CN118159284A - 包含肉毒杆菌衍生肽的用于止痛的组合物 - Google Patents

Abstract

本发明涉及包括肉毒杆菌衍生肽的止痛组合物。包括根据本发明的肉毒杆菌毒素重组蛋白的组合物可以通过将具有肌肉麻痹效果的肉毒杆菌毒素轻链快速递送到疼痛区域而有效地缓解肌肉紧张和减轻疼痛，因此它可以有效地用作预防、改善或治疗疼痛的物质。

Description

包含肉毒杆菌衍生肽的用于止痛的组合物

技术领域

本发明涉及包括肉毒杆菌衍生肽的止痛组合物。

本发明要求基于2021年10月25日提交的韩国专利申请号10-2021-0143024和2022年5月25日提交的韩国专利申请号10-2022-0064311的优先权，并且上述申请的说明书和附图中公开的所有内容都并入本申请。

背景技术

众所周知，疼痛在保护身体免受危险和恢复受损组织方面发挥着重要作用。然而，由病毒或细菌感染、外周神经周围的严重炎症反应或外周神经的直接损伤引起的病理性疼痛对人体有害，并对患者的生活质量产生重大影响。

纵观人类历史，镇痛药早已被开发出来并用于缓解或麻木疼痛，这些镇痛药通常口服给药。然而，口服给药往往会导致肝损伤和肾损伤等问题，而且由于它在进入血液之前必须通过消化系统吸收，因此需要时间才能发挥功效。

为了解决口服给药的这些问题，已经提出了胃肠外给药方法，如血管内注射或肌肉内注射，但这些方法不方便，成本高，并且很难在医院外进行。

因此，正在尝试通过局部给药，特别是透皮给药来给药。

同时，肉毒杆菌毒素是由肉毒梭状芽孢杆菌(Clostridium botulinum)产生的一种神经毒素蛋白，据报道可抑制神经元的神经递质乙酰胆碱和儿茶酚胺的分泌。肉毒杆菌毒素有八种类型：A、B、C、D、E、F、G和H型，A和B型是商用的。

肉毒杆菌毒素是一种神经毒素蛋白，对人类的中位致死剂量在静脉注射或肌肉注射时为1.3ng/kg或更高而在吸入时为10ng/kg或更高，表明它是一种剧毒物质。然而，当调整到适当的量时，它可以用作治疗神经系统疾病、肌肉疾病、多汗症和方颌等的药物，并且它最常用于美容目的，如减少皱纹和小腿肌肉。

由于肉毒杆菌毒素是一种非常大的分子，分子量为150kDa，具有组合的轻链和重链，难以穿透皮肤，因此只能通过注射使用。毒素持续3到6个月，因此毒素需要定期治疗。因此，许多研究正在进行，以寻找另一种能够为用户提供便利的有效递送方式，但成果仍然不足。

发明内容

[技术问题]

作为确认根据本发明的肉毒杆菌毒素重组蛋白(皮肤穿透性肉毒杆菌衍生成分肽)的止痛效果的结果，本发明人确认其对于头痛和慢性疼痛等疼痛的患者表现出优异的疼痛减轻效果，从而完成了本发明。

因此，本发明的目的是提供一种用于止痛的药物组合物，其包含肉毒杆菌毒素重组蛋白作为活性成分，其中在肉毒杆菌毒素重组蛋白中，由SEQ ID NO:1的氨基酸序列组成的细胞穿透性肽与肉毒杆菌毒素轻链的一端或两端融合。

本发明的另一个目的是提供一种用于止痛的准药物组合物，其包含肉毒杆菌毒素重组蛋白作为活性成分。

本发明的又一个目的是提供一种用于止痛的皮肤外用组合物，其包含肉毒杆菌毒素重组蛋白作为活性成分。

本发明的又一个目的是提供一种用于止痛的化妆品组合物，其包含肉毒杆菌毒素重组蛋白作为活性成分。

然而，本发明要解决的技术问题不限于上述问题，本发明所属领域的技术人员可以通过以下描述清楚地理解未提及的其他问题。

[技术方案]

本发明的一个方面提供了一种用于止痛的药物组合物，其包含肉毒杆菌毒素重组蛋白作为活性成分，其中在肉毒杆菌毒素重组蛋白中，由SEQ ID NO:1的氨基酸序列组成的细胞穿透性肽与肉毒杆菌毒素轻链的一端或两端融合。

本发明的另一个方面提供了一种用于止痛的准药物组合物，其包含肉毒杆菌毒素重组蛋白作为活性成分。

本发明的又一方面提供了一种用于止痛的皮肤外用组合物，其包含肉毒杆菌毒素重组蛋白作为活性成分。

本发明的又一方面提供了一种用于止痛的化妆品组合物，其包含肉毒杆菌毒素重组蛋白作为活性成分。

在本发明的一个实施方案中，肉毒杆菌毒素重组蛋白可以由选自由SEQ ID NO:31至SEQ ID NO:58组成的群组的一个或多个氨基酸序列组成。

在本发明的另一个实施方案中，肉毒杆菌毒素轻链可以由选自由SEQ ID NO:3至SEQ ID NO:9组成的群组的一个或多个氨基酸序列组成，但不限于此。

在本发明的又一个实施方案中，肉毒杆菌毒素轻链可以在一端进一步包括六组氨酸标签，但不限于此。

在本发明的又一个实施方案中，肉毒杆菌毒素轻链可以选自由肉毒杆菌毒素血清型A、B、C、D、E、F和G组成的群组，但不限于此。

在本发明的又一个实施方案中，细胞穿透性肽可以与肉毒杆菌毒素轻链的羧基末端、氨基末端或两者融合，但不限于此。

在本发明的又一个实施方案中，融合可以通过肽键或共价键实现，但不限于此。

在本发明的又一个实施方案中，肉毒杆菌毒素重组蛋白可以缓解肌肉紧张，但不限于此。

在本发明的又一个实施方案中，疼痛可选自由以下组成的群组：机械性疼痛、热性疼痛、慢性疼痛、急性疼痛、过敏性疼痛、体源性疼痛、神经性疼痛、心理性疼痛、热量诱导的疼痛、躯体性疼痛、伤害性感觉、过敏、风湿性疼痛、头痛、背部疼痛、骨盆疼痛、肌筋膜疼痛、血管疼痛、偏头痛、创伤相关疼痛、炎性疼痛、关节炎疼痛、糖尿病性疼痛、癌症衍生性疼痛、内脏疼痛、纤维肌痛、术后疼痛、幻肢痛、三叉神经痛、带状疱疹后神经痛、疼痛性糖尿病神经病变、疼痛性糖尿病周围神经病变、糖尿病多发性神经病、坐骨神经痛、神经根病、脊髓神经根病、肌痛和腰部疼痛，但不限于此。

在本发明的又一个实施方案中，用于止痛的组合物可以用于透皮给药，但不限于此。

此外，本发明提供了一种用于缓解疼痛的方法，其包括将包含肉毒杆菌毒素重组蛋白作为活性成分的组合物给药至有需要的受试者。

此外，本发明提供了包含肉毒杆菌毒素重组蛋白的组合物作为活性成分用于止痛的用途。

此外，本发明提供肉毒杆菌毒素重组蛋白用于制备镇痛药物的用途。

[有利效果]

根据本发明的肉毒杆菌毒素重组蛋白可以通过与细胞穿透性肽融合而经皮递送具有肌肉麻痹效果的肉毒杆菌毒轻链，通过方便且快速地将肉毒杆菌毒素轻链递送到疼痛区域而有效地缓解肌肉紧张和减少疼痛，因此它可以有效地用作预防、改善或治疗疼痛的物质。

附图说明

图1是显示在使用包括肉毒杆菌毒素重组蛋白的组合物之后按受试者/时段比较头痛次数的结果的图。

图2是显示在使用包括肉毒杆菌毒素重组蛋白的组合物之后按时段比较头痛次数减少率的结果的图。

图3是显示在使用包括肉毒杆菌毒素重组蛋白的组合物之后按时段比较服用镇痛药的频率的结果的图。

图4是显示为了确认肉毒杆菌毒素重组蛋白的止痛效果而对慢性疼痛患者施用肉毒杆菌乳液后6天的疼痛变化进行确认的结果的图(NRS：

数值评定量表；1至4分：轻度疼痛，5至6分：中度疼痛，并且7至10分：重度疼痛；下同)。

图5是显示为了确认肉毒杆菌毒素重组蛋白的止痛效果而向纤维肌痛和肌痛患者施用肉毒杆菌乳液之后确认持续疼痛减轻6天的结果的图。

图6是显示为了确认肉毒杆菌毒素重组蛋白的止痛效果而使用麻醉性镇痛药向患者施用肉毒杆菌乳液之后确认持续疼痛减轻6天的结果的图。

图7是示意性地显示肉毒杆菌毒素重组蛋白的皮肤渗透过程的图。

具体实施方式

作为确认通过将细胞渗透性肽与肉毒杆菌毒素轻链融合而制备的肉毒杆菌毒素重组蛋白(皮肤穿透性肉毒杆菌衍生成分肽)的止痛效果的结果，本发明人确认其对头痛和慢性疼痛等疼痛患者表现出优异的疼痛减轻效果，从而完成了本发明。

在下文中，将详细描述本发明。

肉毒杆菌毒素作为单一多肽表达，但在表达后，通过重组过程，它被分为约100kDa的重链(H链)和约50kDa的轻链(L链)，并且H链和L链通过二硫键连接。H链与神经元上的受体结合，使肉毒杆菌毒素通过内吞作用进入内部。肉毒杆菌毒素的L链进入细胞后，离开内体，进入细胞质，并在细胞质中切割可溶性N-乙基马来酰亚胺敏感因子附着蛋白受体(SNARE)蛋白，抑制乙酰胆碱分泌，从而表现出肌肉麻痹作用。因此，仅用L链就可抑制神经元的乙酰胆碱分泌，并且H链和L链可以独立发挥作用。

然而，分子量为50kDa的分离的肉毒杆菌毒素轻链不能穿透细胞膜，因此不能独立发挥作用。通常，为了使肉毒杆菌毒素轻链被递送到神经元的细胞质以表现出肉毒杆菌毒素特异性活性，约100kDa的肉毒杆菌毒素重链的帮助是必不可少的。

因此，在本发明的一个实施方案中，为了有效地将肉毒杆菌毒素轻链递送到皮肤和神经元内部，将包括通过融合肉毒杆菌毒素轻链与细胞渗透性肽(大分子转导结构域；MTD)制备的肉毒杆菌毒素重组蛋白作为活性成分的组合物施用于头痛、慢性疼痛、肌痛、纤维肌痛或肌筋膜疼痛患者的疼痛区域，结果，证实显示出优异的疼痛减轻效果。

因此，本发明提供了一种用于止痛的药物组合物，其包含肉毒杆菌毒素重组蛋白作为活性成分，其通过将细胞穿透性肽融合到肉毒杆菌毒素轻链来赋予细胞穿透特性，使得不容易引入细胞的肉毒杆菌毒素轻链能够高效地递送到细胞中，以及

