pro Asn Glu Tyr Ile
820 825 830
Gln Glu Ile Glu Phe Glu Asn Leu Ser Pro Gly Ser Val Ile Ile Phe
835 840 845
Arg Val Ser Leu Asp Pro His Ala Gln Val Ala Val Gly Ile Leu Arg
850 855 860
Asn His Leu Thr Gln Phe Ser Pro His Phe Lys Ser Gly Ser Leu Ala
865 870 875 880
Val Asp Asn Ala Asp Pro Ile Leu Lys Ile Pro Phe Ala Ser Leu Ala
885 890 895
Ser Arg Leu Thr Leu Ala Glu Leu Asn Gln Ile Leu Tyr Arg Cys Glu
900 905 910
Ser Glu Glu Lys Glu Asp Gly Gly Gly Cys Tyr Asp Ile Pro Asn Trp
915 920 925
Ser Ala Leu Lys Tyr Ala Gly Leu Gln Gly Leu Met Ser Val Leu Ala
930 935 940
Glu Ile Arg Pro Lys Asn Asp Leu Gly His Pro Phe Cys Asn Asn Leu
945 950 955 960
Arg Ser Gly Asp Trp Met Ile Asp Tyr Val Ser Asn Arg Leu Ile Ser
965 970 975
Arg Ser Gly Thr Ile Ala Glu Val Gly Lys Trp Leu Gln Ala Met Phe
980 985 990
Phe Tyr Leu Lys Gln Ile Pro Arg Tyr Leu Ile Pro Cys Tyr Phe Asp
995 1000 1005
Ala Ile Leu Ile Gly Ala Tyr Thr Thr Leu Leu Asp Thr Ala Trp
1010 1015 1020
Lys Gln Met Ser Ser Phe Val Gln Asn Gly Ser Thr Phe Val Lys
1025 1030 1035
His Leu Ser Leu Gly Ser Val Gln Leu Cys Gly Val Gly Lys Phe
1040 1045 1050
Pro Ser Leu Pro Ile Leu Ser Pro Ala Leu Met Asp Val Pro Tyr
1055 1060 1065
Arg Leu Asn Glu Ile Thr Lys Glu Lys Glu Gln Cys Cys Val Ser
1070 1075 1080
Leu Ala Ala Gly Leu Pro His Phe Ser Ser Gly Ile Phe Arg Cys
1085 1090 1095
Trp Gly Arg Asp Thr Phe Ile Ala Leu Arg Gly Ile Leu Leu Ile
1100 1105 1110
Thr Gly Arg Tyr Val Glu Ala Arg Asn Ile Ile Leu Ala Phe Ala
1115 1120 1125
Gly Thr Leu Arg His Gly Leu Ile Pro Asn Leu Leu Gly Glu Gly
1130 1135 1140
Ile Tyr Ala Arg Tyr Asn Cys Arg Asp Ala Val Trp Trp Trp Leu
1145 1150 1155
Gln Cys Ile Gln Asp Tyr Cys Lys Met Val Pro Asn Gly Leu Asp
1160 1165 1170
Ile Leu Lys Cys Pro Val Ser Arg Met Tyr Pro Thr Asp Asp Ser
1175 1180 1185
Ala Pro Leu Pro Ala Gly Thr Leu Asp Gln Pro Leu Phe Glu Val
1190 1195 1200
Ile Gln Glu Ala Met Gln Lys His Met Gln Gly Ile Gln Phe Arg
1205 1210 1215
Glu Arg Asn Ala Gly Pro Gln Ile Asp Arg Asn Met Lys Asp Glu
1220 1225 1230
Gly Phe Asn Ile Thr Ala Gly Val Asp Glu Glu Thr Gly Phe Val
1235 1240 1245
Tyr Gly Gly Asn Arg Phe Asn Cys Gly Thr Trp Met Asp Lys Met
1250 1255 1260
Gly Glu Ser Asp Arg Ala Arg Asn Arg Gly Ile Pro Ala Thr Pro
1265 1270 1275
Arg Asp Gly Ser Ala Val Glu Ile Val Gly Leu Ser Lys Ser Ala
1280 1285 1290
Val Arg Trp Leu Leu Glu Leu Ser Lys Lys Asn Ile Phe Pro Tyr
1295 1300 1305
His Glu Val Thr Val Lys Arg His Gly Lys Ala Ile Lys Val Ser
1310 1315 1320
Tyr Asp Glu Trp Asn Arg Lys Ile Gln Asp Asn Phe Glu Lys Leu
1325 1330 1335
Phe His Val Ser Glu Asp Pro Ser Asp Leu Asn Glu Lys His Pro
1340 1345 1350
Asn Leu Val His Lys Arg Gly Ile Tyr Lys Asp Ser Tyr Gly Ala
1355 1360 1365
Ser Ser Pro Trp Cys Asp Tyr Gln Leu Arg Pro Asn Phe Thr Ile
1370 1375 1380
Ala Met Val Val Ala Pro Glu Leu Phe Thr Thr Glu Lys Ala Trp
1385 1390 1395
Lys Ala Leu Glu Ile Ala Glu Lys Lys Leu Leu Gly Pro Leu Gly
1400 1405 1410
Met Lys Thr Leu Asp Pro Asp Asp Met Val Tyr Cys Gly Ile Tyr
1415 1420 1425
Asp Asn Ala Leu Asp Asn Asp Asn Tyr Asn Leu Ala Lys Gly Phe
1430 1435 1440
Asn Tyr His Gln Gly Pro Glu Trp Leu Trp Pro Ile Gly Tyr Phe
1445 1450 1455
Leu Arg Ala Lys Leu Tyr Phe Ser Arg Leu Met Gly Pro Glu Thr
1460 1465 1470
Thr Ala Lys Thr Ile Val Leu Val Lys Asn Val Leu Ser Arg His
1475 1480 1485
Tyr Val His Leu Glu Arg Ser Pro Trp Lys Gly Leu Pro Glu Leu
1490 1495 1500
Thr Asn Glu Asn Ala Gln Tyr Cys Pro Phe Ser Cys Glu Thr Gln
1505 1510 1515
Ala Trp Ser Ile Ala Thr Ile Leu Glu Thr Leu Tyr Asp Leu
1520 1525 1530
<210> 3
<211> 129
<212> RNA
<213> 烟草蚀纹病毒
<400> 3
ucaacacaac auauacaaaa caaacgaauc ucaagcaauc aagcauucua cuucuauugc 60
agcaauuuaa aucauuucuu uuaaagcaaa agcaauuuuc ugaaaauuuu caccauuuac 120
gaacgauag 129
<210> 4
<211> 6
<212> RNA
<213> 未知
<220>
<223> Kozak序列
<400> 4
gccacc 6
<210> 5
<211> 158
<212> RNA
<213> 非洲爪蟾属
<400> 5
cuagugacug acuaggaucu gguuaccacu aaaccagccu caagaacacc cgaauggagu 60
cucuaagcua cauaauacca acuuacacuu acaaaauguu gucccccaaa auguagccau 120
ucguaucugc uccuaauaaa aagaaaguuu cuucacau 158
<210> 6
<211> 114
<212> RNA
<213> 未知
<220>
<223> Poly(A) 114尾
<400> 6
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 60
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaa 114
<210> 7
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 7
augggacacu ccaaacagau ccggauacug cugcugaacg agauggaaaa gcuggaaaag 60
acucuauucc ggcucgagca gggauacgag cugcaguucc gccugggccc cacccuacaa 120
gggaaggccg ugaccgucua caccaacuac ccuuucccgg gcgaaacuuu caaccgggag 180
aaguuccggu cccuugacug ggaaaacccg acugagcgcg aggacgacuc ggacaaauac 240
ugcaagcuga accuccagca gucgggcucu uuccaauauu acuucuugca agggaacgag 300
aaguccggug gcggcuacau cgugguggac ccaauuuugc gcgugggggc ugacaaccac 360
gugcugccac uggauugugu gacccugcaa accuuccugg ccaagugccu cggccccuuc 420
gacgaauggg agucgcgccu gagaguggcg aaagagagcg gauacaacau gauucacuuc 480
acgccgcucc aaacgcuggg ucugagccgg ucaugcuacu cacuggcgaa ccagcucgaa 540
cugaaccccg auuucucccg gccaaacagg aaguacaccu ggaacgacgu gggacagcug 600
gucgagaagu ugaagaagga guggaacgug aucuguauca ccgacgucgu guacaaccac 660
accgcugcaa acucgaagug gauccaggaa cauccggaau gugccuacaa ccucgugaac 720
agcccgcacc ugaagccugc cuggguguug gauagggcac uguggagguu cuccugugac 780
guggcagagg gaaaguacaa ggagaagggu auucccgcac ucaucgaaaa cgaucaccac 840
augaauucga ucagaaagau uaucugggag gauaucuucc cuaagcugaa gcugugggag 900
uucuuucaag uugaugugaa caaggcaguc gaacaguuuc ggcggcuguu aacccaagaa 960
aaccgccgcg ugaccaaguc cgauccgaau cagcaucuga ccauaaucca ggacccggaa 1020
uaccgccggu uuggcugcac cguggacaug aacauugccc ugacuaccuu uaucccgcau 1080
gauaagggcc ccgccgcuau cgaagaaugc ugcaacuggu uccacaagag gauggaggaa 1140
cugaacuccg aaaagcauag gcucauuaac uaccaccagg aacaggcagu gaacugccug 1200
cuggggaacg uguucuacga gcgacuggcu ggacacggac cgaaguuagg acccgugaca 1260
aggaagcacc cacuggucac uagauacuuc accuucccau uugaggaaau cgacuucuca 1320
auggaggagu cgaugaucca cuugccuaac aaggccugcu uucucauggc acauaacgga 1380
ugggucaugg gcgacgaucc ccuacggaau uuugcagaac caggcagcga ggucuaccuu 1440
cggcgggaac ugauuugcug gggcgacucc gucaagcugc gcuacggcaa caagccugag 1500
gacugucccu accuuugggc acacaugaag aaguacacug aaauuacugc gacauacuuc 1560
caaggagucc gcuuagauaa uugucacucc accccgcugc auguggcgga guacaugcug 1620
gaugccgcaa gaaaccucca gccgaaucuc uacgugguag cagagcuguu caccgggagc 1680
gaggaccugg acaauguguu ugucacccgg cuggggaucu ccucccugau ccgggaggcc 1740
auguccgccu acaacucaca cgaggagggg agacuggugu accgcuacgg aggagaaccc 1800
gugggcagcu uugugcagcc uugccuccgg ccgcugaugc ccgcgauugc gcaugcucug 1860
uucauggaua ucacucacga uaacgagugc cccauugugc acagauccgc cuacgacgcc 1920
cuuccuucca caaccaucgu guccauggca ugcugcgccu ccggcuccac ucgggguuac 1980
gaugagcugg ugccacacca gauuuccgug guguccgaag aacgcuucua caccaagugg 2040
aacccggaag cucugccguc aaacaccgga gaagugaacu uccaguccgg gaucaucgca 2100
gcgcgcugug cuauuagcaa gcugcaccag gagcugggag ccaagggguu cauccagguc 2160
uauguggacc aggucgauga ggauaucguc gcugucacga gacacagccc gucuauccau 2220
caaagcgucg uggccguguc ccggacugcg uuccggaacc cuaaaaccuc auucuauucc 2280
aaagaggugc cccagaugug caucccugga aagaucgaag aagucgugcu ggaagcccgg 2340
accaucgagc ggaacaccaa gccguacagg aaggacgaaa acuccaucaa ugguaccccu 2400
gacauuaccg uggaaaucag agaacauauc cagcugaacg aguccaagau cgugaagcag 2460
gccggcgugg cgaccaaggg ucccaacgag uacauucagg aaaucgaguu ugaaaaccug 2520
ucccccggaa gcgugaucau uuuccgggug ucccuggacc cgcaugcgca agucgcuguc 2580
ggaauucugc ggaaucaccu cacccaauuc ucgccgcauu ucaagagugg uucccuggcg 2640
guggauaaug ccgauccgau ccugaagauu cccuucgcgu cccuggcauc gagacucacc 2700
cuggcggagu ugaaccagau ucuguaccgc ugcgaauccg aggaaaagga ggacggaggc 2760
gguugcuacg acauccccaa cugguccgca cuuaaguacg cagggcugca gggucuuaug 2820
agcgugcugg cagaaauucg cccuaagaac gauuugggac accccuucug caacaaccuc 2880
cgguccggag acuggaugau cgauuacgug ucgaacagac ugauuucgag auccggcacc 2940
auugccgagg ucggaaagug gcuucaggcc auguucuucu accugaagca gaucccgaga 3000
uaccugauuc ccugcuacuu cgacgcaauc cugaucgggg cguauaccac ucuucuggac 3060
acugccugga agcagauguc cagcuucgug caaaacggau ccaccuucgu caagcaucuu 3120
agccugggcu cagugcaguu gugcggagug ggaaaauucc cuagccuccc uauucuuuca 3180
ccggcgcuga uggacgugcc uuaccgccug aacgaaauca ccaaagagaa ggagcagugu 3240
ugcgugucgc uggccgcggg ucugccgcau uucuccuccg gcaucuuccg gugcugggga 3300
agggacaccu ucaucgcucu gaggggaauc cugcugauua ccgggcgcua cguggaagcu 3360
cggaacauca uccuggccuu cgcgggaacu cugcgccacg gccugauucc aaacuugcuu 3420
ggcgaaggca ucuacgcgcg cuacaacugc cgcgacgcgg ucugguggug gcuccagugc 3480
auucaagacu acugcaagau ggugccaaac ggccuggaca uccugaagug cccggugucg 3540
agaauguacc ccaccgacga uucugcgccc cugccggccg guacucuuga ccaaccucug 3600
uucgaaguga uccaggaagc aaugcagaag cacaugcagg gcauucaguu ccgggagcgc 3660
aacgcagggc cgcaaaucga caggaacaug aaggacgaag gauucaacau caccgcggga 3720
guggacgaag agacuggcuu cgucuacggu ggaaaucggu ucaacugcgg gaccuggaug 3780
gacaagaugg gcgaaucaga ccgagcccgc aaccgcggaa ucccugccac cccccgggau 3840
gggagcgccg uggagauugu gggacugagc aagagcgcug ugcgcuggcu gcuggaguug 3900
agcaagaaga acauuuuccc cuaucacgaa gugaccguga agcggcacgg aaaagcuauc 3960
aaaguguccu acgacgagug gaacagaaag auccaggaca acuucgagaa gcuguuccac 4020
guguccgagg acccgucgga cuugaaugag aagcacccua accucgugca caagcgggga 4080
aucuacaagg acagcuacgg agcauccucg ccuuggugcg acuaucagcu gaggcccaac 4140
uucacuaucg caaugguggu ggccccagaa cuguucacua ccgaaaaggc cuggaaggca 4200
cuggagaucg cagagaaaaa gcugcugggc ccucugggca ugaaaacccu ggaccccgac 4260
gacauggugu acugcgggau cuacgauaac gcucuugaca augacaacua caaccuggcu 4320
aagggauuca acuaucacca gggccccgag uggcuguggc cgaucgguua cuuccugcgc 4380
gccaagcugu acuuuucccg gcugaugggc ccugagacua ccgcaaagac gaucgugcug 4440
gucaagaacg ugcugucacg gcacuacgug caucuggaac ggagcccgug gaagggguug 4500
cccgaacuga ccaacgaaaa cgcgcaguac uguccguucu cgugcgaaac ucaggccugg 4560
uccaucgcca cuauccucga aacucucuac gaccuguag 4599
<210> 8
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 8
augggacaua gcaaacaaau uaggauccuc cugcugaacg aaauggaaaa acuggaaaag 60
acgcuguuuc ggcuggaaca gggcuacgaa cuccaguuuc gccucgggcc aacgcugcaa 120
gggaaagcag ugaccgugua uacaaacuac ccauucccug gagagacauu caauagggag 180
aaguuuagaa gcuuagacug ggaaaaucca accgaacgag aagaugacag cgauaaguau 240
ugcaagcuga aucugcaaca guccggaagu uuucaguacu acuuuuugca agggaacgaa 300
aagagcggcg gagguuauau ugugguggau ccaauuuuga gaguuggggc cgacaaccau 360
guacugccuc uugacugugu gacacugcaa acguuccugg ccaagugccu gggcccguuu 420
gaugaguggg aaagccguuu gcgcguggcc aaggaauccg gguacaacau gauccauuuu 480
accccacugc aaaccuuagg ccugucaagg agcuguuaca gucuggccaa ccagcuggaa 540
cugaaucccg acuucucccg gccaaauagg aaguacacgu ggaacgacgu uggccagcua 600
guggagaagu ugaagaagga guggaacguu auuugcauca cugacgucgu guauaaucac 660
acugccgcua auagcaaaug gauccaggaa cauccagagu gugccuauaa ccuggucaac 720
agcccgcauc ugaagccagc augggugcug gaccgggcgu uguggcgguu cucgugugac 780
guggccgaag gaaaguauaa ggagaagggu auuccggcuc ugauagagaa cgaccaucau 840
augaauucca ucaggaagau cauuugggag gacaucuuuc caaaguugaa gcugugggag 900
uucuuucaag uggaugugaa caaggccgua gaacaguuuc gccgccuacu aacccaggaa 960
aauaggagag ucacaaaguc cgaucccaac caacaucuga ccauaauuca ggaccccgaa 1020
uaucgucgau uuggcugcac cguggauaug aacaucgcuu ugacaacuuu cauaccacac 1080
gacaaggguc cagccgccau ugaagagugu uguaacuggu uucacaaacg aauggaggag 1140
uugaacagug aaaagcaccg ccugaucaau uaucaucagg agcaggccgu gaacuguuug 1200
cuggggaaug uguucuauga gaggcucgcu ggucauggcc cuaagcuggg uccuguaaca 1260
agaaagcauc cauuagugac gcgcuacuuu acauucccau uugaggagau cgacuuuucg 1320
auggaagagu ccaugauuca ucuccccaac aaggccuguu uucugauggc ccacaacggc 1380
uggguuaugg gggaugaucc guugagaaau uuugcggaac cagguucaga aguguaucug 1440
cgucgggagc ugauuuguug gggcgacucc gugaagcucc gcuaugggaa caagccugag 1500
gauuguccuu aucugugggc gcauaugaag aaguauacag agaucacugc cacauauuuu 1560
cagggcguca ggcuggacaa uugucauagc accccgcuuc auguggcuga guauaugcug 1620
gacgcagcaa ggaaucugca gcccaaucug uauguggugg cggaacuguu uaccgggucc 1680
gaggaccugg acaauguauu uguaacccga uugggcaucu ccagccugau uagggaagca 1740
augagugcau acaacucaca cgaggagggg cggcugguuu aucgauaugg gggggaaccu 1800
gugggcagcu uuguacagcc augucuccgg ccguugaugc cugccauugc ucaugcgcuc 1860
uuuauggaua uaacacauga caacgaaugu ccaaucguuc auaggagugc uuacgacgcc 1920
cugccgagca caacgaucgu guccauggcc uguugugcaa guggcagcac acguggcuau 1980
gacgaauugg uaccccacca aauuagcgug guuuccgagg agagguucua uacaaagugg 2040
aauccagagg cuuugcccuc gaacaccgga gaggucaacu uucaaucugg cauaauugcc 2100
gcucgcugcg ccauaucuaa guugcaucag gaacugggcg cgaagggguu uauacaaguc 2160
uacguugacc aaguggauga ggacaucguu gcugucacgc ggcacucacc uaguauucau 2220
caauccgugg uagcaguguc ucggacggcc uuuagaaacc caaagacauc auuuuacucg 2280
aaagaagugc cucaaaugug uauaccuggc aaaauugaag aggugguccu ggaagcccgc 2340
acgauagaga ggaauacuaa gccguauaga aaggaugaaa auucuaucaa cggcacuccg 2400
gauauuacag uagaaaucag agagcacauu caacuuaaug agucuaagau uguuaagcag 2460
gcugguguug caaccaaagg gccaaacgag uacauccagg agauugaauu ugaaaaucug 2520
ucaccgggcu ccgugaucau cuuuagagua ucuuuggauc cacaugcuca agucgcuguu 2580
ggaauccuga gaaaccaucu gacacaauuu uccccacauu uuaagagcgg cagccuggcc 2640
guggacaacg cagacccaau ccugaagauc ccauuugcau cccuggcuuc ccgccugacu 2700
cuggcugagc ugaaccagau cuuguaucgc ugugaaucug aagaaaaaga ggauggcgga 2760
ggcuguuaug acaucccaaa uuggagcgcc cugaaauaug ccggacucca aggccugaug 2820
uccguucugg ccgagauuag gccaaagaau gaucugggac acccauuuug uaacaacuug 2880
cggucuggag acuggaugau cgacuauguc agcaaccgcc ugauaagcag auccgguacu 2940
aucgcugaag ucggaaaaug gcugcaggcu auguuuuucu auuugaagca gauuccgaga 3000
uauuugaucc ccuguuauuu ugaugccauu cugauuggug cuuauacuac ccugcuggau 3060
accgcuugga aacagaugag uagcuuuguc caaaauggcu ccaccuucgu aaagcaucug 3120
ucgcugggcu ccgugcaauu guguggcguc gggaaguucc cuagccugcc aauccugucc 3180
ccugcucuga uggacgugcc uuaucgccug aacgagauua cgaaggaaaa ggaacagugu 3240
ugcgugucac ucgcugcugg gcugccacac uuuucuucug gaauuuuucg guguuggggg 3300
cgggacaccu uuauugcccu gaggggcauu cugcugauua ccggccgcua ugucgaggcu 3360
aggaacauua uccuggcuuu cgcggguacc cuucggcacg gacucauacc caaccuccug 3420
ggagaaggaa ucuaugcacg guauaacugu cgggacgcag uuugguggug guugcagugu 3480
auucaagauu auugcaagau ggucccgaac ggacuggaca uacugaagug cccagugucc 3540
cggauguauc caacugacga cuccgcacca cugcccgcug ggacacuuga ccagccauug 3600
uuugaaguua uucaggaagc uaugcagaag cauaugcagg gaauucaguu uagggagaga 3660
aacgcuggac cccagauuga ccguaacaug aaggaugagg gguuuaacau uaccgcggga 3720
guggacgaag agacuggcuu cgucuauggu ggcaaucggu ucaacugcgg caccuggaug 3780
gacaaaaugg gcgaaagcga uagagcucga aauaggggca ucccagcaac accacgggau 3840
ggcucugccg uggagauugu gggccugucu aagagcgcag uucggugguu guuggaacuc 3900
agcaagaaga acauuuuucc auaccaugaa guuacaguga agcggcaugg gaaggccauc 3960
aagguuucuu acgacgaaug gaaucggaag auccaggaua acuucgagaa acuguuucau 4020
guauccgagg acccuucuga ccugaacgaa aagcauccaa auuugguaca caaaagggga 4080
aucuauaagg auaguuaugg agccagcagc ccuuggugcg acuaucagcu acgaccaaac 4140
uucacuauug ccaugguagu agcaccagaa cucuuuacua ccgaaaaggc uuggaaggca 4200
cuggagaucg ccgagaagaa gcuguuaggu ccacugggca ugaaaacccu ggaccccgac 4260
gacaugguuu acuguggcau cuaugacaac gcucuggaca augacaacua uaaucuggcc 4320
aagggcuuca acuaccacca gggcccagag uggcuauggc caauuggcua uuuucugcgg 4380
gcgaagcugu acuuuucacg auugaugggc ccagagacaa ccgcuaagac cauaguauug 4440
gucaagaacg ugcugagucg gcacuauguc caccuggaaa ggagccccug gaaggggcug 4500
cccgagcuga ccaacgaaaa ugcacaguau uguccguuuu caugcgaaac ccaggcuugg 4560
uccaucgcga cgauccugga aacccucuau gaucuguag 4599
<210> 9
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 9
augggacacu caaaacagau aaggauccug cugcugaacg aaauggagaa acuggaaaag 60
acccuguucc ggcucgaaca gggauaugaa cuccaguuua ggcugggccc aacucugcaa 120
ggaaaagcug ugaccgucua uacuaacuau ccauuucccg gggaaacguu uaaccgagag 180
aaguuccgau ccuuggacug ggaaaauccc acagagcggg aggaugauuc ggacaaguau 240
ugcaagcuga aucugcaaca aucgggcagc uuucaguacu auuuucuaca aggaaacgaa 300
aagagcggcg ggggauacau ugugguggac ccaauccuga ggguuggggc agacaaucac 360
guacugccuu uggacugugu gacauugcaa acauuccugg caaagugccu gggccccuuc 420
gaugaauggg agucacggcu gcgaguggcu aaggaaucug gcuauaauau gauucacuuu 480
acaccauuac aaacgcuggg ccugagcaga aguugcuacu cgcuggcuaa ucaguuggag 540
uugaacccgg acuuuagccg ucccaaccgg aaguauacau ggaaugaugu uggccaacuc 600
guggagaagc ugaagaagga guggaacgug aucuguauua ccgaugucgu guauaaucau 660
acagccgcga acuccaagug gauucaggag cacccagaau gugcauauaa cuuggucaau 720
agcccccacc ugaagccggc cuggguacug gaucgggccc uguggcgguu uucuugugac 780
guggcagaag gaaaguauaa ggaaaagggg aucccagcac ugauagagaa cgaccaucau 840
augaacagca uuaggaagau cauuugggag gacauuuucc caaaguugaa gcucugggag 900
uuuuuccaag uugacgucaa caaggccgug gaacaauuuc gaaggcugcu gacacaggaa 960
aacagacgug uaacgaaguc cgaccccaau cagcauuuga cgaucauaca ggauccugag 1020
uauaggcgcu uuggcuguac ggucgacaug aauauugcuc ugaccaccuu uaucccucau 1080
gacaaggggc cagccgccau cgaagagugu uguaauuggu uccacaagcg cauggaagag 1140
uugaauuccg aaaagcauag guugaucaac uaccaccagg aacaggcagu gaacugucug 1200
cugggcaacg uguuuuauga gcggcuggcc ggucauggcc cuaagcuggg accagugacc 1260
aggaagcauc cacucguaac cagauauuuu accuucccgu uugaggagau agauuuuagc 1320
auggaagaau ccaugauuca ccuaccgaac aaggcuuguu uucugauggc ucacaacggu 1380
uggguuaugg gcgaugaucc ccugcgcaac uuugcggaac cgggcucuga gguguacuug 1440
agaagggagc ugauauguug gggggacucc gucaaacugc gcuaugggaa uaagccugaa 1500
gauugcccuu aucugugggc ccauaugaag aaguauaccg agauuaccgc gacauauuuu 1560
caaggagugc ggcuggacaa cugccacagc acgccgcugc auguggcaga guauaugcug 1620
gacgcagccc gcaaucugca gccaaaucug uauguugugg cugagcuguu uacgggcucc 1680
gaggaccucg acaacguuuu cguaacccgg cugggcauca gcagccugau ccgggaagcu 1740
augucagcgu auaauagcca ugaggagggc agacuggugu acagauaugg uggcgaacca 1800
guuggcagcu uugugcagcc cuguuugcgc ccucugaugc ccgccaucgc acaugcuuug 1860
uucauggaca ucacgcauga uaacgaaugu cccauuguac aucgcuccgc cuaugacgcu 1920
uugccaucca caacaaucgu guccauggcu uguugugcaa gcggcagcac caggggauau 1980
gacgaauugg uuccgcauca aaucagcgug guaucagagg aaagguuuua uacaaagugg 2040
aauccugaag cccugccauc caacaccggc gaagugaacu uucagucggg uaucauugcc 2100
gcgcguugcg cuauuagcaa acugcaucag gagcugggug cuaaaggcuu uauccaaguu 2160
uauguggauc aaguagacga agauauuguu gccguaacca gacauagccc cagcauucau 2220
caauccgugg uugcuguguc ccggacggcc uucaggaacc cuaaaacauc cuuuuauucc 2280
aaggaagucc cacagaugug uauuccggga aagauagagg aagugguuuu ggaggcucgc 2340
acgauugagc ggaacaccaa gccauauagg aaggacgaga acucgaucaa cggcacgccc 2400
gacauuaccg uugaaauucg cgagcacauu cagcugaacg aaucgaaaau ugucaagcag 2460
gccggcguag cgaccaaggg uccaaacgag uauauccagg aaaucgaguu ugagaaucug 2520
ucgccugggu ccguaaucau uuuuagaguc agccuagacc cucacgcuca aguggccgua 2580
gggauccuga ggaaucaucu gacgcaguuu ucaccccauu uuaaguccgg cagccuggca 2640
guggauaacg ccgaccccau ccugaaaauu cccuuugcuu cccuggccuc gcgccugaca 2700
cuggcagaau ugaaucagau acuguaucgu ugcgagagcg aggagaagga ggacggaggg 2760
ggguguuaug auaucccgaa cugguccgcu cugaaauaug cgggauugca gggcuugaug 2820
aguguacugg ccgaaauuag acccaagaac gaucugggcc auccguucug uaacaaucug 2880
cgauccggcg auuggaugau ugauuauguc agcaaccggu ugauaagccg gagugggaca 2940
auugcagaag ucggaaagug guugcaagcc auguucuuuu accugaagca gaucccucga 3000
uaucugauac cauguuauuu cgaugccauu cugaucgggg ccuauacaac ucuguuagac 3060
acugcuugga aacagauguc uagcuucgug caaaacggcu caacauuugu uaagcacuug 3120
agccuggggu cugugcaguu guguggcgua ggaaaguuuc ccucacugcc cauccugucg 3180
cccgcccuga uggacgugcc cuaccgccuc aacgaaauua ccaaggagaa agagcagugu 3240
uguguuagcu uggcugccgg uuugccgcau uuuucaagcg gaaucuuccg augcugggga 3300
cgcgauacgu ucauagcccu gagggguauu cugcugauua cgggcagaua cguagaggcu 3360
cggaauauua uccuggccuu cgccggcacg cugcggcaug ggcugauucc gaacuugcug 3420
ggagagggca ucuacgcccg guacaacugc agggacgcug ugugguggug gcugcagugc 3480
auccaggacu auugcaaaau ggugccaaac ggccuugaca uacugaagug uccugucucu 3540
cggauguauc caacggauga cuccgcaccc cugcccgcug gaacccugga ucaaccccug 3600
uuugaaguga uacaggaagc aaugcagaag cacaugcagg gaauucaguu uagagagaga 3660
aaugcaggcc cccaaaucga cagaaacaug aaagaugaag gcuuuaacau cacggcugga 3720
guggacgaag aaacuggguu uguguauggc gggaauaggu uuaacugugg gacguggaug 3780
gacaagaugg gcgaauccga uagagcgcgg aacaggggaa ucccggcuac accccgggac 3840
gguagugcug uggagauugu cggcuuaucc aaauccgccg ugcgcuggcu gcuggagcug 3900
uccaagaaga acaucuuucc uuaccaugaa gugaccguga agcggcaugg aaaggccauc 3960
aaagucuccu acgacgaaug gaauaggaag auucaggaca auuuugagaa gcuguuucau 4020
guguccgagg aucccagcga ccugaacgag aagcacccua auuuggugca uaagcggggc 4080
aucuauaagg acuccuacgg cgcuaguucg ccuuggugcg acuaucagcu gcggccaaac 4140
uuuacgauug cgaugguggu agcccccgaa uuguuuacga cggaaaaggc uuggaaggcc 4200
cuggagaucg cagaaaagaa gcugcugggg cccuugggca ugaaaacgcu ggaccccgac 4260
gauaugguuu auuguggcau cuacgacaac gcccucgaca augacaauua caaccuggca 4320
aaggguuuua acuaucauca ggguccugaa uggcuguggc caauuggcua cuuuuugcgg 4380
gccaagcugu acuuuucacg gcugauggga ccugaaacca ccgcuaagac cauuguacug 4440
gucaagaacg ugcugagcag gcauuacgug caucuggaaa gaagcccgug gaagggucug 4500
ccagaguuga cgaacgagaa cgcgcaguau uguccuuucu cuugugaaac gcaggcuugg 4560
aguauugcaa ccauucugga aacucuuuau gaccuguag 4599
<210> 10
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 10
augggccacu caaagcagau uagaaucuua cuccugaacg agauggaaaa gcuggaaaag 60
acccuguucc gccuggaaca gggauaugag cugcaguucc ggcuuggacc aacccuccaa 120
ggaaaggccg ugaccgugua cacgaacuac ccauucccgg gcgaaaccuu caacagagag 180
aaguuccgga gccuggacug ggaaaacccg accgaaaggg aagaugauag cgacaaguac 240
ugcaagcuga acuugcaaca guccggauca uuccaguacu acuuucugca agggaacgag 300
aagucgggcg gcgguuacau cgugguggac ccuauucugc gcgugggagc cgacaaccac 360
guguugccuc uggacugugu cacccugcaa acuuuccugg ccaagugccu ggggccauuc 420
gaugaguggg aaucgcggcu gcggguggca aaagaguccg guuacaacau gauccacuuc 480
accccacucc aaacccuggg ccuguccagg ucgugcuacu cgcuggccaa ccagcuggaa 540
cugaacccgg auuucucgcg gccuaacaga aaguauaccu ggaaugacgu cggacagcuc 600
gucgaaaagc ugaagaagga auggaacgua aucuguauca ccgaugucgu guauaaccac 660
accgcugcca acuccaagug gauccaggaa caccccgagu gcgcuuacaa cuuggugaac 720
ucgccccacc ugaaaccggc uugggugcug gaccgggccc uguggcgguu cagcugcgau 780
guggcugagg gaaaguauaa ggagaagggu aucccagcgc ucauugaaaa cgaccaccau 840
augaacucga uccgcaagau cauuugggag gauauuuucc caaaguugaa acugugggag 900
uuuuuccaag uggacgugaa caaggcgguc gaacaguuca gacggcuguu gacucaggaa 960
aaucggaggg ucaccaaguc ggaucccaac cagcaccuga ccauaaucca ggauccagaa 1020
uaccggcggu ucggcugcac uguggacaug aacauugccc ugaccacuuu uaucccacac 1080
gacaagggcc cggccgccau cgaagagugc ugcaacuggu uccacaagcg gauggaggaa 1140
cugaacucgg agaagcaccg ccugaucaac uaccaccagg agcaggcggu uaacugucug 1200
uugggaaacg uguucuacga gcgguuggcc ggccacggcc cuaagcuggg cccggucacc 1260
cggaagcacc cccuggucac ccgcuacuuc accuuuccau ucgaggagau cgacuucagc 1320
auggaagaau cgaugaucca ucugccgaac aaggccugcu uccugauggc ucacaacgga 1380
ugggugaugg gcgacgaucc ucugagaaac uuugccgagc cgggcucgga gguguaccug 1440
aggagggagc ugaucuguug gggggacagc gugaagcuga gauacggaaa caagccagag 1500
gacugcccgu accugugggc ccauaugaag aaguacaccg agauuaccgc aacauacuuc 1560
caaggggucc ggcuggacaa uugccacucg acuccccugc acguggcgga guacaugcug 1620
gacgcggcca ggaaccucca gcccaaccuc uacgucgugg ccgagcuguu cacugguucc 1680
gaggaccugg auaacguguu cgugaccaga cuggggaucu ccucacugau ucgcgaagcc 1740
auguccgcgu auaacucgca ugaagagggc cgccuggugu accgcuacgg aggcgaaccu 1800
gugggcagcu ucgugcaacc gugucugcgg ccucugaugc cugccauugc gcacgcccug 1860
uucauggaca ucacccacga caacgaaugc cccaucgugc accgguccgc cuacgaugcc 1920
cucccuucga cuacuaucgu guccauggcu ugcugcgcgu ccggcucgac ccgcggcuac 1980
gaugagcucg ugccgcauca gauuagcgug guguccgagg aaagguucua caccaagugg 2040
aauccagaag cgcugccgag caacaccggg gaggucaacu uccagucggg aaucaucgcc 2100
gcccgcugug ccaucuccaa gcugcaccag gaacugggcg cgaaggguuu cauccaaguc 2160
uacgucgauc aggucgacga ggacaucgug gccgugacuc ggcauucccc gagcauucac 2220
caguccgugg uggccguguc gcgcaccgcc uuccgcaacc cuaagaccag cuucuauuca 2280
aaagaagugc cgcagaugug caucccugga aagaucgagg aggugguccu ggaagcgcgg 2340
acuaucgaaa ggaacaccaa gccauaccgc aaggacgaga acuccaucaa cgguaccccg 2400
gacaucacug uggagauccg cgagcauauu caacugaacg aguccaagau cgugaaacag 2460
gccggagugg caaccaaggg accgaacgag uauauccagg aaauugaguu cgagaaccuc 2520
uccccgggaa gcgugauuau cuuccgcgug ucccuggacc cacacgccca aguggccguc 2580
ggcaucuugc ggaaccaccu gacucaguuc uccccgcauu ucaaguccgg aagccuggcg 2640
guggacaacg cggacccaau ccugaagaua cccuucgccu cacuggccuc gcgccugacu 2700