在肉毒杆菌毒素重组蛋白中，将由SEQ ID NO:1的氨基酸序列组成的细胞穿透性肽与肉毒杆菌毒素轻链的一端或两端融合。

在本发明中，“肉毒杆菌毒素重组蛋白”包括细胞穿透性肽和肉毒杆菌毒素轻链，是指由肽键或共价键等化学键形成的复合物。具体地，根据本发明的肉毒杆菌毒素重组蛋白能够通过将细胞穿透性肽融合到肉毒杆菌毒素轻链(其是不容易引入细胞的大分子)来赋予细胞穿透特性，从而高效地将肉毒杆菌毒素轻链递送到细胞中，并且此时，细胞穿透性肽可以融合到肉毒素轻链的羧基末端、氨基末端或两者。

根据本发明的肉毒杆菌毒素重组蛋白可以通过肉毒杆菌毒素轻链和细胞穿透性肽的融合高效地递送到细胞中，并且肉毒杆菌毒素轻链的活性和稳定性得到改善，以使肉毒杆菌毒素在体内的固有功效最大化。

本发明中肉毒杆菌毒素重组蛋白(皮肤穿透性肉毒杆菌衍生成分肽)的组成与韩国专利号10-1882461相同，并且上述文献中公开的所有内容通过引用结合到本申请中。

在本发明中，肉毒杆菌毒素重组蛋白可以通过将具有肌肉麻痹效果的肉毒杆菌毒素轻链高效地递送到细胞中来缓解肌肉紧张并促进肌肉恢复，从而减轻疼痛。

在本发明中，肉毒杆菌毒素重组蛋白可以穿透皮肤并进入SNARE复合物的组分SNAP-25，并且肉毒杆菌毒素轻链可以切割SNAP-25以阻断乙酰胆碱的分泌，从而改善疼痛。

在本发明中，“肉毒杆菌毒素”是指任何已知类型的肉毒杆菌毒素，无论随后是否发现，包括由细菌产生或通过重组技术改造的变体或融合蛋白。

在本发明中，肉毒杆菌毒素轻链可选自由肉毒杆菌毒素血清型A、B、C、D、E、F和G组成的群组，此时，肉毒杆菌毒素轻链可以由选自由SEQ ID NO:3至SEQ ID NO:9组成的群组的一个或多个氨基酸序列组成。此外，肉毒杆菌毒素轻链可以由选自由SEQ ID NO:3至SEQID NO:9组成的群组的一个氨基酸序列组成。此时，肉毒杆菌毒素轻链可以由多核苷酸编码，该多核苷酸由选自由SEQ ID NO:10至16组成的群组的碱基序列组成，但不限于此。

此外，肉毒杆菌毒素轻链还可以在一端包括六组氨酸标签。在本发明中，在肉毒杆菌毒素轻链的一端进一步包括六组氨酸标签的形式可以由选自由SEQ ID NO:17至SEQ IDNO:23组成的群组的氨基酸序列组成，并且它可以由选自由SEQ ID NO:24至SEQ ID NO:30组成的群组的碱基序列编码，但不限于此。

在本发明中，替代地，肉毒杆菌毒素轻链可以是肉毒杆菌毒素衍生物，即具有肉毒杆菌毒素活性但任选在部分或序列中具有一个或多个修饰的化合物。例如，与肉毒杆菌毒素轻链蛋白的七种血清型相比，它可以是以一种方式修饰的形式，该方法维持轻链内肽酶活性，同时通过对氨基酸序列进行缺失、修饰、替换和嵌合融合等方法来增强其特性或减少其副作用。替代地，可以使用通过重组或化学合成产生的肉毒杆菌毒素轻链或肉毒杆菌毒素轻链的一部分。

在本发明中，肉毒杆菌毒素重组蛋白可以由选自由SEQ ID NO:31至SEQ ID NO:58组成的群组的一个或多个氨基酸序列组成，并且编码该氨基酸序列的多核苷酸可以选自由SEQ ID NO:59至SEQ ID NO:86组成的群组，但不限于此。

此外，在本发明中，肉毒杆菌毒素重组蛋白可以由选自由SEQ ID NO:31至SEQ IDNO:58组成的群组的一个氨基酸序列组成。

根据本发明的一个实施方案，肉毒杆菌毒素重组蛋白可以优选由SEQ ID NO:45表示的氨基酸序列组成，并且编码该氨基酸序列的多核苷酸可以是由SEQ ID NO:73表示的核苷酸序列，但不限于此。

在本发明中，由SEQ ID NO:1的氨基酸序列组成的细胞穿透性肽(MTD)可以是可介导生物活性分子细胞内转运的肽，并且可以对人皮肤角质形成细胞和神经元都具有渗透性，但不限于此。

细胞穿透性肽优选不具有确定的酶活性或治疗性生物活性，但用作允许穿过细胞膜的细胞内运输的载体。它可以附着到要转移到细胞中的货物的N末端或C末端以及两个末端，并且它可以在每个末端沿正向或反向附着。此外，根据本发明的肽优选作为单体施用，但不限于此，并且它也可以以二聚体或聚合物的形式使用。此外，根据本发明的肽可以是包括SEQ ID NO:1的氨基酸序列作为最小单元的肽。

在本发明中，细胞穿透性肽可以由SEQ ID NO:2的碱基序列组成的多核苷酸编码，但不限于此。

在本说明书中，“活性成分”是指可单独或与本身无活性的载体组合显示所需活性的成分。

根据本发明的一个实施方案，根据本发明的组合物表现出止痛效果的最合适的给药途径是通过皮肤吸收，因此，根据本发明的组合物可以优选用于透皮给药，但不限于此。

在本发明中，根据本发明的组合物可以进一步包括透皮吸收促进剂，但不限于此。

“透皮吸收促进剂”是乳化剂中影响皮肤渗透并常用于透皮贴剂的一种组分。在本发明中，它增强肉毒杆菌毒素重组蛋白的皮肤渗透和细胞渗透，并且可以是卵磷脂、月桂基吡咯烷酮、甘油单油酸酯、甘油单月桂酸酯、丙二醇单月桂酸酯、聚氧乙烯山梨糖醇单油酸酯、聚氧乙烯山梨糖醇单硬脂酸酯、聚氧乙烯山梨糖醇单月桂酸酯、山梨糖醇单油酸酯、山梨糖醇单硬脂酸酯，或山梨糖醇单月桂酸酯，但不限于此。

在本发明中，“疼痛”是指由于强烈刺激或躯体损伤而感到不舒适情绪的一般概念。为了本发明的目的，疼痛可以意指躯体性疼痛。具体而言，疼痛可选自由以下组成的群组：机械性疼痛、热性疼痛、慢性疼痛、急性疼痛、过敏性疼痛、体源性疼痛、神经性疼痛、心理性疼痛、热量诱导的疼痛、躯体性疼痛、伤害性感觉、过敏、风湿性疼痛、头痛、背部疼痛、骨盆疼痛、肌筋膜疼痛、血管疼痛、偏头痛、创伤相关疼痛、炎性疼痛、关节炎疼痛、糖尿病性疼痛、癌症衍生性疼痛、内脏疼痛、纤维肌痛、术后疼痛、幻肢痛、三叉神经痛、带状疱疹后神经痛、疼痛性糖尿病神经病变、疼痛性糖尿病周围神经病变、糖尿病多发性神经病、坐骨神经痛、神经根病、脊髓神经根病、肌痛和腰部疼痛，但不限于此。根据本发明的一个实施方案，疼痛可以是头痛、慢性疼痛、肌痛、纤维肌痛或肌筋膜疼痛。

本发明中使用的术语“慢性疼痛”是指疼痛的持续时间和强度对患者的功能或生活造成有害影响，或者疼痛持续超过正常组织愈合期(通常为三个月)的情况。

本发明中使用的术语“纤维肌痛”是指肌腱、韧带、筋膜、肌肉和软组织(如脂肪组织)中的疼痛综合征，其长期导致全身肌肉骨骼疼痛、僵硬、感觉异常、睡眠障碍、疲劳，以及全身压痛点(按压时疼痛的部位)

本发明中使用的术语“肌筋膜疼痛综合征”是指当肩部和颈后部周围的肌肉无法休息并长时间保持紧张时，由于不正确的姿势和压力，肌肉中缺乏营养和氧气而导致的肌肉变硬并紧密地挤在一起以引起疼痛的疾病。

由本说明书中描述的特定序列号表示的氨基酸序列组成的蛋白质(肽)不限于相应的氨基酸序列，它们还包括由特定序列号表示的氨基酸序列构成的蛋白质的功能等价物。“功能等价物”是指由于氨基酸的添加、取代或缺失而与特定序列号所表示的氨基酸序列具有至少70％、优选80％或更多、更优选90％或更多、甚至更优选95％或更多的序列同一性的蛋白质，即具有95％或更多的序列同一性且表现出与由特定序列号表示的氨基酸序列组成的蛋白质基本相同的生理活性的蛋白质。例如，它可以包括序列同一性为70％,71％,72％,73％,74％,75％,76％,77％,78％,79％,80％,81％,82％,83％,84％,85％,86％,87％,88％,89％,90％,91％,92％,93％,94％,95％,96％,97％,98％,99％或100％的蛋白质(肽)。

由本说明书中描述的特定序列号表示的碱基序列组成的多核苷酸不限于相应的碱基序列，并且该碱基序列的变体包括在本发明的范围内。本发明的碱基序列的核酸分子是包括构成它的核酸分子的功能等同物的概念，例如由核酸分子形成的变体，其中一些碱基序列通过缺失、取代或插入而修饰，但仍然能够执行与核酸相同的功能。具体地，本发明中公开的多核苷酸可以包括与由特定序列号表示的碱基序列具有70％或更多、更优选80％或更多、甚至更优选90％或更多、最优选95％或更多的序列同一性的碱基序列。例如，它包括序列同一性为70％,71％,72％,73％,74％,75％,76％,77％,78％,79％,80％,81％,82％,83％,84％,85％,86％,87％,88％,89％,90％,91％,92％,93％,94％,95％,96％,97％,98％,99％或100％的多核苷酸。多核苷酸的“序列同一性百分比”是通过比较两个最佳比对序列之间的比较区域来确认的，并且为了两个序列的最佳比对，与参考序列(不包括添加或缺失)相比，比较区域中的多核苷酸序列的一部分可以包括添加或缺失(即间隙)。

在本发明中，“药物组合物”是指为预防或治疗疾病而制备的组合物，它可以按照常规方法以各种形式配制和使用。例如，它可以配制成口服剂型，如粉末、颗粒、片剂、胶囊、悬浮液、乳液和糖浆，并且以外用制剂、栓剂和无菌注射溶液的形式配制和使用。

根据本发明的药物组合物可以进一步包括通常用于制备药物组合物的适当载体、赋形剂和稀释剂。赋形剂可以是例如选自由稀释剂、粘合剂、崩解剂、润滑剂、吸附剂、保湿剂、膜包衣材料和控释添加剂组成的群组的一种或多种。

根据本发明的药物组合物可以通过常规方法以下列形式配制和使用：粉末、颗粒、缓释颗粒、肠溶包衣颗粒、溶液、滴眼液、酏剂、乳液、悬浮液、醑剂、锭剂、香料、柠檬水、片剂、缓释片、肠溶包衣片、舌下片、硬胶囊、软胶囊、缓释胶囊、肠溶包衣胶囊、丸剂、酊剂、软提取物、干提取物、液体提取物、注射剂、囊剂、灌注液、膏药、洗液、糊剂、喷雾剂、吸入剂、贴片、灭菌注射溶液，或外部制剂如气雾剂；并且外部制剂可配制为乳膏、凝胶、贴片、喷雾剂、软膏、膏药、洗液、搽剂、糊剂或巴布剂。

根据本发明的药物组合物中可以包含的载体、赋形剂和稀释剂包含乳糖、葡萄糖、蔗糖、低聚糖、山梨醇、甘露醇、木糖醇、赤藓糖醇、麦芽糖醇、淀粉、阿拉伯胶、藻酸盐、明胶、磷酸钙、硅酸钙、纤维素、甲基纤维素、微晶纤维素、聚乙烯吡咯烷酮、水、羟基苯甲酸甲酯、羟基苯乙酸丙酯、滑石、硬脂酸镁和矿物油。

在制剂的情况下，根据本发明的组合物可以使用稀释剂或赋形剂来制备，例如常用的填料、填充剂、粘合剂、润湿剂、崩解剂和表面活性剂。

作为根据本发明的片剂、粉末、颗粒、胶囊、丸剂和锭剂的添加剂，可以使用赋形剂如玉米淀粉、马铃薯淀粉、小麦淀粉、乳糖、白糖、葡萄糖、果糖、二甘露醇、沉淀碳酸钙、合成硅酸铝、磷酸氢钙、硫酸钙、氯化钠、碳酸氢钠、纯化羊毛脂、微晶纤维素、糊精、藻酸钠、甲基纤维素、羧甲基纤维素钠、高岭土、尿素、胶体硅胶、羟丙基淀粉、羟丙基甲基纤维素(HPMC)1928、HPMC 2208、HPMC 2906、HPMC 2910、丙二醇、酪蛋白、乳酸钙和Primogel；以及粘合剂，如明胶、阿拉伯胶、乙醇、琼脂粉末、乙酸邻苯二甲酸酯纤维、羧甲基纤维素、羧甲基纤维钙、葡萄糖、纯化水、酪蛋白酸钠、甘油、硬脂酸、羧甲基纤维酸钠、甲基纤维素钠、甲基纤维、微晶纤维素、糊精、羟基纤维素、羟丙基淀粉、羟甲基纤维素、精制紫胶、淀粉明胶、羟丙基纤维素、羟丙基甲基纤维素、聚乙烯醇和聚乙烯吡咯烷酮；并且可以使用：崩解剂，如羟丙基甲基纤维素、玉米淀粉、琼脂粉、甲基纤维素、膨润土、羟丙基淀粉、羧甲基纤维素钠、海藻酸钠、羧甲基纤维素钙、柠檬酸钙、十二烷基硫酸钠、硅酸酐、1-羟丙基纤维素、葡聚糖、离子交换树脂、聚乙酸乙烯酯、甲醛处理的酪蛋白和明胶、褐藻酸、直链淀粉、瓜尔胶、碳酸氢钠、聚乙烯吡咯烷酮、磷酸钙、凝胶淀粉、阿拉伯胶、支链淀粉、果胶、多磷酸钠、乙基纤维素、白糖、硅酸镁铝、D-山梨醇溶液和轻质无水硅酸；以及润滑剂，如硬脂酸钙、硬脂酸镁、硬脂酸、氢化植物油、滑石、石松、高岭土、凡士林、硬脂酸钠、可可脂、水杨酸钠、水杨酸镁、聚乙二醇(PEG)4000、PEG 6000、液体石蜡、加氢大豆油(Lubri蜡)、硬脂酸铝、硬脂酸锌、十二烷基硫酸钠、氧化镁、聚乙二醇、合成硅酸铝、硅酸酐、高级脂肪酸、高级醇、硅油、石蜡油、聚乙二醇脂肪酸醚、氯化钠、乙酸钠、油酸钠、DL-亮氨酸和轻质无水硅酸。

作为根据本发明的液体制剂的添加剂，可以使用水、稀盐酸、稀硫酸、柠檬酸钠、蔗糖单硬脂酸酯、聚氧乙烯山梨醇脂肪酸酯(吐温酯)、聚氧乙烯单烷基醚、羊毛脂醚、羊毛脂酯、乙酸、盐酸、氨水、碳酸铵、氢氧化钾、氢氧化钠、脯氨酸、聚乙烯吡咯烷酮、乙基纤维素、羧甲基纤维素钠等。

在根据本发明的糖浆中可以使用白糖、其他糖或甜味剂等的溶液，并且可以根据需要使用调味剂、着色剂、防腐剂、稳定剂、悬浮剂、乳化剂、增稠剂等。

根据本发明，纯化水可用于乳液中，并且可以根据需要使用乳化剂、防腐剂、稳定剂、调味剂等。

根据本发明，在悬浮液中可以使用悬浮剂，例如阿拉伯胶、黄芪胶(tragacanth)、甲基纤维素、羧甲基纤维素、羧基甲基纤维素钠、微晶纤维素、藻酸钠、HPMC、HPMC 1828、HPMC 2906、HPMC 2910等，并且可以根据需要使用表面活性剂、防腐剂、稳定剂、着色剂和调味剂。

根据本发明的注射剂可以包括溶剂，例如注射用蒸馏水、0.9％注射用氯化钠、林格氏溶液、注射用葡萄糖、注射用葡萄糖+氯化钠、PEG、乳酸林格氏溶液、乙醇、丙二醇、非挥发性油芝麻油、棉子油、花生油、大豆油、玉米油、油酸乙酯、肉豆蔻酸异丙酯、苯甲酸苄酯；增溶剂，如苯甲酸钠、水杨酸钠、乙酸钠、尿素、氨基甲酸酯、单乙基乙酰胺、丁唑胺、丙二醇、吐温、烟酰胺、六胺和二甲基乙酰胺；缓冲剂，如弱酸及其盐(乙酸和乙酸钠)、弱碱及其盐(氨和乙酸铵)、有机化合物、蛋白质、白蛋白、蛋白胨和树胶；等渗剂，如氯化钠；稳定剂，如亚硫酸氢钠(NaHSO₃)二氧化碳气体、偏亚硫酸氢钠(Na₂S₂O₅)、亚硫酸钠(Na₂SO₃)、氮气(N₂)和乙二胺四乙酸；抗氧化剂，如0.1％硫酸氢钠、甲醛次硫酸氢钠、硫脲、乙二胺四乙酸二钠和丙酮亚硫酸氢钠；止痛剂，如苄醇、氯丁醇、盐酸普鲁卡因、葡萄糖和葡萄糖酸钙；以及悬浮剂，如羧甲基(CM)钠、海藻酸钠、吐温80和单硬脂酸铝。

对于根据本发明的栓剂，可以使用碱，如可可油、羊毛脂、Witepsol、聚乙二醇、甘油、甲基纤维素、羧甲基纤维素、硬脂酸和油酸的混合物、Subanal、棉籽油、花生油、棕榈油、可可脂+胆固醇、卵磷脂、Lanette蜡、甘油单硬脂酸酯、吐温或Span、Imhausen、单烯(丙二醇单硬脂酸酯)、甘油、Adeps solidus、Buytyrum Tego-G、Cebes Pharma 16、六环内酯碱95、Cotomar、Hydrocote SP、S-70-XXA、S-70-XX75(S-70-XX95)、Hydrokete 25、Hydrokoe 711、Idropostal、Massa estrarium(A,AS,B,C,D,E,I,T)、Mass-MF、Massaupol、Masupol-15、新栓剂-N、Paramount-B、Suposiro(OSI、OSIX、A、B、C、D、H、L)、IV型栓剂基质(AB、B、A、BC、BBG、E、BGF、C、D、299)、Supostal(N、Es)、Wecobi(W、R、S、M、Fs)、Tegestor甘油三酯基质(TG-95、MA、57)。

口服固体制剂包含片剂、丸剂、粉末、颗粒和胶囊。这些固体制剂通过将提取物与至少一种赋形剂(例如淀粉、碳酸钙、蔗糖)、乳糖或明胶等混合来制备。除了简单的赋形剂之外，还使用润滑剂，例如硬脂酸镁和滑石。

悬浮液、口服溶液、乳液、糖浆等对应于口服给药的液体制剂，并且除了常用的简单稀释剂如水和液体石蜡之外，还可以包括各种赋形剂，例如润湿剂、甜味剂、调味剂和防腐剂。用于胃肠外给药的制剂包括无菌水溶液、非水溶液、悬浮液、乳液、冻干制剂和栓剂。可以使用丙二醇、聚乙二醇、植物油如橄榄油和可注射的酯如油酸乙酯作为非水溶剂和悬浮液。

根据本发明的药物组合物以药学有效量给药。在本发明中，“药物有效量”是指足以以适用于医疗的合理获益/风险比治疗疾病的量，并且有效剂量水平可以基于包括患者疾病类型和严重程度、药物活性、对药物的敏感性、给药时间、给药途径和排泄率、治疗持续时间和同时使用的药物等因素及医学领域中众所周知的其他因素来确定。

根据本发明的药物组合物可以作为单独的治疗剂或与另一种治疗剂同时给药，并且它可以顺序地或与常规治疗剂同时给药，并且它可以给药一次或多次。重要的是，通过考虑所有上述因素，给药的量可以以最小的量达到最大效果而没有副作用，这可以由本发明所属领域的技术人员容易地确定。

本发明的药物组合物可以通过各种途径给药至受试者。考虑了所有的给药方式，并且它可以通过例如口服给药、皮下注射、腹膜内给药、静脉注射、肌内注射、椎旁间隙(鞘内)注射、舌下给药、口腔给药、直肠内注射、阴道注射、眼部给药、耳部给药、鼻腔给药、吸入、通过口或鼻喷雾、皮肤给药、透皮给药等给药。

本发明的药物组合物是根据作为活性成分的药物的类型以及各种相关因素来确定的，所述因素例如是要治疗的疾病、给药途径、患者的年龄、性别和体重以及疾病的严重程度。

此外，本发明提供了一种止痛方法，其包括将包含肉毒杆菌毒素重组蛋白作为活性成分的组合物给药至有需要的受试者。

此外，本发明提供了包含肉毒杆菌毒素重组蛋白作为活性成分的组合物用于止痛的用途。

此外，本发明提供肉毒杆菌毒素重组蛋白用于制备镇痛药物的用途。

在本发明中，“受试者”是指需要治疗疾病的受试者，更具体是指人类或非人灵长类动物，以及哺乳动物，如小鼠、大鼠、狗、猫、马和牛。

在本发明中，“给药”是指通过任何适当的方法向受试者提供预定量的本发明组合物。

在本发明中，“改善”可指通过根据本发明的组合物的给药使疼痛症状更好或有益地改变的任何作用，并且还可包括预防或治疗作用。

在本发明中，“预防”是指抑制或延迟靶疾病发作的所有作用，并且“治疗”是指通过根据本发明的药物组合物的给药而更好或有益地改变靶疾病及其相关代谢异常的所有作用。

此外，作为本发明的另一个方面，本发明提供了一种用于止痛的准药物组合物，其包含肉毒杆菌毒素重组蛋白作为活性成分，并且在肉毒杆菌毒素重组蛋白中，由SEQ IDNO:1的氨基酸序列组成的细胞穿透性肽与肉毒杆菌毒素轻链的一端或两端融合。