uuggccgagu ugaaucagau ccuguaccgc ugcgaauccg aagaaaagga ggacggugga 2760
ggcuguuacg acaucccuaa cugguccgca cugaaauacg caggacugca gggacugaug 2820
uccguccucg cugaaaucag accgaagaac gaucucggcc acccauucug caacaaccug 2880
agauccggcg auuggaugau ugauuacgug ucgaaccgac ugaucucccg cucggguacu 2940
auugcggaag ucggaaaaug gcugcaggcc auguucuucu accugaagca gaucccacga 3000
uaccuuaucc cuugcuacuu ugacgcgauu cugauuggug ccuacacgac ccugcuggac 3060
acggccugga agcaaauguc cagcuuugug cagaacggca gcaccuucgu gaaacaccug 3120
ucgcugggau cggugcagcu cugcggcgug gggaaguuuc cgucacugcc gauccugucc 3180
ccggccuuga uggaugugcc cuaccggcug aacgaaauca ccaaggagaa ggagcagugc 3240
ugcgugucgc uggccgccgg gcugccucac uucuccuccg gaauuuuucg augcuggggu 3300
agagacacuu ucauugcgcu gagggggauu cugcuuauua ccggccgcua cguggaagcc 3360
aggaacauca uccuggcauu cgccggaacc cugcggcacg ggcugauucc caaccuccuc 3420
ggagaaggaa ucuacgcucg guacaauugc cgggacgcag ugugguggug gcuccagugc 3480
auccaggacu acugcaagau ggucccuaac ggacuggaca uucugaagug cccagugucc 3540
cggauguacc ccacugacga uucggcaccg cugccggcug guacccugga ccagccgcug 3600
uucgaaguga uccaggaagc caugcaaaag cauaugcagg ggauucaguu ccgcgaaaga 3660
aaugccggac cucagaucga ccgcaacaug aaggaugaag gcuuuaacau cacugccgga 3720
guggacgaag agacuggauu cgucuacggg ggaaacagau ucaauugcgg uacauggaug 3780
gauaagaugg gagaaagcga cagagcucgg aauagaggaa uuccggccac accucgggac 3840
ggcucagccg uggagaucgu ggggcuaucc aagucugccg ugcgcuggcu guuggaacug 3900
ucgaagaaga acauuuuccc auaccacgag gucaccguga agcgccacgg aaaggccauc 3960
aaagugucau acgaugaaug gaaccgcaag auucaggaca acuucgagaa gcuguuccau 4020
gugucggagg acccuuccga ccugaacgaa aagcauccaa accuggugca caagcggggg 4080
aucuacaagg acuccuacgg agcguccucc ccuuggugcg acuaccaacu ccggccaaau 4140
uucacgaucg cgaugguggu ggccccugaa uuguuuacca ccgaaaaggc cuggaaggcc 4200
cucgagaucg cagagaagaa acugcuggga ccccugggca ugaaaacccu ggacccggac 4260
gauauggugu acugcggaau cuacgacaac gcccucgaua augacaacua uaacuuggcc 4320
aaggguuuca auuaccacca ggggccagag uggcuguggc caaucgguua cuuucugcgg 4380
gccaagcugu acuucucgag auugaugggg cccgaaacca ccgcuaagac uaucgugcuc 4440
gucaagaacg ugcugucacg gcauuacgug caccuggaac gcagcccaug gaagggccug 4500
ccggaacuga ccaacgagaa cgcacaguac uguccguucu cgugugaaac ucaggccugg 4560
agcauugcga ccauccugga aacucucuau gaucuguag 4599
<210> 11
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 11
augggccauu cuaagcaaau ucggauacug cugcugaacg aaauggagaa acuugaaaag 60
acccuguucc ggcuggagca gggcuaugag cugcaguuua gauugggccc gacacugcag 120
gggaaggcag ucaccguuua uaccaauuau ccuuuuccug gggaaacguu caaucgugaa 180
aaguuucgga gccucgacug ggaaaacccu accgagcgag aggaugauag cgacaaguau 240
uguaaacuca aucugcagca gucaggcuca uuccaauauu auuuucugca agggaacgag 300
aagucuggcg gaggcuacau ugucguugau ccgauacugc ggguuggcgc cgacaaccac 360
guguugccuc uggacugcgu gacacugcaa acauuucucg cgaaaugcuu aggccccuuu 420
gaugaguggg aaagccgccu gagagucgcc aaggagagcg gcuauaauau gauccauuuu 480
acgccgcugc aaacacuggg ucugucuagg ucauguuacu cauuggcuaa ccagcuggag 540
cugaacccgg acuuuaguag gccuaacagg aaguauacau ggaaugaugu ugggcaacug 600
guggagaagc ugaagaagga auggaauguc auuuguauua ccgaugucgu guacaaccac 660
acggcagcaa auucaaagug gauucaggaa cauccggaau gcgcuuauaa ccucguaaac 720
ucaccgcacc ugaagccggc cuggguacug gacagggcac uguggagauu cucuugugau 780
guagccgagg ggaaguacaa ggagaaaggc auucccgcuc uuauugaaaa ugaccaccau 840
augaauagca uuaggaagau caucugggaa gauauuuuuc caaaacugaa gcugugggaa 900
uucuuucaag uugacgugaa caaggcuguc gagcaguuca ggcggcuguu gacccaggag 960
aacagacggg uuacuaagag ugaccccaac caacaucuga cgaucaucca ggacccagag 1020
uaucggcgcu uuggcugcac aguggacaug aacaucgccc ugaccacguu uaucccccau 1080
gauaagggcc cggccgcuau ugaagagugu uguaacuggu uucauaagcg cauggaagaa 1140
cucaacagcg aaaagcauag acugaucaac uaucaucaag aacaggcugu gaauugucug 1200
cugggcaacg uguuuuauga gcgguuggcu ggccauggcc ccaaguuggg ccccgugacg 1260
cgaaaacauc cccucgugac aagauauuuc accuuuccau uugaagagau cgacuucuca 1320
auggaggaaa gcaugauuca ucugccaaac aaggcuuguu uucugauggc ucauaauggc 1380
uggguuaugg gcgacgaccc ccugagaaac uuugcggaac cagggucuga aguguaucug 1440
cggagggaau ugauuugcug gggagacucu guaaagcugc gauaugggaa caagccggag 1500
gauuguccgu aucugugggc ccauaugaag aaguauacag aaauuaccgc aacguauuuu 1560
cagggcguac gccuggauaa uugucauuca acgccacugc auguggcaga guacaugcua 1620
gacgcggcua ggaaucugca gccgaaucug uaugucgugg ccgagcuguu caccggcucc 1680
gaagauuugg acaauguguu ugugacccgc cucgguauua gcucacugau uagagaagca 1740
augucggcuu acaauucgca ugaggaaggu cgccuggugu accguuacgg cggcgaaccc 1800
gugggcaguu uugugcaacc cugucugagg ccacuaaugc ccgcuauugc acaugcccug 1860
uuuauggaua uaacacauga uaacgaaugu ccaaucguac accggagugc uuaugaugcu 1920
cugcccucaa cgaccaucgu aucaauggca ugcugugccu ccggcucaac acggggcuau 1980
gaugaacugg uaccacacca gaucagcgug guuucugagg aacgguuuua cacaaagugg 2040
aauccugagg cguugccauc caacacuggc gaagugaacu uucagagcgg cauuauugcu 2100
gcccgaugug cuauuucaaa gcuacaccag gaguugggcg caaagggguu uauccagguu 2160
uauguggacc agguugacga agauaucgua gcugugacgc ggcacucucc gucuauccau 2220
caauccgugg uagccguguc uaggacggcu uuucgcaacc ccaagacaag uuucuauucc 2280
aaagaaguac cccagaugug uaucccgggg aagauugagg aagugguccu ggaggccagg 2340
acgauagaac ggaacacgaa gccguaucgc aaggacgaaa acagcauaaa cgggacgccu 2400
gacaucacgg uggagauucg cgagcacauc caguugaacg aaucaaagau agugaaacag 2460
gcuggggugg cuaccaaggg cccaaaugag uacauucagg agaucgaguu cgaaaaccug 2520
uccccgggca gugugaucau cuuuagaguc ucuuuggauc cgcaugccca ggucgcugug 2580
ggcaucuugc gaaaucaccu gacgcaguuu ucuccacauu uuaagagugg cucccuggcu 2640
guggacaacg ccgacccaau cuugaagauu ccguucgcuu cccuggcuuc acgucugacu 2700
cucgcugaau ugaaucaaau ucuguauaga ugcgaaucag aggagaagga agauggaggc 2760
gguuguuacg acaucccgaa uuggucggca cugaaauacg ccggucugca aggcuugaug 2820
agcguacugg ccgagauaag accaaagaac gaucugggcc acccauuuug caacaacuug 2880
cgguccggcg auuggaugau ugacuauguc ucuaaccgac ucaucaguag aagcggaacc 2940
auagcugagg ucggcaaaug gcugcaagcc auguucuucu auuugaagca gaucccccga 3000
uaucugaucc ccuguuauuu ugacgcaauc cucauuggcg ccuauaccac acuccucgac 3060
accgcuugga agcagauguc cucauuugug caaaacggua gcaccuuugu gaagcaucug 3120
ucgcugggua gugugcagcu cugcggcgua gggaaguucc ccaguuugcc gauccugagu 3180
ccagcucuaa uggacgugcc auauaggcug aacgagauua ccaaggaaaa ggagcagugu 3240
ugugucuccc uugcugccgg ucugccccau uuuuccucgg gcauuuuuag auguuggggg 3300
cgggacaccu ucaucgcucu gcggggcauc cugcugauca ccggcagaua uguggaggcu 3360
cggaauauua uucuggcuuu ugcugggacg cugcgccaug gacugauucc caaucuauug 3420
ggcgaaggca uauacgcuag guauaacugu cgggacgcug uuugguggug gcugcagugu 3480
auccaagauu auugcaagau gguccccaac ggccuggaca uacugaaaug uccaguaucu 3540
aggauguacc cgacugacga cagugcuccc cugcccgcug gaacuuugga ucaaccccug 3600
uuugagguua uucaagaggc uaugcagaag cacaugcagg gcauucaguu ucgcgaaagg 3660
aacgccggcc cccaaauuga ccguaacaug aaggacgaag gcuucaauau uacggcuggc 3720
guagaugaag aaacaggcuu uguguacggc ggcaaccgcu ucaacugugg gacauggaug 3780
gacaagaugg gcgaaagcga cagggcucgg aacaggggaa uuccagccac gccccgcgau 3840
ggcagugcug uugaaauugu cggcuuguca aaaucagccg uacgcugguu auuagagcuc 3900
uccaagaaga acaucuuucc guaucaugaa gucacgguua agcggcaugg aaaagcaauc 3960
aaagugucuu acgacgaaug gaauaggaaa auucaggaca acuuugagaa gcuguuccau 4020
gugucugagg acccguccga ucuuaacgag aagcauccca auuuggugca caagaggggc 4080
aucuauaagg acucauacgg ggcuagcuca ccuuggugug acuaucagcu gcgaccgaau 4140
uucacaauag cuauggucgu ggcuccggag cuuuuuacca ccgagaaggc uuggaaggcc 4200
cuggagauug ccgagaagaa gcuguuaggc ccguugggca ugaaaacguu ggauccagac 4260
gacauggucu auuguggcau cuaugacaac gcucucgaca augacaacua uaaccuagcu 4320
aagggcuuca auuaccauca gggcccggag uggcuguggc caauuggaua uuuccugagg 4380
gcaaagcugu auuucucccg gcugaugggc cccgagacaa cggcaaaaac cauagucuug 4440
guuaagaacg ugcugucccg ccacuaugug caccuugaaa ggaguccgug gaagggccug 4500
ccggagcuga caaacgaaaa ugcccaguac ugcccauuuu cuugcgaaac ucaggcuugg 4560
agcaucgcca caaucuugga aacccuguau gacuuguag 4599
<210> 12
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 12
augggacacu cgaagcagau ccgcauccuc cugcugaacg aaauggaaaa acuggaaaag 60
acacucuucc ggcuggaaca gggauaugag cugcaguuua gacuggggcc caccuuacaa 120
ggaaaggccg ugacugucua uaccaacuac ccguuccccg gagaaacuuu caaccgggaa 180
aaguuccggu cacuggacug ggaaaacccg accgagcggg aggacgauag cgacaaguac 240
ugcaagcuga accuucagca guccggaucg uuccaguacu acuuccugca agggaacgag 300
aagucgggcg gaggauacau uguggucgau ccuauccugc gcgugggagc ugacaaucac 360
gugcucccuc uggacugcgu gacgcugcaa accuuccucg caaagugccu cggaccuuuc 420
gacgaauggg aauccagacu ccgcguggcc aaggagucag gauacaacau gauccacuuc 480
acgccgcugc agaccuuggg acucucucgg agcuguuacu cccuggccaa ccagcuggaa 540
cugaacccag acuucucacg cccgaauaga aaguacaccu ggaacgacgu gggacagcuu 600
guggagaagc ucaagaagga auggaacguc aucugcauaa ccgacgucgu guacaaccac 660
accgcagcca auucuaagug gauucaggaa caccccgaau gcgccuacaa ucugguuaac 720
ucaccccacc uuaagcccgc cuggguccua gacagagccc ucuggcgguu cucgugugac 780
guggccgagg gaaaguacaa ggagaaggga auccccgcuc ugauugaaaa cgaccaccac 840
augaacucca uccgcaagau uaucugggag gacaucuucc cgaagcucaa gcugugggaa 900
uucuuccaag uggacgugaa caaggcgguc gagcaguuca gacgccugcu gacccaggaa 960
aaucggagag ugaccaagag cgauccuaac cagcaccuga ccaucaucca agacccagag 1020
uaccggcggu ucggaugcac uguggauaug aacaucgccc ugaccacuuu caucccucac 1080
gacaagggac cggccgcaau ugaggaaugc ugcaacuggu uccauaagcg gauggaggaa 1140
cugaacagcg agaagcaucg acugaucaau uaucaucaag agcaggcugu gaauugccug 1200
cugggaaacg uguucuacga acggcuggcc ggacauggcc cuaagcuggg gccugugacc 1260
cggaagcacc cucuugugac ccgauacuuc accuucccgu uugaagaaau ugauuucucc 1320
auggaagaau ccaugaucca ucugccaaac aaggccugcu uccuuauggc ccacaauggc 1380
ugggucaugg gggacgaccc ucuucggaac uucgccgaac cagggagcga gguguaccuc 1440
agaagggagc ucaucuguug gggggauucc gugaagcuca gauacggaaa caagccagaa 1500
gauugccccu accuuugggc ccacaugaag aaguacaccg aaaucaccgc cacauacuuc 1560
caaggagugc ggcuggacaa cugccauuca acuccccugc acgucgccga guacaugcug 1620
gacgcugcga gaaacuugca gcccaaccuu uacguggugg ccgagcuguu caccgggagc 1680
gaggaccugg acaacguguu ugugaccagg cucggaaucu cgucgcugau ucgcgaagcc 1740
augagcgccu acaacuccca cgaagagggu agacuggugu acagauacgg aggagagcca 1800
gugggauccu ucguccaacc gugccugcgg ccgcucaugc cugcgaucgc acacgcgcug 1860
uucauggaca ucacccacga uaacgaaugu ccuaucgugc auaggagcgc cuaugaugcc 1920
cuucccucca ccaccaucgu guccauggcg ugcugugccu cggguagcac caggggauac 1980
gacgagcugg ugccgcacca gaucucggug guguccgaag aacgguuuua cacuaagugg 2040
aacccugagg cgcugccuuc caacaccgga gaagugaacu uccaguccgg uaucauugcc 2100
gcucgcugcg caaucagcaa acugcaccag gagcuuggug ccaaaggauu cauccaaguu 2160
uacgucgauc aaguggacga ggauauugug gccgucacua ggcacucucc aagcauucac 2220
caguccguag uggcaguguc gaggaccgcc uuccggaacc ccaagacuuc auuuuacucg 2280
aaagaggucc cacagaugug caucccugga aagaucgaag aaguggugcu ggaagcccgg 2340
accaucgaga ggaacacaaa gcccuaccgg aaggacgaga acuccaucaa cggaaccccc 2400
gacauuaccg uggaaauuag agagcacauc caguugaacg agucgaagau cgugaagcag 2460
gccggagugg ccacuaaggg accaaacgag uacauccagg agaucgaguu ugaaaaccug 2520
uccccgggcu ccgugaucau cuuccgggug ucccuugacc cccaugccca aguggccguc 2580
ggaauccuua ggaaccaccu gacccaguuu ucgccccauu ucaaguccgg auccuuggcu 2640
gucgacaaug ccgaucccau ccugaaaauu ccguucgccu cccucgcuuc ccggcucacc 2700
cuugccgaac ucaaccagau ucuguaccgc ugcgagucag aagaaaagga agauggaggg 2760
ggaugcuacg acaucccgaa cugguccgcc cuuaaauacg ccgggcugca aggccugaug 2820
uccgugcugg cggagauuag accgaagaac gacuuggguc acccuuuuug caacaacuug 2880
cggagcggag acuggaugau cgacuacgug ucgaaccggc ugauuagucg guccggaacc 2940
aucgccgaag ucggaaagug gcuccaagcc auguucuucu accugaagca aauuccccgc 3000
uaccugauac cgugcuacuu cgacgccauc cucauuggag ccuacaccac gcugcuggac 3060
accgccugga agcaaaugag cuccuucgug caaaacggau caaccuucgu gaagcaccug 3120
ucccugggua gcguccagcu cuguggcgug ggaaaguuuc cgucccugcc aauucugagc 3180
ccggcucuga uggaugugcc guaucgccug aacgagauca cgaaggagaa ggagcagugu 3240
ugcgugucgc uggcugcggg acugccucac uucucguccg gaaucuuccg cuguugggga 3300
cgcgacacgu uuaucgcguu gagggguauu cuccucauua ccggacgcua cguggaagcg 3360
cggaacauua uccuggcguu cgccggaacc cugcgccacg gucugauucc uaaucugcug 3420
ggagagggaa ucuacgcgcg guacaacugc cgggaugcug ugugguggug guugcagugc 3480
auccaggacu auuguaaaau ggugccgaac ggccuggaca uccugaagug cccggugucc 3540
cggauguacc cgaccgauga uucagcacca cugcccgccg ggacccugga ccagccccug 3600
uucgaaguca uucaggaagc gaugcagaag cauaugcagg gaauccaguu ccgcgaaaga 3660
aacgccggac cucagaucga ccgcaacaug aaggaugagg gcuucaacau caccgcggga 3720
guggaugagg aaaccggcuu cgucuacgga gggaaccggu ucaacugcgg aaccuggaug 3780
gacaagaugg gagaguccga ccgcgcaaga aaccgcggua uuccugccac cccgcgggac 3840
ggaagcgcgg uggagaucgu cggacugagc aagagcgcag ugcgcuggcu gcuggaguug 3900
uccaagaaga auaucuuccc cuaucacgag gucaccguga aacgccacgg gaaggccauc 3960
aaaguguccu acgaugagug gaaccgcaag auucaggaua acuucgaaaa gcuguuucau 4020
guguccgagg aucccuccga ccucaacgaa aagcacccga accucgugca uaagaggggg 4080
aucuacaagg acagcuacgg ugccuccuca ccuuggugcg auuaucagcu ccgcccgaac 4140
uucaccauug ccaugguggu cgccccugaa cuguuuacua ccgagaaggc cuggaaggcg 4200
cuggagauug ccgaaaagaa gcuguuggga ccccugggca ugaaaacccu cgaccccgac 4260
gacauggugu acugcggaau cuacgacaac gcccucgaca acgacaacua caaccucgcg 4320
aagggguuca acuaccacca gggccccgaa uggcuguggc ccaucggaua cuuccuccgg 4380
gcgaaguugu acuucucccg ccugaugggc ccugaaacca cggccaaaac gaucgugcuc 4440
gugaagaacg ugcugucgag gcauuacgug caccucgagc ggagcccgug gaaggggcug 4500
ccggaacuga ccaacgagaa ugcacaguac ugccccuucu ccugcgaaac gcaggcuugg 4560
uccauugcca cuauuuugga aacgcuguac gaccuguag 4599
<210> 13
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 13
augggacacu cgaaacagau cagaauccug uugcucaacg aaauggagaa gcuggaaaag 60
acccucuuuc ggcucgagca gggcuacgag cugcaguucc gccugggucc gacacuacaa 120
ggaaaggcag ugacugugua caccaacuac ccauuucccg gagaaaccuu caaccgggag 180
aaguuccggu cccuggacug ggaaaaccca accgaacgag aggaugacuc cgacaaguac 240
ugcaagcuga accuccaaca guccgguuca uuccaguacu acuuucugca agggaacgag 300
aaguccggag gcggcuacau cguggucgac ccgauacuua gagugggagc cgacaaucau 360
guccugccuc uggacugcgu gacccugcaa accuucuugg ccaaaugucu gggcccguuc 420
gaugaguggg aaagccgccu cagagucgca aaggaguccg gauacaacau gauucacuuc 480
acuccgcugc aaacccucgg ucugucccgg ucgugcuauu cucuggcgaa ccagcuggag 540
cuuaaccccg acuucucgcg cccaaaccgg aaguacaccu ggaacgacgu cgggcagcuu 600
guggaaaagc ugaagaagga guggaacgug aucugcauca cugacguggu guacaaccau 660
acggccgcca acucgaagug gauccaagag caucccgagu gcgcguacaa ccucgugaac 720
agcccgcauc ugaagccugc uugggugcug gauagagccc ucuggagauu cagcugcgac 780
guggccgagg ggaaguacaa agaaaaggga auuccggccu ugauugagaa cgaccaucac 840
augaacucaa uccgcaagau caucugggag gauaucuucc cuaagcuuaa guugugggag 900
uucuuccaag uggacguaaa caaggcagug gagcaguuca gaagguugcu gacucaagaa 960
aacagacgcg ucacuaaguc cgauccuaac cagcaccuua ccaucauuca agacccugag 1020
uaccgccggu uuggcugcac cgucgacaug aacaucgccc ugaccacuuu caucccgcau 1080
gacaagggcc cggcggcaau cgaggaaugc uguaacuggu uucauaagag gauggaggaa 1140
cugaauuccg agaagcaccg gcugauuaac uaccaccagg aacaggcagu gaauugccuc 1200
cugggcaacg uguucuacga acggcuggcu ggacacggac cgaagcuggg ucccgugacu 1260
cgcaagcauc cgcucgugac ucgcuacuuc accuucccgu uugaggagau ugacuucucc 1320
auggaagaau ccaugaucca ccucccgaac aaggcuugcu uccugauggc gcacaacgga 1380
ugggucaugg gcgacgaccc acugcgcaac uucgcugagc cuggcucgga ggucuaccug 1440
agaagggaau ugauuugcug gggagacucc gucaagcugc gcuauggaaa caagcccgaa 1500
gauugccccu accugugggc ucacaugaag aaguacacgg aaaucacugc cacguacuuc 1560
cagggagucc ggcuggacaa uugccacucc accccccucc auguggccga guacaugcuc 1620
gaugcagcga ggaaucugca gcccaaucug uacgugguug cagaacuguu cacuggcucc 1680
gaggaccucg acaacguguu cgugaccaga cuggggaucu ccucccugau ccgggaagcc 1740
augucggccu acaacuccca ugaagagggc cgccuggugu accgcuacgg cggagaaccc 1800
gugggaagcu ucgugcagcc uugccuccgg ccgcugaugc cugcgaucgc ccacgcccug 1860
uucauggaua ucacucacga caacgaaugc cccauugugc aucgcucggc cuacgacgca 1920
cugccuucga ccacuaucgu guccauggcc ugcugcgccu ccgggagcac ccgcggauac 1980
gaugaacucg ugccgcacca gaucagcgug guguccgaag aaagauucua uaccaagugg 2040
aaccccgaag cccugccgag caauaccggg gaagugaacu uccaguccgg uauuaucgcc 2100
gcucgcugug ccaucagcaa acuccaccaa gagcucggug ccaagggauu cauucaaguc 2160
uacguggauc aggucgacga agauauugug gccgugacca ggcacucacc uuccauccac 2220
caauccgucg ucgccguguc ccggacugcg uuucggaacc ccaagacuuc guucuacucg 2280
aaagaagugc cacagaugug uaucccggga aaaaucgaag aggucgugcu cgaagcccgg 2340
accauugaga ggaacaccaa gccuuaccgg aaagacgaga acucuaucaa cgguaccccu 2400
gauauuacug uggagauccg cgaacacauc cagcugaacg aaucaaagau cgucaagcag 2460
gcuggagugg ccaccaaggg acccaacgag uacauccagg agaucgaauu ugaaaaccuc 2520
uccccuggcu ccgugauuau cuuccgggug ucccuggacc cucacgccca aguggccgug 2580
ggaauucuca gaaaccaccu gacccaguuc ucaccccacu uuaaguccgg uucccuggcg 2640
guggacaacg ccgauccgau cuugaagauc cccuucgcau cgcuggccuc ccgccugacu 2700
cucgcggaac ugaaccagau ccuguaccgc ugugaaucag aggaaaagga ggacggcggc 2760
ggcuguuacg auauccccaa uuggucggcu uugaaauacg cgggacuuca ggggcugaug 2820
ucugugcugg cggaaauccg gccgaagaac gaccugggac acccauucug caacaacuug 2880
cggagcggag acuggaugau cgauuacguc agcaacagau ugaucagccg gagcggcacu 2940
aucgccgagg ucggaaagug gcuccaggcc auguucuucu accugaagca gaucccccga 3000
uaccucaucc ccuguuacuu cgacgccauu cugaucgggg ccuacaccac ccugcuggac 3060
accgccugga agcagaugag caguuuugug caaaacgggu ccaccuucgu gaagcaccuu 3120
ucacugggcu cagugcagcu cugcggcgug ggaaaguucc ccucgcugcc cauucugagc 3180
ccggcccuga uggacguccc uuaccggcug aacgagauca ccaaggagaa ggagcagugc 3240
uguguuuccc uggcugccgg gcugccacac uucucguccg gcaucuuccg gugcuggggc 3300
cgggauaccu ucauugcccu gcggggaauc cugcuuauca ccggucgcua cguggaggcu 3360
cggaacauua uucuggcguu cgccggcacc cuuagacacg gucugauucc gaaucuuuug 3420
ggcgaaggaa ucuacgccag auacaacugu cgggacgccg ugugguggug gcuccagugc 3480
auucaggacu auugcaagau ggugccgaac ggccuggaca uccugaagug cccagugucg 3540
aggauguauc caaccgacga cagcgcaccu cugccggccg ggacccucga ccaaccccug 3600
uucgaaguca uucaagaggc uaugcagaag cacaugcagg guauucaguu ccgggagcgg 3660
aacgcggggc cccagauuga uaggaacaug aaagacgagg gcuucaacau cacugccggc 3720
guggacgaag aaaccgguuu uguguacgga ggaaacagau ucaacugcgg uaccuggaug 3780
gacaagaugg gagaguccga ucgcgcgcgc aacagaggga ucccggcaac cccgcgggac 3840
ggauccgcgg uggaaauugu gggacugagc aagagcgccg ugcgguggcu ccuggaacug 3900
agcaaaaaga acaucuuccc cuaccacgaa gugaccguga agcggcacgg aaaggccauc 3960
aaagucucau acgaugaaug gaauaggaag auccaggaua acuucgagaa gcuguuucac 4020
guguccgagg aucccuccga ucugaacgaa aagcauccga aucucgugca caagcgcggg 4080
aucuacaagg acucguacgg agcguccucc ccuuggugcg acuaucagcu gcggccuaac 4140
uucaccauug ccauggucgu ggccccggag cuguucacaa cugagaaggc cuggaaggcc 4200
cuugaaauug ccgagaagaa gcugcugggg ccuuugggga ugaaaacccu ggauccggac 4260
gacauggugu acugcggaau cuacgacaac gcccuggaca acgauaacua caaucuggcg 4320
aagggcuuca auuaccacca gggcccggaa uggcucuggc cuauugggua cuuccugcgc 4380
gccaagcugu acuucucacg gcugauggga ccagagacua ccgccaagac uaucguccuc 4440
gugaagaacg ugcugucccg gcacuacgug caucuggaga ggagcccuug gaagggacuu 4500
ccugagcuga cgaacgaaaa cgcgcaguac ugccccuucu ccugcgaaac ccaggcuugg 4560
uccauugcca cuauacugga aaccuuauau gaccuguag 4599
<210> 14
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 14
augggucacu ccaagcaaau cagaaucuua cuccuuaacg aaauggaaaa gcuugaaaag 60
acacucuucc gcuuggagca gggcuacgag cugcaguucc ggcugggucc gacacuucag 120
gggaaagccg ugaccgugua uaccaacuac cccuuuccgg gagaaacauu caaucgcgag 180
aaguuccggu cgcuggacug ggagaacccu accgagaggg aggacgacuc ugauaaguac 240
ugcaagcuga aucuccagca aucagguucu uuccaauauu acuuccuuca aggaaacgaa 300
aaguccgggg gcggcuacau uguaguggac cccauccuuc gggucggcgc ggauaaccau 360
gugcuucccc ucgacugcgu gacucuccaa acuuuccucg ccaagugccu gggaccauuc 420
gaugaauggg aaucccgccu gcgcguggcc aaggagagcg gcuacaacau gauucacuuc 480
acuccgcuuc aaacccuugg gcugucccgc uccugcuauu cccuugcgaa ccagcuggaa 540
cuuaacccgg acuucucucg gccgaacaga aaguacacuu ggaacgacgu gggccagcuu 600
gucgagaagc ugaagaagga auggaacgug aucugcauca ccgacguggu guacaaccac 660
accgcggcaa acuccaagug gauccaggaa cauccugagu gcgcauacaa ccucgugaac 720
aguccgcauc ugaagccugc cugggugcug gauagagccc uguggcgcuu cuccugcgac 780
guggcugagg gaaaguacaa ggaaaagggc aucccugcgc ucauugaaaa cgaucaccac 840
augaacagca uccgcaagau uauuugggag gacaucuucc cuaagcucaa gcucugggag 900
uuuuuccaag uugacgugaa caaggccgug gagcaguuca gacggcuccu gacucaagaa 960
aacagacgcg ucaccaaguc cgacccuaac caacaccuca ccaucaucca ggaucccgaa 1020
uaccgccggu uuggcugcac uguggacaug aacauugccc ugaccaccuu cauuccgcac 1080
gacaaaggcc cggccgcgau ugaagagugc uguaacuggu uccacaagag aauggaggag 1140
cugaacuccg aaaagcauag acuuaucaac uaccaccagg aacaggccgu gaacugccug 1200
cugggaaacg uguuuuacga gagacucgcc ggacacgguc caaaguuggg uccugugacc 1260
cgcaaacacc cgcucgucac ccgcuacuuu accuucccgu ucgaggagau cgacuucagu 1320
auggaggaga gcaugaucca ucuccccaac aaggccuguu uccucauggc ccacaacggc 1380
ugggucaugg gcgacgaccc ccugcgcaac uucgcugaac ccggcucgga aguguaccuu 1440
cggagggagc uuaucuguug gggcgacagc gugaagcuua gauacggcaa caagccagaa 1500
gauuguccgu accugugggc gcacaugaag aaguacaccg agaucaccgc gacguacuuu 1560
caaggagugc ggcucgauaa cugccacucc accccucugc acguggcaga guacaugcua 1620
gacgcggccc ggaaccucca gcccaaccuu uacguggugg ccgaacucuu cacugguucu 1680
gaggaucuug auaacguguu cgugacuagg cucggcauuu ccucccucau ccgggaagcc 1740
augucggccu auaacuccca cgaggagggg cggcuggugu accgcuacgg aggcgaaccg 1800
gucggcagcu ucgugcagcc gugccuccgc ccucugaugc ccgcuauugc ucacgcccuu 1860
uucauggaua ucacucacga uaaugagugc ccuaucgugc aucggagcgc cuacgacgcu 1920
cucccuucca ccaccaucgu guccauggcg ugcugcgccu ccgguucaac caggggcuac 1980
gaugaacuug ugccgcacca gaucucaguc gucagcgagg aaagguucua cacuaagugg 2040
aacccugaag cccugcccuc uaacacgggc gaagugaacu uucagagcgg uaucauugcc 2100
gcuagaugcg caaucuccaa guugcaccag gaacugggag ccaagggguu cauccagguc 2160
uacguggacc aggucgacga ggacaucguc gccgugaccc ggcauucccc gagcauccau 2220
caguccgugg ucgccguguc acggaccgcc uuccgcaacc ccaagaccuc cuucuacucc 2280
aaggaagugc cgcaaaugug uaucccuggc aaaaucgagg aaguggugcu cgaagcgcgg 2340
acgauugaga ggaauacuaa gccguacaga aaggacgaaa acuccaucaa cggcaccccg 2400
gacaucacug uggagauccg ggagcacauc cagcucaacg agagcaaaau ugugaagcag 2460
gccggcgucg cuacuaaggg cccaaacgag uacauccagg agauugaguu cgaaaaccuu 2520
agcccugggu cugugaucau cuuucgcgug ucccucgacc cgcacgcaca ggucgcaguc 2580
gggauucucc ggaaccaucu gacucaguuc agcccccacu ucaagagcgg cagccuugcc 2640
gucgacaacg ccgaucccau ccucaaaauc ccuuucgcau cccuugcguc gaggcuuacc 2700
cuggcggaau ugaaccagau ucuguaccgc ugcgagucgg aagaaaaaga ggauggcggc 2760
ggcugcuacg acauuccgaa cugguccgcc cugaaauacg cgggccuuca gggccuuaug 2820
agcgucuugg ccgagauccg ccccaagaac gaccuggggc accccuuuug caacaaccuc 2880
agaagcggcg auuggaugau cgacuacgug ucgaacaggc ucaucagccg auccggcacu 2940
auagccgagg ucggaaagug gcugcaggcc auguucuuuu accucaaaca gaucccgcgg 3000
uaccugaucc cgugcuacuu cgacgcuauu cucauuggcg ccuacacuac ccugcucgau 3060
accgcuugga agcagaugag cucauucgug caaaacggaa gcaccuucgu gaagcaccuc 3120
ucccugggau cagugcagcu gugcggcgug ggaaaguucc cauccuuacc aauucucucg 3180
ccugcccuga uggacguccc uuaucgccug aacgaaauca cgaaggagaa ggaacagugu 3240
ugugucucac uggcugccgg ccucccgcac uucucauccg gcaucuuccg gugcuggggu 3300
agagacacuu ucauugcgcu ccggggaauu cugcuuauca cuggccgcua cguggaagcc 3360
cgcaacauca uccuugccuu ugccgggacc cugcggcacg gccugauccc uaaccuucuc 3420
ggggagggca ucuaugcgcg auacaauugc cgggacgccg ucugguggug gcugcagugc 3480
auucaggacu auugcaagau ggugccaaau ggucuggaca uucugaagug uccagugucc 3540
cggauguacc cuaccgacga uagcgcccca cugcccgccg gaacccucga ucagccccug 3600
uucgaaguca uucaggaagc gaugcagaag cauaugcagg guauccaauu ccgcgaaagg 3660
aaugccggcc cacaaaucga cagaaauaug aaggaugagg gcuuuaacau caccgccggc 3720
guggacgagg agacugguuu cgucuacggc ggcaaucggu ucaauugcgg gaccuggaug 3780
gacaagaugg gcgaaagcga ccgggccaga aaccggggca uuccggcuac cccccgcgau 3840
ggcucggccg uggaaaucgu gggccucucc aagucugccg ugcgguggcu uuuggagcuc 3900
uccaagaaaa acaucuuccc guaccacgaa gugaccguga agagacacgg gaaggccauc 3960
aaaguguccu acgacgaaug gaaccggaag auccaggaca acuucgaaaa gcuguuccac 4020
guguccgagg aucccuccga cuugaacgaa aagcacccca accucgugca caagcgcggc 4080
aucuacaagg auuccuacgg agcguccuca ccuuggugcg acuaucagcu gaggcccaac 4140
uucaccaucg caaugguggu ggccccugag uuguucacca cugagaaggc uuggaaggcc 4200
cuugagaucg ccgaaaagaa gcugcucggc ccgcugggga ugaaaacccu cgaccccgau 4260
gacauggugu acugcgggau auacgacaau gcacuagaca acgacaacua caaccuggcc 4320
aagggcuuca auuaccauca gggcccggag uggcuuuggc cuaucggcua cuuccugcgg 4380