本发明的术语“准药物”是指用于诊断、治疗、改善、减轻、治疗或预防人类或动物疾病的目的的物品中效果比药物温和的物品。例如，根据《药物事务法》(thePharmaceutical Affairs Act)，准药物不包括用于医药用途的物品，包括用于治疗或预防人类/动物疾病的产品以及对人体有轻微或没有直接影响的产品。

本发明的准药物组合物可以制备为选自身体清洁剂、泡沫、肥皂、面膜、软膏、乳膏、乳液、精华液和喷雾剂中的一种或多种，但不限于此。此外，该组合物可以以绷带、卫生巾等的形式制造，但不限于此。

当根据本发明的肉毒杆菌毒素重组蛋白用作准药物添加剂时，该组合物可以按原样添加或与另一种准药物或准药物组分一起使用，并且可以根据常规方法适当使用。活性成分的混合量可以根据使用目的适当地确定。

此外，作为本发明的又一方面，本发明提供了一种用于止痛的皮肤外用组合物，其包含肉毒杆菌毒素重组蛋白作为活性成分，并且在肉毒杆菌毒素重组蛋白中，将由SEQ IDNO:1的氨基酸序列组成的细胞穿透性肽与肉毒杆菌毒素轻链的一端或两端融合。

本发明的皮肤外用组合物包含肉毒杆菌毒素重组蛋白作为活性成分，并且可以包括药学上可接受的载体。此外，本发明的皮肤外用组合物可以进一步包括皮肤科领域中常用的佐剂，例如脂肪物质、有机溶剂、增溶剂、增稠剂和胶凝剂、柔软剂、抗氧化剂、悬浮剂、稳定剂、发泡剂、调味剂、表面活性剂、水、离子或非离子乳化剂、填料、掩蔽剂和螯合剂、防腐剂、维生素、阻断剂、润湿剂、精华液、染料、颜料、亲水性或亲脂性活性剂、脂质囊泡或任何其他常用于皮肤外用制剂的成分。此外，所述成分可以以皮肤科领域中常用的量引入。

用于皮肤外用的组合物的剂量可以根据给药受试者的体重、年龄、性别、健康状况、给药时间、清除率、疾病的严重程度等而变化。

用于皮肤外用的组合物的制剂形式包括但不限于例如液体包衣剂、喷雾剂、乳液、凝胶、糊状物、软膏、气雾剂、粉末、贴片、乳膏、走珠和透皮吸收剂。

本发明的皮肤外用组合物中的药学上可接受的载体根据剂型而变化，但它们可以包括烃类，如凡士林、液体石蜡和胶凝烃类(Plastibase)；动物和植物油，如中链脂肪酸甘油三酯、猪肉脂肪、硬脂肪和可可脂；高级脂肪醇和脂肪酸，以及它们的酯，如十六醇、硬脂醇、硬脂酸和棕榈酸异丙酯；水溶性碱，如聚乙二醇、1,3-丁二醇、甘油、明胶、白糖和糖醇；乳化剂，如甘油脂肪酸酯、聚氧化硬脂酸酯和聚氧乙烯氢化蓖麻油；粘合剂，如丙烯酸酯和海藻酸钠；推进剂，如液化石油气和二氧化碳；以及防腐剂，如对羟基苯甲酸酯。此外，除此之外，可以根据需要添加稳定剂、香料、着色剂、pH调节剂、稀释剂、表面活性剂、防腐剂、抗氧化剂等。当使用根据本发明的皮肤外用组合物时，优选通过常规方法将其施用于受影响的区域或发生疼痛的区域的皮肤。

此外，根据本发明的皮肤外用组合物可以通过粘附到固体载体例如传统绷带的伤口剥离覆盖物上来使用。粘附可以通过使本发明的组合物在固体载体中饱和然后将其脱水来实现。优选地，可以首先在固体载体上涂覆粘合剂，以提高本发明组合物与固体载体的粘合性。粘合剂的实例包括聚丙烯酸酯和氰基丙烯酸酯。

此外，作为本发明的又一方面，本发明提供了一种用于止痛的化妆品组合物，其包含肉毒杆菌毒素重组蛋白作为活性成分，并且在肉毒杆菌毒素重组蛋白中，由SEQ ID NO:1的氨基酸序列组成的细胞穿透性肽与肉毒杆菌毒素轻链的一端或两端融合。

根据本发明的化妆品组合物可以以本领域中通常制备的任何制剂形式制备，并且可以以用于局部或全身施用的佐剂的形式制备，除了本发明的组合物之外，所述佐剂在皮肤科领域中与皮肤科可接受的介质、基质、佐剂、赋形剂等一起常规制备。例如，它可以以基本化妆品组合物(爽肤水、面霜、精华液、清洁剂如清洁泡沫和清洁水、面膜、身体油)、彩色化妆品组合物(粉底、唇膏、睫毛膏、化妆基底)、头发产品组合物(洗发水、护发乳、护发素、发胶)和肥皂的形式制备。

此外，本发明的化妆品组合物还可包括皮肤科领域常用的佐剂，例如脂肪物质、有机溶剂、增溶剂、增稠剂和胶凝剂、柔软剂、抗氧化剂、悬浮剂、稳定剂、发泡剂、调味剂、表面活性剂、水、离子或非离子乳化剂、填料、掩蔽剂和螯合剂、防腐剂、维生素、阻断剂、润湿剂、精华液、染料、颜料、亲水性或亲脂性活性剂、脂质囊泡或任何其他常用于皮肤外用制剂的成分。此外，所述成分可以以皮肤科领域中常用的量引入。

赋形剂可包括但不限于润肤剂、皮肤渗透促进剂、着色剂、调味剂、乳化剂、柔软剂、增稠剂、浓缩剂、胶凝剂、悬浮剂、稳定剂、发泡剂、表面活性剂、填料和溶剂。此外，本发明的化妆品组合物可以进一步包括香料、颜料、杀菌剂、抗氧化剂、防腐剂、佐剂和保湿剂，并且可以包括增稠剂、无机盐、合成聚合物材料等用于改善物理性能的目的。例如，当使用本发明的化妆品组合物制备洗面奶或皂时，可以通过将本发明的提取物添加到传统的洗面奶或皂基中来容易地制备。当制备乳膏时，它可以通过将本发明的肉毒杆菌毒素重组蛋白添加到普通水包油(O/W)乳膏基质中来制备。这里，为了改善物理性质的目的，可以添加香料、螯合剂、颜料、抗氧化剂、防腐剂等以及合成或天然材料，如蛋白质、矿物质和维生素。

化妆品组合物的合适的制剂形式包括例如溶液、凝胶、固体或糊状无水产品、通过将油相分散在水相中获得的乳液、悬浮液、微乳液、微胶囊、微粒、离子(脂质体)或非离子泡状分散剂形式、乳膏、爽肤水、乳液、粉末、软膏、喷雾剂或遮瑕棒。此外，它可以以泡沫的形式或进一步包括压缩推进剂的气溶胶组合物的形式制备。

可添加本发明化妆品组合物的产品包括但不限于，皮肤乳液、皮肤柔软剂、皮肤爽肤水、收敛爽肤水、软化爽肤水和滋养爽肤水、收敛剂、乳液、牛奶乳液、保湿乳液、营养乳液、身体霜、按摩霜、营养霜、保湿霜、护手霜、精华液、营养精华液、面膜、肥皂、洗发水、清洁泡沫、清洁乳液、清洁霜、身体乳液、身体清洁剂、护理液、美容精华液、乳液、压制粉末、散粉和眼影。

根据本发明的组合物可以进一步包括提取物，例如薄荷醇、乳木果油、苏格兰松叶、芦荟、薄荷和迷迭香，但不限于此。

在下文中，提供优选的实施例以帮助理解本发明。然而，提供以下实施例只是为了更容易理解本发明，并且本发明的内容不受以下实施例的限制。

[实施例]

实施例1.确认头痛改善效果

1-1.实验材料和方法

每天早晨将2.5cc含有根据韩国专利号10-1882461制备的肉毒杆菌毒素重组蛋白(皮肤穿透性肉毒杆菌衍生成分肽)的溶液直接施用到在日常生活中由于头痛而感到不适且正服用镇痛药的23至53岁的总共9名成年男性和女性(7名女性，2名男性，平均年龄36岁)的疼痛的头皮区域和前额。

通过在测试前对受试者进行调查，然后要求受试者通过韩国头痛协会(KoreanHeadache Society)提供的“头痛日记应用程序”直接输入头痛次数和头痛发生时每天服用镇痛药的频率来进行评估。试验期共12周，根据受试者直接输入的信息分四个时间段对头痛次数进行比较和评估：使用前、使用后1至4周、使用后5至8周和使用后9至12周。以三个时间段比较和评估镇痛药的服用频率：使用后1至4周、使用后5至8周和使用后9至12周。

1-2.头痛的次数

作为比较和评估头痛次数的结果，如图1所示，证实了所有9名受试者的头痛都得到了改善。具体而言，使用后时段的平均头痛次数：

使用前4周时间段为14.3次，使用后1至4周时间段为8.5次，使用5至8周时间段为6.9次，使用后9至12周时间段为2.5次，证实了与使用前4周时间段相比较，使用含有皮肤渗透性肉毒杆菌衍生成分肽的溶液后9至12周时间段期间，受试者平均头痛次数减少84％。

此外，如图2所示，在使用后前1至4周(对应于使用的早期阶段)和使用后最后9至12周之间比较使用后时段的头痛次数减少率，结果，证实两名受试者经历了超过20％的改善(7→9)；四名受试者经历了超过50％的改善(4→8)，四名受试者经历了超过70％的改善(1→5)；并且三名受试者经历了100％的改善(0→3)。

1-3.服用镇痛药的频率

作为比较使用后1至4周与9至12周服用镇痛药的频率的结果，发现9名受试者中有8名服用镇痛药的频率减少，而其中1名受试者服用镇痛药的频率增加。

作为按使用后的时段比较镇痛药服用频率的结果，如图3所示，在使用后1至4周的时间段内，3名受试者服用2次镇痛药，3名受试者服用1次，3名受试者服用0次，并且在使用后5至8周的时间段内，1名受试者服用3次镇痛药、2名受试者服药2次、4名受试者服药1次并且3名受试验者服药0次，表明有1名受试者的镇痛药服用频率增加了1次而有1名受试者的镇痛药服用频率减少了1次。经证实，在使用后9至12周的时间段内，1名受试者服用了3次镇痛药，0名受试者服药2次，2名受试者服药1次，并且6名受试者服药0次，这表明与使用早期阶段相对应的使用后1至4周的时间段相比，不需要服用镇痛药的受试者人数增加了3人。

根据以上结果，当将表现出类似于肉毒杆菌毒素注射的皱纹改善效果的皮肤穿透性肉毒杆菌衍生成分肽施用于头部和颈部时，头痛的总次数和服用镇痛药的总频率降低，因此判断将具有头痛改善效果。

实施例2.止痛效果的确认

2-1.实验材料和方法

该实验对24名年龄在30至75岁之间的患有慢性疼痛、纤维肌痛、肌筋膜疼痛或肌痛的成年人进行，通过将皮肤穿透性肉毒杆菌衍生成分肽施用于他们的皮肤来确认该肽是否可以通过缓解肌肉张力来减轻疼痛，减轻肌肉痉挛疼痛，并确认皮肤穿透性肉毒衍生成分肽对由重复运动和机械冲击引起的慢性疼痛的改善程度。

选择实验受试者以满足下表1中的三个标准之一，并允许他们每天两次将适当量的含有皮肤穿透性肉毒杆菌衍生成分肽的肉毒杆菌乳液直接施用到疼痛区域。

[表1]

为了进行分析，24名受试者被分为如下三组：

A组：慢性疼痛患者

B组：纤维肌痛和肌痛患者

C组：麻醉镇痛药使用组(C组与A组和B组有样本重叠)。

2-2.A组

作为确认14名慢性疼痛患者在使用肉毒杆菌乳液后6天内的疼痛变化(将满足表1中的每个标准的患者适当混合)的结果，如图4所示，证实了78.57％的患者在使用了肉毒杆菌乳液之后疼痛减轻(14名受试者中有11名疼痛减轻)。使用肉毒杆菌乳液时疼痛保持减轻。在使用肉毒杆菌乳液后立即表现出即刻疼痛减轻效果，并且证实了在施用该乳液时通常保持疼痛减轻效果。

2-3.B组

作为确认在7名纤维肌痛和肌痛患者中使用肉毒杆菌乳液后疼痛减轻是否继续的结果(将满足表1中的每一个标准的患者适当混合)，证实了71.43％的患者在使用肉毒杆菌乳液后立即疼痛减轻(7名受试者中有5名疼痛减轻，现在如图所示)，并且如图5所示，57.14％的患者在施用该乳液时保持了疼痛减轻状态(7名患者中有4名保持了疼痛减轻状态)。通常，发现在使用肉毒杆菌乳液后立即体验到疼痛减轻效果，并且在施用肉毒杆菌乳液时保持该效果。

2-4.C组

作为确认在使用麻醉性镇痛药的15名患者中使用肉毒杆菌乳液之后疼痛减轻是否继续的结果，如图6所示，80％患者的应答为疼痛立即减轻6(图中未示出)，并且如图6中所示，66.67％患者(15名受试者中有10名)的应答为在使用肉毒杆菌乳液时存在持续的疼痛减轻效果。研究发现，在使用肉毒杆菌乳液后，通常立即体验到疼痛减轻效果，并且在施用肉毒杆菌乳液时保持了该效果。

上述对本发明的描述是为了说明的目的，本领域技术人员将理解，在不改变本发明的技术思想或基本特征的情况下，本发明可以容易地修改为其他特定形式。因此，上文所描述的实例应在所有方面被理解为说明性的而非限制性的。

工业适用性

根据本发明的肉毒杆菌毒素重组蛋白可以通过与细胞穿透性肽融合而经皮递送具有肌肉麻痹效果的肉毒杆菌毒轻链，通过方便且快速地将肉毒杆菌毒素轻链递送到疼痛区域而有效地缓解肌肉紧张和减少疼痛，因此它可以有效地用作预防、改善或治疗疼痛的物质。因此，根据本发明的肉毒杆菌毒素重组蛋白具有工业实用性。