gccaagcugu acuucucacg gcuuauggga ccggagacua cugcaaagac cauugugcuu 4440
gugaagaacg ugcuuucgcg ccacuacgug caucuggaac ggagccccug gaaggggcuc 4500
cccgagcuga ccaacgagaa cgcccaguac ugucccuucu ccugugaaac ccaggcuugg 4560
uccauugcca ccauucugga aacccuguac gaccucuag 4599
<210> 15
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 15
augggacacu caaagcaaau ucggauccuc cugcucaacg agauggaaaa gcucgagaaa 60
acccucuuuc gccuggagca ggguuaugag cuccaguucc gccugggucc gacccuccaa 120
gggaaggccg ucacugugua cacuaacuau ccauuccccg gagagacuuu caaccgggaa 180
aaguuccgca gccucgauug ggagaacccu acugaacggg aggacgauuc ggacaaguac 240
uguaaacuga accuccagca gagcggcuca uuucaauacu acuuucugca agggaacgag 300
aaguccggag ggggguacau cgucguggau ccuauccuuc gcgugggcgc cgacaaccau 360
gugcugccgu uggacugcgu gacccugcaa accuuccucg ccaaaugccu cggaccuuuc 420
gaugaguggg aauccaggcu gagaguggcc aaggaaucgg gguacaacau gauucacuuc 480
accccucucc aaacccuggg ccugucucgg agcugcuacu cccuggccaa ccagcuggag 540
cugaaucccg acuucucccg gcccaaccgg aaguacacuu ggaaugacgu gggccaacug 600
guggagaagc ucaagaagga guggaacgug aucugcauua ccgacguggu guacaaccau 660
accgccgcca acuccaagug gauccaggaa cacccagaau gugccuacaa ccucgucaac 720
ucaccccacc ugaaaccagc augggugcug gaucgcgccc ucuggcgguu cucgugugac 780
guggccgaag gaaaguacaa agagaagggc aucccugccc ugaucgaaaa ugaccaccac 840
augaauucca uuagaaagau cauuugggag gacauuuucc cuaaguugaa gcucugggag 900
uucuuccaag uggacgucaa caaggccgug gaacaguuuc gccggcuccu gacccaagaa 960
aaccgccgcg ugaccaaguc cgacccuaac cagcaccuga ccauaaucca ggacccugag 1020
uacagacggu ucgguugcac ugucgacaug aacaucgccu ugacuacuuu caucccgcac 1080
gacaaggguc cugccgccau ugaggagugc ugcaacuggu uccacaagcg gauggaagaa 1140
cugaacuccg aaaagcaccg ccucaucaac uaccaccagg agcaggccgu gaacugccug 1200
cuggggaacg uauucuacga gaggcuggcc ggacacggac ccaagcuggg acccgugacc 1260
agaaagcauc cucucgucac ccgcuacuuu acuuucccgu uugaagagau cgacuucuca 1320
auggaagagu cgaugaucca ucucccuaac aaggccugcu uccugauggc ccacaacggc 1380
ugggugaugg gcgacgaucc ucugagaaac uucgcugagc ccgguucgga aguguaccuu 1440
agacgggaac uuauuugcug gggcgauucc gugaaguuac gcuacggaaa caagccugag 1500
gacugcccuu accugugggc ccauaugaag aaguacaccg agauuaccgc caccuacuuc 1560
caaggggucc ggcuggacaa cugccacuca acuccucugc acguggcuga guacaugcug 1620
gaugcggccc ggaauuugca gccuaaccuu uacguggucg ccgaacucuu uaccgggucc 1680
gaggaccugg acaacguguu cgugacucgg cucggaaucu ccucacugau uagagaggcc 1740
auguccgcau acaacucgca cgaagaaggc cggcuggucu aucgauacgg cggcgaaccu 1800
gucggaagcu ucguccagcc cugccugcgg ccgcugaugc cagcgaucgc ccacgcccuc 1860
uucauggaca ucacccauga caacgaaugu cccaucgugc accgcuccgc cuacgaugcc 1920
uugccaagca ccaccaucgu guccauggcg ugcugcgcca gcgguagcac uaggggcuac 1980
gaugaacuug ugccgcacca gaucagcgug guguccgagg aaagguuuua cacgaagugg 2040
aaccccgaag cccugcccuc caacacuggg gaagugaacu uccaguccgg gaucauugcg 2100
gcccgcugcg cgaucucgaa gcuccaccag gaacucggcg cgaaaggauu cauucaaguu 2160
uacguggacc aggucgacga ggacaucguc gccgugacuc gccacucccc uucaauccau 2220
caauccgugg uggcgguguc gcggaccgcu uuccggaacc cuaagacuuc guucuacucg 2280
aaagaagugc cucagaugug uauccccgga aagaucgagg aaguggugcu cgaggccagg 2340
acuauugaga ggaauaccaa gccuuaccgg aaggaugaga acuccauuaa cgguacuccu 2400
gacaucaccg ucgagauccg ggaacacauc cagcucaacg aaagcaaaau cgugaagcag 2460
gccggcgugg ccacuaaggg uccgaacgag uacauccagg aaaucgaauu ugagaaccug 2520
ucgcccggaa gcgugauuau uuuucgggug ucccuggacc cgcacgccca agucgccgug 2580
gguauccugc gcaaucaucu gacucaguuc agcccucacu ucaaguccgg cagccuugcc 2640
guggacaacg ccgauccgau ccugaagauc ccauucgccu cacucgccuc acggcucaca 2700
cuggccgaac ucaaucagau ccucuaucgc ugugaauccg aagagaagga ggacggcgga 2760
gguugcuacg auauuccgaa uugguccgca cugaaauacg ccggacugca gggccucaug 2820
uccguguugg ccgaaauccg cccuaagaac gaccucggcc acccguucug caacaaccuc 2880
agaucuggag acuggaugau cgauuacgug ucaaaccgcc ugaucucgag guccggcacu 2940
aucgccgaag ucggaaagug gcuccaagca auguucuucu accugaagca gaucccucgc 3000
uaccugauac cuuguuacuu cgacgccauc cucauuggcg ccuacacuac ucugcuggau 3060
acugccugga agcaaaugag cagcuucgug cagaacggaa gcacuuucgu caagcaucug 3120
ucgcucggga gcgugcagcu gugcggcguc ggaaaguuuc cuucccugcc cauucugucc 3180
ccugcccuca uggaugugcc guaccgccuu aacgagauca cuaaggagaa ggagcagugu 3240
ugcgugagcc uggcugccgg gcucccucac uucucguccg gaaucuucag augcuggggc 3300
cgcgacaccu ucauugcccu gcgagggauc cuguugauua cuggccgcua cguggaggcc 3360
aggaauauca uucucgccuu cgcgggaacc cugcggcacg gccugauccc uaaccuccug 3420
ggagaaggaa ucuaugcgag auauaacugc agggacgccg ugugguggug gcuccagugc 3480
auccaggacu acugcaagau ggugcccaac gggcuggaca uccucaagug ccccguguca 3540
cggauguacc caacugauga cagugcuccu cugccggccg guacucucga ccagccacuc 3600
uuugaaguca uccaggaggc caugcagaag cacaugcagg gcauucaauu ccgggagagg 3660
aacgccggac cucagauuga ccggaacaug aaggacgagg guuucaauau cacugccggc 3720
guggacgaag aaaccggcuu cgucuacgga ggaaacagau ucaacugcgg aaccuggaug 3780
gauaagaugg gagaaagcga cagagcgcgg aacaggggua ucccugccac uccccgggac 3840
ggaucggccg uggaaauugu gggacuuucc aaguccgccg ugcgguggcu ccuggagcuu 3900
uccaagaaga acaucuuccc uuaccaugaa gugaccguga agcggcacgg gaaggccauc 3960
aaagucuccu acgaugaaug gaaccgcaag auucaggaca auuucgagaa gcuguuccau 4020
gucuccgagg acccuucuga ccugaaugag aagcauccca accuggugca caagagaggc 4080
aucuacaagg acagcuacgg agcuuccucc ccuuggugcg auuaccagcu ccggccaaac 4140
uucacuaucg ccaugguggu ggcgccugaa cuguucacca cugaaaaggc cuggaaggcg 4200
cucgaaaucg cggagaagaa gcugcucggg ccucucggga ugaaaacccu cgacccugau 4260
gauauggugu acugcggaau cuacgauaac gcccuagaca acgacaacua caaccuggcc 4320
aagggauuca acuaccauca ggggcccgag ugguuguggc cuauuggcua cuuccugaga 4380
gccaagcucu acuucucccg ccucaugggc cccgaaacca cugccaagac caucgugcuc 4440
gugaagaacg ugcucucccg gcacuaugug caccuugagc gcucgccaug gaagggacug 4500
cccgaacuga cuaacgagaa cgcccaguac ugccccuucu ccugugaaac ucaggccugg 4560
uccaucgcca caauuuugga aacccucuac gaucuguag 4599
<210> 16
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 16
augggacauu caaagcaaau ccggauucug uuacuuaaug agauggagaa guuggagaaa 60
acauuguuua gacucgagca gggcuacgaa cugcaguuuc gccugggccc uacacuacaa 120
gggaaagccg ucaccgucua uacaaacuau cccuuccccg gcgaaacguu uaaucgggag 180
aaguucagau cucuggauug ggagaacccc accgaacggg aggaugauag cgacaaguau 240
ugcaaacuga acuugcagca gucuggcagu uuucaguauu auuuucugca agggaacgag 300
aaguccggcg gcggcuauau aguggucgac cccauccuuc gugugggagc ugacaaccau 360
guguugcccc uggacugugu uacacugcaa accuuucugg cuaagugucu cgguccuuuu 420
gaugaauggg aaucgcggcu caggguggca aaagaauccg gcuauaacau gauucauuuu 480
acaccccugc aaacccuggg ccucucaagg aguuguuaca gccuugccaa ucagcuggag 540
cugaaccccg acuuuucccg ucccaaccgg aaguauacuu ggaacgacgu gggccagcuc 600
guagaaaagc ugaaaaagga guggaacgug aucuguauaa ccgauguggu cuauaaccac 660
accgccgcca auucgaagug gauccaggag caucccgagu gugcuuacaa ccucgugaac 720
agcccccauc uuaagcccgc uuggguacuu gaccgcgcac uuuggagguu uuccugugac 780
guggcugagg ggaaguauaa ggaaaagggc auccccgccc ugaucgaaaa cgaccaucac 840
augaacagua uacggaagau cauuugggag gauauuuuuc ccaaacugaa gcugugggag 900
uuuuuucaag uggaugugaa caaggcuguc gaacaguuua ggaggcugcu gacccaagaa 960
aauagacgcg ugacuaagag cgacccaaau cagcauuuga caauuauaca ggaccccgaa 1020
uaucgccgau ucggcuguac cguggauaug aauaucgcau ugacuacuuu caucccccac 1080
gacaagggcc ccgccgcuau ugaggaaugc uguaauuggu uucauaagcg aauggaggag 1140
cugaacucug agaagcacag gcugauuaac uaucaucagg aacaggccgu uaacuguuug 1200
cugggcaacg uguuuuauga acggcuugcc ggccauggcc cgaaguuggg cccgguuaca 1260
agaaagcauc cacuugugac ccgcuacuuc accuuuccgu ucgaagaaau ugacuuuagc 1320
auggaggagu caaugaucca ccuccccaac aaggcuugcu uucuuauggc ucacaacggc 1380
ugggugaugg gcgacgaccc ccugcggaac uuugcggaac cgggcucaga ggucuaucua 1440
agaagggaac ugauuugcug gggcgacucc gugaagcuga gauacgggaa caagccugag 1500
gauugccccu accucugggc ucacaugaaa aaguauacag aaauuacugc uacguauuuc 1560
caaggcguac ggcuugacaa cugucacucc accccacugc augucgccga guauauguua 1620
gaugcugcuc gaaacuugca gcccaauuug uauguggucg cugaacuguu uacggggagu 1680
gaagaucucg acaacgucuu uguaacccga cugggcauca gcucgcuaau ucgggaggcc 1740
auguccgcuu acaacuccca ugaggaaggc cgccuuguau aucguuaugg cggcgaaccc 1800
guggggagcu uuguucaacc gugucuacgc ccccugaugc ccgccaucgc ucaugcccua 1860
uucauggaua ucacucauga caaugagugu ccuauugugc auaggagugc uuaugacgcc 1920
cucccaagca caacgaucgu guccauggcu uguugugcua guggcuccac aagaggcuau 1980
gacgaacugg ugccccauca aaucuccgua gugucugaag agagauuuua caccaagugg 2040
aaccccgagg cucucccuuc aaacacugga gagguuaacu uucaauccgg gauuauugcu 2100
gcuagaugug cuaucagcaa gcuccaucag gaacugggcg caaagggcuu cauucaaguu 2160
uauguagacc agguugacga ggacauaguu gcaguaacuc ggcauucccc uucgauacau 2220
cagucugucg uggccguguc caggacagca uuucgcaauc ccaagacuag cuuuuacucc 2280
aaagaaguac cacaaaugug uauccccggg aagaucgagg aagugguacu ggaagcccgg 2340
acuauugaga gaaacaccaa gccguaucgg aaggacgaga auucuaucaa cggcacaccu 2400
gauauaacag uggaaauacg cgaacacauu caguugaaug aguccaagau cgugaagcag 2460
gccggcgucg ccaccaaggg ccccaacgag uauauccagg agaucgaauu ugaaaaccug 2520
agccccggcu cuguuauuau auuucggguu ucauuagauc cucaugcuca aguggcugua 2580
ggcauccucc ggaaucaucu gacucaguuc ucuccccacu ucaagagcgg cagccuggcu 2640
gucgauaaug ccgaccccau acuuaagauu cccuuugcuu cccuggcuuc aaggcugacc 2700
cuggcugaac uuaaucaaau ccuauaccga ugcgaaagcg aagagaagga agauggcggc 2760
ggcuguuacg auaucccgaa cuggagcgca cugaaauaug cuggcuuaca aggccucaug 2820
aguguguugg cugagauuag acccaagaau gacuugggcc auccauuuug uaacaaccug 2880
agaagcggcg acuggaugau cgauuacgug ucuaaccgac ucaucucccg aagcggcacc 2940
auugcugagg uuggcaaaug gcugcaggcu auguucuuuu aucugaaaca gauuccucgg 3000
uaccugauuc ccuguuauuu cgacgcuauu cugauuggcg cauauaccac gcucuuggac 3060
acugcaugga agcaaaugag cucuuuuguc caaaauggcu ccacuuuugu uaaacaucug 3120
aguuugggca gcgugcaguu auguggcguu ggcaaauuuc caagccugcc cauacugucc 3180
cccgcucuga uggacguccc cuaccgacug aacgagauca ccaaggaaaa ggaacagugc 3240
ugugugucuu uagcugccgg cuugccgcau uuuucaagcg gcauuuuccg guguuggggu 3300
cgggacaccu ucaucgcacu gagaggcauu cugcugauca cuggccgcua uguggaagcu 3360
aggaauauca uuuuagccuu ugcggguacc cuucggcacg ggcugauccc caaucuuuug 3420
ggcgaaggca ucuacgcacg uuacaauugc cgggaugccg uaugguggug gcuccaaugu 3480
auccaggacu auugcaagau gguucccaac ggccucgaua uccugaagug ccccguguca 3540
aggauguauc cuaccgauga cagugcuccc cuuccugcug gcacccugga ucagccgcuc 3600
uuugaaguua uucaggaagc aaugcaaaag cauaugcagg gcauccaguu ucgggaaaga 3660
aacgcuggcc cgcaaauaga caggaacaug aaggaugaag gcuucaauau uacugcuggc 3720
guagacgaag agacaggguu cgucuacggc ggcaauaggu uuaacugugg cacuuggaug 3780
gacaagaugg gcgaaucuga ccgcgcgagg aacagaggca ucccagcaac accgagggac 3840
ggcagcgcug uggagauugu gggccugucu aagucugccg ugcgcuggcu acucgaacug 3900
uccaagaaga auaucuuucc cuaucaugaa gucaccguaa agcggcaugg caaagcuauu 3960
aagguuuccu augacgagug gaacaggaag auucaggaca auuuugagaa gcuguuccau 4020
gugucggagg acccuagcga ucucaacgag aagcacccca acuuaguaca uaagaggggc 4080
aucuauaagg auagcuaugg cgcuagcagc ccuuggugug acuaucagcu ccgucccaac 4140
uuuaccauug cuaugguagu ggcgcccgag uuguuuacca cugagaaggc uuggaaggcu 4200
cuugaaauag ccgagaagaa auugcugggc ccccugggca ugaaaacucu ggaucccgau 4260
gacaugguau acuguggcau cuaugacaau gcccuggaca augacaauua caaucuggcc 4320
aaggguuuua acuaucacca gggcccugaa uggcuguggc cuauuggcua uuuucuucgc 4380
gccaagcuau auuuuaguag gcugaugggu ccagaaacaa cugcaaagac aauugugcuc 4440
gugaagaacg ugcuuucccg gcacuaugug caucuggaaa ggaguccaug gaagggcuug 4500
ccggaauuga cuaacgagaa cgcccaguau ugucccuuuu cuugugaaac ucaggccugg 4560
uccauugcua cuauucugga gacucuauau gacuuguag 4599
<210> 17
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 17
augggccauu ccaagcaaau ucgaauucug cugcugaacg aaauggaaaa guuggaaaag 60
acauuguucc gucuggagca gggcuaugag cugcaguuua ggcuuggccc gacguugcag 120
ggcaaagccg ugaccguaua uacuaacuau cccuucccug gcgaaacguu uaaccgggag 180
aaguuucgcu cacuggacug ggagaauccc acggaaaggg aggacgauag cgauaaguac 240
uguaaacuga aucugcaaca gucuggcucu uuccaauauu auuuucugca agggaacgag 300
aaguccgggg gcggcuauau uguaguggac cccaucuuga gagugggcgc ugacaaccau 360
guauuacccc uugauugcgu gacgcugcag accuuucugg cuaaguguuu gggcccuuuc 420
gaugaguggg aaagccggcu gaggguggcc aaggagagcg gcuauaauau gauucauuuu 480
accccuuugc aaacucuggg gcucucuagg agcuguuauu ccuuggcuaa ucagcuggaa 540
cugaaccccg auuuuagccg gcccaaccga aaguauacau ggaaugacgu ugggcaauug 600
guugagaagc ugaagaagga auggaauguu auauguauaa cagacguggu guauaaccau 660
accgcugcua auucgaaaug gauucaggaa cauccugagu gcgccuacaa ccucguaaau 720
aguccucauc ugaagccggc uugggugcug gacagggcuc ucuggagguu uagcugugau 780
guggccgaag ggaaguauaa ggagaagggu aucccggcuc ugauugagaa cgaccaccau 840
augaacagua uaaggaagau aaucugggag gacauuuuuc ccaaacugaa gcugugggaa 900
uucuuucaag uggacgugaa caaggccguu gagcaguuua gacgcuugcu gacacaagag 960
aaccgaaggg ucaccaagag ugacccuaac cagcaucuua caauuaucca agaccccgag 1020
uaucgucggu uuggcuguac uguggacaug aacauugcuc ugaccaccuu uauaccccac 1080
gauaagggcc ccgcugcuau cgaagagugu uguaacuggu uccacaagag gauggaagag 1140
uugaacucug aaaagcaucg gcugauuaac uaucaucagg aacaggcugu gaacugccug 1200
cucggcaacg uauucuacga gagauuggcu ggccauggac cuaagcuggg cccgguuacg 1260
agaaagcauc cccugguuac gcgguauuuc accuucccgu uugaggagau ugauuuuagc 1320
auggaagaau ccaugaucca ccuccccaac aaggcuugcu uccugauggc acauaacgga 1380
uggguuaugg gcgacgaccc ccugaggaau uuugcugaac ccggcuccga agucuaucug 1440
agaagggaac ugaucuguug gggcgacucc gucaaguugc gcuauggcaa uaagccugag 1500
gacuguccau aucugugggc ucacaugaag aaguacacag agaucaccgc uacauacuuu 1560
cagggcguca ggcucgacaa uugucacucc acaccccugc acguggcuga guauaugcug 1620
gaugccgcua ggaaucucca gccuaaccua uauguggucg cugaacuguu caccggcucu 1680
gaggaucugg auaauguguu ugugacacgc cugggcauca gcucccugau ccgcgaggcu 1740
augagugccu acaauaguca cgaggaaggc cggcuugugu accgcuaugg cggcgaaccc 1800
gugggcagcu uuguacaacc uugucugcgg ccucugaugc ccgcuauugc ucacgcuuug 1860
uuuauggaca ucacccauga caacgaaugu cccauuguac accggucugc uuacgacgcu 1920
cugcccucca caacuauugu caguauggcu uguugugcau cuggcucaac ucggggcuac 1980
gacgaauuag uuccgcauca aaucucugua guguccgaag aacgguuuua uacaaaaugg 2040
aaccccgaag cgcugcccuc caauaccggc gaagugaacu uucagucugg gaucaucgcc 2100
gcuagaugug cuauuucuaa acugcaucag gaacugggcg cuaagggcuu uauucaagua 2160
uauguggacc aggucgacga ggacaucgua gcuguaaccc ggcacagccc cagcauacau 2220
cagagugucg uggcuguguc uagaacagcu uuuaggaacc caaagaccuc uuuuuacucc 2280
aaagagguuc cacagaugug uaucccgggc aaaauugagg aaguggucuu ggaggccagg 2340
acaauugagc guaauacgaa gcccuauaga aaggaugaga auagcaucaa cggcacuccu 2400
gauaucacug uggaaauccg ggagcauaua caguugaacg agucuaagau cguuaagcag 2460
gcugguguag cuaccaaggg ucccaaugag uauauucagg aaauugaguu ugaaaaccug 2520
uccccgggca gcguaauuau cuuuagaguc agucucgacc cccaugcuca aguggcagug 2580
gggauccugc ggaaccauuu gacucaguuc uccccccauu uuaagucugg cagucuggcc 2640
guugacaacg cugaccccau auugaagauu cccuuugcuu cccuggcuuc acgguugacc 2700
cuggccgaac ugaaccagau uuuauaucgc ugcgaguccg aggaaaagga ggauggcggc 2760
ggcuguuaug acauucccaa cuggucugcu cuaaaauacg cugggcugca gggucugaug 2820
agugugcugg cugaaauucg ccccaagaac gaccugggcc auccauuuug caacaaucug 2880
cgcaguggcg acuggaugau ugauuaugug uccaaccggc ugauuagucg gagcggcacg 2940
aucgcagaag ucggcaaaug gcuccaggcu auguuuuuuu accuaaagca gauacccaga 3000
uaccugaucc cuuguuauuu ugacgcuaua cugauuggcg cuuacacaac cuugcuggac 3060
accgccugga agcagaugag cagcuuuguc caaaauggca gcacauucgu gaagcauuug 3120
ucucugggca gugugcagcu guguggcgug ggcaaauuuc caaguuugcc gauccugucu 3180
cccgcuuuga uggaugugcc auaucgacug aacgagauaa ccaaggaaaa ggaacagugu 3240
uguguuucac ucgcugcugg cuugccccau uuuuccucug gcauuuuccg auguuggggg 3300
agggacacau ucauugcccu gcgugggauc cugcugauua ccggccgcua cguugaggcu 3360
agaaacauua uuuuggcuuu cgccggcaca uugagacaug gucugauacc caaucugcug 3420
ggcgaaggca uauaugcuag guauaacugu cgcgacgcag ugugguggug gcugcaaugc 3480
auccaggauu auugcaagau gguuccuaac ggccuagaca uccugaagug cccagugucc 3540
cggauguauc caaccgauga uucagcuccc cugcccgccg guacacugga ccaaccccug 3600
uucgaaguua uucaagaagc caugcagaag cacaugcagg guauccaguu ucgagagagg 3660
aaugcaggcc cccaaauuga ccggaacaug aaggaugaag gcuucaacau uacugcuggc 3720
gucgacgaag aaacaggguu uguguacggc ggcaaccggu uuaacugcgg cacauggaug 3780
gacaagaugg gcgaaagcga ucgcgcacgg aaccggggca uccccgccac gccgcgugau 3840
ggcucugcug uggagaucgu cggccucagc aaauccgcug uccgauggcu guuagagcug 3900
ucgaagaaga acaucuuucc cuaccaugag guuaccguua agagacacgg caaagcuauc 3960
aaagugucuu augacgaaug gaauagaaag auucaagaca acuucgagaa acuguuucau 4020
guguccgagg aucccagcga ccucaacgaa aagcauccca aucucguaca uaagaggggc 4080
aucuauaagg acucauaugg cgcaucaagu ccuuggugcg acuaucagcu ucggcccaau 4140
uucacgaucg caauggucgu ggcacccgag cuguuuacua cggagaaggc uuggaaggcu 4200
cuggaaaucg cugagaagaa gcugcugggc cccuugggca ugaaaacccu ggaucccgac 4260
gacaugguau acuguggcau auaugacaac gcucuggaua acgauaacua uaauuuggcu 4320
aaaggcuuua auuaucacca gggcccagaa uggcuauggc ccauuggcua cuuucugcga 4380
gcuaagcuau auuuuucucg ccugaugggc ccugaaacca ccgcuaagac uaucgugcuc 4440
gucaagaacg uacugaguag gcauuacgug cacuuggagc gcagcccaug gaagggcuug 4500
ccggaacuga cgaacgaaaa cgcucaguau uguccguuuu cuugugaaac acaggcuugg 4560
aguauugcua cuauccugga aacucuguac gaucuguag 4599
<210> 18
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 18
augggucauu ccaaacaaau uaggauccug cuguugaacg aaauggaaaa gcucgaaaag 60
accuuauuuc gccuggagca gggcuaugag uugcaguuuc ggcugggacc aacucugcaa 120
ggaaaggcug uaaccgugua cacaaauuau ccuuuucccg gcgaaacguu uaacagggag 180
aaguuucgau cucuggauug ggagaacccc acugaaaggg aggacgauag cgacaaguau 240
ugcaaacuga aucugcaaca guccggcagc uuucaauauu auuuucugca agggaacgaa 300
aagagcggcg ggggcuauau uguaguagac ccuauacuga gagucggagc ugacaaccau 360
guucugcccu uggacugcgu gacacugcag accuuuuugg cuaagugucu gggccccuuu 420
gaugaauggg agagucggcu ccguguggcu aaggagagcg gguacaauau gauucacuuu 480
accccccugc aaacuuuggg acuuucccgg agcuguuaua gccuggcaaa ccagcuggag 540
uugaauccug acuuuucccg cccaaauagg aaguauaccu ggaacgacgu gggccaacug 600
guggagaagc ugaagaagga guggaacgug aucugcauua ccgacgucgu guauaaucau 660
accgccgcca acuccaagug gauacaggaa cauccugagu gugcuuacaa ccucguaaac 720
uccccgcauc ugaagccagc uuggguucug gauagagcuu uguggcgguu cagcugugac 780
gucgcugagg gaaaguacaa ggaaaaaggg auuccugcuc ucaucgagaa ugaccaccac 840
augaacagua uuaggaagau uaucugggag gacaucuucc cgaagcuaaa gcugugggaa 900
uucuuccaag uggacgugaa caaggccgug gagcaguucc gacggcuguu aacucaagag 960
aaccguaggg ucacaaaauc ggauccuaac cagcaccuga ccauuaucca agacccggaa 1020
uaucggcggu ucggcuguac uguggacaug aacauugcuu ugacaacuuu cauuccccac 1080
gacaaggguc cagcagcuau cgaggaaugu uguaauuggu uucauaagcg gauggaggaa 1140
cucaacuccg agaagcauag gcugauuaau uaucaucagg aacaggcugu gaauugucug 1200
uugggcaaug uuuucuacga acggcuggcu ggccauggcc ccaaacucgg accuguuaca 1260
aggaagcauc cacugguuac ccgauacuuu accuuuccau uugaagaaau ugauuuuagc 1320
auggaagaau ccaugaucca ucuaccuaac aaggccuguu uucugauggc ccauaacggc 1380
uggguuaugg gagaugaucc cuugcggaac uuugcugaac ccggaucuga aguauaucug 1440
cggagagagu ugauuuguug gggcgacucc guaaagcugc gcuauggcaa caagccagag 1500
gacugucccu auuugugggc ucauaugaag aaguauaccg agaucaccgc uacguauuuu 1560
caaggcguca ggcucgauaa uugucauucc acuccgcugc auguugcuga guauauguug 1620
gacgcugcuc gaaaucugca gcccaauuug uauguugugg ccgagcuguu uaccggcucc 1680
gaggaccucg auaacguguu cgucacgcga cuaggcauca gcagcuugau ccgggaagcc 1740
auguccgcuu auaacuccca cgaggagggc cgacuggucu accgcuaugg cggcgaacca 1800
gugggcaguu uuguacagcc uugucugagg ccccucaugc ccgcuauugc ucaugcucug 1860
uucauggaca uuacucauga uaacgagugc ccuauagugc aucgguccgc cuacgacgcc 1920
cugccgagca cuacaauagu guccauggcu uguugugcaa gcgggagcac ccgcggcuau 1980
gacgagcugg ugccgcauca aauaucugua gucagcgaag aaagguuuua uaccaagugg 2040
aacccggaag cuuuaccuuc caauacuggg gaagugaacu uucagagcgg cauuaucgcu 2100
gcgagaugug cuauauccaa guugcaucag gaacuggggg caaagggguu uauucaagua 2160
uauguagacc agguugacga agauauagug gcugugacac gccacagccc aagcauccac 2220
caguccgugg uggcugucag ccggacugcu uuucgcaacc caaagacaag cuuuuauagc 2280
aaagaagugc cgcagaugug cauucccggc aaaauugagg aggucgugcu ggaggcuagg 2340
acuaucgaaa ggaacaccaa gccguacagg aaggacgaga auuccaucaa cgggacucca 2400
gauauuacag uugagauccg ggaacauauu caguugaaug agucgaagau uguuaagcag 2460
gcuggcguag cuacaaaggg gccaaacgag uauauucaag agauagaauu cgaaaaccug 2520
agccccgguu ccgugaucau uuuucgagug ucccuggauc cucaugcuca aguggccguu 2580
ggcauucuga gaaaccaucu cacacaauuu uccccucacu uuaaaagugg cagccuggcu 2640
guggacaacg ccgauccgau ccugaagauu ccauucgcuu cccuggcuag ucgccugaca 2700
cuggcugaac uaaaccagau ucuuuaccgc ugugaaucug aagagaagga ggacggcggc 2760
gguugcuaug acaucccaaa uuggagcgcu cugaaauacg cugggcugca gggccucaug 2820
agcguacugg cagaaaucag acccaagaac gaccugggcc accccuucug uaacaaccug 2880
agguccggcg auuggaugau cgacuaugug ucgaacagac ucaucucaag aagcggcacu 2940
auagcugagg ucggcaaaug guugcaggcu auguucuuuu aucugaagca aauuccgcgg 3000
uaucugaucc cgugcuauuu ugacgcuaua uugaucggcg cuuauacgac uuugcuggac 3060
acagccugga agcagauguc cagcuucgug cagaacggca gcacuuuugu gaagcaccug 3120
agucugggcu caguacagcu guguggcgug ggcaaauucc ccagccuacc aauucugucc 3180
cccgcucuga uggauguucc cuaucggcug aacgagauua ccaaggaaaa ggagcaaugu 3240
ugcgugagcc uggcagccgg ccugccccac uuuucuuccg gcauuuuucg cuguuggggu 3300
cgggacaccu ucauugcacu gcggggcauc cuguuaauca ccggccgcua uguggaagcu 3360
agaaauauua ucuuggcuuu ugcuggcacu cugcgacacg gccugauccc caaccuucug 3420
ggcgaaggca ucuaugcucg cuacaauugu cgcgacgccg ucugguggug gcugcagugu 3480
auacaggacu auugcaagau ggugccaaau ggccuggaca uccuuaaaug uccaguuucc 3540
cggauguacc caacugacga cagcgcuccu uugccugccg gcacacugga ucagccgcug 3600
uuugaaguga uccaggaagc uaugcagaag cauaugcagg gcauucaguu ucgggaacga 3660
aaugcuggcc cacaaauuga cagaaacaug aaggaugagg guuuuaacau uacugcuggc 3720
gucgaugaag agacaggguu uguguacggc ggcaacaggu ucaacugcgg cacauggaug 3780
gacaagaugg gcgaaucuga ccgugcuagg aacaggggaa uccccgcuac cccccgggac 3840
ggcagcgcug uggaaaucgu aggccugucc aaguccgccg uucgcuggcu ucuggagcug 3900
agcaagaaaa acaucuuucc cuaucacgag gucacaguga agagacaugg caaagcuauc 3960
aaagucucuu acgaugagug gaauagaaag auucaagaua acuuugaaaa guuguuccac 4020
guuucggagg accccagcga ucugaaugag aagcauccca acuuggugca caagaggggc 4080
auauacaagg acucuuaugg ugcuagcagc ccuuggugcg acuaucagcu gaggcccaac 4140
uuuacuaucg ccaugguggu ggcaccggaa cuguuuacaa cggagaaggc uuggaaggcu 4200
cuggaaauug cugagaagaa guugcugggg ccccugggga ugaaaacgcu ggauccagac 4260
gacauggucu acuguggcau auaugauaau gcucuggaua acgauaauua caaucuggcu 4320
aaggguuuca acuaucauca ggggccagag uggcucuggc cuauuggcua uuuccuccgu 4380
gcuaagcugu acuuuucucg ccugaugggg ccagaaacua ccgcaaagac cauuguauug 4440
gucaagaacg ugcugagccg gcauuaugua cauuuggaac gguccccuug gaaaggccug 4500
cccgaacuca cuaacgagaa cgcucaguau uguccguuuu ccugcgaaac ccaggcuugg 4560
uccaucgcaa cuauuuugga aacccuguac gaccuguag 4599
<210> 19
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 19
augggccaca gcaagcaaau cagaauccuc cugcucaacg agauggaaaa gcucgaaaag 60
acccuguuca gacuggagca gggauacgag cugcaguucc gccucggacc aacccuccaa 120
gggaaggccg ugaccgugua caccaacuac ccauucccgg gagaaaccuu caaccgggaa 180
aaguucagga gccuggacug ggaaaacccc accgagagag aggacgacag cgacaaauac 240
ugcaagcuga accuccaaca gagcggcagc uuccaguacu acuuccucca aggaaacgaa 300
aagagcgggg gcggcuacau cgugguggac ccaauccugc gggucggggc agacaaccac 360
gugcugccac uggacugcgu gacacugcag accuuccugg ccaagugccu gggaccguuc 420
gacgaauggg agagccggcu gcggguggcc aaggagagcg gcuacaacau gauccacuuc 480
acaccgcucc aaacccucgg ccugagcaga agcugcuaca gccuagccaa ccagcuggag 540
cucaacccgg acuucagcag gcccaacaga aaguacaccu ggaacgacgu gggacagcuc 600
guggaaaagc ugaaaaagga auggaacgug aucugcauca ccgacguggu guacaaccac 660
accgccgcca acagcaaaug gauccaggaa cacccggaau gcgcguacaa ccucgugaac 720
agcccccacc ugaagccggc cuggguacug gacagggcac uguggcgcuu cagcugcgac 780
guggccgaag gaaaguacaa ggagaagggc auccccgccc ugaucgagaa cgaccaccac 840
augaacagca uccggaagau caucugggag gacaucuucc caaagcugaa gcugugggag 900
uucuuccaag ucgacgugaa caaggccgug gaacaguuca gacgccugcu gacgcaggag 960
aaccggaggg ucaccaagag cgaccccaac cagcaccuga ccaucaucca agacccggaa 1020
uaccggagau ucggaugcac cguggacaug aacaucgccc ugaccaccuu caucccacac 1080
gacaaggggc ccgcagccau cgaggagugc ugcaacuggu uccacaagag aauggaggaa 1140
cugaacagcg aaaagcaccg ccucaucaac uaccaccagg aacaggccgu gaacugccuc 1200
cucggaaacg uguucuacga gcggcuggcc ggccacgggc ccaagcuggg ccccgugacc 1260
cgcaagcacc cgcucgugac gcgguacuuc accuucccgu ucgaagaaau cgacuucagc 1320
auggaggaga gcaugaucca ccucccgaac aaggccugcu uccucauggc ccacaacggc 1380