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<213> 人工序列

<220>

<223> BoNT/B轻链氨基酸序列

<400> 4

Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn

1 5 10 15

Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg

20 25 30

Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu

35 40 45

Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly

50 55 60

Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn

65 70 75 80

Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe

85 90 95

Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile

100 105 110

Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu

115 120 125

Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn

130 135 140

Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile

145 150 155 160

Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly

165 170 175

Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln

180 185 190

Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu

195 200 205

Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro

210 215 220

Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr

225 230 235 240

Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe

245 250 255

Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe

260 265 270

Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile

275 280 285

Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn

290 295 300

Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr

305 310 315 320

Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly

325 330 335

Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu

340 345 350

Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys

355 360 365

Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys

370 375 380

Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile

385 390 395 400

Ser Asp Lys Asp Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile

405 410 415

Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr

420 425 430

Lys Ile Gln Met Cys Lys Ser Val Lys

435 440

<210> 5

<211> 449

<212> PRT

<213> 人工序列

<220>

<223> BoNT/C轻链氨基酸序列

<400> 5

Met Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro Val Asp Asn

1 5 10 15

Lys Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu

20 25 30

Pro Glu Lys Ala Phe Arg Ile Thr Gly Asn Ile Trp Val Ile Pro Asp

35 40 45

Arg Phe Ser Arg Asn Ser Asn Pro Asn Leu Asn Lys Pro Pro Arg Val

50 55 60

Thr Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp

65 70 75 80

Ser Asp Lys Asp Pro Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg

85 90 95

Ile Asn Ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ser Thr

100 105 110

Asp Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp

115 120 125

Phe Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn

130 135 140

Asn Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly

145 150 155 160

Pro Arg Glu Asn Ile Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr

165 170 175

Asn Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile

180 185 190

Ser Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asp

195 200 205

Val Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile

210 215 220

Leu Ile Leu Met His Glu Leu Asn His Ala Met His Asn Leu Tyr Gly

225 230 235 240

Ile Ala Ile Pro Asn Asp Gln Thr Ile Ser Ser Val Thr Ser Asn Ile

245 250 255

Phe Tyr Ser Gln Tyr Asn Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala

260 265 270

Phe Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser Ala Arg Lys Tyr

275 280 285

Phe Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu

290 295 300

Asn Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly

305 310 315 320

Glu Tyr Lys Gln Lys Leu Ile Arg Lys Tyr Arg Phe Val Val Glu Ser

325 330 335

Ser Gly Glu Val Thr Val Asn Arg Asn Lys Phe Val Glu Leu Tyr Asn

340 345 350

Glu Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn

355 360 365

Val Gln Asn Arg Lys Ile Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr

370 375 380

Ala Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn

385 390 395 400

Ile Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser

405 410 415

Arg Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu

420 425 430

Phe Thr Lys Phe Cys His Lys Ala Ile Asp Gly Arg Ser Leu Tyr Asn

435 440 445

Lys

<210> 6

<211> 442

<212> PRT

<213> 人工序列

<220>

<223> BoNT/D轻链氨基酸序列

<400> 6

Met Thr Trp Pro Val Lys Asp Phe Asn Tyr Ser Asp Pro Val Asn Asp

1 5 10 15

Asn Asp Ile Leu Tyr Leu Arg Ile Pro Gln Asn Lys Leu Ile Thr Thr

20 25 30

Pro Val Lys Ala Phe Met Ile Thr Gln Asn Ile Trp Val Ile Pro Glu

35 40 45

Arg Phe Ser Ser Asp Thr Asn Pro Ser Leu Ser Lys Pro Pro Arg Pro

50 55 60

Thr Ser Lys Tyr Gln Ser Tyr Tyr Asp Pro Ser Tyr Leu Ser Thr Asp

65 70 75 80

Glu Gln Lys Asp Thr Phe Leu Lys Gly Ile Ile Lys Leu Phe Lys Arg

85 90 95

Ile Asn Glu Arg Asp Ile Gly Lys Lys Leu Ile Asn Tyr Leu Val Val

100 105 110

Gly Ser Pro Phe Met Gly Asp Ser Ser Thr Pro Glu Asp Thr Phe Asp

115 120 125

Phe Thr Arg His Thr Thr Asn Ile Ala Val Glu Lys Phe Glu Asn Gly

130 135 140

Ser Trp Lys Val Thr Asn Ile Ile Thr Pro Ser Val Leu Ile Phe Gly

145 150 155 160

Pro Leu Pro Asn Ile Leu Asp Tyr Thr Ala Ser Leu Thr Leu Gln Gly

165 170 175

Gln Gln Ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser Ile Leu

180 185 190

Lys Val Ala Pro Glu Phe Leu Leu Thr Phe Ser Asp Val Thr Ser Asn

195 200 205

Gln Ser Ser Ala Val Leu Gly Lys Ser Ile Phe Cys Met Asp Pro Val

210 215 220

Ile Ala Leu Met His Glu Leu Thr His Ser Leu His Gln Leu Tyr Gly

225 230 235 240

Ile Asn Ile Pro Ser Asp Lys Arg Ile Arg Pro Gln Val Ser Glu Gly

245 250 255

Phe Phe Ser Gln Asp Gly Pro Asn Val Gln Phe Glu Glu Leu Tyr Thr

260 265 270

Phe Gly Gly Leu Asp Val Glu Ile Ile Pro Gln Ile Glu Arg Ser Gln

275 280 285

Leu Arg Glu Lys Ala Leu Gly His Tyr Lys Asp Ile Ala Lys Arg Leu

290 295 300

Asn Asn Ile Asn Lys Thr Ile Pro Ser Ser Trp Ile Ser Asn Ile Asp

305 310 315 320

Lys Tyr Lys Lys Ile Phe Ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn

325 330 335

Thr Gly Asn Phe Val Val Asn Ile Asp Lys Phe Asn Ser Leu Tyr Ser

340 345 350

Asp Leu Thr Asn Val Met Ser Glu Val Val Tyr Ser Ser Gln Tyr Asn

355 360 365

Val Lys Asn Arg Thr His Tyr Phe Ser Arg His Tyr Leu Pro Val Phe

370 375 380

Ala Asn Ile Leu Asp Asp Asn Ile Tyr Thr Ile Arg Asp Gly Phe Asn

385 390 395 400

Leu Thr Asn Lys Gly Phe Asn Ile Glu Asn Ser Gly Gln Asn Ile Glu

405 410 415

Arg Asn Pro Ala Leu Gln Lys Leu Ser Ser Glu Ser Val Val Asp Leu

420 425 430

Phe Thr Lys Val Cys Leu Arg Leu Thr Lys

435 440

<210> 7

<211> 422

<212> PRT

<213> 人工序列

<220>

<223> BoNT/E轻链氨基酸序列

<400> 7

Met Pro Thr Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asn Arg

1 5 10 15

Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser

20 25 30

Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile

35 40 45

Gly Thr Ile Pro Gln Asp Phe Leu Pro Pro Thr Ser Leu Lys Asn Gly

50 55 60

Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Gln Glu Lys

65 70 75 80

Asp Lys Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asp

85 90 95

Asn Leu Ser Gly Arg Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro

100 105 110

Tyr Leu Gly Asn Asp Asn Thr Pro Asp Gly Asp Phe Ile Ile Asn Asp

115 120 125

Ala Ser Ala Val Pro Ile Gln Phe Ser Asn Gly Ser Gln Ser Ile Leu

130 135 140

Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr

145 150 155 160

Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His

165 170 175

Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe

180 185 190

Arg Phe Lys Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu

195 200 205

Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala

210 215 220

Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Lys Gln Asn Pro Leu

225 230 235 240

Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly

245 250 255

Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Ser Asn Asp Ile Tyr

260 265 270

Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys

275 280 285

Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp Val Phe Glu

290 295 300

Ala Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn

305 310 315 320

Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu

325 330 335

Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile

340 345 350

Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile

355 360 365

Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe

370 375 380

Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr

385 390 395 400

Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val

405 410 415

Ser Val Lys Gly Ile Arg

420

<210> 8

<211> 436

<212> PRT

<213> 人工序列

<220>

<223> BoNT/F轻链氨基酸序列

<400> 8

Met Pro Val Ala Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp

1 5 10 15

Asp Thr Ile Leu Tyr Met Gln Ile Pro Tyr Glu Glu Lys Ser Lys Lys

20 25 30

Tyr Tyr Lys Ala Phe Glu Ile Met Arg Asn Val Trp Ile Ile Pro Glu

35 40 45

Arg Asn Thr Ile Gly Thr Asn Pro Ser Asp Phe Asp Pro Pro Ala Ser

50 55 60

Leu Lys Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr

65 70 75 80

Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr Ile Lys Leu Phe Lys

85 90 95

Arg Ile Asn Ser Asn Pro Ala Gly Lys Val Leu Leu Gln Glu Ile Ser

100 105 110

Tyr Ala Lys Pro Tyr Leu Gly Asn Asp His Thr Pro Ile Asp Glu Phe

115 120 125

Ser Pro Val Thr Arg Thr Thr Ser Val Asn Ile Lys Leu Ser Thr Asn

130 135 140

Val Glu Ser Ser Met Leu Leu Asn Leu Leu Val Leu Gly Ala Gly Pro

145 150 155 160

Asp Ile Phe Glu Ser Cys Cys Tyr Pro Val Arg Lys Leu Ile Asp Pro

165 170 175

Asp Val Val Tyr Asp Pro Ser Asn Tyr Gly Phe Gly Ser Ile Asn Ile

180 185 190

Val Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp Ile Ser Gly

195 200 205

Gly His Asn Ser Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser

210 215 220

Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Arg

225 230 235 240

Gly Val Thr Tyr Glu Glu Thr Ile Glu Val Lys Gln Ala Pro Leu Met

245 250 255

Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly

260 265 270

Gln Asp Leu Asn Ile Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn

275 280 285

Asn Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Ser Glu Val

290 295 300

Asn Ser Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe

305 310 315 320

Gln Trp Lys Tyr Gly Leu Asp Lys Asn Ala Asp Gly Ser Tyr Thr Val

325 330 335

Asn Glu Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr

340 345 350

Glu Ser Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr

355 360 365

Phe Ile Lys Tyr Glu Phe Leu Lys Val Pro Asn Leu Leu Asp Asp Asp

370 375 380

Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn

385 390 395 400

Asn Arg Gly Gln Ser Ile Lys Leu Asn Pro Lys Ile Ile Asp Ser Ile

405 410 415

Pro Asp Lys Gly Leu Val Glu Lys Ile Val Lys Phe Cys Lys Ser Val

420 425 430

Ile Pro Arg Lys

435

<210> 9

<211> 441

<212> PRT

<213> 人工序列

<220>

<223> BoNT/G轻链氨基酸序列

<400> 9

Met Pro Val Asn Ile Lys Phe Asn Tyr Asn Asp Pro Ile Asn Asn Asp

1 5 10 15

Asp Ile Ile Met Met Glu Pro Phe Asn Asp Pro Gly Pro Gly Thr Tyr

20 25 30

Tyr Lys Ala Phe Arg Ile Ile Asp Arg Ile Trp Ile Val Pro Glu Arg

35 40 45

Phe Thr Tyr Gly Phe Gln Pro Asp Gln Phe Asn Ala Ser Thr Gly Val

50 55 60

Phe Ser Lys Asp Val Tyr Glu Tyr Tyr Asp Pro Thr Tyr Leu Lys Thr

65 70 75 80

Asp Ala Glu Lys Asp Lys Phe Leu Lys Thr Met Ile Lys Leu Phe Asn