ugggucaugg gcgacgaccc gcugcggaac uucgcagagc ccggaagcga aguguaccuc 1440
agaagggagc ugaucugcug gggagacagc gugaagcugc gcuacggaaa caagcccgag 1500
gacugcccgu accugugggc gcacaugaag aaguacaccg agaucaccgc caccuacuuc 1560
caaggagugc ggcuggacaa cugccacagc accccgcugc acguggccga guacaugcug 1620
gacgccgcca gaaaccucca gcccaaccuc uacguggugg cagagcuguu caccgggagc 1680
gaggaccucg acaacguguu cgugacccga cucggcauca gcagccugau ccgggaagca 1740
augagcgccu acaacagcca cgaggaaggg aggcuggugu acagauacgg aggagaaccc 1800
gugggcagcu ucgugcagcc gugccugagg ccccugaugc cagccaucgc gcacgcgcug 1860
uucauggaca ucacccacga caacgaaugc ccgaucgugc accggagcgc auacgacgcc 1920
cugccgagca ccacgaucgu cagcauggcc ugcugcgcca gcggcagcac ccgaggauac 1980
gacgagcugg ugccccacca aaucagcguc gucagcgaag aacgcuucua caccaagugg 2040
aacccggaag cacugccgag caacaccgga gaagugaacu uccagagcgg aaucaucgcc 2100
gcgcgcugcg cgaucagcaa acugcaccag gaacugggag ccaagggguu cauccagguc 2160
uacgucgacc aaguggacga ggacaucgug gcagugaccc gccacagccc cagcauccac 2220
caaagcgugg uggccgugag ccggacagcg uuccggaacc ccaagacgag cuucuacagc 2280
aaagaggugc cccagaugug caucccggga aagaucgagg aaguggugcu cgaagcgcgc 2340
accaucgaac gcaacaccaa accguaccgc aaggacgaaa acagcauaaa cgggaccccc 2400
gacaucaccg uggagaucag ggaacacauc cagcugaacg agagcaagau cgugaagcag 2460
gccggggucg ccaccaaggg cccgaacgag uacauccagg agaucgaguu cgagaaccuc 2520
agccccggga gcgugaucau cuucagaguc agccuggacc cacacgccca aguggccgug 2580
ggcauccucc ggaaccaccu gacccaguuc agcccgcacu ucaagagcgg gagccucgcc 2640
guggacaacg ccgacccgau ccugaagauc ccguucgcga gccuggccag ccggcucacc 2700
cuggccgaac ugaaccagau ccuguaccgc ugcgagagcg aagaaaagga ggacggagga 2760
gggugcuacg acaucccaaa cuggagcgcg cugaaauacg cgggccugca gggccugaug 2820
agcgugcugg ccgaaauccg ccccaagaac gaccugggac acccauucug caacaaccug 2880
aggagcggag acuggaugau cgacuacguc agcaacagac ugaucagccg cagcggcacc 2940
aucgccgaag ucggaaaaug gcuccaggcc auguucuucu accugaagca gaucccgcgg 3000
uaccugaucc cgugcuacuu cgacgccauc cugaucggcg ccuacaccac ccugcucgac 3060
accgccugga agcagaugag cagcuucguc caaaacggga gcaccuucgu gaagcaccuc 3120
agccugggca gcgugcagcu gugcgggguc ggaaaauucc cgagccugcc cauccugagc 3180
ccggcgcuga uggacgugcc guacagacug aacgagauca ccaaggaaaa ggaacagugc 3240
ugcgugagcc uggccgccgg acugccgcac uucagcagcg gcaucuuccg gugcuggggg 3300
cgcgacaccu ucaucgcccu gcggggaauc cugcugauca cgggccgcua cguggaggcc 3360
cggaacauca uccuggccuu cgccggcacc cugcggcacg gacugauccc gaaccuccug 3420
ggagagggga ucuacgcccg guacaacugc cgggacgccg ugugguggug gcugcagugc 3480
auccaggacu acugcaagau ggugcccaac ggccuggaca uccugaagug ccccgucagc 3540
cggauguacc cgaccgacga cagcgccccg cuccccgccg gaacccucga ccagccacug 3600
uucgaaguga uccaggaggc caugcaaaag cacaugcagg gaauccaguu cagagaacgg 3660
aacgccggac cccagaucga ccggaacaug aaggacgagg gauucaacau caccgccgga 3720
gucgacgagg aaaccggcuu cgucuacgga gggaaccggu ucaacugcgg aacauggaug 3780
gacaagaugg gagagagcga ccgcgccagg aaccgcggaa ucccagcaac cccgcgggac 3840
gggagcgcag uggaaaucgu ggggcugagc aaaagcgccg ugcgguggcu gcucgaacuc 3900
agcaagaaga acaucuuccc cuaccacgaa gucaccguga agagacacgg aaaggccauc 3960
aaagucagcu acgacgaaug gaacaggaag auacaggaca acuucgagaa gcuguuccac 4020
gucagcgagg acccgagcga ccugaacgag aagcacccca accuggugca caagcgcggg 4080
aucuacaagg acagcuacgg cgcgagcagc ccguggugcg acuaccaacu gcgccccaac 4140
uucaccaucg ccauggucgu ggcccccgaa cucuucacga ccgagaaagc guggaaggcg 4200
cuggagaucg cggaaaagaa gcuccucgga ccccugggga ugaaaacccu ggaccccgac 4260
gacauggugu acugcggcau cuacgacaac gcgcuggaca acgacaacua caaccuggcc 4320
aagggcuuca acuaccacca gggccccgaa uggcuauggc cgaucggcua cuuccugcgg 4380
gccaagcugu acuucagccg ccucaugggc ccagaaacca ccgcaaagac caucgugcug 4440
gucaagaacg uccugagccg ccacuacgug caccucgaga gaagcccaug gaagggacug 4500
cccgagcuga ccaacgaaaa cgcgcaguac ugcccguuca gcugcgaaac ccaggccugg 4560
agcaucgcca ccauccugga aacacuguac gaccucuag 4599
<210> 20
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 20
augggccaca gcaaacagau ccggauccuc cuacucaacg aaauggaaaa gcucgaaaag 60
acucuguucc ggcuggagca aggauacgag cuucaguucc gguugggccc gacgcugcag 120
gggaaggccg ugacagucua cacuaacuac ccauuccccg gcgaaaccuu caacagagag 180
aaguucaggu cccuggacug ggagaacccc acugaacggg aagaugauuc cgacaaguac 240
ugcaagcuga accuccagca auccgguuca uuccaguacu auuuccugca aggaaacgaa 300
aagagcggag gaggcuacau ugugguugau ccaauccuuc gcgugggagc cgauaaucau 360
gugcugccgc uggauugcgu cacccugcaa accuuccugg cgaagugccu gggcccguuc 420
gacgaauggg aaucccgccu gcgggucgcc aaagaguccg gguauaacau gauccacuuc 480
accccacucc aaacucuugg ccuguccaga uccugcuacu cguuggccaa ccaguuggag 540
cugaacccgg acuucucacg gccuaaccgg aaguacacuu ggaacgacgu cggacaacuc 600
guggagaagc ugaagaagga auggaauguc aucugcauua cugaugucgu guacaaccac 660
acggcggcga auucaaagug gauccaggag cacccagaau gcgccuacaa ccuggucaau 720
uccccucacc ugaagccggc cugggugcuc gaccgggccc uguggcgguu uucgugcgau 780
guggcggagg gaaaguacaa ggagaagggg auuccggcuc ucaucgaaaa cgaccaccac 840
augaacucca uccggaaaau uaucugggag gacaucuucc cgaaacuuaa gcugugggaa 900
uucuuccaag uggacgucaa caaggcugug gagcaguucc ggagauugcu gacacaagaa 960
aaccgccgcg ugaccaaauc cgauccgaac cagcaccuga cuaucauuca agacccggaa 1020
uaccggaggu uuggcugcac uguggacaug aauaucgcgc ugaccaccuu caucccgcac 1080
gacaagggac cggcagccau cgaagagugc uguaacuggu uccauaagag gauggaagaa 1140
uugaacucug aaaagcaccg gcugauuaac uaccaucagg aacaggccgu gaacugucuc 1200
cugggaaacg uguucuacga gcggcuggcg ggacacggac ccaagcucgg ccccgugacc 1260
cgcaagcauc cucucgugac uagauacuuu acuuucccau ucgaggagau cgacuucucc 1320
auggaagaau caaugaucca ccucccuaac aaggcuugcu uucugauggc acacaacggc 1380
ugggugaugg gcgaugaccc ccugaggaac uuugccgagc ccggcuccga aguguaccug 1440
aggagagagc ugauuugcug gggggacagc gucaagcugc gcuauggaaa caagccggaa 1500
gauugcccuu accucugggc ccacaugaag aaguacacug agauuacugc caccuacuuu 1560
caaggagugc gccuggauaa cugccacuca accccgcugc augucgcuga guacauguug 1620
gacgcagccc ggaaucugca accgaaccuc uacguggugg cggagcuguu caccggcucg 1680
gaggaucucg acaacguguu cgugacucgc cugggcaucu caucgcugau ccgggaagca 1740
auguccgccu acaacuccca ugaggagggu cggcuggugu accgcuacgg cggagaaccc 1800
gugggguccu ucgugcaacc gugccugcga ccccugaugc ccgccaucgc ccaugcccuc 1860
uuuauggaua uuacccacga uaacgaaugc ccuaucgugc accgcucagc cuaugaugcc 1920
cuccccucca ccaccaucgu guccauggcc ugcugcgccu cggggagcac ccgggguuau 1980
gacgagcugg ugccgcacca gauuucggug guguccgagg agagauucua caccaagugg 2040
aacccagaag cucugccguc aaacacugga gagguuaacu uucaguccgg uauuaucgcc 2100
gcuagaugug cuauuagcaa acugcaccaa gagcugggcg ccaaggguuu cauccaaguc 2160
uacgucgauc aggucgacga ggacaucgug gcugucacua ggcacucacc uagcauccac 2220
cagagcgugg uggccgugag ccgcacugcc uuccgcaacc caaagaccag cuuuuacucc 2280
aaggaggucc cucagaugug cauuccugga aagaucgaag aagugguccu ggaagcccgg 2340
accaucgaac gcaacaccaa gccuuaccgg aaggacgaaa acucgaucaa cgguaccccg 2400
gauauuacug uggagauucg cgaacacauc cagcucaaug aguccaagau cgugaagcag 2460
gccggagugg caaccaaggg gccgaacgag uacauccagg agaucgaauu cgagaaucuc 2520
agcccuggca gcgugaucau cuuccgagug ucguuggacc cacaugcuca ggucgccgug 2580
ggcauccuga ggaaccaccu gacccaguuu uccccgcauu ucaaguccgg uucgcuggcc 2640
guggacaacg cagauccgau ccugaagauc cccuucgccu ccuuggcuuc gcgccucacg 2700
cuggccgagc ugaaccagau ccuguauaga ugcgaauccg aagaaaagga agauggaggc 2760
gguuguuacg acaucccgaa cugguccgcc uugaaauacg ccggacugca gggauugaug 2820
uccgugcugg ccgaaauuag accgaagaac gaccuggggc acccguucug caacaaccuc 2880
cggucggggg acuggaugau ugauuacgug ucgaaccgcc ucaucucccg guccgguacu 2940
auugcggagg ucggcaaaug gcuccaggcc auguucuucu acuugaagca aaucccccgc 3000
uaccugaucc ccugcuacuu cgacgccauc cugaucggag ccuacaccac ccugcuggac 3060
accgcgugga agcagauguc uagcuucguc cagaacggcu ccaccuucgu gaagcaccug 3120
ucacugggcu cagugcagcu gugcggaguu ggaaaguucc ccagccugcc uauccugucc 3180
ccggccuuga uggacgugcc guacagacug aacgaaauca cuaaggagaa ggagcagugu 3240
ugcgugucgu uggcggccgg gcugccccac uuuucaagcg ggauuuuccg gugcugggga 3300
agggacaccu ucaucgcgcu gcggggaauc cugcugauua ccggacgcua cguggaggcu 3360
cggaacauca uucucgcguu cgccggcacc cugcgccaug gacugauccc uaaucugcuu 3420
ggagagggca ucuacgcucg guacaacugc agagaugccg ugugguggug gcuccaaugc 3480
auccaggauu acugcaagau ggugccuaac ggucuggaca uucugaagug cccugugucc 3540
cgcauguacc ccaccgacga cuccgccccc cugccugccg gaacccucga ccagccucug 3600
uucgaaguca uccaagaggc caugcagaag cauaugcagg gcauucaguu ccgcgaacgg 3660
aacgcagggc cccagaucga ccggaacaug aaggaugaag gguucaacau caccgccgga 3720
guggaugaag agacuggauu ugucuacgga ggaaaccgcu uuaacugcgg gaccuggaug 3780
gacaagaugg gagagucaga cagggccaga aauagaggaa uccccgcgac cccgcgcgac 3840
ggauccgccg uggagauugu gggccuguca aagagcgccg uccgcuggcu gcuggaacuu 3900
agcaagaaga acaucuuccc uuaccacgaa gugaccguga aaagacaugg caaagccauc 3960
aaagucuccu acgacgagug gaaccgcaag auucaggaca acuucgaaaa gcuguuccac 4020
guguccgaag aucccuccga ucugaacgag aagcauccga accuggucca uaagcgggga 4080
aucuacaagg auagcuacgg ugccagcucg ccguggugug acuaccagcu gcggccaaau 4140
uucacgauug cgauggucgu cgccccugag uuguucacca cugagaaggc cuggaaagcc 4200
cuggaaauag ccgagaagaa auuacugggg ccacugggca ugaaaacucu cgacccugac 4260
gacauggugu acuguggaau cuacgacaac gcacuggaca acgauaacua caauuuggcc 4320
aagggauuca acuaccacca ggggcccgaa uggcuguggc cuaucggaua cuuccuucgg 4380
gcaaagcuuu acuucucccg ccugauggga ccugaaacua cugccaagac cauugugcuu 4440
gugaagaacg ugcucucacg gcacuacgug caccuggaga gauccccgug gaaggggcuc 4500
ccggagcuga ccaacgagaa cgcccaguac ugcccuuucu ccugcgaaac ccaagccugg 4560
uccaucgcca ccauacuuga aacucuguac gaccuguag 4599
<210> 21
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 21
augggacacu ccaagcagau ccgcauucuc cuccugaacg aaauggaaaa gcuggaaaag 60
accuuauuuc gccuugaaca aggauacgag cugcaguucc ggcuuggacc gacccuccaa 120
gggaaagcug ucaccgucua caccaacuac cccuucccgg gggagacuuu caacagagag 180
aaguucagau ccuuggacug ggaaaauccc acggaacggg aggacgacuc cgauaaguac 240
uguaaacuga accuccagca gucgggaucg uuccaguacu acuuccuaca agggaacgaa 300
aaguccggug gugguuacau cgugguggac ccgauccugc gcgugggggc cgacaaccac 360
gugcugccgc uggacugcgu gacucugcaa accuuccugg ccaagugccu cggcccguuc 420
gaugaauggg agucgagacu gcggguggcc aaggaaagcg gauacaauau gauucacuuu 480
acuccgcugc aaacccuggg gcugucgcgc uccugcuacu cgcucgcuaa ccagcuggaa 540
cugaauccgg auuuuagccg gccgaaccgg aaguauacgu ggaacgacgu cggucagcug 600
gucgagaagc ucaagaagga guggaacguc auuugcauca ccgacguggu cuacaaccac 660
accgcggcca auuccaagug gauccaggaa cacccggagu gcgccuauaa cuuggugaac 720
uccccucauu ugaaacccgc augggugcuc gaccgggccc uguggcgcuu uucaugcgac 780
guggcggaag ggaaguacaa ggaaaagggg auucccgccc ugaucgagaa cgaucaccac 840
augaacucaa uucgcaaaau caucugggag gauaucuucc cuaagcucaa gcugugggaa 900
uucuuccaag ucgacgucaa caaggccgug gaacaauuca gaaggcugcu gacccaagag 960
aaccgcagag ugaccaaguc ggacccgaac cagcaccuga cuauaaucca ggacccggaa 1020
uaccggaggu ucgguugcac cguggacaug aacauugcac ucaccaccuu caucccgcac 1080
gauaaggguc cggcggcgau ugaggagugc ugcaacuggu uccacaagcg gauggaggaa 1140
cugaacuccg agaagcaccg gcugauuaac uaccaccagg aacaggcugu gaauugccug 1200
cuggggaacg uguucuacga acggcucgcc ggccacggcc ccaagcuggg gcccgucacc 1260
cgcaagcacc cgcucgugac ucgauauuuc accuucccgu ucgaggaaau ugauuucucc 1320
auggaggaau caaugaucca ucugccgaac aaggccuguu ucuugauggc ccacaacggc 1380
ugggugaugg gagaugaccc gcugagaaau uucgccgagc cgggguccga gguguaccug 1440
aggagagaac ucaucugcug gggggauucc gugaaacugc gcuacgggaa caagcccgag 1500
gacugccccu accugugggc acauaugaag aaguacaccg aaaucaccgc caccuacuuc 1560
caaggggucc ggcuggauaa cugccauuca acuccgcucc auguggccga auacaugcug 1620
gacgccgcac ggaacuugca gcccaaccuc uacguggugg ccgaacucuu cacugggagc 1680
gaagaucucg auaacguguu cgugacccgg cucggaauuu cgagccugau ccgggaagcg 1740
augucugcuu acaacucgca cgaagaggga aggcuggugu acagauacgg gggggagccc 1800
gugggauccu ucgugcaacc gugccugagg ccgcuuaugc ccgcgaucgc ccacgcucug 1860
uucauggaca ucacccacga caacgaaugu ccgauugucc accggagugc cuacgaugcc 1920
cuuccgagca cuaccaucgu gucgauggcg ugcugugcgu ccgggucuac ccgcggcuac 1980
gacgaacucg ucccgcacca aaucagcgug guguccgaag aacgguuuua cacuaagugg 2040
aacccggagg ccuugcccuc gaacaccggg gaggucaacu uccagucggg aaucauugcc 2100
gcccgaugug ccaucucaaa guugcaccaa gaacucgggg caaagggguu cauccaagug 2160
uacguggacc aaguggacga ggauauugug gccgugacca ggcacucccc gucgauccac 2220
caguccgugg uggcagucuc aagaacugcc uuccggaacc ccaagaccuc cuuuuacucg 2280
aaggaagugc cccagaugug caucccgggc aaaaucgaag aagucgugcu ggaagccaga 2340
acuaucgagc ggaacaccaa gcccuaccgg aaggaugaga acagcaucaa cggcacgccc 2400
gacaucaccg ucgagaucag agagcacauc cagcugaacg aauccaagau ugucaagcag 2460
gccggcguag cgacuaaggg acccaacgaa uacauccagg agaucgaauu cgaaaaccug 2520
ucgccgggau ccgugauuau cuuccgggug ucccuggacc cgcacgccca aguggccgug 2580
ggaauccugc ggaaccaccu gacccaguuc uccccgcauu ucaaguccgg cucccuggcg 2640
guggacaaug cagacccgau ucucaagauc ccguucgccu ccuuggccuc ccgccugacc 2700
cucgccgaac ucaaccagau ccuguaccgc ugcgagucug aggaaaagga ggacggggga 2760
ggaugcuacg auauuccgaa cuggucugcc cuuaaauacg cgggacugca gggucugaug 2820
ucggugcucg ccgagaucag accgaagaac gaccuggggc acccguuuug caacaaccug 2880
agaucggggg auuggaugau cgacuacgug ucgaaccggc uuaucucucg cuccggcacc 2940
aucgccgagg ucgggaagug gcugcaggcc auguucuucu accugaagca aauuccgcgc 3000
uaccugaucc cgugcuacuu ugaugccauc cugaucggcg ccuacacgac cuugcuggac 3060
accgccugga agcagaugag cagcuucgug cagaacggcu ccacguucgu gaagcaucug 3120
ucccugggau ccgugcagcu uugcgguguc ggaaaguucc cuucgcugcc uauucugucc 3180
ccggcgcuga uggacgugcc guaccggcug aacgagauca cuaaggagaa ggaacaaugc 3240
ugcgugucac uggcggcggg acucccccac uucuccuccg ggauuuuccg guguugggga 3300
agagauaccu ucauugcccu gagggggaua cugcugauca ccggaagaua cguggaggcu 3360
agaaacauca uccucgccuu cgccggcacu cuccgccacg ggcugauccc caaccucuug 3420
ggcgaaggaa ucuacgcccg guauaacugc cgcgacgcgg ucugguggug gcuucagugc 3480
auccaggauu acugcaaaau ggugccgaau ggacuggaua uucugaagug ccccguuagc 3540
cgcauguacc ccaccgauga cuccgcaccc uugccggcgg gcaccuugga ccagccgcuc 3600
uuugaaguga uccaggaggc caugcagaag cacaugcagg guauccaguu cagagagcgg 3660
aacgcugggc cgcagauuga ccggaacaug aaggacgaag guuucaacau uacugccggg 3720
guggacgaag agacuggguu uguguacggg gggaaccgcu ucaacugugg gaccuggaug 3780
gauaagaugg gggagucgga ucgggcucgg aacaggggga uccccgcuac cccccgggac 3840
gggucggcug uggagauugu cgggcugagc aagagcgcag ugcgcuggcu gcuggagcuc 3900
agcaaaaaga acaucuuccc cuaccacgaa gucacuguga agagacacgg aaaggccauc 3960
aagguguccu acgaugagug gaacagaaag auccaagaca acuucgagaa gcuguuccau 4020
guguccgagg auccgagcga ucugaacgaa aagcauccga accucgugca caagcgcggg 4080
aucuacaagg acucguacgg cgcauccucg ccguggugcg acuaccagcu gcgccccaau 4140
uucaccauug ccaugguggu ggcgcccgag cuguucacua ccgagaaggc cuggaaggcc 4200
cuggagauug cugagaagaa gcugcuggga ccgcucggga ugaaaacuuu ggacccugac 4260
gacauggugu acuguggaau cuacgacaau gcgcucgaca acgacaacua caaucuugcg 4320
aagggauuca acuaccauca ggggcccgaa uggcuguggc cgaucgggua cuuccugcgc 4380
gccaagcugu acuucucccg gcugaugggg cccgagacaa ccgccaagac cauugugcuc 4440
gucaagaacg uucucucccg gcacuacgug caccucgaaa gauccccgug gaaggggcug 4500
cccgagcuca ccaacgagaa cgcacaguac ugcccguucu ccugugaaac ccaggccugg 4560
agcaucgcca ccauauugga aacucuguau gaccucuag 4599
<210> 22
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 22
augggacacu cgaaacagau cagaauccug uugcugaacg aaauggagaa gcuggaaaag 60
acccucuuuc ggcucgaaca gggcuacgag cugcaguucc gccugggucc gacucuacaa 120
ggaaaggcag ugacugugua caccaacuac ccauuucccg gagaaaccuu caaccgggag 180
aaguuccggu cccuggacug ggaaaaccca accgaacgag aggaugauuc cgacaaguac 240
ugcaagcuga accuccaaca guccggauca uuccaguacu acuuuuugca aggaaacgag 300
aaguccggag gaggcuacau cguggucgac ccgauucuga gagugggagc ugacaaucau 360
guccugccuc uggacugcgu aacccugcaa accuucuugg ccaaaugccu gggcccguuc 420
gaugaguggg aaagccgccu ccgggucgca aaggaaagcg gcuacaauau gauucacuuc 480
acuccucugc aaacccucgg ucugucccgg uccuguuauu cccuggcgaa ccagcuggag 540
cuuaaccccg acuucucgcg cccaaaccgg aaguacaccu ggaacgacgu cgggcagcuu 600
guggaaaagc ugaagaagga guggaacgug aucugcauca cugacguggu guacaaccac 660
accgcggcca acuccaagug gauccaagaa caucccgaau gcgcguacaa ccucgugaac 720
agcccgcauc ugaagccugc cugggugcug gauagggccc ucuggagauu cagcugcgac 780
guggccgagg ggaaguacaa agaaaaggga auucccgccc ugauugagaa cgaccaucac 840
augaacucaa uccgcaagau caucugggag gauaucuucc cuaagcuuaa guugugggag 900
uucuuccaag uggacgugaa caaggcagug gagcaguuca gaaggcugcu gacucaagaa 960
aacagacggg ucacuaaguc cgacccuaac cagcaccuua ccaucauuca agacccggag 1020
uaccgccggu uuggcugcac ugucgacaug aacaucgccc ugaccacuuu caucccgcau 1080
gacaagggcc cggcggcaau cgaggaaugc uguaacuggu uucauaagag gauggaggaa 1140
cugaauuccg agaagcaccg gcugauuaac uaccaccagg aacaggcagu gaauugccuc 1200
cugggaaacg uguucuacga acggcuggcu ggacacggac cgaagcuggg ucccgugacu 1260
cgcaagcauc cgcucgugac ucgcuacuuc accuucccgu uugaggagau ugacuuuucc 1320
auggaagaau ccaugaucca ccucccgaac aaggcuugcu uccugauggc gcacaacgga 1380
ugggucaugg gcgacgaccc acugcgcaac uucgcugagc cuggcagcga ggucuaccug 1440
agaagggaac ugauuugcug gggagacucc gucaagcugc gcuauggaaa caagcccgaa 1500
gauugccccu accugugggc ucacaugaag aaguacacug aaaucacugc cacguacuuc 1560
cagggagucc ggcuggacaa uugccacucc accccccucc auguggccga guacaugcuc 1620
gaugcagcga ggaaucugca gcccaaucug uacgugguug ccgaacuguu caccggcucc 1680
gaggaccucg acaacguguu cgugaccaga cuugggauuu ccagccugau ccgggaagcc 1740
augucggccu acaacuccca ugaagagggc cgccuggugu accgcuacgg cggagaaccc 1800
gugggaagcu ucgugcagcc uugccuccgg ccgcugaugc cugcgaucgc ccacgcccug 1860
uucauggaua uuacccacga caacgaaugc cccaucgugc aucgcucggc cuacgacgca 1920
cugccuucga ccaccaucgu guccauggcc ugcugcgccu ccgguagcac ccgcggauac 1980
gaugaacucg ugccgcacca gaucagcgug guguccgaag aaagauucua caccaagugg 2040
aacccugagg cacugccgag caacaccgga gaagugaacu uccagucggg uauuaucgcc 2100
gcucgcugug ccaucuccaa acuccaccaa gagcucggug ccaagggauu cauucaaguc 2160
uacguggauc aggucgacga agauauugug gccgugacca ggcacucacc uuccauccac 2220
caauccgucg ucgccguguc acggacugcc uuccggaacc ccaagacuuc guucuacucg 2280
aaagaggugc cacagaugug uauccccgga aagaucgaag aggucguccu ggaagcccgg 2340
accauugaga ggaacaccaa gccuuaccgg aaggacgaga acagcaucaa cgguaccccu 2400
gauauuacug uggagauccg cgaacacauc cagcucaacg aaucaaagau ugucaagcag 2460
gccggagugg ccaccaaggg acccaacgag uacauccagg agaucgaauu ugaaaaccuc 2520
uccccuggcu ccgugauuau cuuccgggug ucccuggacc cucacgccca aguggccguc 2580
ggcauccuca gaaaccaccu gacccaguuc ucaccccauu ucaaguccgg uucccuggcg 2640
guggacaacg ccgauccgau cuugaagauc cccuuugcau cccuggccuc ccgccugacu 2700
cucgcggaac ugaaccagau ccuguaccgc ugcgaaucag aggaaaagga ggacggcggc 2760
ggcuguuacg auaucccgaa uugguccgcu cugaaauaug cgggacuuca ggggcugaug 2820
agcgugcugg cggagauccg gccgaagaau gaccugggac acccauucug caacaacuug 2880
cggagcggag acuggaugau cgauuacguc agcaacagau ugaucucccg gagcggcacu 2940
aucgcggagg ucggaaagug gcuccaggcc auguucuucu accugaagca gaucccccga 3000
uaccucaucc cguguuacuu cgacgccauu cugaucgggg ccuacaccac ccugcuggac 3060
accgccugga agcagaugag cuccuuugug caaaacgggu ccaccuucgu gaagcaccuu 3120
ucacugggau cagugcagcu cugcggcgug ggaaaauucc ccucgcugcc cauucugagc 3180
ccggcccuca uggacguccc uuaccggcug aacgagauca ccaaggagaa ggagcagugc 3240
uguguuuccc uggcugccgg acuuccacac uucucguccg gcaucuuccg gugcuggggc 3300
cgggauaccu ucauugcccu gcggggcauc uuguugauca ccggucgcua cguggaggcu 3360
cggaacauua uucuggcauu cgccggcacu cugagacacg gucugauucc gaaucuuuug 3420
ggcgaaggaa ucuacgcccg cuacaacugu cgggacgccg ugugguggug gcuccagugc 3480
auucaggacu auugcaagau ggugccgaac ggccuggaca uccugaagug cccagugucg 3540
aggauguauc caaccgacga cagcgcgccu cugccggccg ggacccucga ccaaccccug 3600
uucgaaguca uucaagaggc uaugcagaag cacaugcagg guauucaguu ccgggagaga 3660
aacgcggggc cccagauuga uaggaacaug aaagacgagg gcuucaacau cacugccggc 3720
guggacgaag aaaccgguuu uguguacgga ggaaaccggu ucaacugcgg uaccuggaug 3780
gacaagaugg gagaguccga ucgcgcgcgc aacagaggga ucccggcaac cccgcgggac 3840
ggaucagcgg uggaaauugu gggacugagc aagagcgccg ugcgcuggcu ccuggaacug 3900
agcaaaaaga acaucuuccc cuaccacgaa gugaccguga agcggcacgg aaaggccauc 3960
aaagucucau acgaugaaug gaauaggaag auccaggaua acuucgagaa gcuguuucac 4020
guguccgagg aucccuccga ucugaacgaa aagcauccga aucucgugca caagcgcggc 4080
aucuacaagg acucguacgg agccuccucc ccuuggugcg auuaucagcu gcggccuaac 4140
uucaccauug ccauggucgu ggcuccggag cuguucacua cugagaaggc cuggaaggca 4200
cuugaaauug ccgagaagaa gcugcuuggg ccuuugggga ugaaaacccu ggauccggac 4260
gacauggugu acugcggaau cuacgacaac gcccuggaca acgacaacua caaucuggcg 4320
aagggcuuca auuaccacca gggcccggaa uggcuguggc cuauugggua cuuccugcgc 4380
gccaagcugu acuucucacg gcugauggga ccagagacua ccgccaagac uaucguccuc 4440
gugaagaacg ugcugucccg gcacuacgug caucuggaga ggagcccuug gaagggccuu 4500
ccugagcuga ccaacgaaaa cgcccaguac ugccccuucu ccugcgaaac ccaggcuugg 4560
uccauugcca cuauacugga aaccuuauau gaccuguag 4599
<210> 23
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 23
augggacacu caaagcagau ccggauccuc cugcugaacg aaauggaaaa gcucgaaaag 60
acucuguucc ggcuggaaca gggcuaugag cugcaguucc ggcucggacc gacgcugcaa 120
ggaaaggccg ugacugugua caccaacuac cccuucccgg gagaaacuuu uaacagagaa 180
aaguuuaggu cccuggacug ggagaacccg accgagcggg aagaugacuc cgauaaguac 240
ugcaagcuga accuccagca gucgggaucg uuccaguacu acuuccuuca aggaaaugag 300
aagucuggug gaggauacau cgugguggac cccauccuga gagugggagc cgacaaucac 360
guccugccgc uugacugcgu gacucugcaa accuuccugg cuaagugucu cgggccguuc 420
gacgaauggg aguccaggcu gcgcguggcu aaggagagcg gcuacaauau gauccacuuc 480
accccgcucc aaacucucgg acucucgcgc uccuguuacu cccucgccaa ccaacuggag 540
cugaacccgg acuucucccg cccgaaccgg aaguacaccu ggaacgacgu cggucagcuc 600
guggagaaau ugaagaagga guggaacgug aucugcauua cggacgucgu guacaaccau 660
acugcggcca acuccaagug gauucaagag cacccggaau gcgcuuacaa ccuugucaac 720
uccccgcacc ugaagccggc cugggugcug gacagagccc uguggcgguu cucgugcgac 780
guggcagagg gaaaguacaa ggagaagggc auccccgccc uuauugagaa ugaccaucac 840
augaacucca uccggaagau caucugggag gauaucuucc cgaagcucaa gcugugggaa 900
uucuuccaag uggacgugaa caaggccgug gaacaguucc ggagacuccu gacccaagaa 960
aaccggagag ugaccaagag cgacccgaac cagcaucuga cuaucauuca ggauccggag 1020
uaucggcggu uuggcugcac uguggacaug aacaucgccc ucaccaccuu caucccgcac 1080
gacaagggcc ccgccgcgau cgaggaaugc ugcaacuggu uccacaagcg cauggaggag 1140
cugaauuccg aaaagcaccg ccugaucaac uaccaucagg agcaggcagu gaacugucuc 1200
cugggaaacg uguuuuacga acggcuggcc ggacacggcc cgaagcuggg ucccgugacc 1260
cgcaagcauc cccucgugac gcgguacuuc accuucccgu ucgaggagau cgacuucagc 1320
auggaagagu ccaugaucca ucugccgaac aaggccugcu uccucauggc gcauaauggu 1380
ugggucaugg gagaugaucc ccuccgaaac uuugcggagc cggguucgga aguguaucug 1440
aggagggagc ucaucugcug gggagauagc gugaaacuga gauacgggaa caagccggaa 1500
gauuguccgu accugugggc acacaugaag aaguacaccg aaaucacugc cacuuacuuc 1560
caaggaguuc gccuggauaa cugccauuca accccucugc augucgccga guacaugcug 1620
gacgccgcuc gcaaccuuca gccgaaucuc uacguggucg ccgagcuguu caccgguucc 1680
gaagaucugg acaacguguu cgugacuaga cugggaauca gcagccugau ccgggaagcg 1740
augagcgccu acaacuccca cgaagagggc cggcucgugu auagauacgg cggagagccg 1800
gucgggagcu ucgugcaacc cugccugcgg ccgcugaugc ccgccauugc ccacgccuug 1860
uucauggaua ucacccacga caacgagugu ccgaucgugc accggagcgc guacgacgcg 1920
uuaccgucca ccacgauugu gucgauggcc ugcugcgccu ccggaucgac ccgcggcuac 1980
gaugagcugg ucccgcauca aaucagcguc gucagcgaag aacgguucua cacuaagugg 2040
aaccccgagg cgcuccccuc caacaccgga gaagugaacu uccaauccgg cauuaucgcu 2100
gcacgcugcg cgauuagcaa gcugcaucag gagcuuggcg cuaagggguu cauacagguc 2160
uacguggauc aggucgacga ggacauugug gccgugacuc gccacucacc guccauucac 2220
caaagcgugg uggcuguguc ccggaccgcu uuccggaauc ccaagaccuc auucuacucc 2280
aaggagguuc cgcagaugug uaucccggga aagauugagg aagugguccu agaggcucgc 2340
acuauugaac gcaacaccaa gccguacaga aaggacgaga auuccauuaa cgggaccccg 2400
gauauuaccg uagaaauuag agaacauauu cagcugaacg agucgaagau cgugaagcag 2460
gccggggugg cuaccaaggg cccgaacgaa uacauccagg agauugaguu cgaaaaccuc 2520
ucccccggcu cggugaucau cuuccgggug ucccucgauc cccacgccca aguggccgug 2580
ggaaucuuga gaaaccaccu gacucaguuc agcccgcauu ucaaguccgg aucacucgcc 2640
guggacaacg ccgacccgau ccuuaagauc cccuucgcau cacuggccag ccgccugacc 2700
cuugccgaac ugaaccaaau ccucuaucgg ugcgaauccg aggagaagga agaugggggu 2760
ggaugcuaug acauuccuaa cuggucugcc cugaaauacg caggccugca gggacugaug 2820
uccguccugg ccgaaauccg cccgaagaau gaccuggggc accccuuuug caacaaucug 2880
agguccggag auuggaugau ugauuacgug uccaaccgcc ugauuucgcg gagcggcacc 2940
aucgcggaag ugggaaagug gcugcaagcc auguuuuucu accugaaaca gaucccccgg 3000
uaccucaucc cgugcuauuu cgaugcgauu uugauuggcg cguacaccac ccuccucgac 3060