85 90 95

Arg Ile Asn Ser Lys Pro Ser Gly Gln Arg Leu Leu Asp Met Ile Val

100 105 110

Asp Ala Ile Pro Tyr Leu Gly Asn Ala Ser Thr Pro Pro Asp Lys Phe

115 120 125

Ala Ala Asn Val Ala Asn Val Ser Ile Asn Lys Lys Ile Ile Gln Pro

130 135 140

Gly Ala Glu Asp Gln Ile Lys Gly Leu Met Thr Asn Leu Ile Ile Phe

145 150 155 160

Gly Pro Gly Pro Val Leu Ser Asp Asn Phe Thr Asp Ser Met Ile Met

165 170 175

Asn Gly His Ser Pro Ile Ser Glu Gly Phe Gly Ala Arg Met Met Ile

180 185 190

Arg Phe Cys Pro Ser Cys Leu Asn Val Phe Asn Asn Val Gln Glu Asn

195 200 205

Lys Asp Thr Ser Ile Phe Ser Arg Arg Ala Tyr Phe Ala Asp Pro Ala

210 215 220

Leu Thr Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr Gly

225 230 235 240

Ile Lys Ile Ser Asn Leu Pro Ile Thr Pro Asn Thr Lys Glu Phe Phe

245 250 255

Met Gln His Ser Asp Pro Val Gln Ala Glu Glu Leu Tyr Thr Phe Gly

260 265 270

Gly His Asp Pro Ser Val Ile Ser Pro Ser Thr Asp Met Asn Ile Tyr

275 280 285

Asn Lys Ala Leu Gln Asn Phe Gln Asp Ile Ala Asn Arg Leu Asn Ile

290 295 300

Val Ser Ser Ala Gln Gly Ser Gly Ile Asp Ile Ser Leu Tyr Lys Gln

305 310 315 320

Ile Tyr Lys Asn Lys Tyr Asp Phe Val Glu Asp Pro Asn Gly Lys Tyr

325 330 335

Ser Val Asp Lys Asp Lys Phe Asp Lys Leu Tyr Lys Ala Leu Met Phe

340 345 350

Gly Phe Thr Glu Thr Asn Leu Ala Gly Glu Tyr Gly Ile Lys Thr Arg

355 360 365

Tyr Ser Tyr Phe Ser Glu Tyr Leu Pro Pro Ile Lys Thr Glu Lys Leu

370 375 380

Leu Asp Asn Thr Ile Tyr Thr Gln Asn Glu Gly Phe Asn Ile Ala Ser

385 390 395 400

Lys Asn Leu Lys Thr Glu Phe Asn Gly Gln Asn Lys Ala Val Asn Lys

405 410 415

Glu Ala Tyr Glu Glu Ile Ser Leu Glu His Leu Val Ile Tyr Arg Ile

420 425 430

Ala Met Cys Lys Pro Val Met Tyr Lys

435 440

<210> 10

<211> 1347

<212> DNA

<213> 人工序列

<220>

<223> BoNT/A轻链cDNA序列

<400> 10

atgccctttg tcaacaaaca gttcaactac aaggacccag ttaatggagt agacatcgca 60

tatatcaaga ttcccaacgc tggccagatg caacccgtta aggcatttaa aatccataac 120

aaaatctggg ttatcccaga gcgggatacc ttcaccaacc ccgaggaggg cgatctgaac 180

cccccgccgg aggcgaagca ggtcccagtg agctactacg atagcaccta cctcagcacc 240

gacaacgaga aggacaacta cctcaaagga gtcacgaagt tgttcgagag aatctactcc 300

acagacctcg gccgcatgct tctaaccagc attgtgcgtg gcattccctt ttggggcggc 360

tctaccatcg acacagagct gaaggtgata gacaccaact gcatcaacgt aatccagcct 420

gacggcagct accgaagcga ggagcttaac ctggtgatca tcggcccttc cgccgatatc 480

atccaattcg agtgcaagag cttcggccac gaggtcctga acctcacccg gaacggctat 540

ggaagcaccc agtacataag attcagccct gacttcacct tcgggtttga ggagagcttg 600

gaggtcgaca caaaccccct gctgggagcc gggaagttcg ccactgaccc agccgtgact 660

ctggcacacg agctgatcca cgccggtcac cgcctgtacg gcatagctat aaacccaaac 720

agggtgttca aagtgaacac caacgcttac tatgaaatga gcggcctgga ggtgagcttc 780

gaggagctga gaacgttcgg gggacatgat gctaaattta tcgacagcct gcaggagaac 840

gagttcaggc tgtactacta caataagttc aaggatatag cgagcactct gaacaaggcc 900

aagtccatcg taggcactac tgcatccctc cagtatatga agaatgtgtt caaagagaaa 960

tacctgctga gcgaggatac cagcggtaag ttcagcgtgg ataagcttaa gttcgacaag 1020

ctgtataaga tgctcaccga aatctacacc gaggataatt tcgttaagtt cttcaaggtc 1080

ctgaaccgga agacctacct gaacttcgac aaggccgtgt tcaagatcaa catcgtgcct 1140

aaagtgaact acaccatcta cgacgggttt aacctgagga acaccaacct ggccgctaac 1200

ttcaacgggc agaacacaga gatcaacaac atgaatttca cgaagttgaa gaacttcacc 1260

ggactgtttg agttctacaa attgctgtgt gtgcgcggga tcatcactag caagaccaag 1320

agccttgaca aaggctacaa caagtga 1347

<210> 11

<211> 1323

<212> DNA

<213> 人工序列

<220>

<223> BoNT/B轻链cDNA序列

<400> 11

atgccagtta caataaataa ttttaattat aatgatccta ttgataataa taatattatt 60

atgatggagc ctccatttgc gagaggtacg gggagatatt ataaagcttt taaaatcaca 120

gatcgtattt ggataatacc ggaaagatat acttttggat ataaacctga ggattttaat 180

aaaagttccg gtatttttaa tagagatgtt tgtgaatatt atgatccaga ttacttaaat 240

actaatgata aaaagaatat atttttacaa acaatgatca agttatttaa tagaatcaaa 300

tcaaaaccat tgggtgaaaa gttattagag atgattataa atggtatacc ttatcttgga 360

gatagacgtg ttccactcga agagtttaac acaaacattg ctagtgtaac tgttaataaa 420

ttaatcagta atccaggaga agtggagcga aaaaaaggta ttttcgcaaa tttaataata 480

tttggacctg ggccagtttt aaatgaaaat gagactatag atataggtat acaaaatcat 540

tttgcatcaa gggaaggctt cgggggtata atgcaaatga agttttgccc agaatatgta 600

agcgtattta ataatgttca agaaaacaaa ggcgcaagta tatttaatag acgtggatat 660

ttttcagatc cagccttgat attaatgcat gaacttatac atgttttaca tggattatat 720

ggcattaaag tagatgattt accaattgta ccaaatgaaa aaaaattttt tatgcaatct 780

acagatgcta tacaggcaga agaactatat acatttggag gacaagatcc cagcatcata 840

actccttcta cggataaaag tatctatgat aaagttttgc aaaattttag agggatagtt 900

gatagactta acaaggtttt agtttgcata tcagatccta acattaatat taatatatat 960

aaaaataaat ttaaagataa atataaattc gttgaagatt ctgagggaaa atatagtata 1020

gatgtagaaa gttttgataa attatataaa agcttaatgt ttggttttac agaaactaat 1080

atagcagaaa attataaaat aaaaactaga gcttcttatt ttagtgattc cttaccacca 1140

gtaaaaataa aaaatttatt agataatgaa atctatacta tagaggaagg gtttaatata 1200

tctgataaag atatggaaaa agaatataga ggtcagaata aagctataaa taaacaagct 1260

tatgaagaaa ttagcaagga gcatttggct gtatataaga tacaaatgtg taaaagtgtt 1320

aaa 1323

<210> 12

<211> 1347

<212> DNA

<213> 人工序列

<220>

<223> BoNT/C轻链cDNA序列

<400> 12

atgccaataa caattaacaa ctttaattat tcagatcctg ttgataataa aaatatttta 60

tatttagata ctcatttaaa tacactagct aatgagcctg aaaaagcctt tcgcattaca 120

ggaaatatat gggtaatacc tgatagattt tcaagaaatt ctaatccaaa tttaaataaa 180

cctcctcgag ttacaagccc taaaagtggt tattatgatc ctaattattt gagtactgat 240

tctgacaaag atacattttt aaaagaaatt ataaagttat ttaaaagaat taattctaga 300

gaaataggag aagaattaat atatagactt tcgacagata taccctttcc tgggaataac 360

aatactccaa ttaatacttt tgattttgat gtagatttta acagtgttga tgttaaaact 420

agacaaggta acaactgggt taaaactggt agcataaatc ctagtgttat aataactgga 480

cctagagaaa acattataga tccagaaact tctacgttta aattaactaa caatactttt 540

gcggcacaag aaggatttgg tgctttatca ataatttcaa tatcacctag atttatgcta 600

acatatagta atgcaactaa tgatgtagga gagggtagat tttctaagtc tgaattttgc 660

atggatccaa tactaatttt aatgcatgaa cttaatcatg caatgcataa tttatatgga 720

atagctatac caaatgatca aacaatttca tctgtaacta gtaatatttt ttattctcaa 780

tataatgtga aattagagta tgcagaaata tatgcatttg gaggtccaac tatagacctt 840

attcctaaaa gtgcaaggaa atattttgag gaaaaggcat tggattatta tagatctata 900

gctaaaagac ttaatagtat aactactgca aatccttcaa gctttaataa atatataggg 960

gaatataaac agaaacttat tagaaagtat agattcgtag tagaatcttc aggtgaagtt 1020

acagtaaatc gtaataagtt tgttgagtta tataatgaac ttacacaaat atttacagaa 1080

tttaactacg ctaaaatata taatgtacaa aataggaaaa tatatctttc aaatgtatat 1140

actccggtta cggcgaatat attagacgat aatgtttatg atatacaaaa tggatttaat 1200

atacctaaaa gtaatttaaa tgtactattt atgggtcaaa atttatctcg aaatccagca 1260

ttaagaaaag tcaatcctga aaatatgctt tatttattta caaaattttg tcataaagca 1320

atagatggta gatcattata taataaa 1347

<210> 13

<211> 1326

<212> DNA

<213> 人工序列

<220>

<223> BoNT/D轻链cDNA序列

<400> 13

atgacatggc cagtaaaaga ttttaattat agtgatcctg ttaatgacaa tgatatatta 60

tatttaagaa taccacaaaa taagttaatt actacacctg taaaagcttt tatgattact 120

caaaatattt gggtaatacc agaaagattt tcatcagata ctaatccaag tttaagtaaa 180

ccgcccagac ctacttcaaa gtatcaaagt tattatgatc ctagttattt atctactgat 240

gaacaaaaag atacattttt aaaagggatt ataaaattat ttaaaagaat taatgaaaga 300

gatataggaa aaaaattaat aaattattta gtagttggtt caccttttat gggagattca 360

agtacgcctg aagatacatt tgattttaca cgtcatacta ctaatattgc agttgaaaag 420

tttgaaaatg gtagttggaa agtaacaaat attataacac caagtgtatt gatatttgga 480

ccacttccta atatattaga ctatacagca tcccttacat tgcaaggaca acaatcaaat 540

ccatcatttg aagggtttgg aacattatct atactaaaag tagcacctga atttttgtta 600

acatttagtg atgtaacatc taatcaaagt tcagctgtat taggcaaatc tatattttgt 660

atggatccag taatagcttt aatgcatgag ttaacacatt ctttgcatca attatatgga 720

ataaatatac catctgataa aaggattcgt ccacaagtta gcgagggatt tttctctcaa 780

gatggaccca acgtacaatt tgaggaatta tatacatttg gaggattaga tgttgaaata 840

atacctcaaa ttgaaagatc acaattaaga gaaaaagcat taggtcacta taaagatata 900

gcgaaaagac ttaataatat taataaaact attccttcta gttggattag taatatagat 960

aaatataaaa aaatattttc tgaaaagtat aattttgata aagataatac aggaaatttt 1020

gttgtaaata ttgataaatt caatagctta tattcagact tgactaatgt tatgtcagaa 1080

gttgtttatt cttcgcaata taatgttaaa aacaggactc attatttttc aaggcattat 1140

ctacctgtat ttgcaaatat attagatgat aatatttata ctataagaga tggttttaat 1200

ttaacaaata aaggttttaa tatagaaaat tcgggtcaga atatagaaag gaatcctgca 1260

ctacaaaagc ttagttcaga aagtgtagta gatttattta caaaagtatg tttaagatta 1320

acaaaa 1326

<210> 14

<211> 1266

<212> DNA

<213> 人工序列

<220>

<223> BoNT/E轻链cDNA序列

<400> 14

atgccaacaa ttaatagttt taattataat gatcctgtta ataatagaac aattttatat 60

attaaaccag gcggttgtca acaattttat aaatcattta atattatgaa aaatatttgg 120

ataattccag agagaaatgt aattggtaca attccccaag attttcttcc gcctacttca 180

ttgaaaaatg gagatagtag ttattatgac cctaattatt tacaaagtga tcaagaaaag 240

gataaatttt taaaaatagt cacaaaaata tttaatagaa taaatgataa tctttcagga 300

aggattttat tagaagaact gtcaaaagct aatccatatt taggaaatga taatactcca 360

gatggtgact tcattattaa tgatgcatca gcagttccaa ttcaattctc aaatggtagc 420

caaagcatac tattacctaa tgttattata atgggagcag agcctgattt atttgaaact 480

aacagttcca atatttctct aagaaataat tatatgccaa gcaatcacgg ttttggatca 540

atagctatag taacattctc acctgaatat tcttttagat ttaaagataa tagtatgaat 600

gaatttattc aagatcctgc tcttacatta atgcatgaat taatacattc attacatgga 660

ctatatgggg ctaaagggat tactacaaag tatactataa cacaaaaaca aaatccccta 720

ataacaaata taagaggtac aaatattgaa gaattcttaa cttttggagg tactgattta 780

aacattatta ctagtgctca gtccaatgat atctatacta atcttctagc tgattataaa 840

aaaatagcgt ctaaacttag caaagtacaa gtatctaatc cactacttaa tccttataaa 900

gatgtttttg aagcaaagta tggattagat aaagatgcta gcggaattta ttcggtaaat 960

ataaacaaat ttaatgatat ttttaaaaaa ttatacagct ttacggaatt tgatttagca 1020

actaaatttc aagttaaatg taggcaaact tatattggac agtataaata cttcaaactt 1080

tcaaacttgt taaatgattc tatttataat atatcagaag gctataatat aaataattta 1140

aaggtaaatt ttagaggaca gaatgcaaat ttaaatccta gaattattac accaattaca 1200

ggtagaggac tagtaaaaaa aatcattaga ttttgtaaaa atattgtttc tgtaaaaggc 1260

ataagg 1266

<210> 15

<211> 1308

<212> DNA

<213> 人工序列

<220>

<223> BoNT/F轻链cDNA序列

<400> 15

atgccagttg caataaatag ttttaattat aatgaccctg ttaatgatga tacaatttta 60

tacatgcaga taccatatga agaaaaaagt aaaaaatatt ataaagcttt tgagattatg 120

cgtaatgttt ggataattcc tgagagaaat acaataggaa cgaatcctag tgattttgat 180

ccaccggctt cattaaagaa cggaagcagt gcttattatg atcctaatta tttaaccact 240

gatgctgaaa aagatagata tttaaaaaca acgataaaat tatttaagag aattaatagt 300

aatcctgcag ggaaagtttt gttacaagaa atatcatatg ctaaaccata tttaggaaat 360

gaccacacgc caattgatga attctctcca gttactagaa ctacaagtgt taatataaaa 420

ttatcaacta atgttgaaag ttcaatgtta ttgaatcttc ttgtattggg agcaggacct 480

gatatatttg aaagttgttg ttaccccgtt agaaaactaa tagatccaga tgtagtttat 540

gatccaagta attatggttt tggatcaatt aatatcgtga cattttcacc tgagtatgaa 600

tatactttta atgatattag tggagggcat aatagtagta cagaatcatt tattgcagat 660

cctgcaattt cactagctca tgaattgata catgcactgc atggattata cggggctagg 720

ggagttactt atgaagagac tatagaagta aagcaagcac ctcttatgat agccgaaaaa 780

cccataaggc tagaagaatt tttaaccttt ggaggtcagg atttaaatat tattactagt 840

gctatgaagg aaaaaatata taacaatctt ttagctaact atgaaaaaat agctactaga 900

cttagtgaag ttaatagtgc tcctcctgaa tatgatatta atgaatataa agattatttt 960

caatggaagt atgggctaga taaaaatgct gatggaagtt atactgtaaa tgaaaataaa 1020

tttaatgaaa tttataaaaa attatatagt tttacagaga gtgacttagc aaataaattt 1080