acugccugga agcaaaugag cuccuucgug caaaacgguu caaccuucgu caagcacuug 3120
ucgcuggggu cgguccaacu guguggaguc ggaaaguucc cgagccugcc cauccugucg 3180
ccugcccuga uggacgugcc cuaccggcug aacgagauca ccaaagaaaa ggagcagugc 3240
ugcguguccc uggcggccgg acucccccau uucucguccg ggaucuuuag auguugggga 3300
cgggauacuu ucauugcgcu gcgcggcauc uuguugauua ccggacgcua cguggaagcg 3360
cggaacauca uacucgccuu cgccggcacc cucagacacg gccugauccc gaaccuccug 3420
ggagaaggca ucuacgcacg auacaacugu cgggacgccg ucugguggug gcugcagugc 3480
auucaggacu acugcaagau ggugccgaac ggucuggaca uccugaagug ccccguguca 3540
aggauguacc cgaccgacga uagcgcaccg cugcccgcgg ggacccugga ucagccgcug 3600
uucgaaguga uccaagaagc caugcaaaag cacaugcagg gaauucaguu cagagaaagg 3660
aacgcaggcc cccagauuga ccggaacaug aaggacgagg guuucaacau caccgccggc 3720
gucgacgaag aaacgggguu uguguacggg ggcaaccgcu ucaacugcgg uacuuggaug 3780
gacaagaugg gggaaucaga ccgcgcccgc aaccgcggaa uuccggcgac cccgcgcgau 3840
ggauccgcag uggaaaucgu gggacugucc aagucggcug ugcgguggcu gcuggagcug 3900
uccaagaaga acaucuuccc guaccacgag gucaccguga agagacacgg gaaggccauc 3960
aagguguccu acgacgaaug gaaccggaaa auccaggaua acuucgaaaa acuguuucac 4020
guguccgagg acccgucuga ccugaacgaa aaacacccga acuuggugca uaagcgggga 4080
aucuacaagg acaguuacgg agccucaagc ccguggugcg acuaccagcu gcggcccaac 4140
uucacaaucg cgaugguggu ggcgcccgag cuuuucacca cggagaaagc cuggaaggca 4200
cuggagaucg cugaaaagaa gcugcucggu ccccucggca ugaaaacccu ggacccggac 4260
gauauggugu acugcggaau cuacgacaac gcccuggaca augauaacua caaccuggcc 4320
aagggauuca auuaccacca ggggccggaa uggcuguggc cgaucggcua cuuucugcgg 4380
gcuaagcucu acuucucgcg gcugauggga ccggaaacca cagccaagac cauuguccuc 4440
gucaagaacg ugcugucccg ccacuacgug caucuggagc ggucaccuug gaagggccug 4500
cccgagcuga ccaacgaaaa cgcccaguac ugccccuucu ccugugaaac ucaagccugg 4560
ucaaucgcca cuauucucga aacucuguac gaucuguag 4599
<210> 24
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 24
augggucaca gcaaacaaau ccggauccug cugcucaacg aaauggaaaa gcuggaaaag 60
acucuguuuc gccuggagca gggcuacgaa cuccaauucc ggcugggacc aacgcugcaa 120
gggaaggccg ucaccgugua caccaacuac cccuuucccg gagagacuuu uaaccgcgaa 180
aaguucagaa gccuggacug ggaaaacccc accgaacggg aagaugauuc ggacaaguac 240
ugcaagcuga accuccagca guccggcagc uuccaguacu acuuccucca agggaacgaa 300
aagagcggag gaggcuacau cgugguggau ccgauccuuc ggguuggagc ggacaaccau 360
gugcugccuc uggacugcgu gacucugcaa accuuccuag ccaagugccu cggcccguuc 420
gaugaguggg aaucccggcu gcgaguggcg aaggagucag guuacaacau gauccacuuc 480
acaccgcugc agacccuggg acucucccgg uccugcuacu cauuggccaa ccagcucgaa 540
cugaacccgg acuucucaag gccgaaccgc aaguacacuu ggaaugacgu gggacaguug 600
guggagaagc ugaagaaaga guggaacgug aucugcauua ccgacguggu guacaaccac 660
acugcggcga auucgaagug gauucaggaa cauccggaau gcgccuacaa cuuggugaac 720
ucaccucacc ugaagccugc cugggugcug gacagagccc uguggcgcuu uagcugugac 780
gucgcggagg ggaaguacaa ggaaaaggga auccccgccc ugaucgagaa cgaccaccau 840
augaauagca uucgcaagau caucugggag gacauuuucc cgaagcugaa gcugugggag 900
uucuuccaag ucgacgucaa caaggccguc gaacaguucc ggcgguugcu gacccaagaa 960
aaccggagag ugacgaaaag cgacccuaac cagcaccuga caaucauuca ggacccugaa 1020
uaccggagau ucggcuguac cgucgauaug aacaucgccc ugaccaccuu uauuccccac 1080
gauaagggcc cggcggccau cgaggagugu ugcaacuggu uccacaagag aauggaggaa 1140
cucaacuccg aaaaacaccg guugauuaau uaccaccagg agcaggcggu caacugccug 1200
cugggcaacg uguucuacga aaggcucgcu gggcacggcc cuaagcuggg accugucacu 1260
cggaaacacc cccuggugac ccgguacuuu acuuucccgu ucgaagagau cgacuucucc 1320
auggaggaaa gcaugaucca ccucccuaac aaggccugcu uccugauggc ccacaacgga 1380
ugggucaugg gcgacgaccc uuugcggaac uuugccgagc cgggcucaga gguguaccuu 1440
agacgcgaac ugaucugcug gggagacucc gugaagcugc gcuacgggaa caagcccgaa 1500
gauuguccuu accugugggc acacaugaag aaguacaccg aaaucacugc caccuacuuc 1560
caaggagugc gcuuggauaa cugucauagc acuccucugc acgucgccga guacaugcug 1620
gacgcagcca ggaaucucca gccuaaucug uacguggugg ccgaacuguu caccgggagc 1680
gaggaccugg acaacguguu cgugacccgg cugggcauca gcucccuuau ccgggaggcc 1740
auguccgcau acaacucgca cgaggagggu cgccuggugu accgcuacgg aggagagccg 1800
gucggaucau uuguccaacc augccuccgg ccccugaugc cagcgaucgc gcacgcucuc 1860
uucauggaca ucacccacga uaacgaaugc ccuaucgugc accggagcgc uuacgacgcc 1920
cucccuucga ccaccauugu guccauggcc ugcugcgccu ccgguuccac ucggggcuac 1980
gaugaacugg ugccacacca gauuuccgug gugucagagg agcgguucua caccaagugg 2040
aacccagagg cgcugccgag caacacuggg gaagugaacu uccaguccgg aaucauagcg 2100
gcuagaugug caauuuccaa gcuucaucag gagcugggcg ccaaaggauu cauccagguc 2160
uacguggacc aaguggauga ggacauugug gccgugacua gacauucacc gucaauccac 2220
caaucggucg uggcuguguc cagaaccgca uuccgcaacc ccaagacuuc cuucuacucc 2280
aaggaggucc cucagaugug caucccggga aagaucgagg agguggugcu ggaggccagg 2340
accaucgaaa gaaacacuaa gccuuaccgg aaagacgaaa acuccaucaa ugggacuccc 2400
gauauuaccg uggagauuag ggagcacauu cagcugaacg aaucaaagau ugugaaacag 2460
gccggggucg caacuaaagg gccuaaugag uacauccaag agaucgaguu cgaaaaccug 2520
ucgccggguu ccgugaucau uuuccgcgug uccuuggacc cucacgccca aguggccgug 2580
ggcauccuga ggaaccaccu gacccaguuc agcccacacu ucaaguccgg auccuuggcu 2640
guggacaacg ccgauccaau ccucaagauc ccuuuugcgu cgcuggccuc acggcuuacc 2700
cuggccgagc ugaaccagau ccuguaccgc ugcgagagcg aagaaaagga ggacggugga 2760
gguugcuacg acauuccaaa cugguccgcg cuuaaguacg ccggacucca gggucugaug 2820
uccgugcuug ccgagaucag accgaaaaac gaccuggggc accccuuuug caacaaccug 2880
agaagcggag acuggaugau cgacuaugug uccaaccggc ugauuucgag aagcgguacu 2940
auugccgagg ucggaaagug gcuccaagca auguuuuucu accugaagca aauuccccgc 3000
uaccugaucc cgugcuacuu ugacgccauu cugaucggug cauacacuac ccugcuggac 3060
accgcgugga agcagauguc cagcuucgug cagaauggcu ccaccuucgu caagcaucug 3120
ucccucggaa gcgugcagcu gugcggagug ggaaaguuuc cgucgcugcc aauccugucc 3180
cccgcgcuga uggauguccc guaccggcug aacgaaauca cgaaggaaaa agaacagugc 3240
ugcguguccc uugccgccgg acugccgcac uucuccuccg gcauuuuccg gugcugggga 3300
cgggauaccu ucaucgcgcu gagagguauu cugcugauua ccggcagaua uguggaagcc 3360
cgcaacauca uccuggccuu ugccggcacu cugcggcacg ggcucauccc uaaccucuug 3420
ggagaaggca ucuacgcgcg cuacaacugc cgggaugcug ugugguggug gcugcagugc 3480
auccaggacu acuguaaaau ggugcccaau ggccuugaca uccucaagug cccagugucc 3540
cggauguacc cgaccgauga cuccgcgccc cugcccgccg gcacccuuga ucaaccucug 3600
uucgaaguca uccaagaggc caugcagaag cacaugcagg gaauccaguu cagggaaaga 3660
aacgccgggc cucagaucga ccggaacaug aaggacgagg gauucaacau uaccgccgga 3720
guggacgagg aaacuggcuu cguguacggu ggaaaccgcu ucaacugcgg caccuggaug 3780
gauaagaugg gcgaauccga ucgcgcccgc aaccggggaa ucccagcaac uccuagggac 3840
ggaagcgcag ucgagaucgu ggggcugucc aaguccgccg ugcgguggcu cuuagaacug 3900
uccaagaaga auauuuuccc cuaccacgag gucaccguga agcgccaugg aaaggccauc 3960
aaaguguccu augacgaaug gaaccgcaag auccaggaca acuucgaaaa guuguuccac 4020
guguccgagg accccagcga ucugaacgaa aagcacccca accucgugca caagaggggc 4080
aucuacaagg acuccuacgg agcuagcucc ccuuggugcg auuaccaacu gcggccuaau 4140
uucaccaucg ccauggucgu cgcacccgaa cucuucacca ccgagaaggc cuggaaggcu 4200
cuggaaaucg ccgaaaagaa gcuucugggc ccgcugggca ugaaaacucu cgauccugac 4260
gauauggucu acuguggcau cuacgacaac gcccuugaca acgacaacua caaccuggcc 4320
aagggcuuua acuaccacca gggcccagag uggcucuggc ccaucggaua uuuccugcgg 4380
gccaaacugu acuucucgcg guugaugggu ccggaaacca cagcuaagac caucgugcuc 4440
gugaagaaug ugcugucgcg gcacuauguc caucuggagc gcucccccug gaagggacuc 4500
ccugagcuga cuaaugagaa cgcccaguac ugcccuuucu ccugcgaaac ccaggccugg 4560
agcaucgcua ccauucucga aacgcucuac gaucuguag 4599
<210> 25
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 25
augggacauu cuaaacagau ucggauccuc cugcucaacg aaauggagaa gcucgaaaag 60
acccuguucc ggcucgaaca gggauacgaa cugcaauucc ggcugggucc caccuuacaa 120
ggaaaggccg ugacugucua caccaacuac ccguuccccg gcgaaaccuu caauagggag 180
aaguuccggu cccucgauug ggagaacccu acagagcgcg aagaugauuc agacaaguau 240
ugcaagcuga auuugcagca gagcggcuca uuccaauacu acuuccugca aggaaacgaa 300
aagucgggcg gcggcuacau cgugguggac ccuauucuga gagugggcgc cgacaaccau 360
guccugccgc uggacugcgu cacccugcaa acuuuccugg ccaagugccu ggguccuuuc 420
gaugaguggg aaucgagacu ucgcguggca aaggaaucgg guuauaauau gauucacuuc 480
accccgcugc aaacccuggg ccugucgcgg ucuugcuacu cacuggccaa ccagcuggaa 540
cugaacccag auuucucacg ccccaaccgg aaguacaccu ggaacgaugu gggccagcuu 600
guggaaaagc ucaaaaagga guggaacgug aucugcauua cugacguggu guacaaccau 660
acugcugcua auuccaagug gauccaggaa cauccugagu gugcuuacaa ccuggucaac 720
uccccacacc ugaagcccgc cugggugcug gaucgggcuc ucuggcgguu cuccugcgau 780
gucgccgaag gaaaguacaa ggagaagggg aucccugcgc ugaucgaaaa cgaccaucac 840
augaauucca ucaggaaaau caucugggag gacaucuucc cuaagcucaa gcugugggag 900
uucuuucaag ucgacgugaa caaggccgug gagcaguuca gacggcugcu gacacaagag 960
aaccgccgcg ucaccaagag cgacccgaau cagcaccuga cgauuaucca ggacccugag 1020
uacagaagau ucggcugcac cguggauaug aacauugcuc ugaccaccuu caucccccac 1080
gauaaggguc ccgcagccau ugaggagugc ugcaacuggu uccacaagag gauggaagaa 1140
cugaauuccg agaagcaccg gcugaucaau uaucaccaag aacaagccgu gaacugccuu 1200
cugggaaacg ucuuuuacga gcgccuggcg ggacaugggc caaagcucgg gcccgugacc 1260
agaaagcacc cacuggucac ccgcuacuuc accuucccgu ucgaggaaau cgacuucagc 1320
auggaagagu cgaugaucca cuugccgaac aaggccugcu uccucauggc ccauaacggu 1380
ugggucaugg gcgaugaccc ucuucgcaac uucgcggagc ccgguucuga agucuaucuu 1440
cggcgggaac ugauuugcug gggagacucc gucaagcugc gcuacggaaa caagcccgag 1500
gauugcccgu acuugugggc ccauaugaag aaguacacug agaucacugc cacuuacuuc 1560
caaggagugc gccucgacaa uugucacagc acuccgcugc acguggcgga guacaugcug 1620
gacgccgcuc gcaacuugca gccuaaucug uaugucgugg ccgaguuguu caccggcucg 1680
gaagaucugg acaacguguu cgucacucgc cugggaaucu ccucccugau ccgggaagcg 1740
augagcgccu acaacuccca cgaagagggg cgguuggugu accgcuacgg cggagagccu 1800
gucggaaguu ucgugcagcc cugucugagg ccccugaugc ccgcuaucgc ccaugcgcug 1860
uuuauggaca ucacccacga caacgaaugc ccuauugugc accgcuccgc cuaugaugcc 1920
cugcccucca ccacuauugu cagcauggcc ugcugcgccu cgggguccac ccggggauac 1980
gacgaacugg ugccccacca aauuuccgug guguccgagg aacgguucua caccaagugg 2040
aacccugaag cgcugccauc gaacacugga gaagugaacu uucagucggg aauuaucgca 2100
gcccgaugcg ccaucagcaa gcugcaccag gaacucggcg caaaggguuu uauccaaguc 2160
uacguggacc aggucgacga ggacauuguc gccgugaccc ggcacucccc auccauccac 2220
cagucugugg uggcuguguc aaggacggcu uuccggaacc caaagaccag cuucuacagc 2280
aaggaagugc cucagaugug caucccgggg aagaucgaag aaguggugcu ggaggccaga 2340
accaucgaaa gaaacaccaa gcccuaucgg aaggacgaga acucgaucaa cgguacuccg 2400
gacauuaccg ucgagauacg cgagcacauu cagcugaacg aguccaaaau cgugaagcag 2460
gccggggugg ccacgaaggg ucccaacgag uacauucagg agaucgaguu cgagaaccug 2520
agccccgggu ccgugaucau cuuccgcgug ucccuggacc cccacgccca aguggccgug 2580
ggcauccugc ggaaccaucu gacccaguuc uccccgcacu ucaagagcgg cucgcucgcc 2640
guggacaacg cggacccgau ccucaagauc ccuuucgcau cgcuggccuc ccgccugacc 2700
cuggccgaac ugaaucagau cuuguaccga ugcgaaucag aagagaagga ggacgggggg 2760
ggcugcuacg auauccccaa cuggagcgcg uugaaguacg caggauugca gggauugaug 2820
uccguccucg cugaaauccg cccgaagaac gaccugggac acccguuuug caacaaccug 2880
agaucagggg auuggaugau cgauuacgug ucgaacagac ugaucucgcg cagcggcacu 2940
auugccgaag uggggaagug gcuccaggcc auguucuucu accugaagca gaucccucgg 3000
uacuugaucc cuuguuacuu cgacgccauc cugaucggag ccuacaccac ccugcuugac 3060
acugcaugga agcagauguc cagcuucgug caaaacggaa gcaccuucgu gaagcaccug 3120
ucccugggau ccgugcagcu cugcggcgug ggaaaguuuc cgucccuccc cauccugagc 3180
ccugcccuua uggacgugcc guacaggcug aacgaaauua ccaaagagaa ggagcaaugc 3240
uguguguccu uggcggccgg auugccgcau uucuccuccg ggauuuuccg gugcugggga 3300
cgggacaccu uuaucgcacu gagggguauu cuccugauca ccggucgcua cguggaggcu 3360
cgcaacauua uucuggccuu cgcgggcacg cuuagacaug gauugauccc uaaccuucug 3420
ggagaaggga ucuacgcgcg guacaacugc cgcgaugccg ugugguggug gcugcagugc 3480
auccaggacu acugcaaaau ggugccgaau ggucuggaua uccugaagug uccgguuucg 3540
cggauguacc cuaccgacga cagcgccccu cucccggccg gcacucucga ccagccccua 3600
uuugaaguaa uccaggaggc caugcaaaag cacaugcagg gcauacaguu cagagagagg 3660
aacgccggac cgcagauuga ccggaacaug aaggacgagg gauucaacau uaccgcggga 3720
guggaugagg aaacugguuu cguguacggc ggaaaccggu uuaacugcgg cacuuggaug 3780
gacaagaugg gagaauccga ccgcgcccga aaccgcggaa uucccgccac uccccgcgac 3840
ggcuccgccg uggaaauugu gggacuguca aaguccgcag uccgcuggcu gcuggaacuc 3900
ucaaagaaga acaucuuccc guaccacgag gucaccguga agcggcacgg caaagcgauc 3960
aaagugucgu acgacgagug gaaccggaag auucaggaua acuucgagaa gcuguuucac 4020
guguccgaag auccaagcga ccugaacgag aagcauccca acuuggugca caagcgcggc 4080
aucuacaagg auuccuacgg agccagcagc ccguggugcg acuaccaacu ccgccccaac 4140
uucaccaucg ccaugguggu ggcgccggag cuguucacga cggagaaagc uuggaaggcu 4200
cucgaaaucg cggagaagaa gcugcugggu ccucugggga ugaaaacccu ggacccggac 4260
gauauggugu acugugggau cuacgacaac gcccuggaca acgacaacua caaccucgcc 4320
aagggguuca acuaccacca gggacccgaa uggcucuggc caaucggaua cuuccugaga 4380
gcgaagcuuu acuucucgcg gcugaugggu ccugaaacca cggccaagac caucgugcuc 4440
gugaaaaaug ugcugucaag gcacuacgug caucuggaga ggucgccaug gaagggucug 4500
ccggaacuga ccaacgaaaa cgcacaguac ugccccuuuu cgugcgagac ucaggccugg 4560
uccaucgcca ccauucucga aacucucuac gaccuguag 4599
<210> 26
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 26
augggacacu cgaaacagau cagaauccug uugcucaacg aaauggagaa gcuggaaaag 60
acccucuuuc ggcucgaaca gggcuacgag cugcaguucc gccugggucc gacacuacaa 120
ggaaaggcag ucacugugua caccaacuac ccauuucccg gagaaaccuu caaccgggag 180
aaguuccggu cccuggacug ggaaaaccca accgaacgag aggaugacuc cgacaaguac 240
ugcaagcuga accuccaaca guccggauca uuccaguacu acuuucugca agggaacgag 300
aaguccggag gcggcuacau cguggucgac ccaauacuua gagugggagc cgacaaucau 360
guccugccuc uggacugcgu gacccugcaa accuucuugg cgaaaugucu gggcccguuc 420
gaugaguggg aaagccgccu cagagucgca aaggaguccg gauacaacau gauucacuuc 480
acuccgcugc aaacccucgg ucugucccgg ucgugcuauu cucuggcgaa ccagcuggag 540
cuuaaccccg acuucucgcg cccaaaccgg aaguacaccu ggaacgacgu cgggcagcuu 600
guggaaaagc ugaagaagga guggaacgug aucugcauca cugacguggu guacaaccau 660
acggccgcca acucgaagug gauccaagag caucccgagu gcgcguacaa ccucgugaau 720
agcccgcauc ugaagccugc uugggugcug gauagagccc ucuggagauu cagcugcgac 780
guggccgagg ggaaguacaa agaaaaggga auuccggccu ugauugagaa cgaccaucac 840
augaacucaa uccgcaagau caucugggag gauaucuucc cuaagcuuaa guugugggag 900
uucuuccaag uggacguaaa caaggcagug gagcaguuca gaagguugcu gacucaagaa 960
aacagacggg ucacuaaguc cgauccuaac cagcaccuua ccaucauuca agacccugag 1020
uaccgccggu uuggcugcac cgucgacaug aacaucgccc ugaccacuuu caucccgcau 1080
gacaagggcc cggcggcaau cgaggaaugc uguaacuggu uucauaagag gauggaggaa 1140
cugaauuccg agaagcaccg gcugauuaac uaccaccagg aacaggcagu caacugccuc 1200
cugggcaacg uguucuacga acggcuggcu ggacacggac cgaagcuggg ucccgugacu 1260
cgcaagcauc cgcucgugac ucgcuacuuc accuucccgu uugaggagau ugacuucucc 1320
auggaagaau ccaugaucca ccucccgaac aaggcuugcu uccugauggc gcacaacgga 1380
ugggucaugg gcgacgaccc acugcgcaau uucgcugagc cuggcucgga ggucuaccug 1440
agaagggaau ugauuugcug gggagacucc gugaagcugc gcuauggcaa caagccugaa 1500
gauugccccu accugugggc ucacaugaag aaguacacgg aaaucacugc cacguacuuc 1560
cagggagucc ggcuggacaa uugccacucg accccgcucc auguggccga guacaugcug 1620
gaugcagcga ggaaucugca gcccaaucug uacgugguug cagaacuguu cacuggcucc 1680
gaggaccucg acaacguguu cgugaccaga cuggggauuu ccucacugau ccgggaagcc 1740
augucggccu acaacuccca ugaagagggc cgccuggugu accgcuaugg gggagaaccc 1800
gugggaagcu ucgugcagcc uugccuccgg ccgcugaugc cugcgaucgc ccacgcccug 1860
uucauggaua ucacucacga caacgaaugc cccauugugc aucgcucggc cuacgacgca 1920
cugccuucga ccacuaucgu guccauggcc ugcugcgccu ccgggagcac ccgcggauac 1980
gaugaacucg ugccgcacca gaucagcgug guguccgaag aacgguucua caccaagugg 2040
aaccccgaag cccugccuag caauaccggg gaagugaacu uccaguccgg uauuaucgcc 2100
gcucgcugcg ccaucuccaa acuccaccaa gagcucggug ccaagggauu cauucaaguc 2160
uacguggauc aggucgacga agauauugug gccgugacca ggcacucacc uuccauccac 2220
caauccgucg ucgccguguc ccggacugcg uuucggaacc ccaagacuuc guucuacucg 2280
aaagaagugc cacagaugug uaucccggga aaaaucgagg aggucgugcu cgaagcccgg 2340
accauugaga ggaacaccaa gccuuaccgg aaagacgaga acucuaucaa cgguaccccu 2400
gauauuacug uggagauccg cgaacacauc cagcugaacg aaucaaagau cgucaagcag 2460
gcuggagugg ccaccaaggg acccaacgag uacauccagg agaucgaauu ugaaaaccuc 2520
uccccuggcu ccgugauuau cuuccgggug ucccuggacc cucacgccca aguggccgug 2580
ggaauucuca gaaaccaccu gacccaguuc ucaccacacu uuaaguccgg uucccuggcg 2640
guggacaacg ccgauccgau cuugaagauc cccuucgcau cgcucgccuc ccgccugacu 2700
cucgcggaac ugaaccagau ccuguaccgc ugugaauccg aggaaaagga ggacggcggc 2760
ggcuguuacg auauccccaa uuggucggcu uugaaauacg cgggacuuca ggggcugaug 2820
ucugugcugg cggaaauccg gccgaagaac gaccugggac acccauucug caacaacuug 2880
cggagcggag acuggaugau cgauuacguc agcaacagau ugaucagccg gagcggcacu 2940
aucgccgagg ucggaaagug gcuccaggcc auguucuucu accugaagca gaucccccga 3000
uaccucaucc ccuguuacuu cgacgccauu cugaucgggg ccuacaccac ccugcuggac 3060
accgccugga agcagaugag caguuuugug caaaacgggu ccaccuucgu gaagcaccuu 3120
ucacugggcu cagugcagcu cugcggcgug ggaaaguucc ccucgcugcc cauucugagc 3180
cccgcccuga uggacguccc uuaccggcug aacgagauca ccaaggagaa ggagcagugc 3240
uguguuuccc uggcugccgg gcugccacac uucucguccg gcaucuuccg guguuggggc 3300
cgggauaccu ucauugcccu gcggggaauc cugcuuauca ccggucgcua cguggaggcu 3360
cggaacauua uucucgcguu cgccggcacc cugagacacg gucugauucc gaaucuguug 3420
ggcgaaggaa ucuacgccag auacaacugu cgggacgccg ugugguggug gcuccagugc 3480
auucaggacu auugcaagau ggugccgaac ggccuggaca uccugaagug cccagugucg 3540
aggauguauc caacugacga cagcgcaccu cugccggccg ggacccucga ccaaccccug 3600
uucgaaguca uucaagaggc uaugcagaag cacaugcagg guauucaguu ccgggagcgg 3660
aacgcggggc cccagauuga uaggaacaug aaagacgagg gcuucaacau cacugccggc 3720
guggacgaag aaaccgguuu uguguacgga ggaaaccgcu ucaacugcgg uaccuggaug 3780
gacaagaugg gagaauccga ucgcgcgcgc aacagaggga ucccggcaac cccucgggac 3840
ggauccgcgg uggaaauugu gggacugagc aagagcgccg ugcgguggcu ccuggaacug 3900
uccaaaaaga acaucuuccc cuaccacgaa gugaccguga agagacacgg aaaggccauc 3960
aaagucucau acgaugaaug gaacaggaag auccaggaua acuucgagaa gcuguuucac 4020
guguccgagg aucccuccga ucugaacgag aagcauccga aucuggugca caagcgcggg 4080
aucuacaagg acucguacgg agcguccucc ccuuggugcg acuaucagcu gcggccuaac 4140
uucaccauug ccauggucgu ggccccggag cuguucacaa cugagaaggc cuggaaggcc 4200
cuugaaauug ccgagaagaa gcugcugggg ccuuugggga ugaaaacccu ggauccggac 4260
gacauggugu acugcggaau cuacgacaac gcccuggaca acgauaacua caaucucgcg 4320
aagggcuuca auuaccacca aggccccgaa uggcucuggc cuauugggua cuuccugcgc 4380
gccaagcugu acuucucacg gcugauggga ccagagacua ccgccaagac uaucguccuc 4440
gugaagaacg ugcugucccg gcacuacgug caucuggaga ggagcccuug gaagggacuu 4500
ccugagcuga cgaacgaaaa cgcgcaguac ugccccuucu ccugcgaaac ccaggcuugg 4560
uccauugcca cuauacugga aaccuuauau gaccuguag 4599
<210> 27
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 27
augggacacu cgaaacagau ccggauccug uugcugaacg aaauggagaa gcuugaaaag 60
acccuguuuc ggcuggagca gggcuacgag cugcaguucc gccugggucc gacacuacaa 120
ggaaaggccg ugacugugua caccaacuac ccauuucccg gagaaacuuu caaccgggag 180
aaguuccggu cccuggacug ggaaaaccca accgaacgag aggacgacuc ggacaaguac 240
ugcaagcuga accuccaaca guccggauca uuccaguacu acuuucugca agggaacgag 300
aaguccggag gcggcuacau cguggucgac ccgauacuua gagugggcgc cgacaaucau 360
guccugccuc uggauugcgu gacccugcaa accuucuugg ccaaaugucu gggcccguuc 420
gaugaguggg aaagccgccu cagagucgca aaggaguccg gauacaacau gauucauuuc 480
accccgcugc aaacccuggg ucugucccgg ucgugcuauu cucuggcgaa ccagcuggag 540
cuuaaccccg acuucucgcg cccaaaccgg aaguacaccu ggaacgacgu cgggcagcuu 600
guggaaaagc ugaagaagga guggaacgug aucugcauca cugacguggu guacaaccau 660
acggccgcca acucgaagug gauccaggag caucccgagu gcgcauacaa ccucgugaac 720
agcccgcacc uuaagccugc uugggugcug gacagagccc ucuggagauu caguugcgac 780
guggccgagg ggaaguacaa ggaaaaggga auuccggccu ugaucgagaa cgaccaucac 840
augaacucaa ucaggaagau caucugggag gacaucuucc cuaagcuuaa guugugggag 900
uucuuccaag uggacguaaa caaggcagug gagcaguuca gaagguugcu gacccaagaa 960
aacagacgcg ucacuaaguc cgauccuaac caacaccuua ccaucauuca agacccugaa 1020
uaccgccggu uuggcugcac cgucgauaug aacaucgccc ugaccacuuu caucccgcau 1080
gacaagggcc cggcggcaau cgaggaaugc uguaacuggu uucauaagag aauggaagaa 1140
cugaauuccg agaagcaccg gcugauuaac uaccaccagg aacaggcagu gaacugccug 1200
cugggcaaug uguucuacga acggcuggcu ggacacggac cgaagcuggg ucccgugacu 1260
aggaagcauc cgcucgugac ucgcuacuuc accuucccgu uugaggagau ugacuucucg 1320
auggaagaau ccaugaucca ucugccuaac aaggcuugcu uccugauggc gcacaacgga 1380
ugggucaugg gcgacgaccc ccugcgcaac uucgccgagc cuggcucgga ggucuaccug 1440
agaagggaac uuaucuguug gggagacucc gucaagcugc gcuacggaaa caagcccgaa 1500
gauugccccu accugugggc ucacaugaag aaguacacgg aaaucacugc aacguacuuc 1560
cagggagucc ggcuggacaa uugccacucc accccccuuc auguggccga guacaugcuc 1620
gaugcagcga ggaaucugca gccgaaucug uacgugguug ccgaacuguu cacuggcucc 1680
gaggacuugg acaacguguu cgugaccaga cuggggaucu ccucccugau ccgggaagcc 1740
augucggccu acaacuccca ugaagagggc cgccuggugu accgcuacgg gggagaaccc 1800
gugggaagcu ucgugcaacc uugccugcgg ccgcugaugc cugcgaucgc ccacgcccug 1860
uucauggaca ucacucacga uaacgaaugc ccgauugugc aucgcucggc cuacgacgca 1920
cugccgagca ccacuaucgu guccauggcc ugcugcgccu ccgggagcac ucgcggauac 1980
gaugaacucg ugccgcacca gaucagcgug guguccgaag aacgcuucua uaccaagugg 2040
aaccccgaag cgcugccauc gaauacuggc gaagugaacu uccaguccgg uauuaucgcc 2100
gcucgcugug ccaucagcaa acugcaccaa gagcuuggug ccaagggauu cauucaaguc 2160
uacguggauc aggucgacga agauauugug gccgugacca ggcacucacc uuccauccac 2220
caaucagucg uggccguguc ccggaccgcg uuccggaacc ccaagaccag cuucuacucg 2280
aaagaagugc cucagaugug uaucccggga aaaaucgaag aggucgugcu ggaagcccgg 2340
accauugaga ggaacaccaa gccuuaccgg aaagacgaga acucuaucaa cgguaccccu 2400
gauauuacug uggagauccg cgaacacauc cagcugaacg aaucaaagau cgucaagcag 2460
gcuggagugg ccaccaaggg acccaacgag uacauccagg agaucgaauu ugaaaaccuc 2520
uccccuggcu ccgugauuau cuuucgggug ucccuggacc cucacgccca aguggccgug 2580
ggaauucuca gaaaccaccu gacccaguuc ucaccccacu ucaaguccgg uucccuggcg 2640
guggauaacg ccgacccgau uuugaagauc cccuucgccu cgcuggccuc ccgccugacu 2700
cucgcggaac ugaaccagau ccuguaccgc ugugaaucag aggaaaaaga ggacggcggc 2760
ggcuguuacg auauucccaa uuggucggcu uugaaauacg cgggacuuca ggggcugaug 2820
ucugugcugg cggaaauccg gccgaagaac gaccugggac acccauucug caacaacuug 2880
cgguccggag acuggaugau cgauuacguc agcaacagau ugaucagccg gagcggcacu 2940
aucgcugagg ucggaaagug gcugcaggcc auguucuucu aucugaagca gaucccccga 3000
uaccucaucc ccuguuacuu cgacgccauu cugaucgggg ccuacaccac ccugcuggac 3060
accgccugga agcagaugag caguuuugug caaaacgggu ccaccuucgu gaagcaccuu 3120
ucacugggcu cagugcagcu cugcggcgug ggaaaguucc ccucucugcc cauucugagc 3180
ccggcccuga uggacguccc uuaccggcug aacgagauca ccaaggagaa ggagcagugc 3240
ugcguuuccc uggcugccgg gcugccacac uucucguccg gcaucuuccg gugcuggggc 3300
cgggauaccu ucaucgcccu gcgggguauc cugcuuauca ccggucgcua cguggaggcu 3360
cggaacauua uucuggcguu cgccggcacc cuuagacacg gacugauucc gaaccuuuug 3420
ggcgaaggaa ucuacgccag auacaacugu cgggacgccg ugugguggug gcuucagugc 3480
auucaggacu auugcaagau ggugccgaac ggccuggaca uccugaagug cccagugucg 3540
aggauguauc caaccgacga cagcgcaccu cugccggccg ggacccucga ccaaccccug 3600
uucgaaguca uucaggaggc uaugcagaag cacaugcagg guauucaguu ccgggagcgg 3660
aacgcggggc cgcagauuga uaggaacaug aaagacgagg gcuucaacau cacugccggc 3720
guggacgaag aaaccgguuu uguguacgga ggaaacagau ucaacugcgg uaccuggaug 3780
gacaagaugg gagaguccga ucgcgcgcgc aacagaggga ucccggcaac cccgcgggac 3840
ggauccgcgg uggaaauugu gggacugagc aagagcgccg ugcgguggcu gcuggaacug 3900
agcaaaaaga acaucuuccc cuaccacgaa gugaccguga agcggcacgg aaaggccauc 3960
aaagucucau acgaugaaug gaauaggaag auccaggaua acuucgagaa gcuguuucac 4020
guguccgagg aucccuccga ucugaacgaa aagcacccga aucucgugca caagcgcggg 4080
aucuauaagg acucguacgg agcguccucc ccuuggugcg acuaucagcu gcggccuaac 4140
uucaccauug ccauggucgu ggccccggag cuguucacaa cugagaaggc cuggaaggcc 4200
cuugaaauug ccgagaagaa gcugcugggg ccuuugggga ugaaaacccu ggauccggac 4260
gacauggugu acugcggaau cuacgacaac gcccuggaca acgauaacua caaucuggcc 4320
aagggcuuca auuaccacca gggcccggaa uggcuguggc ccauugggua cuuccugcgc 4380
gccaagcugu acuucucacg gcugauggga ccagagacua ccgccaagac uaucguccuc 4440
gugaagaacg ugcugucccg gcacuacgug caucuggaga ggagcccuug gaagggacuu 4500
ccugaacuga cuaacgagaa cgcgcaguac ugccccuucu ccugcgaaac ccaggcuugg 4560
uccauugcca ccauacugga aacccuuuau gaccuguag 4599
<210> 28
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 28
augggacacu cgaaacagau uagaauccug uugcucaacg aaauggagaa gcuggaaaag 60
acccucuuuc ggcuggagca gggcuacgag cuccaguucc gccugggucc gacacuacaa 120
gggaaggcag ugaccgugua caccaacuac ccauuucccg gagaaaccuu caaccgggag 180
aaguuccggu cccuggacug ggaaaaccca accgaacgag aggaugacag cgacaaguac 240
ugcaagcuga accuccaaca gucgggaucg uuccaguacu acuuucugca agggaacgag 300
aaguccggag gaggcuacau cguggucgac ccgauacuua gagugggagc cgacaaccau 360
guccugccuc uggacugcgu gacccugcaa accuucuugg ccaaaugucu gggcccguuc 420
gaugaguggg aaagccgccu cagaguggcc aaggaguccg gguacaacau gauucacuuc 480
acuccgcugc aaacccucgg ucugucccgg ucgugcuauu cucuggcgaa ccagcuggag 540
cugaaccccg acuucucccg cccaaaccgg aaguacaccu ggaacgacgu cgggcagcuu 600
guggaaaagc ugaagaagga guggaacgug aucugcauua cugacguggu guacaaccac 660
accgccgcca acucgaagug gauccaagaa caucccgagu gcgcguacaa ccucgugaac 720
agcccgcauc ugaagccggc uugggugcuc gauagagccc ucuggagauu cuccugugac 780
gucgccgagg ggaaguacaa agagaagggu auuccggccc ucauugagaa cgaccaucac 840
augaacucaa uccggaagau caucugggag gauaucuucc cuaagcuuaa guugugggag 900
uucuuccaag uggacguaaa caaggccgug gaacaguuca gaaggcugcu gacucaagaa 960
aaccgccgcg ucacuaaguc cgauccuaac cagcaucuua ccaucaucca agacccugag 1020
uaucgccggu uugggugcac cgucgacaug aacaucgcac ugaccacuuu caucccgcau 1080
gacaaggggc cggcggccau cgaggaaugc uguaacuggu uucauaagag gauggaggaa 1140
cugaauuccg agaagcaccg gcugauuaac uaccaccagg aacaggcagu gaauugccuc 1200
cuggggaacg uguucuacga aaggcuggcu ggacacggac cgaagcuggg gcccgugacu 1260
cgcaagcauc cgcucgugac ucgcuacuuc accuucccgu uugaggagau ugacuucuca 1320
auggaagaau ccaugaucca ccucccgaac aaggcuugcu ucuugauggc gcacaaugga 1380
ugggucaugg gcgacgaccc acugcgcaac uucgcugagc cuggcucgga ggucuaccug 1440
agaagggaau ugauuugcug gggggacucc gucaagcugc gcuauggaaa caagccggaa 1500
gauugccccu accugugggc ucacaugaag aaguacacgg aaauuacugc cacguacuuc 1560
cagggcgucc ggcuggacaa cugccacucc acuccccucc auguggccga guacaugcuc 1620
gacgcagcga ggaaucugca gcccaaucug uacgugguug cagaacuguu cacugggucc 1680
gaggaccucg acaauguguu cgugaccaga cuggggaucu ccucccugau ccgggaagcc 1740
augucggccu acaacuccca ugaagagggc cgccuggugu accgcuacgg gggagaaccc 1800
guggggagcu ucgugcagcc uugccuccgc ccgcugaugc cugccaucgc ccacgcccug 1860
uucauggaua ucacucauga caacgaaugu cccauugugc aucgcucggc cuacgacgca 1920
cugccuucga ccacuaucgu guccauggcc ugcugcgccu ccgggagcac caggggauac 1980
gacgaacucg ugccgcacca gaucagcgug guguccgaag agagauucua uaccaagugg 2040
aaccccgaag cgcugcccag caauaccggg gaagugaacu uccaguccgg uauuaucgcc 2100
gcucgcugug cgaucagcaa gcuccaccag gagcucggug ccaagggauu cauucaaguc 2160
uacguggacc aggucgacga agauaucgug gccgugacca ggcacucacc uuccauucac 2220
caauccgugg ucgccguguc ccggacugcu uuucggaacc ccaagacuuc guucuacucg 2280
aaagaagugc cacagaugug uaucccgggg aaaaucgaag aggucguccu cgaagcccgg 2340
accauugaga ggaacaccaa gccuuaccgg aaagacgaga acucuaucaa cgggaccccu 2400
gauauuacug uggagauccg cgaacacauc cagcugaacg aaucaaagau cgucaagcag 2460
gcuggagugg cgaccaaggg acccaacgag uacauccagg agaucgaauu ugaaaaccuc 2520
uccccuggcu ccgugauuau cuuccgggug ucccuggacc cucacgccca aguggccgug 2580
ggaauucuca gaaaccaccu gacucaguuc ucaccccacu uuaagucugg gucccuggcg 2640
guggacaacg ccgauccgau cuugaaaauc ccuuucgcaa gccuggccuc ccgccugacu 2700
uuggccgagc ugaaccagau ccuguaccgc ugugaaucag aggaaaagga ggacgggggu 2760
ggcuguuacg auauccccaa cugguccgcu uugaaauacg cgggacugca ggggcugaug 2820
uccgugcugg cggaaaucag accgaagaac gaucuggggc accccuucug caacaacuug 2880
cgguccggag acuggaugau cgauuacguc agcaaccggu ugaucagcag aagcgguacu 2940
aucgccgagg ucggaaagug gcugcaggcc auguucuucu accugaagca gaucccucga 3000
uaccucaucc ccuguuacuu cgacgccauu uugaucgggg ccuacaccac ccugcuggac 3060
acugccugga agcagaugag caguuuugug caaaaugggu cgaccuucgu gaagcaccuu 3120
uccuugggcu cagugcagcu cugcggcgug gggaaguucc ccucgcugcc cauucugucc 3180
ccggcccuga uggacguccc uuaccggcug aacgagauua ccaaggagaa ggagcagugc 3240
uguguuuccc uggcugccgg gcugccacac uucucguccg ggaucuuccg gugcuggggc 3300
cgcgauaccu ucauugcgcu gcgggguauc cugcuuauca ccggucgcua cguggaggcu 3360
cggaacauua uccuugcauu cgccgguacc cugagacacg gucugauccc gaaucuucuc 3420
ggggaaggaa ucuacgcaag auacaacugc cgggacgccg ugugguggug gcuccagugc 3480
auucaggacu auugcaagau ggugccgaac ggacuugaca uccugaagug cccagugucg 3540
aggauguacc cuaccgacga cagcgcuccu cugccggccg ggacccugga ccaaccccug 3600
uucgaaguca uccaagaggc uaugcagaag cacaugcagg guauucaguu ccgggaacgg 3660
aacgcggggc cccagauuga uaggaacaug aaggacgagg guuucaacau cacugccggc 3720
guggacgaag aaacuggguu uguguacgga ggaaacagau ucaacugcgg uaccuggaug 3780
gacaagaugg gagaauccga ucgcgcgcgc aacagaggga ucccggcaac cccgcgggac 3840
ggauccgcgg uggaaauugu gggacugagc aagagcgccg ugcgguggcu ccuggaacug 3900
uccaaaaaga acaucuuccc cuaccacgaa gugaccguga agcggcacgg aaaggccauc 3960
aaagucucau acgaugaaug gaaucggaag auccaggaua acuucgagaa gcuguuucac 4020
guguccgagg aucccuccga ucucaacgaa aagcauccga aucucgugca caagcgcggg 4080
aucuacaagg acucguacgg ggcguccuca ccuuggugcg acuaucagcu gcggccuaac 4140
uucacuauug cgauggucgu ggccccggag uuauucacaa cggagaaggc cuggaaggcc 4200
cuugaaauug cggagaagaa gcugcugggg ccucucggga ugaaaacccu ggauccggac 4260
gacauggugu acugcggaau cuacgacaac gcccuugaca acgauaacua caaucuggcc 4320
aagggguuca auuaccacca ggggccggaa uggcucuggc ccauugggua cuuccugcgc 4380
gccaagcugu acuucucacg gcugaugggg ccagagacua ccgccaagac uaucguccuc 4440
gugaaaaacg ugcugucccg gcacuacgug caucuggaga ggagcccuug gaagggacug 4500
ccagagcuga cgaacgagaa cgcgcaguac ugccccuucu ccugcgaaac ccaggcuugg 4560
ucgauugcca cuauacugga aaccuuauau gaccuguag 4599
<210> 29
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 29
augggacacu caaagcaaau ccgcauucug cugcugaacg agauggaaaa gcuggaaaag 60
acucuguucc ggcuggaaca ggguuacgag cugcaguuuc ggcucggccc aaccuugcaa 120
gggaaggccg ucacugucua caccaacuac ccuuuuccgg gggagacuuu caaccgcgaa 180
aaguuccgcu cccuggauug ggaaaacccc acugaacggg aggaugacag cgacaaguac 240
ugcaaacuga accuucagca guccggcagc uuccaauacu acuuccugca aggaaacgaa 300
aaguccggcg gcgguuacau cguggucgac cccauucuga gagugggagc cgauaaucau 360
gugcugcccc uggacugcgu cacccugcaa acuuuccugg cuaagugccu gggaccguuc 420
gaugaguggg aauccaggcu ccgcguggcg aaggagucgg gcuacaacau gauucauuuc 480
acuccccuuc aaacucuggg gcugagucgc agcuguuacu cccuggccaa ccagcuugaa 540
cugaacccag acuuuucccg gccaaacaga aaguacaccu ggaacgacgu gggacaauug 600
guggagaagc uuaagaagga auggaacgug aucugcauca ccgauguggu guauaaccac 660
acugccgcga auuccaagug gauccaggaa caccccgagu gugcuuacaa ccuugugaac 720
agcccucacc uuaagccggc cugggugcug gaucgggcgc uguggagauu cuccugcgac 780
guggccgaag ggaaguacaa agaaaaggga auccccgccc ugauugaaaa ugaccaccau 840
augaacucca uucggaaaau cauuugggaa gauaucuuuc ccaagcugaa gcucugggag 900
uucuuucaag uggaugugaa caaggcugug gagcaguucc ggcggcugcu gacccaggag 960
aaccgccgcg ugaccaaguc cgacccuaac cagcacuuga ccaucauaca ggacccggaa 1020
uaccggagau ucggcugcac ugucgacaug aacauugccc ucacuaccuu caucccgcau 1080
gacaagggac ccgcugccau cgaagagugc ugcaacuggu uccacaagcg gauggaagaa 1140
cugaacucug aaaagcacag gcugaucaac uaccaccagg aacaggcugu gaacugccug 1200
cugggcaacg uguucuacga gagacucgca ggacacggac cgaagcuggg cccugugacc 1260
cggaagcauc cucuggucac ccgcuacuuc accuucccgu ucgaagagau cgacuuuucg 1320
auggaagaau cgaugaucca ccucccuaac aaggccugcu uccucauggc gcacaacggc 1380
ugggucaugg gcgacgaccc gcugagaaac uucgccgagc ccgggagcga aguguaccuc 1440
cggcgggaac uuauuugcug gggagauagc gugaagcuua gauauggcaa caagccugag 1500
gacugcccau accugugggc gcacaugaag aaguacacug aaauuaccgc gaccuacuuc 1560
caaggagucc gacucgacaa cugccacagc accccacuuc acgucgcgga guacauguug 1620
gaugccgcac ggaaucucca gcccaaucug uaugucgugg cugaacuguu cacuggaucc 1680
gaggaccuug acaauguguu cgugacuaga cuggggaucu ccagccugau ccgggaagcu 1740
auguccgcgu acaacuccca cgaagaggga cggcuggugu accgcuacgg cggagagccc 1800
gugggaagcu ucgugcagcc cugccugcgg ccucugaugc cggccaucgc ucacgcccug 1860
uucauggaua ucacucacga caaugagugu ccuaucgugc acaggagcgc guacgacgcc 1920
cugcccucca cuacuaucgu gucgauggcc ugcugcgcaa gcgguucuac ccgcgguuac 1980
gacgagcuug ucccgcacca aauauccgug gugucagagg aacgguucua caccaagugg 2040
aacccggagg cccugccuuc aaacaccggc gaagugaacu uccaguccgg aaucauugcc 2100
gcccgcugug ccauuucaaa guugcaccag gagcugggcg ccaagggauu cauucagguc 2160
uacguugacc aggucgacga agauaucgug gccguuacua gacauucacc gagcauccau 2220
cagagcgugg ucgcagucag caggacugcc uuccgcaacc cgaaaaccuc guucuacucc 2280
aaggaagugc cccagaugug uaucccggga aaaauugagg aaguggugcu ggaggcccgg 2340
accaucgagc ggaacacuaa gcccuaccgg aaggacgaga auucaaucaa cggaaccccu 2400
gacaucaccg uggagauccg cgagcauauc caacugaacg agucgaagau cgucaagcag 2460
gcuggggugg caacuaaggg cccuaacgag uacauucagg agauugaauu cgagaaccug 2520
ucccccgggu ccgugaucau uuuccgcgug ucccuggacc cacaugcuca aguagcagug 2580
gggauccuga gaaaccaccu gacccaguuu agcccgcacu ucaaguccgg aucccuggcc 2640
guggauaacg ccgacccgau ccugaagauc cccuuugcau cccuggcuuc ccggcugacc 2700
uuggccgaac ugaaccagau ucuguaccgc ugcgaaucag aggaaaagga ggacggaggc 2760
ggaugcuacg auauccccaa uuggucggcg cugaaguacg ccggccuuca aggacugaug 2820
uccgugcugg ccgagaucag gccgaagaau gaccugggac acccguucug caacaacuug 2880
agaagcgggg acuggaugau ugauuacgug ucgaaccggc ugaucucccg gagcggcacc 2940
aucgcggaag ucggaaagug gcuccaggcc auguucuucu accugaagca gaucccccgc 3000
uaccugaucc ccugcuacuu cgacgcgauc uugauugggg cauacaccac ucugcuugac 3060
accgcuugga agcagauguc cuccuucgug caaaacggau ccaccuucgu gaagcaccug 3120
ucccuuggau cagugcagcu gugcggcgug ggaaaguucc cuucccuucc cauccugucc 3180
ccugcgcuga uggacgugcc guaccggcuc aacgagauca ccaaggaaaa ggaacagugc 3240
ugcgugucac uggcagccgg ccuuccgcau uuuucgagcg gaauuuucag auguuggggc 3300
agagacaccu ucauugcgcu gcgcgguauc cugcugauua ccggcagaua cguggaagcc 3360
agaaacauua uccuggcguu ugcugguacu cugcggcacg gacugauucc uaaccuguug 3420
ggagagggga ucuacgcccg guacaauugc agagaugccg ugugguggug gcugcagugc 3480
auccaggacu acugcaagau ggugcccaac ggacuugaca uucugaagug cccggugucg 3540
cgcauguacc ccaccgacga cucugcgccc cugccggccg guacccugga ucagccgcug 3600
uucgaaguga uccaggaagc caugcaaaag cacaugcagg gaauucaguu cagggaacgg 3660
aacgcaggcc cgcaaaucga ccggaacaug aaggacgaag gcuucaacau uaccgccggc 3720
guggacgagg aaaccggcuu cguguacggc ggcaaccggu ucaauugcgg uacuuggaug 3780
gacaagaugg gagaaagcga ccgcgccagg aaucggggca uuccugccac cccgcgggac 3840
gguagcgcgg uggagaucgu gggccuuucg aaguccgcgg uccgcuggcu ucuggagcug 3900
ucuaagaaaa acaucuuccc uuaccacgag gucaccguga aacgccacgg aaaggccauc 3960
aagguguccu acgacgaaug gaacaggaag auccaggaca acuuugagaa gcuguuucac 4020
gugucggaag auccguccga ccugaacgaa aagcacccca accuugucca uaagcgcggu 4080
aucuacaaag auucguaugg ugcauccucc ccuuggugcg acuaccaacu ccggccgaac 4140
uucaccaucg caaugguggu ggccccggag cuguucacca cugaaaaggc cuggaaggcc 4200
cuggaaaucg ccgaaaagaa gcugcuggga ccgcugggga ugaaaacccu ggaccccgau 4260
gauauggugu acugcgggau cuacgacaac gcccuggaua acgacaacua caaccuggcc 4320
aagggcuuca acuaccauca ggguccggag uggcuguggc caaucggaua cuuccugagg 4380
gccaagcugu acuucucccg cuugaugggc cccgaaacua ccgcaaagac uaucgugcuc 4440
gugaagaacg uccugucccg gcacuacgug caucuggaac ggucgccgug gaaaggccug 4500
ccagaguuga ccaacgagaa ugcccaguau ugcccguucu caugcgaaac ccaagccugg 4560
agcauugcca cuauucugga aacccucuac gaccuguag 4599
<210> 30
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 30
augggacacu cgaaacagau cagaauccug uugcucaacg aaauggagaa gcucgaaaag 60
acccuguucc ggcuggagca ggguuacgag cugcaguucc gccugggucc gacucuacaa 120
gggaaagcag ugacggucua caccaacuac ccguuucccg gagaaaccuu caaccgggag 180
aaguuccggu cccuggacug ggagaacccg accgaacgag aggaugacuc agauaaguac 240
ugcaagcuga acuugcaaca guccggguca uuccaguacu acuuucugca agggaacgag 300
aagucaggag ggggcuacau cguggucgac ccgauacugc gcgugggagc cgacaaucac 360
guccugccgc uggacugcgu gacccugcaa accuuccucg ccaaaugucu ggggccguuc 420
gaugaguggg aaagccgccu cagagucgca aaggaguccg gauacaacau gauucacuuc 480
acuccgcugc aaacacucgg ucugucccgg ucgugcuauu cucuggcgaa ccagcuggag 540
cuuaaucccg acuuuucccg cccgaaccgg aaguacaccu ggaacgacgu cgggcagcuu 600
guggaaaagc ugaagaagga guggaacgug aucugcauca cugauguggu guacaaccac 660
acggcugcca acucgaagug gauccaagag caucccgagu gcgcguacaa cuuggugaac 720
agcccgcauc ugaagccugc uugggugcug gacagagccc ucuggagauu cagcugcgac 780
guggccgagg ggaaguacaa agaaaagggg auuccggccu ugauugagaa cgaccaucac 840
augaacucaa uccgcaagau cauuugggag gauaucuucc cuaagcuuaa guugugggag 900
uucuuccaag uggacguaaa caaggcagug gagcaguuca gaaggcugcu gacucaagaa 960
aacagacgcg ucacuaaguc cgacccgaac cagcaccuua ccaucaucca agacccggag 1020
uaccgccggu ucggcugcac cgucgauaug aacauagccc ugaccacuuu caucccccau 1080
gacaaggggc cggcggcaau cgaggaaugc uguaacuggu uucauaagcg gauggaggaa 1140
cugaauuccg agaagcaccg gcugauuaac uaccaccagg aacaggcagu gaacugccug 1200
cugggcaacg uguucuacga acggcuggcu ggacacggac cgaagcuggg ucccgugacu 1260
agaaagcauc cgcucgucac ucgcuacuuc accuucccgu uugaggagau ugacuucucc 1320
auggaagaau ccaugaucca cuugccgaac aaggcuugcu uccugauggc gcacaacgga 1380
ugggucaugg gcgacgaccc gcugaggaau uucgcggagc cggguucgga aguguaccug 1440
agaagggaac ucauuugcug gggagacucc gucaagcugc gcuaugggaa caagcccgag 1500
gauugccccu accugugggc ucacaugaag aaguacacgg aaaucaccgc cacguacuuc 1560
cagggagucc ggcuggacaa uugccacucc accccccucc auguggccga guacaugcug 1620
gaugcagcgc gcaaucugca gccgaaucug uacgugguug cagagcuguu cacugggucc 1680
gaggaccucg acaacguguu cgugacuaga uuggggaucu ccucccucau ccgggaagcc 1740
augucggccu acaacuccca ugaggagggg aggcuggugu acagauacgg cggcgaaccc 1800
gugggaagcu ucgugcagcc gugccuccgg ccgcugaugc cggccaucgc ccacgcccug 1860
uucauggaua ucacucacga caacgaaugc ccgaucgugc aucgcucggc cuacgacgca 1920
cugccgucca ccacuaucgu guccauggca ugcugcgccu ccgggagcac ccgcggauac 1980
gaugagcucg ugccgcacca gauuagcgug guguccgaag aacgcuucua uaccaagugg 2040
aaccccgaag cccugccguc caauaccggg gaagugaacu uccaguccgg uauuaucgcc 2100
gcucgcugug cgaucucgaa acuccaccaa gagcucggug ccaagggguu cauucagguc 2160
uacguggauc aggucgacga ggauauugug gcagugacca ggcacucacc uagcauccac 2220
caauccgugg ucgccguguc acgcacugcg uuucggaacc ccaagaccuc guucuacucg 2280
aaagaagugc cgcagaugug uaucccggga aagaucgaag aggucgugcu ggaagcacgg 2340
accauugaga ggaacaccaa gccuuaccgg aaagacgaga acucuaucaa cgggaccccg 2400
gauaucacug uggagauucg cgaacacauc cagcugaacg aaucaaagau cgucaagcag 2460
gcuggagugg ccaccaaggg acccaacgag uacauccagg agaucgaauu ugaaaaccuc 2520
uccccggggu ccgugaucau cuuccgggug ucccuggacc cccacgccca aguggccgug 2580
ggaauucuca gaaaccaccu gacccaguuc ucaccccacu uuaagucggg uucccuggcc 2640
guggacaacg ccgauccgau ccucaagauc ccguucgcgu cgcuggccuc ccgccucacu 2700
cucgcggaac ugaaccagau ccuguaccgc ugugaaucag aggaaaagga ggacggcggc 2760
ggcuguuacg auauuccgaa uuggucggcu uugaaauacg cgggacuuca ggggcugaug 2820
ucuguccugg cggaaauccg gccgaagaac gaccuggggc acccguucug caacaacuug 2880
cggagcggag auuggaugau cgacuacguc agcaacagac ugaucagccg gagcggcacu 2940
aucgccgagg ucggaaagug guugcaggcc auguucuucu accugaagca gaucccccga 3000
uaccucaucc cguguuacuu cgacgccauc cugaucgggg ccuacaccac ccuccuggac 3060
accgccugga agcagaugag caguuuugug caaaacgggu ccaccuucgu gaagcaccuu 3120
ucccugggcu cagugcagcu gugcggcgug ggaaaguucc ccucgcugcc cauucugagc 3180
ccggcccuga uggacguccc cuaccggcug aacgagauca ccaaggagaa ggagcagugc 3240
uguguguccc uggcugccgg gcugccgcac uuuucguccg gcaucuuccg gugcuggggg 3300
cgggauaccu ucauugcccu gcggggaauc cuucuuauca ccggucgcua uguggaggcu 3360
cggaacauua uucuggcguu cgccggaacc cugagacacg ggcugauucc gaaccucuug 3420
ggggaaggga ucuacgcccg cuacaacugu cgggacgccg ugugguggug gcuccagugc 3480
auucaggacu auugcaagau ggugcccaac gggcuggaca uccugaagug cccggugucg 3540
aggauguauc cgaccgacga cagcgcaccc cugccggccg ggacccucga ccaaccccug 3600
uucgaaguca uucaagaggc uaugcagaag cacaugcagg guauucaguu ccgggaacgg 3660
aacgcggggc cccagauuga uaggaacaug aaagacgagg guuucaacau cacugccggc 3720
guggacgaag aaaccgguuu uguguacgga ggaaaccggu ucaacugcgg uaccuggaug 3780
gauaagaugg gagaaucaga ucgcgcgcgc aacagaggga ucccggcaac cccgcgggac 3840
ggaucggcug uggaaauugu gggacugagc aagagcgccg ugcgguggcu gcuggaacug 3900
agcaaaaaga acaucuuccc cuaccacgaa gugaccguga agcggcacgg aaaggccauc 3960
aaagucuccu acgaugaaug gaauaggaag auccaggaua acuucgagaa gcuuuuucac 4020
guguccgagg aucccuccga ucugaacgag aagcauccga aucucgugca uaagaggggg 4080
aucuacaagg acuccuacgg agcguccucc ccuuggugcg acuaucagcu gcggccuaac 4140
uucaccauug ccauggucgu ggccccggag cucuuuacaa ccgagaaggc cuggaaggcc 4200
cucgaaauug ccgaaaagaa gcugcugggg cccuugggga ugaaaacccu ggauccggac 4260
gacauggugu acugcggaau cuacgacaac gcccuggaca acgacaacua caaucuggcg 4320
aagggcuuca auuaccacca ggggccggaa uggcucuggc cgauugggua cuuccugcgc 4380
gccaagcugu acuucucacg gcugauggga ccggagacua ccgccaagac caucguccuc 4440
gugaagaacg ugcugucgcg gcacuacgug caccuggaga ggagccccug gaaggggcuu 4500
cccgagcuga cgaacgagaa cgcgcaguac ugucccuucu ccugcgaaac ccaagccugg 4560
uccauugcca cuauacugga aaccuuauau gaccuguag 4599
<210> 31
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 31
augggccaca gcaagcagau ccggaucuug cugcugaacg agauggagaa gcuggagaag 60
acccuguuca ggcuggagca gggcuacgag cugcaguucc gguugggccc caccuugcag 120
ggcaaggccg ugaccgugua caccaacuac cccuuccccg gcgaaaccuu caacagggag 180
aaguuccggu cccuggacug ggagaacccc accgagaggg aggacgacuc cgacaaguac 240
ugcaagcuga accugcagca guccggcucc uuccaguacu acuuccugca gggcaacgag 300
aaaagcggcg gcggcuacau cgugguggac cccaucuugc gggugggcgc cgacaaccac 360
gugcugcccu uggacugcgu gacccugcag accuucuugg ccaagugcuu gggccccuuc 420
gacgaguggg agagcaggcu gaggguggcc aaggaguccg gcuacaacau gauccacuuc 480
acccccuugc agacccuggg ccuguccagg uccugcuacu cccuggccaa ccaguuggag 540
uugaaccccg acuucuccag gcccaacagg aaguacaccu ggaacgacgu gggccagcug 600
guggagaagu ugaagaagga guggaacgug aucugcauca ccgacguggu guacaaccac 660
accgccgcca acagcaagug gauccaggag caccccgagu gcgccuacaa ccuggugaac 720
uccccccacu ugaagcccgc cuggguguug gacagggccc uguggcgguu cuccugcgac 780
guggccgagg gcaaguacaa ggagaagggc auccccgccu ugaucgagaa cgaccaccac 840
augaacucca uccggaagau caucugggag gacaucuucc ccaagcugaa gcugugggag 900
uucuuccagg uggacgugaa caaggccgug gagcaguuca ggaggcugcu gacccaggag 960
aacaggcggg ugaccaaguc cgaccccaac cagcaccuga ccaucaucca ggaccccgag 1020
uacaggcggu ucggcugcac cguggacaug aacaucgccc ugaccaccuu caucccccac 1080
gacaagggcc ccgccgccau cgaggagugc ugcaacuggu uccacaagag gauggaggag 1140
uugaacuccg agaagcaccg gcugaucaac uaccaccagg agcaggccgu gaacugccug 1200
uugggcaacg uguucuacga gcggcuggcc ggccacggcc ccaagcuggg ccccgugacc 1260
aggaagcacc ccuuggugac cagguacuuc accuuccccu ucgaggagau cgacuucucc 1320
auggaggagu ccaugaucca ccugcccaac aaggccugcu uccugauggc ccacaacggc 1380
ugggugaugg gcgacgaccc ccugcggaac uucgccgagc ccggcuccga gguguaccug 1440
aggagggagc ugaucugcug gggcgacagc gugaaguugc gguacggcaa caagcccgag 1500
gacugccccu accugugggc ccacaugaag aaguacaccg agaucaccgc caccuacuuc 1560
cagggcgugc ggcuggacaa cugccacucc accccccugc acguggccga guacauguug 1620
gacgccgcca ggaacuugca gcccaacuug uacguggugg ccgagcuguu caccggcagc 1680
gaggaccugg acaacguguu cgugaccagg cugggcauca gcuccuugau cagggaggcc 1740
augagcgccu acaacagcca cgaggagggc agguuggugu accgguacgg cggcgagccc 1800
gugggcuccu ucgugcagcc cugcuugagg cccuugaugc ccgccaucgc ccacgcccug 1860
uucauggaca ucacccacga caacgagugc cccaucgugc acagguccgc cuacgacgcc 1920
cugcccagca ccaccaucgu guccauggcc ugcugcgcca gcggcagcac caggggcuac 1980
gacgaguugg ugccccacca gaucuccgug guguccgagg agcgguucua caccaagugg 2040
aaccccgagg ccuugcccuc caacaccggc gaggugaacu uccagagcgg caucaucgcc 2100
gccaggugcg ccaucagcaa gcugcaccag gagcugggcg ccaagggcuu cauccaggug 2160
uacguggacc agguggacga ggacaucgug gccgugacca ggcacucccc cagcauccac 2220
caguccgugg uggccguguc caggaccgcc uucaggaacc ccaagaccuc cuucuacagc 2280
aaggaggugc cccagaugug cauccccggc aagaucgagg agguggugcu ggaggccagg 2340
accaucgaga ggaacaccaa gcccuacagg aaggacgaga acuccaucaa cggcaccccc 2400
gacaucaccg uggagaucag ggagcacauc cagcugaacg agagcaagau cgugaagcag 2460
gccggcgugg ccaccaaggg ccccaacgag uacauccagg agaucgaguu cgagaacuug 2520
ucccccggca gcgugaucau cuucagggug agccuggacc cccacgccca gguggccgug 2580
ggcauccugc ggaaccaccu gacccaguuc agcccccacu ucaaguccgg cagccuggcc 2640
guggacaacg ccgaccccau cuugaagauc cccuucgccu cccuggccuc cagguugacc 2700
uuggccgagc ugaaccagau ccuguaccgg ugcgaguccg aggagaagga ggacggcggc 2760
ggcugcuacg acauccccaa cugguccgcc cugaaguacg ccggccugca gggcuugaug 2820
uccguguugg ccgagaucag gcccaagaac gacuugggcc accccuucug caacaacuug 2880
agguccggcg acuggaugau cgacuacgug agcaaccggc ugaucucccg guccggcacc 2940
aucgccgagg ugggcaagug guugcaggcc auguucuucu accugaagca gaucccccgg 3000
uaccugaucc ccugcuacuu cgacgccauc uugaucggcg ccuacaccac ccugcuggac 3060
accgccugga agcagauguc cagcuucgug cagaacggcu ccaccuucgu gaagcaccug 3120
uccuugggcu ccgugcagcu gugcggcgug ggcaaguucc ccucccugcc cauccugucc 3180
cccgcccuga uggacgugcc cuacagguug aacgagauca ccaaggagaa ggagcagugc 3240
ugcguguccc uggccgccgg cuugccccac uucuccuccg gcaucuuccg gugcuggggc 3300
agggacaccu ucaucgcccu gaggggcauc cugcugauca ccggccggua cguggaggcc 3360
aggaacauca ucuuggccuu cgccggcacc cugaggcacg gccugauccc caaccugcug 3420
ggcgagggca ucuacgccag guacaacugc cgggacgccg ugugguggug gcugcagugc 3480
auccaggacu acugcaagau ggugcccaac ggccuggaca uccugaagug ccccgugucc 3540
aggauguacc ccaccgacga cuccgccccc uugcccgccg gcacccugga ccagcccuug 3600
uucgagguga uccaggaggc caugcagaag cacaugcagg gcauccaguu ccgggagagg 3660
aacgccggcc cccagaucga ccggaacaug aaggacgagg gcuucaacau caccgccggc 3720
guggacgagg aaaccggcuu cguguacggc ggcaaccggu ucaacugcgg caccuggaug 3780
gacaagaugg gcgagagcga cagggccagg aacaggggca uccccgccac ccccagggac 3840
ggcuccgccg uggagaucgu gggccugagc aaguccgccg ugcggugguu gcuggaguug 3900
uccaagaaga acaucuuccc cuaccacgag gugaccguga agaggcacgg caaggccauc 3960
aagguguccu acgacgagug gaacaggaag auccaggaca acuucgagaa gcuguuccac 4020
guguccgagg accccuccga cuugaacgag aagcacccca accuggugca caagcggggc 4080
aucuacaagg acagcuacgg cgccuccagc cccuggugcg acuaccagcu gaggcccaac 4140
uucaccaucg ccaugguggu ggcccccgag cuguucacca ccgagaaggc cuggaaggcc 4200
uuggagaucg ccgagaagaa guugcugggc ccccugggca ugaagaccuu ggaccccgac 4260
gacauggugu acugcggcau cuacgacaac gccuuggaca acgacaacua caaccuggcc 4320
aagggcuuca acuaccacca gggccccgag uggcuguggc ccaucggcua cuuccugcgg 4380
gccaaguugu acuucuccag guugaugggc cccgaaacca ccgccaagac caucguguug 4440
gugaagaacg ugcugucccg gcacuacgug caccuggaga ggucccccug gaagggccug 4500
cccgagcuga ccaacgagaa cgcccaguac ugccccuuca gcugcgaaac ccaggccugg 4560
uccaucgcca ccauccugga aacccuguac gacuuguag 4599
<210> 32
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 32
augggacaua gcaagcagau ccggaucuug cugcugaaug aaauggaaaa gcuggaaaag 60
acccuguuca gacuggaaca gggauacgaa cugcaguucc gguugggacc uaccuugcag 120
ggaaaggcug ugaccgugua caccaauuac ccuuucccug gagaaaccuu caauagagaa 180
aaguucagau cucuagauug ggaaaauccu accgaaagag aagaugauuc ugauaaguac 240
ugcaagcuga aucugcagca gucuggaucu uuccaguacu acuuccugca gggaaaugaa 300
aagagcggag gaggauacau cgugguggau ccuaucuugc gggugggagc ugauaaucau 360
gugcugccuu uggauugcgu gacccugcag accuucuugg cuaagugcuu gggaccuuuc 420
gaugaauggg aaagcagacu gagaguggcu aaggaaucug gauacaauau gauccauuuc 480
accccuuugc agacccuggg acugucuaga ucuugcuacu cucuggcuaa ucaguuggaa 540
uugaauccug auuucucuag accuaauaga aaguacaccu ggaaugaugu gggacagcug 600
guggaaaagu ugaagaagga auggaaugug aucugcauca ccgauguggu guacaaucau 660
accgcugcua auagcaagug gauccaggaa cauccugaau gcgcuuacaa ucuggugaau 720
ucuccucauu ugaagccugc uuggguguug gauagagcuc uguggcgguu cucuugcgau 780
guggcugaag gaaaguacaa ggaaaaggga aucccugcuu ugaucgaaaa ugaucaucau 840
augaauucua uccggaagau caucugggaa gauaucuucc cuaagcugaa gcugugggaa 900
uucuuccagg uggaugugaa uaaggcugug gaacaguuca gaagacugcu gacccaggaa 960
aauagacggg ugaccaaguc ugauccuaau cagcaucuga ccaucaucca ggauccugaa 1020
uacagacggu ucggaugcac cguggauaug aauaucgcuc ugaccaccuu caucccucau 1080
gauaagggac cugcugcuau cgaagaaugc ugcaauuggu uccauaagag aauggaagaa 1140
uugaauucug aaaagcaucg gcugaucaau uaccaucagg aacaggcugu gaauugccug 1200
uugggaaaug uguucuacga acggcuggcu ggacauggac cuaagcuggg accugugacc 1260
agaaagcauc cuuuggugac cagauacuuc accuucccuu ucgaagaaau cgauuucucu 1320
auggaagaau cuaugaucca ucugccuaau aaggcuugcu uccugauggc ucauaaugga 1380
ugggugaugg gagaugaucc ucugcggaau uucgcugaac cuggaucuga aguguaccug 1440
agaagagaac ugaucugcug gggagauagc gugaaguugc gguacggaaa uaagccugaa 1500
gauugcccuu accugugggc ucauaugaag aaguacaccg aaaucaccgc uaccuacuuc 1560
cagggagugc ggcuggauaa uugccauucu accccucugc auguggcuga auacauguug 1620
gaugcugcua gaaauuugca gccuaauuug uacguggugg cugaacuguu caccggaagc 1680
gaagaucugg auaauguguu cgugaccaga cugggaauca gcucuuugau cagagaagcu 1740
augagcgcuu acaauagcca ugaagaagga agauuggugu accgguacgg aggagaaccu 1800
gugggaucuu ucgugcagcc uugcuugagg ccuuugaugc cugcuaucgc ucaugcucug 1860
uucauggaua ucacccauga uaaugaaugc ccuaucgugc auagaucugc uuacgaugcu 1920
cugccuagca ccaccaucgu gucuauggcu ugcugcgcua gcggaagcac cagaggauac 1980
gaugaauugg ugccucauca gaucucugug gugucugaag aacgguucua caccaagugg 2040
aauccugaag cuuugccuuc uaauaccgga gaagugaauu uccagagcgg aaucaucgcu 2100
gcuagaugcg cuaucagcaa gcugcaucag gaacugggag cuaagggauu cauccaggug 2160
uacguggauc agguggauga agauaucgug gcugugacca gacauucucc uagcauccau 2220
cagucugugg uggcuguguc uagaaccgcu uucagaaauc cuaagaccuc uuucuacagc 2280
aaggaagugc cucagaugug caucccugga aagaucgaag aaguggugcu ggaagcuaga 2340
accaucgaaa gaaauaccaa gccuuacaga aaggaugaaa auucuaucaa uggaaccccu 2400
gauaucaccg uggaaaucag agaacauauc cagcugaaug aaagcaagau cgugaagcag 2460
gcuggagugg cuaccaaggg accuaaugaa uacauccagg aaaucgaauu cgaaaauuug 2520
ucuccuggaa gcgugaucau cuucagagug agccuggauc cucaugcuca gguggcugug 2580
ggaauccugc ggaaucaucu gacccaguuc agcccucauu ucaagucugg aagccuggcu 2640
guggauaaug cugauccuau cuugaagauc ccuuucgcuu cucuggcuuc uagauugacc 2700
uuggcugaac ugaaucagau ccuguaccgg ugcgaaucug aagaaaagga agauggagga 2760
ggaugcuacg auaucccuaa uuggucugcu cugaaguacg cuggacugca gggauugaug 2820
ucuguguugg cugaaaucag accuaagaau gauuugggac auccuuucug caauaauuug 2880
agaucuggag auuggaugau cgauuacgug agcaaucggc ugaucucucg gucuggaacc 2940
aucgcugaag ugggaaagug guugcaggcu auguucuucu accugaagca gaucccucgg 3000
uaccugaucc cuugcuacuu cgaugcuauc uugaucggag cuuacaccac ccugcuggau 3060
accgcuugga agcagauguc uagcuucgug cagaauggau cuaccuucgu gaagcaucug 3120
ucuuugggau cugugcagcu gugcggagug ggaaaguucc cuucucugcc uauccugucu 3180
ccugcucuga uggaugugcc uuacagauug aaugaaauca ccaaggaaaa ggaacagugc 3240
ugcgugucuc uggcugcugg auugccucau uucucuucug gaaucuuccg gugcugggga 3300
agagauaccu ucaucgcucu gagaggaauc cugcugauca ccggacggua cguggaagcu 3360
agaaauauca ucuuggcuuu cgcuggaacc cugagacaug gacugauccc uaaucugcug 3420
ggagaaggaa ucuacgcuag auacaauugc cgggaugcug ugugguggug gcugcagugc 3480
auccaggauu acugcaagau ggugccuaau ggacuggaua uccugaagug cccugugucu 3540
agaauguacc cuaccgauga uucugcuccu uugccugcug gaacccugga ucagccuuug 3600
uucgaaguga uccaggaagc uaugcagaag cauaugcagg gaauccaguu ccgggaaaga 3660
aaugcuggac cucagaucga ucggaauaug aaggaugaag gauucaauau caccgcugga 3720
guggaugaag aaaccggauu cguguacgga ggaaaucggu ucaauugcgg aaccuggaug 3780
gauaagaugg gagaaagcga uagagcuaga aauagaggaa ucccugcuac cccuagagau 3840
ggaucugcug uggaaaucgu gggacugagc aagucugcug ugcggugguu gcuggaauug 3900
ucuaagaaga auaucuuccc uuaccaugaa gugaccguga agagacaugg aaaggcuauc 3960
aaggugucuu acgaugaaug gaauagaaag auccaggaua auuucgaaaa gcuguuccau 4020
gugucugaag auccuucuga uuugaaugaa aagcauccua aucuggugca uaagcgggga 4080
aucuacaagg auagcuacgg agcuucuagc ccuuggugcg auuaccagcu gaggccuaau 4140
uucaccaucg cuaugguggu ggcuccugaa cuguucacca ccgaaaaggc uuggaaggcu 4200
uuggaaaucg cugaaaagaa guugcuggga ccucugggaa ugaagaccuu ggauccugau 4260
gauauggugu acugcggaau cuacgauaau gcuuuggaua augauaauua caaucuggcu 4320
aagggauuca auuaccauca gggaccugaa uggcuguggc cuaucggaua cuuccugcgg 4380
gcuaaguugu acuucucuag auugauggga ccugaaacca ccgcuaagac caucguguug 4440
gugaagaaug ugcugucucg gcauuacgug caucuggaaa gaucuccuug gaagggacug 4500
ccugaacuga ccaaugaaaa ugcucaguac ugcccuuuca gcugcgaaac ccaggcuugg 4560
ucuaucgcua ccauccugga aacccuguac gauuuguag 4599
<210> 33
<211> 158
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 经过修饰的非洲爪蟾β-珠蛋白3' UTR
<220>
<221> misc_feature
<222> (2)..(2)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (157)..(157)
<223> n为解锁核单体UNA-A
<400> 33
cnagugacug acuaggaucu gguuaccacu aaaccagccu caagaacacc cgaauggagu 60
cucuaagcua cauaauacca acuuacacuu acaaaauguu gucccccaaa auguagccau 120
ucguaucugc uccuaauaaa aagaaaguuu cuucacnu 158
<210> 34
<211> 158
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 经过修饰的非洲爪蟾β-珠蛋白3' UTR
<220>
<221> misc_feature
<222> (2)..(2)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (3)..(3)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (4)..(4)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (155)..(155)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (156)..(156)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (157)..(157)
<223> n为解锁核单体UNA-A
<400> 34
cnnnugacug acuaggaucu gguuaccacu aaaccagccu caagaacacc cgaauggagu 60
cucuaagcua cauaauacca acuuacacuu acaaaauguu gucccccaaa auguagccau 120
ucguaucugc uccuaauaaa aagaaaguuu cuucnnnu 158
<210> 35
<211> 158
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 经过修饰的非洲爪蟾β-珠蛋白3' UTR
<220>
<221> misc_feature
<222> (2)..(2)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (8)..(8)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (14)..(14)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (20)..(20)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (26)..(26)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (32)..(32)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (38)..(38)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (44)..(44)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (50)..(50)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (56)..(56)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (62)..(62)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (68)..(68)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (74)..(74)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (80)..(80)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (86)..(86)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (92)..(92)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (98)..(98)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (104)..(104)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (110)..(110)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (116)..(116)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (122)..(122)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (128)..(128)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (134)..(134)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (140)..(140)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (146)..(146)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (152)..(152)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (157)..(157)
<223> n为解锁核单体UNA-A
<400> 35
cnaguganug acunggaucn gguuancacu anaccagncu caanaacacn cgaaungagu 60
cncuaagnua caunauaccn acuuanacuu anaaaaunuu gucncccaan auguanccau 120
unguaucngc uccnaauaan aagaanguuu cnucacnu 158
<210> 36
<211> 158
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 经过修饰的非洲爪蟾β-珠蛋白3' UTR
<220>
<221> misc_feature
<222> (2)..(2)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (3)..(3)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (8)..(8)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (9)..(9)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (14)..(14)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (15)..(15)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (20)..(20)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (21)..(21)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (26)..(27)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (32)..(33)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (38)..(39)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (44)..(44)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (45)..(45)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (50)..(51)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (56)..(57)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (62)..(62)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (63)..(63)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (68)..(68)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (69)..(69)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (74)..(75)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (80)..(81)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (86)..(86)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (87)..(87)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (92)..(92)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (93)..(93)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (98)..(98)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (99)..(99)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (104)..(105)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (110)..(111)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (116)..(116)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (117)..(117)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (122)..(122)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (123)..(123)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (128)..(128)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (129)..(129)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (134)..(134)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (135)..(135)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (140)..(141)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (146)..(146)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (147)..(147)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (152)..(153)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (157)..(157)
<223> n为解锁核单体UNA-A
<400> 36
cnnguganng acunngaucn nguuannacu annccagnnu caannacacn ngaaunnagu 60
cnnuaagnna caunnuaccn ncuuanncuu annaaaunnu gucnnccaan nuguanncau 120
unnuaucnnc uccnnauaan nagaannuuu cnncacnu 158
<210> 37
<211> 158
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 经过修饰的非洲爪蟾β-珠蛋白3' UTR
<220>
<221> misc_feature
<222> (1)..(1)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (2)..(2)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (3)..(3)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (4)..(4)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (5)..(5)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (6)..(6)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (7)..(7)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (8)..(8)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (9)..(9)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (10)..(10)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (11)..(11)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (12)..(12)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (13)..(13)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (14)..(14)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (15)..(16)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (17)..(17)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (18)..(18)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (19)..(19)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (20)..(20)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (21)..(22)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (23)..(24)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (25)..(25)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (26)..(27)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (28)..(28)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (29)..(29)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (30)..(30)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (31)..(33)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (34)..(35)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (36)..(36)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (37)..(37)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (38)..(39)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (40)..(40)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (41)..(41)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (42)..(43)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (44)..(44)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (45)..(46)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (47)..(47)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (48)..(48)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (49)..(51)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (52)..(52)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (53)..(54)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (55)..(55)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (56)..(57)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (58)..(58)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (59)..(59)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (60)..(60)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (61)..(61)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (62)..(62)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (63)..(63)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (64)..(64)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (65)..(66)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (67)..(67)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (68)..(68)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (69)..(69)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (70)..(70)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (71)..(71)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (72)..(72)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (73)..(73)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (74)..(75)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (76)..(76)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (77)..(77)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (78)..(79)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (80)..(81)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (82)..(82)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (83)..(84)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (85)..(85)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (86)..(86)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (87)..(87)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (88)..(88)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (89)..(90)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (91)..(91)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (92)..(92)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (93)..(96)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (97)..(97)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (98)..(98)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (99)..(100)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (101)..(101)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (102)..(102)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (103)..(107)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (108)..(111)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (112)..(112)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (113)..(113)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (114)..(114)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (115)..(115)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (116)..(116)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (117)..(118)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (119)..(119)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (120)..(121)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (122)..(122)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (123)..(123)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (124)..(124)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (125)..(125)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (126)..(126)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (127)..(127)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (128)..(128)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (129)..(129)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (130)..(130)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (131)..(131)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (132)..(133)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (134)..(134)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (135)..(136)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (137)..(137)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (138)..(142)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (143)..(143)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (144)..(146)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (147)..(147)
<223> n为解锁核单体UNA-G
<220>
<221> misc_feature
<222> (148)..(149)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (150)..(150)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (151)..(151)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (152)..(153)
<223> n为解锁核单体UNA-U
<220>
<221> misc_feature
<222> (154)..(154)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (155)..(155)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (156)..(156)
<223> n为解锁核单体UNA-C
<220>
<221> misc_feature
<222> (157)..(157)
<223> n为解锁核单体UNA-A
<220>
<221> misc_feature
<222> (158)..(158)
<223> n为解锁核单体UNA-U
<400> 37
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 60
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnn 158
<210> 38
<211> 100
<212> RNA
<213> 未知
<220>
<223> Poly(A) 100尾
<400> 38
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 60
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 100
<210> 39
<211> 110
<212> RNA
<213> 未知
<220>
<223> Poly(A) 110尾
<400> 39
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 60
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 110
<210> 40
<211> 9
<212> RNA
<213> 未知
<220>
<223> 三联终止密码子
<400> 40
auaagugaa 9
<210> 41
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 41
augggccaca gcaagcagau ccggauccug cugcugaacg agauggagaa gcuggagaag 60
acccuguucc ggcuggagca gggcuacgag cugcaguucc ggcugggccc cacccugcag 120
ggcaaggccg ugaccgugua caccaacuac cccuuccccg gcgagaccuu caaccgggag 180
aaguuccgga gccuggacug ggagaacccc accgagcggg aggacgacag cgacaaguac 240
ugcaagcuga accugcagca gagcggcagc uuccaguacu acuuccugca gggcaacgag 300
aagagcggcg gcggcuacau cgugguggac cccauccugc gggugggcgc cgacaaccac 360
gugcugcccc uggacugcgu gacccugcag accuuccugg ccaagugccu gggccccuuc 420
gacgaguggg agagccggcu gcggguggcc aaggagagcg gcuacaacau gauccacuuc 480
accccccugc agacccuggg ccugagccgg agcugcuaca gccuggccaa ccagcuggag 540
cugaaccccg acuucagccg gcccaaccgg aaguacaccu ggaacgacgu gggccagcug 600
guggagaagc ugaagaagga guggaacgug aucugcauca ccgacguggu guacaaccac 660
accgccgcca acagcaagug gauccaggag caccccgagu gcgccuacaa ccuggugaac 720
agcccccacc ugaagcccgc cugggugcug gaccgggccc uguggcgguu cagcugcgac 780
guggccgagg gcaaguacaa ggagaagggc auccccgccc ugaucgagaa cgaccaccac 840
augaacagca uccggaagau caucugggag gacaucuucc ccaagcugaa gcugugggag 900
uucuuccagg uggacgugaa caaggccgug gagcaguucc ggcggcugcu gacccaggag 960
aaccggcggg ugaccaagag cgaccccaac cagcaccuga ccaucaucca ggaccccgag 1020
uaccggcggu ucggcugcac cguggacaug aacaucgccc ugaccaccuu caucccccac 1080
gacaagggcc ccgccgccau cgaggagugc ugcaacuggu uccacaagcg gauggaggag 1140
cugaacagcg agaagcaccg gcugaucaac uaccaccagg agcaggccgu gaacugccug 1200
cugggcaacg uguucuacga gcggcuggcc ggccacggcc ccaagcuggg ccccgugacc 1260
cggaagcacc cccuggugac ccgguacuuc accuuccccu ucgaggagau cgacuucagc 1320
auggaggaga gcaugaucca ccugcccaac aaggccugcu uccugauggc ccacaacggc 1380
ugggugaugg gcgacgaccc ccugcggaac uucgccgagc ccggcagcga gguguaccug 1440
cggcgggagc ugaucugcug gggcgacagc gugaagcugc gguacggcaa caagcccgag 1500
gacugccccu accugugggc ccacaugaag aaguacaccg agaucaccgc caccuacuuc 1560
cagggcgugc ggcuggacaa cugccacagc accccccugc acguggccga guacaugcug 1620
gacgccgccc ggaaccugca gcccaaccug uacguggugg ccgagcuguu caccggcagc 1680
gaggaccugg acaacguguu cgugacccgg cugggcauca gcagccugau ccgggaggcc 1740
augagcgccu acaacagcca cgaggagggc cggcuggugu accgguacgg cggcgagccc 1800
gugggcagcu ucgugcagcc cugccugcgg ccccugaugc ccgccaucgc ccacgcccug 1860
uucauggaca ucacccacga caacgagugc cccaucgugc accggagcgc cuacgacgcc 1920
cugcccagca ccaccaucgu gagcauggcc ugcugcgcca gcggcagcac ccggggcuac 1980
gacgagcugg ugccccacca gaucagcgug gugagcgagg agcgguucua caccaagugg 2040
aaccccgagg cccugcccag caacaccggc gaggugaacu uccagagcgg caucaucgcc 2100
gcccggugcg ccaucagcaa gcugcaccag gagcugggcg ccaagggcuu cauccaggug 2160
uacguggacc agguggacga ggacaucgug gccgugaccc ggcacagccc cagcauccac 2220
cagagcgugg uggccgugag ccggaccgcc uuccggaacc ccaagaccag cuucuacagc 2280
aaggaggugc cccagaugug cauccccggc aagaucgagg agguggugcu ggaggcccgg 2340
accaucgagc ggaacaccaa gcccuaccgg aaggacgaga acagcaucaa cggcaccccc 2400
gacaucaccg uggagauccg ggagcacauc cagcugaacg agagcaagau cgugaagcag 2460
gccggcgugg ccaccaaggg ccccaacgag uacauccagg agaucgaguu cgagaaccug 2520
agccccggca gcgugaucau cuuccgggug agccuggacc cccacgccca gguggccgug 2580
ggcauccugc ggaaccaccu gacccaguuc agcccccacu ucaagagcgg cagccuggcc 2640
guggacaacg ccgaccccau ccugaagauc cccuucgcca gccuggccag ccggcugacc 2700
cuggccgagc ugaaccagau ccuguaccgg ugcgagagcg aggagaagga ggacggcggc 2760
ggcugcuacg acauccccaa cuggagcgcc cugaaguacg ccggccugca gggccugaug 2820
agcgugcugg ccgagauccg gcccaagaac gaccugggcc accccuucug caacaaccug 2880
cggagcggcg acuggaugau cgacuacgug agcaaccggc ugaucagccg gagcggcacc 2940
aucgccgagg ugggcaagug gcugcaggcc auguucuucu accugaagca gaucccccgg 3000
uaccugaucc ccugcuacuu cgacgccauc cugaucggcg ccuacaccac ccugcuggac 3060
accgccugga agcagaugag cagcuucgug cagaacggca gcaccuucgu gaagcaccug 3120
agccugggca gcgugcagcu gugcggcgug ggcaaguucc ccagccugcc cauccugagc 3180
cccgcccuga uggacgugcc cuaccggcug aacgagauca ccaaggagaa ggagcagugc 3240
ugcgugagcc uggccgccgg ccugccccac uucagcagcg gcaucuuccg gugcuggggc 3300
cgggacaccu ucaucgcccu gcggggcauc cugcugauca ccggccggua cguggaggcc 3360
cggaacauca uccuggccuu cgccggcacc cugcggcacg gccugauccc caaccugcug 3420
ggcgagggca ucuacgcccg guacaacugc cgggacgccg ugugguggug gcugcagugc 3480
auccaggacu acugcaagau ggugcccaac ggccuggaca uccugaagug ccccgugagc 3540
cggauguacc ccaccgacga cagcgccccc cugcccgccg gcacccugga ccagccccug 3600
uucgagguga uccaggaggc caugcagaag cacaugcagg gcauccaguu ccgggagcgg 3660
aacgccggcc cccagaucga ccggaacaug aaggacgagg gcuucaacau caccgccggc 3720
guggacgagg agaccggcuu cguguacggc ggcaaccggu ucaacugcgg caccuggaug 3780
gacaagaugg gcgagagcga ccgggcccgg aaccggggca uccccgccac cccccgggac 3840
ggcagcgccg uggagaucgu gggccugagc aagagcgccg ugcgguggcu gcuggagcug 3900
agcaagaaga acaucuuccc cuaccacgag gugaccguga agcggcacgg caaggccauc 3960
aaggugagcu acgacgagug gaaccggaag auccaggaca acuucgagaa gcuguuccac 4020
gugagcgagg accccagcga ccugaacgag aagcacccca accuggugca caagcggggc 4080
aucuacaagg acagcuacgg cgccagcagc cccuggugcg acuaccagcu gcggcccaac 4140
uucaccaucg ccaugguggu ggcccccgag cuguucacca ccgagaaggc cuggaaggcc 4200
cuggagaucg ccgagaagaa gcugcugggc ccccugggca ugaagacccu ggaccccgac 4260
gacauggugu acugcggcau cuacgacaac gcccuggaca acgacaacua caaccuggcc 4320
aagggcuuca acuaccacca gggccccgag uggcuguggc ccaucggcua cuuccugcgg 4380
gccaagcugu acuucagccg gcugaugggc cccgagacca ccgccaagac caucgugcug 4440
gugaagaacg ugcugagccg gcacuacgug caccuggagc ggagccccug gaagggccug 4500
cccgagcuga ccaacgagaa cgcccaguac ugccccuuca gcugcgagac ccaggccugg 4560
agcaucgcca ccauccugga gacccuguac gaccuguag 4599
<210> 42
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 42
augggccaua gcaagcagau ccggauccuc cugcucaaug agauggaaaa gcucgagaaa 60
acccucuuua ggcuggagca aggcuacgaa cuccaguuuc ggcucgggcc cacucuccag 120
ggcaaggccg ugaccgucua caccaacuau cccuucccug gcgagaccuu caacagggag 180
aaguuucggu cccucgacug ggagaacccc accgagaggg aggacgacuc cgacaaguau 240
ugcaagcuga accuccaaca auccgggucc uuccaguacu acuuucugca aggcaacgaa 300
aagagcgggg gcggguauau uguggucgau cccauccucc gggugggggc ugacaaccac 360
guccuccccc uggauugcgu gacucugcag accuuccucg cuaaaugccu gggcccuuuc 420
gacgaguggg agagcaggcu gcggguggcc aaggaguccg gguauaauau gauccacuuc 480
accccccugc agacucuggg ccucucccgg uccuguuaua gccuggccaa ccagcucgag 540
cucaacccug auuucuccag gccuaacagg aaauacaccu ggaacgacgu gggccagcuc 600
gucgagaagc ucaagaaaga guggaacgug aucugcauca cugacguggu cuauaaccac 660
acugcugcua acagcaagug gauucaggag caccccgagu gcgccuacaa ccuggucaac 720
uccccccauc ucaagccugc cuggguccuc gauagggccc uguggcgguu uagcugcgac 780
guggccgagg gcaaguauaa ggagaaaggc auuccugcuc ugaucgagaa cgaccaucac 840
augaacagca uucggaagau uaucugggaa gacaucuucc ccaaacugaa gcucugggag 900
uucuuucaag ucgacgucaa uaaggcugug gaacaauuca ggaggcugcu gacccaagag 960
aaccggcggg ucaccaaauc cgaccccaau caacaucuga cuaucaucca agacccugag 1020
uauaggcggu ucgggugcac ugucgacaug aauaucgccc ucacuacuuu uauuccccac 1080
gauaaaggcc ccgccgccau cgaggagugu ugcaacuggu uccacaagag gauggaagag 1140
cucaacuccg aaaaacaccg gcucaucaau uaccaccagg agcaggccgu gaacugucug 1200
cugggcaacg ucuucuacga gcggcucgcu gggcacgggc ccaagcucgg cccugucacu 1260
aggaaacacc cucucgugac ccgguacuuc acuuuucccu ucgaggaaau ugauuuuagc 1320
auggaggagu ccaugaucca ccuccccaac aaggcuugcu uccucauggc ccauaacggc 1380
ugggucaugg gcgacgaccc ucugcggaau uucgcugagc ccggguccga gguguaucuc 1440
cggagggagc ugaucuguug gggcgauagc gugaagcucc gguacggcaa caagcccgaa 1500
gauugcccuu accucugggc ccauaugaag aaguauacug agauuacugc cacuuacuuu 1560
cagggcgucc ggcuggacaa uugucauucc accccucugc augucgcuga auauaugcug 1620
gacgcugcuc ggaaccugca acccaaccuc uacgucgucg cugagcucuu uaccggcagc 1680
gaggaccucg auaacgucuu cgugacuagg cucgggauca gcagccucau uagggaagcc 1740
augagcgccu acaacagcca cgaggaaggg aggcucgugu aucgguacgg cggcgagccu 1800
gugggcagcu ucgugcagcc cugccugcgg ccucucaugc ccgcuaucgc ccacgcccuc 1860
uucauggaca ucacucacga caacgaaugc ccuaucgucc acagguccgc uuacgacgcu 1920
cugcccagca cuaccaucgu guccauggcc ugcugcgcua gcggcagcac caggggguac 1980
gacgagcucg ucccucacca gaucuccguc guguccgagg agcgguucua uacuaaaugg 2040
aacccugagg cccugccuuc caauaccggg gaggugaacu uucaaagcgg gaucaucgcc 2100
gcccggugcg cuauuagcaa gcugcaccag gaacugggcg ccaaaggguu cauccagguc 2160
uacgucgacc aaguggacga ggauauuguc gccgucacca ggcauucccc uagcauucac 2220
caguccgugg ucgcugucuc caggacugcu uuucggaacc ccaaaacuuc cuucuauagc 2280
aaagaggucc cucaaaugug uauuccuggg aagaucgagg aggucgugcu ggaggcuagg 2340
acuaucgaaa ggaauacuaa gccuuaccgg aaagacgaaa acuccauuaa cggcaccccc 2400
gacauuaccg ucgagaucag ggagcacauc cagcugaacg agagcaagau cgugaagcaa 2460
gcuggcgugg ccaccaaggg ccccaacgag uacauccaag agauugaguu cgagaaucug 2520
ucccccggca gcgugaucau uuuuagggug agccuggacc cccacgccca agucgcugug 2580
ggcauccugc ggaaccaccu cacccaauuu agcccucauu ucaaguccgg gagccucgcu 2640
guggauaacg ccgacccuau ucucaagauc ccuuucgcuu cccuggccag caggcucacu 2700
cuggcugaac ucaaccagau ucuguaucgg ugcgaguccg aggagaaaga ggacgggggc 2760
ggcuguuacg auauucccaa uugguccgcc cugaaguacg ccgggcugca agggcucaug 2820
uccguccugg ccgagauuag gcccaaaaac gaucugggcc acccuuucug caacaaccug 2880
agguccggcg acuggaugau cgauuacguc agcaaucggc ugaucucccg guccggcacu 2940
aucgcugagg uggggaagug gcugcaggcu auguuuuuuu aucucaaaca gauuccccgg 3000
uaucugauuc ccugcuauuu cgacgcuauu cucaucgggg ccuacaccac ucugcucgac 3060
accgccugga aacagauguc cagcuucgug cagaacgggu ccaccuucgu caagcaucug 3120
ucccuggggu ccgugcaacu cugcggcguc ggcaaguuuc cuagccuccc cauccugucc 3180
ccugcccuca uggacguccc uuaccggcuc aacgaaauua ccaaggagaa agaacaaugc 3240
ugcguguccc ucgcugccgg gcucccucac uucuccuccg ggaucuuucg guguuggggc 3300
cgggacacuu ucaucgcccu gagggggauu cuccucauca cuggccggua cgucgaggcc 3360
cggaacauca uccucgccuu cgcugggacc cuccggcacg ggcucauccc uaaccuccuc 3420
ggggagggca ucuacgccag guauaacugc cgggacgcug ucugguggug gcuccagugc 3480
auucaggacu auugcaagau ggugcccaac gggcucgaua uccucaaaug ucccgugagc 3540
aggauguacc cuaccgacga cuccgcuccu cugccugcug ggacccucga ccagccccug 3600
uucgagguca uucaggaggc caugcaaaag cauaugcagg ggauucaguu ucgggagcgg 3660
aacgcugggc cccagauuga ccggaauaug aaagaugagg gguucaacau uacugccggc 3720
guggacgagg agaccggcuu cguguacggc ggcaaccggu uuaacugcgg gaccuggaug 3780
gacaagaugg gcgagagcga uagggcuagg aauaggggca uucccgccac ccccagggac 3840
ggcuccgcug ucgagaucgu cgggcucagc aaguccgcug ugcgguggcu gcucgagcuc 3900
agcaagaaga acaucuuucc uuaccacgag gucaccguca agaggcacgg caaagcuauu 3960
aaagucuccu acgacgaaug gaauaggaag auucaagaua auuucgagaa acucuuccac 4020
gugagcgagg auccuuccga ccucaacgag aaacacccca accucgugca uaagcggggg 4080
auuuauaagg acagcuacgg cgcuuccagc ccuuggugcg auuaccagcu ccggccuaac 4140
uucaccauug ccaugguggu cgccccugaa cucuucacua ccgagaaggc cuggaaggcu 4200
cuggaaaucg ccgagaagaa gcugcugggg ccccugggga ugaagacucu cgaccccgac 4260
gacauggugu auugcggcau cuacgauaac gcccucgaua acgauaauua uaaccuggcu 4320
aagggguuua acuaccauca aggcccugaa uggcucuggc cuaucggcua cuuccugcgg 4380
gccaagcucu acuucagcag gcugaugggg cccgaaacua cugccaaaac uauugugcug 4440
gugaagaacg ugcugagccg gcacuacgug caccuggaaa ggagcccuug gaagggccug 4500
cccgagcuca ccaacgaaaa cgcccaguau ugcccuuuua gcugcgagac ccaagccugg 4560
uccaucgcua cuauccugga aacccuguac gaccucuag 4599
<210> 43
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 43
auggggcaca gcaaacaaau caggauccuc cuccucaacg aaauggaaaa acucgaaaaa 60
acccucuuca ggcucgaaca aggguacgaa cuccaauuca ggcucgggcc gacgcuccaa 120
gggaaagcgg ugacggugua cacgaacuac ccguucccgg gggaaacguu caacagggaa 180
aaauucagga gccucgacug ggaaaacccg acggaaaggg aagacgacag cgacaaauac 240
ugcaaacuca accuccaaca aagcgggagc uuccaauacu acuuccucca agggaacgaa 300
aaaagcgggg ggggguacau cgugguggac ccgauccuca gggugggggc ggacaaccac 360
gugcucccgc ucgacugcgu gacgcuccaa acguuccucg cgaaaugccu cgggccguuc 420
gacgaauggg aaagcaggcu caggguggcg aaagaaagcg gguacaacau gauccacuuc 480
acgccgcucc aaacgcucgg gcucagcagg agcugcuaca gccucgcgaa ccaacucgaa 540
cucaacccgg acuucagcag gccgaacagg aaauacacgu ggaacgacgu ggggcaacuc 600
guggaaaaac ucaaaaaaga auggaacgug aucugcauca cggacguggu guacaaccac 660
acggcggcga acagcaaaug gauccaagaa cacccggaau gcgcguacaa ccucgugaac 720
agcccgcacc ucaaaccggc gugggugcuc gacagggcgc ucuggagguu cagcugcgac 780
guggcggaag ggaaauacaa agaaaaaggg aucccggcgc ucaucgaaaa cgaccaccac 840
augaacagca ucaggaaaau caucugggaa gacaucuucc cgaaacucaa acucugggaa 900
uucuuccaag uggacgugaa caaagcggug gaacaauuca ggaggcuccu cacgcaagaa 960
aacaggaggg ugacgaaaag cgacccgaac caacaccuca cgaucaucca agacccggaa 1020
uacaggaggu ucgggugcac gguggacaug aacaucgcgc ucacgacguu caucccgcac 1080
gacaaagggc cggcggcgau cgaagaaugc ugcaacuggu uccacaaaag gauggaagaa 1140
cucaacagcg aaaaacacag gcucaucaac uaccaccaag aacaagcggu gaacugccuc 1200
cucgggaacg uguucuacga aaggcucgcg gggcacgggc cgaaacucgg gccggugacg 1260
aggaaacacc cgcucgugac gagguacuuc acguucccgu ucgaagaaau cgacuucagc 1320
auggaagaaa gcaugaucca ccucccgaac aaagcgugcu uccucauggc gcacaacggg 1380
ugggugaugg gggacgaccc gcucaggaac uucgcggaac cggggagcga aguguaccuc 1440
aggagggaac ucaucugcug gggggacagc gugaaacuca gguacgggaa caaaccggaa 1500
gacugcccgu accucugggc gcacaugaaa aaauacacgg aaaucacggc gacguacuuc 1560
caagggguga ggcucgacaa cugccacagc acgccgcucc acguggcgga auacaugcuc 1620
gacgcggcga ggaaccucca accgaaccuc uacguggugg cggaacucuu cacggggagc 1680
gaagaccucg acaacguguu cgugacgagg cucgggauca gcagccucau cagggaagcg 1740
augagcgcgu acaacagcca cgaagaaggg aggcucgugu acagguacgg gggggaaccg 1800
guggggagcu ucgugcaacc gugccucagg ccgcucaugc cggcgaucgc gcacgcgcug 1860
uucauggaca ucacgcacga caacgaaugc ccgaucgugc acaggagcgc guacgacgcg 1920
cucccgagca cgacgaucgu gagcauggcg ugcugcgcga gcgggagcac gaggggguac 1980
gacgaacucg ugccgcacca aaucagcgug gugagcgaag aaagguucua cacgaaaugg 2040
aacccggaag cgcucccgag caacacgggg gaagugaacu uccaaagcgg gaucaucgcg 2100
gcgaggugcg cgaucagcaa acuccaccaa gaacucgggg cgaaaggguu cauccaagug 2160
uacguggacc aaguggacga agacaucgug gcggugacga ggcacagccc gagcauccac 2220
caaagcgugg uggcggugag caggacggcg uucaggaacc cgaaaacgag cuucuacagc 2280
aaagaagugc cgcaaaugug caucccgggg aaaaucgaag aaguggugcu cgaagcgagg 2340
acgaucgaaa ggaacacgaa accguacagg aaagacgaaa acagcaucaa cgggacgccg 2400
gacaucacgg uggaaaucag ggaacacauc caacucaacg aaagcaaaau cgugaaacaa 2460
gcgggggugg cgacgaaagg gccgaacgaa uacauccaag aaaucgaauu cgaaaaccuc 2520
agcccgggga gcgugaucau cuucagggug agccucgacc cgcacgcgca aguggcggug 2580
gggauccuca ggaaccaccu cacgcaauuc agcccgcacu ucaaaagcgg gagccucgcg 2640
guggacaacg cggacccgau ccucaaaauc ccguucgcga gccucgcgag caggcucacg 2700
cucgcggaac ucaaccaaau ccucuacagg ugcgaaagcg aagaaaaaga agacgggggg 2760
gggugcuacg acaucccgaa cuggagcgcg cucaaauacg cggggcucca agggcucaug 2820
agcgugcucg cggaaaucag gccgaaaaac gaccucgggc acccguucug caacaaccuc 2880
aggagcgggg acuggaugau cgacuacgug agcaacaggc ucaucagcag gagcgggacg 2940
aucgcggaag uggggaaaug gcuccaagcg auguucuucu accucaaaca aaucccgagg 3000
uaccucaucc cgugcuacuu cgacgcgauc cucaucgggg cguacacgac gcuccucgac 3060
acggcgugga aacaaaugag cagcuucgug caaaacggga gcacguucgu gaaacaccuc 3120
agccucggga gcgugcaacu cugcggggug gggaaauucc cgagccuccc gauccucagc 3180
ccggcgcuca uggacgugcc guacaggcuc aacgaaauca cgaaagaaaa agaacaaugc 3240
ugcgugagcc ucgcggcggg gcucccgcac uucagcagcg ggaucuucag gugcuggggg 3300
agggacacgu ucaucgcgcu cagggggauc cuccucauca cggggaggua cguggaagcg 3360
aggaacauca uccucgcguu cgcggggacg cucaggcacg ggcucauccc gaaccuccuc 3420
ggggaaggga ucuacgcgag guacaacugc agggacgcgg ugugguggug gcuccaaugc 3480
auccaagacu acugcaaaau ggugccgaac gggcucgaca uccucaaaug cccggugagc 3540
aggauguacc cgacggacga cagcgcgccg cucccggcgg ggacgcucga ccaaccgcug 3600
uucgaaguga uccaagaagc gaugcaaaaa cacaugcaag ggauccaauu cagggaaagg 3660
aacgcggggc cgcaaaucga caggaacaug aaagacgaag gguucaacau cacggcgggg 3720
guggacgaag aaacgggguu cguguacggg gggaacaggu ucaacugcgg gacguggaug 3780
gacaaaaugg gggaaagcga cagggcgagg aacaggggga ucccggcgac gccgagggac 3840
gggagcgcgg uggaaaucgu ggggcucagc aaaagcgcgg ugagguggcu ccucgaacuc 3900
agcaaaaaaa acaucuuccc guaccacgaa gugacgguga aaaggcacgg gaaagcgauc 3960
aaagugagcu acgacgaaug gaacaggaaa auccaagaca acuucgaaaa acucuuccac 4020
gugagcgaag acccgagcga ccucaacgaa aaacacccga accucgugca caaaaggggg 4080
aucuacaaag acagcuacgg ggcgagcagc ccguggugcg acuaccaacu caggccgaac 4140
uucacgaucg cgaugguggu ggcgccggaa cucuucacga cggaaaaagc guggaaagcg 4200
cucgaaaucg cggaaaaaaa acuccucggg ccgcucggga ugaaaacgcu cgacccggac 4260
gacauggugu acugcgggau cuacgacaac gcgcucgaca acgacaacua caaccucgcg 4320
aaaggguuca acuaccacca agggccggaa uggcucuggc cgaucgggua cuuccucagg 4380
gcgaaacucu acuucagcag gcucaugggg ccggaaacga cggcgaaaac gaucgugcuc 4440
gugaaaaacg ugcucagcag gcacuacgug caccucgaaa ggagcccgug gaaagggcuc 4500
ccggaacuca cgaacgaaaa cgcgcaauac ugcccguuca gcugcgaaac gcaagcgugg 4560
agcaucgcga cgauccucga aacgcucuac gaccucuga 4599
<210> 44
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 44
augggccaca gcaagcagau caggauccuc cuccucaacg aaauggaaaa gcucgaaaag 60
acacucuuca ggcucgaaca gggcuacgaa cuccaguuca ggcucggccc aacacuccag 120
ggcaaggccg ugacagugua cacaaacuac ccauucccag gcgaaacauu caacagggaa 180
aaguucagga gccucgacug ggaaaaccca acagaaaggg aagacgacag cgacaaguac 240
ugcaagcuca accuccagca gagcggcagc uuccaguacu acuuccucca gggcaacgaa 300
aagagcggcg gcggcuacau cgugguggac ccaauccuca gggugggcgc cgacaaccac 360
gugcucccac ucgacugcgu gacacuccag acauuccucg ccaagugccu cggcccauuc 420
gacgaauggg aaagcaggcu caggguggcc aaggaaagcg gcuacaacau gauccacuuc 480
acaccacucc agacacucgg ccucagcagg agcugcuaca gccucgccaa ccagcucgaa 540
cucaacccag acuucagcag gccaaacagg aaguacacau ggaacgacgu gggccagcuc 600
guggaaaagc ucaagaagga auggaacgug aucugcauca cagacguggu guacaaccac 660
acagccgcca acagcaagug gauccaggaa cacccagaau gcgccuacaa ccucgugaac 720
agcccacacc ucaagccagc cugggugcuc gacagggccc ucuggagguu cagcugcgac 780
guggccgaag gcaaguacaa ggaaaagggc aucccagccc ucaucgaaaa cgaccaccac 840
augaacagca ucaggaagau caucugggaa gacaucuucc caaagcucaa gcucugggaa 900
uucuuccagg uggacgugaa caaggccgug gaacaguuca ggaggcuccu cacacaggaa 960
aacaggaggg ugacaaagag cgacccaaac cagcaccuca caaucaucca ggacccagaa 1020
uacaggaggu ucggcugcac aguggacaug aacaucgccc ucacaacauu caucccacac 1080
gacaagggcc cagccgccau cgaagaaugc ugcaacuggu uccacaagag gauggaagaa 1140
cucaacagcg aaaagcacag gcucaucaac uaccaccagg aacaggccgu gaacugccuc 1200
cucggcaacg uguucuacga aaggcucgcc ggccacggcc caaagcucgg cccagugaca 1260
aggaagcacc cacucgugac aagguacuuc acauucccau ucgaagaaau cgacuucagc 1320
auggaagaaa gcaugaucca ccucccaaac aaggccugcu uccucauggc ccacaacggc 1380
ugggugaugg gcgacgaccc acucaggaac uucgccgaac caggcagcga aguguaccuc 1440
aggagggaac ucaucugcug gggcgacagc gugaagcuca gguacggcaa caagccagaa 1500
gacugcccau accucugggc ccacaugaag aaguacacag aaaucacagc cacauacuuc 1560
cagggcguga ggcucgacaa cugccacagc acaccacucc acguggccga auacaugcuc 1620
gacgccgcca ggaaccucca gccaaaccuc uacguggugg ccgaacucuu cacaggcagc 1680
gaagaccucg acaacguguu cgugacaagg cucggcauca gcagccucau cagggaagcc 1740
augagcgccu acaacagcca cgaagaaggc aggcucgugu acagguacgg cggcgaacca 1800
gugggcagcu ucgugcagcc augccucagg ccacucaugc cagccaucgc ccacgcccuc 1860
uucauggaca ucacacacga caacgaaugc ccaaucgugc acaggagcgc cuacgacgcc 1920
cucccaagca caacaaucgu gagcauggcc ugcugcgcca gcggcagcac aaggggcuac 1980
gacgaacucg ugccacacca gaucagcgug gugagcgaag aaagguucua cacaaagugg 2040
aacccagaag cccucccaag caacacaggc gaagugaacu uccagagcgg caucaucgcc 2100
gccaggugcg ccaucagcaa gcuccaccag gaacucggcg ccaagggcuu cauccaggug 2160
uacguggacc agguggacga agacaucgug gccgugacaa ggcacagccc aagcauccac 2220
cagagcgugg uggccgugag caggacagcc uucaggaacc caaagacaag cuucuacagc 2280
aaggaagugc cacagaugug caucccaggc aagaucgaag aaguggugcu cgaagccagg 2340
acaaucgaaa ggaacacaaa gccauacagg aaggacgaaa acagcaucaa cggcacacca 2400
gacaucacag uggaaaucag ggaacacauc cagcucaacg aaagcaagau cgugaagcag 2460
gccggcgugg ccacaaaggg cccaaacgaa uacauccagg aaaucgaauu cgaaaaccuc 2520
agcccaggca gcgugaucau cuucagggug agccucgacc cacacgccca gguggccgug 2580
ggcauccuca ggaaccaccu cacacaguuc agcccacacu ucaagagcgg cagccucgcc 2640
guggacaacg ccgacccaau ccucaagauc ccauucgcca gccucgccag caggcucaca 2700
cucgccgaac ucaaccagau ccucuacagg ugcgaaagcg aagaaaagga agacggcggc 2760
ggcugcuacg acaucccaaa cuggagcgcc cucaaguacg ccggccucca gggccucaug 2820
agcgugcucg ccgaaaucag gccaaagaac gaccucggcc acccauucug caacaaccuc 2880
aggagcggcg acuggaugau cgacuacgug agcaacaggc ucaucagcag gagcggcaca 2940
aucgccgaag ugggcaagug gcuccaggcc auguucuucu accucaagca gaucccaagg 3000
uaccucaucc caugcuacuu cgacgccauc cucaucggcg ccuacacaac acuccucgac 3060
acagccugga agcagaugag cagcuucgug cagaacggca gcacauucgu gaagcaccuc 3120
agccucggca gcgugcagcu cugcggcgug ggcaaguucc caagccuccc aauccucagc 3180
ccagcccuca uggacgugcc auacaggcuc aacgaaauca caaaggaaaa ggaacagugc 3240
ugcgugagcc ucgccgccgg ccucccacac uucagcagcg gcaucuucag gugcuggggc 3300
agggacacau ucaucgcccu caggggcauc cuccucauca caggcaggua cguggaagcc 3360
aggaacauca uccucgccuu cgccggcaca cucaggcacg gccucauccc aaaccuccuc 3420
ggcgaaggca ucuacgccag guacaacugc agggacgccg ugugguggug gcuccagugc 3480
auccaggacu acugcaagau ggugccaaac ggccucgaca uccucaagug cccagugagc 3540
aggauguacc caacagacga cagcgcccca cucccagccg gcacacucga ccagccacuc 3600
uucgaaguga uccaggaagc caugcagaag cacaugcagg gcauccaguu cagggaaagg 3660
aacgccggcc cacagaucga caggaacaug aaggacgaag gcuucaacau cacagccggc 3720
guggacgaag aaacaggcuu cguguacggc ggcaacaggu ucaacugcgg cacauggaug 3780
gacaagaugg gcgaaagcga cagggccagg aacaggggca ucccagccac accaagggac 3840
ggcagcgccg uggaaaucgu gggccucagc aagagcgccg ugagguggcu ccucgaacuc 3900
agcaagaaga acaucuuccc auaccacgaa gugacaguga agaggcacgg caaggccauc 3960
aaggugagcu acgacgaaug gaacaggaag auccaggaca acuucgaaaa gcuguuccac 4020
gugagcgaag acccaagcga ccucaacgaa aagcacccaa accucgugca caagaggggc 4080
aucuacaagg acagcuacgg cgccagcagc ccauggugcg acuaccagcu caggccaaac 4140
uucacaaucg ccaugguggu ggccccagaa cucuucacaa cagaaaaggc cuggaaggcc 4200
cucgaaaucg ccgaaaagaa gcuccucggc ccacucggca ugaagacacu cgacccagac 4260
gacauggugu acugcggcau cuacgacaac gcccucgaca acgacaacua caaccucgcc 4320
aagggcuuca acuaccacca gggcccagaa uggcucuggc caaucggcua cuuccucagg 4380
gccaagcucu acuucagcag gcucaugggc ccagaaacaa cagccaagac aaucgugcuc 4440
gugaagaacg ugcucagcag gcacuacgug caccucgaaa ggagcccaug gaagggccuc 4500
ccagaacuca caaacgaaaa cgcccaguac ugcccauuca gcugcgaaac acaggccugg 4560
agcaucgcca caauccucga aacacucuac gaccucuga 4599
<210> 45
<211> 4599
<212> RNA
<213> PatentIn版本3.5
<220>
<223> 密码子优化的AGL编码序列
<400> 45
augggccaca gcaagcagau ccggaucuug cugcugaacg agauggagaa gcuggagaag 60
acccuguuca ggcuggagca gggcuacgag cugcaguucc gguugggccc caccuugcag 120
ggcaaggccg ugaccgugua caccaacuac cccuuccccg gcgagacguu caacagggag 180
aaguuccggu cccuggacug ggagaacccc accgagaggg aggacgacuc cgacaaguac 240
ugcaagcuga accugcagca guccggcucc uuccaguacu acuuccugca gggcaacgag 300
aagaguggcg gcggcuacau cgugguggac cccaucuugc gggugggcgc cgacaaccac 360
gugcugcccu uggacugcgu gacccugcag accuucuugg ccaagugcuu gggccccuuc 420
gacgaguggg agagcaggcu gaggguggcc aaggaguccg gcuacaacau gauccacuuc 480
acccccuugc agacccuggg ccuguccagg uccugcuacu cccuggccaa ccaguuggag 540
uugaaccccg acuucuccag gcccaacagg aaguacaccu ggaacgacgu gggccagcug 600
guggagaagu ugaagaagga guggaacgug aucugcauca ccgacguggu guacaaccac 660
accgccgcca acagcaagug gauccaggag caccccgagu gcgccuacaa ccuggugaac 720
uccccccacu ugaagcccgc cuggguguug gacagggccc uguggcgguu cuccugcgac 780
guggccgagg gcaaguacaa ggagaagggc auccccgccu ugaucgagaa cgaccaccac 840
augaacucca uccggaagau caucugggag gacaucuucc ccaagcugaa gcugugggag 900
uucuuccagg uggacgugaa caaggccgug gagcaguuca ggaggcugcu gacccaggag 960
aacaggcggg ugaccaaguc cgaccccaac cagcaccuga ccaucaucca ggaccccgag 1020
uacaggcggu ucggcugcac cguggacaug aacaucgccc ugaccaccuu caucccccac 1080
gacaagggcc ccgccgccau cgaggagugc ugcaacuggu uccacaagag gauggaggag 1140
uugaacuccg agaagcaccg gcugaucaac uaccaccagg agcaggccgu gaacugccug 1200
uugggcaacg uguucuacga gcggcuggcc ggccacggcc ccaagcuggg ccccgugacc 1260
aggaagcacc ccuuggugac cagguacuuc accuuccccu ucgaggagau cgacuucucc 1320
auggaggagu ccaugaucca ccugcccaac aaggccugcu uccugauggc ccacaacggc 1380
ugggugaugg gcgacgaccc ccugcggaac uucgccgagc ccggcuccga gguguaccug 1440
aggagggagc ugaucugcug gggcgacagc gugaaguugc gguacggcaa caagcccgag 1500
gacugccccu accugugggc ccacaugaag aaguacaccg agaucaccgc caccuacuuc 1560
cagggcgugc ggcuggacaa cugccacucc accccccugc acguggccga guacauguug 1620
gacgccgcca ggaacuugca gcccaacuug uacguggugg ccgagcuguu caccggcagc 1680
gaggaccugg acaacguguu cgugaccagg cugggcauca gcuccuugau cagggaggcc 1740
augagcgccu acaacagcca cgaggagggc agguuggugu accgguacgg cggcgagccc 1800
gugggcuccu ucgugcagcc cugcuugagg cccuugaugc ccgccaucgc ccacgcccug 1860
uucauggaca ucacccacga caacgagugc cccaucgugc acagguccgc cuacgacgcc 1920
cugcccagca ccaccaucgu guccauggcc ugcugcgcca gcggcagcac caggggcuac 1980
gacgaguugg ugccccacca gaucuccgug guguccgagg agcgguucua caccaagugg 2040
aaccccgagg ccuugcccuc caacaccggc gaggugaacu uccagagcgg caucaucgcc 2100
gccaggugcg ccaucagcaa gcugcaccag gagcugggcg ccaagggcuu cauccaggug 2160
uacguggacc agguggacga ggacaucgug gccgugacca ggcacucccc cagcauccac 2220
caguccgugg uggccguguc caggaccgcc uucaggaacc ccaagaccuc cuucuacagc 2280
aaggaggugc cccagaugug cauccccggc aagaucgagg agguggugcu ggaggccagg 2340
accaucgaga ggaacaccaa gcccuacagg aaggacgaga acuccaucaa cggcaccccc 2400
gacaucaccg uggagaucag ggagcacauc cagcugaacg agagcaagau cgugaagcag 2460
gccggcgugg ccaccaaggg ccccaacgag uacauccagg agaucgaguu cgagaacuug 2520
ucccccggca gcgugaucau cuucagggug agccuggacc cccacgccca gguggccgug 2580
ggcauccugc ggaaccaccu gacccaguuc agcccccacu ucaaguccgg cagccuggcc 2640
guggacaacg ccgaccccau cuugaagauc cccuucgccu cccuggccuc cagguugacc 2700
uuggccgagc ugaaccagau ccuguaccgg ugcgaguccg aggagaagga ggacggcggc 2760
ggcugcuacg acauccccaa cugguccgcc cugaaguacg ccggccugca gggcuugaug 2820
uccguguugg ccgagaucag gcccaagaac gacuugggcc accccuucug caacaacuug 2880
agguccggcg acuggaugau cgacuacgug agcaaccggc ugaucucccg guccggcacc 2940
aucgccgagg ugggcaagug guugcaggcc auguucuucu accugaagca gaucccccgg 3000
uaccugaucc ccugcuacuu cgacgccauc uugaucggcg ccuacaccac ccugcuggac 3060
accgccugga agcagauguc cagcuucgug cagaacggcu ccaccuucgu gaagcaccug 3120
uccuugggcu ccgugcagcu gugcggcgug ggcaaguucc ccucccugcc cauccugucc 3180
cccgcccuga uggacgugcc cuacagguug aacgagauca ccaaggagaa ggagcagugc 3240
ugcguguccc uggccgccgg cuugccccac uucuccuccg gcaucuuccg gugcuggggc 3300
agggacaccu ucaucgcccu gaggggcauc cugcugauca ccggccggua cguggaggcc 3360
aggaacauca ucuuggccuu cgccggcacc cugaggcacg gccugauccc caaccugcug 3420
ggcgagggca ucuacgccag guacaacugc cgggacgccg ugugguggug gcugcagugc 3480
auccaggacu acugcaagau ggugcccaac ggccuggaca uccugaagug ccccgugucc 3540
aggauguacc ccaccgacga cuccgccccc uugcccgccg gcacccugga ccagcccuug 3600
uucgagguga uccaggaggc caugcagaag cacaugcagg gcauccaguu ccgggagagg 3660
aacgccggcc cccagaucga ccggaacaug aaggacgagg gcuucaacau caccgccggc 3720
guggacgagg agacuggcuu cguguacggc ggcaaccggu ucaacugcgg caccuggaug 3780
gacaagaugg gcgagagcga cagggccagg aacaggggca uccccgccac ccccagggac 3840
ggcuccgccg uggagaucgu gggccugagc aaguccgccg ugcggugguu gcuggaguug 3900
uccaagaaga acaucuuccc cuaccacgag gugaccguga agaggcacgg caaggccauc 3960
aagguguccu acgacgagug gaacaggaag auccaggaca acuucgagaa gcuguuccac 4020
guguccgagg accccuccga cuugaacgag aagcacccca accuggugca caagcggggc 4080
aucuacaagg acagcuacgg cgccuccagc cccuggugcg acuaccagcu gaggcccaac 4140
uucaccaucg ccaugguggu ggcccccgag cuguucacca ccgagaaggc cuggaaggcc 4200
uuggagaucg ccgagaagaa guugcugggc ccccugggca ugaagaccuu ggaccccgac 4260
gacauggugu acugcggcau cuacgacaac gccuuggaca acgacaacua caaccuggcc 4320
aagggcuuca acuaccacca gggccccgag uggcuguggc ccaucggcua cuuccugcgg 4380
gccaaguugu acuucuccag guugaugggc cccgagacga ccgccaagac caucguguug 4440
gugaagaacg ugcugucccg gcacuacgug caccuggaga ggucccccug gaagggccug 4500
cccgagcuga ccaacgagaa cgcccaguac ugccccuuca gcugcgagac gcaggccugg 4560
uccaucgcca ccauccugga gacgcuguac gacuuguag 4599

Claims (41)

1.一种用于表达SEQ ID NO:2的人淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(amylo-alpha-1,6-glucosidase,4-alpha-glucanotransferase)(AGL)或其片段的多核苷酸，其中所述多核苷酸包括天然核苷酸和经过化学修饰的核苷酸并且能够表达为提供所述人AGL或其具有AGL活性的片段，其中所述多核苷酸包含与选自SEQ ID NO:7-32或SEQ IDNO:41-45的核碱基序列至少99％一致性的核碱基序列。
2.根据权利要求1所述的多核苷酸，其中与人AGL野生型mRNA相比，所述多核苷酸是密码子优化的。
3.根据权利要求1所述的多核苷酸，其中所述经过化学修饰的核苷酸选自5-羟基胞苷、5-甲基胞苷、5-羟甲基胞苷、5-羧基胞苷、5-甲酰基胞苷、5-甲氧基胞苷、5-丙炔基胞苷、2-硫代胞苷；
5-羟基尿苷、5-甲基尿苷、5,6-二氢-5-甲基尿苷、2'-O-甲基尿苷、2'-O-甲基-5-甲基尿苷、2'-氟代-2'-脱氧尿苷、2'-氨基-2'-脱氧尿苷、2'-叠氮基-2'-脱氧尿苷、4-硫代尿苷、5-羟甲基尿苷、5-羧基尿苷、5-羧甲基酯尿苷、5-甲酰基尿苷、5-甲氧基尿苷、5-丙炔基尿苷、5-溴代尿苷、5-碘代尿苷、5-氟代尿苷；
假尿苷、2'-O-甲基-假尿苷、N1-羟基假尿苷、N1-甲基假尿苷、2'-O-甲基-N1-甲基假尿苷、N1-乙基假尿苷、N1-羟甲基假尿苷和阿糖尿苷(Arauridine)；
N6-甲基腺苷、2-氨基腺苷、3-甲基腺苷、7-脱氮腺苷、8-氧代腺苷、肌苷；
噻吩并鸟苷、7-脱氮鸟苷、8-氧代鸟苷和6-O-甲基鸟嘌呤。
4.根据权利要求1所述的多核苷酸，其中所述经过化学修饰的核苷酸是N1-甲基假尿苷。
5.根据权利要求1所述的多核苷酸，其中所述经过化学修饰的核苷酸是5-甲氧基尿苷。
6.根据权利要求1所述的多核苷酸，其中所述经过化学修饰的核苷酸是假尿苷和N1-甲基假尿苷的组合。
7.根据权利要求1所述的多核苷酸，其中所述经过化学修饰的核苷酸是5-甲基胞苷和N1-甲基假尿苷的组合。
8.根据权利要求1所述的多核苷酸，其中所述经过化学修饰的核苷酸是5-甲氧基尿苷和N1-甲基假尿苷的组合。
9.根据权利要求1所述的多核苷酸，其中所述经过化学修饰的核苷酸是5-甲氧基尿苷、5-甲基胞苷和N1-甲基假尿苷的组合。
10.根据权利要求1-9中任一项所述的多核苷酸，其中与人AGL野生型mRNA相比，所述多核苷酸的翻译效率增加了至少50％。
11.根据权利要求1-9中任一项所述的多核苷酸，其中与人AGL野生型mRNA相比，所述多核苷酸的所述翻译效率增加了至少三倍。
12.根据权利要求1-9中任一项所述的多核苷酸，其中所述多核苷酸包括200个到12,000个核苷酸。
13.根据权利要求1-9中任一项所述的多核苷酸，其中经过化学修饰的核苷酸占所述核苷酸的1-99％。
14.根据权利要求1-9中任一项所述的多核苷酸，其中经过化学修饰的核苷酸占所述核苷酸的50-99％。
15.根据权利要求1所述的多核苷酸，其中所述多核苷酸包括5'端帽、5'非翻译区、编码区、3'非翻译区和尾区。
16.根据权利要求1所述的多核苷酸，其中所述多核苷酸包括翻译增强子。
17.根据权利要求1所述的多核苷酸，其中所述多核苷酸在哺乳动物细胞中能够翻译以表达所述人AGL或其具有AGL活性的片段。
18.根据权利要求1所述的多核苷酸，其中所述多核苷酸在受试者中能够在体内翻译以表达所述人AGL或其具有AGL活性的片段。
19.根据权利要求1所述的多核苷酸，其中所述多核苷酸的翻译产物是活性人AGL或其具有AGL活性的片段。
20.根据权利要求1所述的多核苷酸，其中与人AGL野生型mRNA相比，所述多核苷酸具有减少的免疫原性。
21.根据权利要求1所述的多核苷酸，其中所述多核苷酸包括选自SEQ ID NO:7-32或SEQ ID NO:41-45的核碱基序列。
22.根据权利要求1所述的多核苷酸，其中所述核碱基序列在全长人AGL编码序列SEQID NO:1上与野生型人AGL编码序列的一致性小于80％。
23.根据权利要求1所述的多核苷酸，其中所述多核苷酸包含一个或多个UNA单体。
24.一种组合物，其包括一种或多种根据权利要求1到23中任一项所述的多核苷酸以及药学上可接受的载剂。
25.根据权利要求24所述的组合物，其中所述载剂包括转染试剂、纳米颗粒或脂质体。
26.根据权利要求24或25所述的组合物，其用于医学疗法。
27.根据权利要求24或25所述的组合物，其用于治疗人类或动物身体。
28.一种根据权利要求24或25所述的组合物的用途，其用于制备或制造用于改善、预防、延缓发作或治疗与有需要的受试者的淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)活性降低相关联的疾病或病症的药物。
29.根据权利要求28所述的用途，其中，所述疾病是III型糖原贮积病。
30.根据权利要求29所述的用途，其中，所述III型糖原贮积病选自IIIa型糖原贮积病、IIIb型糖原贮积病、IIIc型糖原贮积病和IIId型糖原贮积病。
31.根据权利要求24-27中任一项所述的组合物在制备用于改善、预防、延缓发作或治疗与有需要的受试者的淀粉-α-1,6-葡萄糖苷酶,4-α-葡聚糖转移酶(AGL)活性降低相关联的疾病或病症的药物中的用途。
32.根据权利要求31所述的用途，其中所述疾病是III型糖原贮积病。
33.根据权利要求32所述的用途，其中所述III型糖原贮积病选自IIIa型糖原贮积病、IIIb型糖原贮积病、IIIc型糖原贮积病和IIId型糖原贮积病。
34.根据权利要求31-33中任一项所述的用途，其中所述施用是静脉内、皮下、肺部、肌内、腹膜内、真皮、口服、鼻内或吸入。
35.根据权利要求31-33中任一项所述的用途，其中所述施用是每天、每周、每两周或每月一次。
36.根据权利要求31-33中任一项所述的用途，其中所述施用包括0.01到10mg/kg的有效剂量。
37.根据权利要求31-33中任一项所述的用途，其中所述施用增加了AGL在所述受试者的肝脏、血清、血浆、肾脏、心脏、肌肉、大脑、脑脊液或淋巴结中的表达。
38.根据权利要求31所述的用途，其中在施用后，所述受试者的所述肝脏中的AGL水平为10到1500ng/mg肝脏总蛋白。
39.根据权利要求31所述的用途，其中在施用后，所述受试者的所述肝脏中的AGL水平为20到150ng/mg肝脏总蛋白。
40.一种用于在体内表达人AGL的试剂盒，所述试剂盒包括剂量为0.1到500mg的一种或多种根据权利要求1到23中任一项所述的多核苷酸以及用于施用所述剂量的装置。
41.根据权利要求40所述的试剂盒，其中所述装置是注射针、静脉针或吸入装置。
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