aaagtaaaat gtagaaatac ttattttatt aaatatgaat ttttaaaagt tccaaatttg 1140

ttagatgatg atatttatac tgtatcagag gggtttaata taggtaattt agcagtaaac 1200

aatcgcggac aaagtataaa gttaaatcct aaaattattg attccattcc agataaaggt 1260

ctagtagaaa agatcgttaa attttgtaag agcgttattc ctagaaaa 1308

<210> 16

<211> 1326

<212> DNA

<213> 人工序列

<220>

<223> BoNT/G轻链cDNA序列

<400> 16

atgccagtta atataaaaan ctttaattat aatgacccta ttaataatga tgacattatt 60

atgatggaac cattcaatga cccagggcca ggaacatatt ataaagcttt taggattata 120

gatcgtattt ggatagtacc agaaaggttt acttatggat ttcaacctga ccaatttaat 180

gccagtacag gagtttttag taaagatgtc tacgaatatt acgatccaac ttatttaaaa 240

accgatgctg aaaaagataa atttttaaaa acaatgatta aattatttaa tagaattaat 300

tcaaaaccat caggacagag attactggat atgatagtag atgctatacc ttatcttgga 360

aatgcatcta caccgcccga caaatttgca gcaaatgttg caaatgtatc tattaataaa 420

aaaattatcc aacctggagc tgaagatcaa ataaaaggtt taatgacaaa tttaataata 480

tttggaccag gaccagttct aagtgataat tttactgata gtatgattat gaatggccat 540

tccccaatat cagaaggatt tggtgcaaga atgatgataa gattttgtcc tagttgttta 600

aatgtattta ataatgttca ggaaaataaa gatacatcta tatttagtag acgcgcgtat 660

tttgcagatc cagctctaac gttaatgcat gaacttatac atgtgttaca tggattatat 720

ggaattaaga taagtaattt accaattact ccaaatacaa aagaattttt catgcaacat 780

agcgatcctg tacaagcaga agaactatat acattcggag gacatgatcc tagtgttata 840

agtccttcta cggatatgaa tatttataat aaagcgttac aaaattttca agatatagct 900

aataggctta atattgtttc aagtgcccaa gggagtggaa ttgatatttc cttatataaa 960

caaatatata aaaataaata tgattttgtt gaagatccta atggaaaata tagtgtagat 1020

aaggataagt ttgataaatt atataaggcc ttaatgtttg gctttactga aactaatcta 1080

gctggtgaat atggaataaa aactaggtat tcttatttta gtgaatattt gccaccgata 1140

aaaactgaaa aattgttaga caatacaatt tatactcaaa atgaaggctt taacatagct 1200

agtaaaaatc tcaaaacgga atttaatggt cagaataagg cggtaaataa agaggcttat 1260

gaagaaatca gcctagaaca tctcgttata tatagaatag caatgtgcaa gcctgtaatg 1320

tacaaa 1326

<210> 17

<211> 468

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/A轻链氨基酸序列

<400> 17

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro

1 5 10 15

Arg Gly Ser His Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp

20 25 30

Pro Val Asn Gly Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly

35 40 45

Gln Met Gln Pro Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val

50 55 60

Ile Pro Glu Arg Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn

65 70 75 80

Pro Pro Pro Glu Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr

85 90 95

Tyr Leu Ser Thr Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr

100 105 110

Lys Leu Phe Glu Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu

115 120 125

Thr Ser Ile Val Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp

130 135 140

Thr Glu Leu Lys Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro

145 150 155 160

Asp Gly Ser Tyr Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro

165 170 175

Ser Ala Asp Ile Ile Gln Phe Glu Cys Lys Ser Phe Gly His Glu Val

180 185 190

Leu Asn Leu Thr Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe

195 200 205

Ser Pro Asp Phe Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr

210 215 220

Asn Pro Leu Leu Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr

225 230 235 240

Leu Ala His Glu Leu Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala

245 250 255

Ile Asn Pro Asn Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu

260 265 270

Met Ser Gly Leu Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly

275 280 285

His Asp Ala Lys Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu

290 295 300

Tyr Tyr Tyr Asn Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala

305 310 315 320

Lys Ser Ile Val Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val

325 330 335

Phe Lys Glu Lys Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser

340 345 350

Val Asp Lys Leu Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile

355 360 365

Tyr Thr Glu Asp Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys

370 375 380

Thr Tyr Leu Asn Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro

385 390 395 400

Lys Val Asn Tyr Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn

405 410 415

Leu Ala Ala Asn Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn

420 425 430

Phe Thr Lys Leu Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu

435 440 445

Leu Cys Val Arg Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Lys

450 455 460

Gly Tyr Asn Lys

465

<210> 18

<211> 461

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/B轻链氨基酸序列

<400> 18

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro

1 5 10 15

Arg Gly Ser His Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp

20 25 30

Pro Ile Asp Asn Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg

35 40 45

Gly Thr Gly Arg Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp

50 55 60

Ile Ile Pro Glu Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn

65 70 75 80

Lys Ser Ser Gly Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro

85 90 95

Asp Tyr Leu Asn Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met

100 105 110

Ile Lys Leu Phe Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu

115 120 125

Leu Glu Met Ile Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val

130 135 140

Pro Leu Glu Glu Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys

145 150 155 160

Leu Ile Ser Asn Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala

165 170 175

Asn Leu Ile Ile Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr

180 185 190

Ile Asp Ile Gly Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly

195 200 205

Gly Ile Met Gln Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn

210 215 220

Asn Val Gln Glu Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr

225 230 235 240

Phe Ser Asp Pro Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu

245 250 255

His Gly Leu Tyr Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn

260 265 270

Glu Lys Lys Phe Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu

275 280 285

Leu Tyr Thr Phe Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr

290 295 300

Asp Lys Ser Ile Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val

305 310 315 320

Asp Arg Leu Asn Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn

325 330 335

Ile Asn Ile Tyr Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu

340 345 350

Asp Ser Glu Gly Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu

355 360 365

Tyr Lys Ser Leu Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn

370 375 380

Tyr Lys Ile Lys Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro

385 390 395 400

Val Lys Ile Lys Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu

405 410 415

Gly Phe Asn Ile Ser Asp Lys Asp Met Glu Lys Glu Tyr Arg Gly Gln

420 425 430

Asn Lys Ala Ile Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His

435 440 445

Leu Ala Val Tyr Lys Ile Gln Met Cys Lys Ser Val Lys

450 455 460

<210> 19

<211> 469

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/C轻链氨基酸序列

<400> 19

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro

1 5 10 15

Arg Gly Ser His Met Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp

20 25 30

Pro Val Asp Asn Lys Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr

35 40 45

Leu Ala Asn Glu Pro Glu Lys Ala Phe Arg Ile Thr Gly Asn Ile Trp

50 55 60

Val Ile Pro Asp Arg Phe Ser Arg Asn Ser Asn Pro Asn Leu Asn Lys

65 70 75 80

Pro Pro Arg Val Thr Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr

85 90 95

Leu Ser Thr Asp Ser Asp Lys Asp Pro Phe Leu Lys Glu Ile Ile Lys

100 105 110

Leu Phe Lys Arg Ile Asn Ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr

115 120 125

Arg Leu Ser Thr Asp Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile

130 135 140

Asn Thr Phe Asp Phe Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr

145 150 155 160

Arg Gln Gly Asn Asn Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val

165 170 175

Ile Ile Thr Gly Pro Arg Glu Asn Ile Ile Asp Pro Glu Thr Ser Thr

180 185 190

Phe Lys Leu Thr Asn Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala

195 200 205

Leu Ser Ile Ile Ser Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn

210 215 220

Ala Thr Asn Asp Val Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys

225 230 235 240

Met Asp Pro Ile Leu Ile Leu Met His Glu Leu Asn His Ala Met His

245 250 255

Asn Leu Tyr Gly Ile Ala Ile Pro Asn Asp Gln Thr Ile Ser Ser Val

260 265 270

Thr Ser Asn Ile Phe Tyr Ser Gln Tyr Asn Val Lys Leu Glu Tyr Ala

275 280 285

Glu Ile Tyr Ala Phe Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser

290 295 300

Ala Arg Lys Tyr Phe Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile

305 310 315 320

Ala Lys Arg Leu Asn Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn

325 330 335

Lys Tyr Ile Gly Glu Tyr Lys Gln Lys Leu Ile Arg Lys Tyr Arg Phe

340 345 350

Val Val Glu Ser Ser Gly Glu Val Thr Val Asn Arg Asn Lys Phe Val

355 360 365

Glu Leu Tyr Asn Glu Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala

370 375 380

Lys Ile Tyr Asn Val Gln Asn Arg Lys Ile Tyr Leu Ser Asn Val Tyr

385 390 395 400

Thr Pro Val Thr Ala Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln

405 410 415

Asn Gly Phe Asn Ile Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly

420 425 430

Gln Asn Leu Ser Arg Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn

435 440 445

Met Leu Tyr Leu Phe Thr Lys Phe Cys His Lys Ala Ile Asp Gly Arg

450 455 460

Ser Leu Tyr Asn Lys

465

<210> 20

<211> 462

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/D轻链氨基酸序列

<400> 20

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro

1 5 10 15

Arg Gly Ser His Met Thr Trp Pro Val Lys Asp Phe Asn Tyr Ser Asp

20 25 30

Pro Val Asn Asp Asn Asp Ile Leu Tyr Leu Arg Ile Pro Gln Asn Lys

35 40 45

Leu Ile Thr Thr Pro Val Lys Ala Phe Met Ile Thr Gln Asn Ile Trp

50 55 60

Val Ile Pro Glu Arg Phe Ser Ser Asp Thr Asn Pro Ser Leu Ser Lys

65 70 75 80

Pro Pro Arg Pro Thr Ser Lys Tyr Gln Ser Tyr Tyr Asp Pro Ser Tyr

85 90 95

Leu Ser Thr Asp Glu Gln Lys Asp Thr Phe Leu Lys Gly Ile Ile Lys

100 105 110

Leu Phe Lys Arg Ile Asn Glu Arg Asp Ile Gly Lys Lys Leu Ile Asn

115 120 125

Tyr Leu Val Val Gly Ser Pro Phe Met Gly Asp Ser Ser Thr Pro Glu

130 135 140

Asp Thr Phe Asp Phe Thr Arg His Thr Thr Asn Ile Ala Val Glu Lys

145 150 155 160

Phe Glu Asn Gly Ser Trp Lys Val Thr Asn Ile Ile Thr Pro Ser Val

165 170 175

Leu Ile Phe Gly Pro Leu Pro Asn Ile Leu Asp Tyr Thr Ala Ser Leu

180 185 190

Thr Leu Gln Gly Gln Gln Ser Asn Pro Ser Phe Glu Gly Phe Gly Thr

195 200 205

Leu Ser Ile Leu Lys Val Ala Pro Glu Phe Leu Leu Thr Phe Ser Asp

210 215 220

Val Thr Ser Asn Gln Ser Ser Ala Val Leu Gly Lys Ser Ile Phe Cys

225 230 235 240

Met Asp Pro Val Ile Ala Leu Met His Glu Leu Thr His Ser Leu His

245 250 255

Gln Leu Tyr Gly Ile Asn Ile Pro Ser Asp Lys Arg Ile Arg Pro Gln

260 265 270

Val Ser Glu Gly Phe Phe Ser Gln Asp Gly Pro Asn Val Gln Phe Glu

275 280 285

Glu Leu Tyr Thr Phe Gly Gly Leu Asp Val Glu Ile Ile Pro Gln Ile

290 295 300

Glu Arg Ser Gln Leu Arg Glu Lys Ala Leu Gly His Tyr Lys Asp Ile

305 310 315 320

Ala Lys Arg Leu Asn Asn Ile Asn Lys Thr Ile Pro Ser Ser Trp Ile

325 330 335

Ser Asn Ile Asp Lys Tyr Lys Lys Ile Phe Ser Glu Lys Tyr Asn Phe

340 345 350

Asp Lys Asp Asn Thr Gly Asn Phe Val Val Asn Ile Asp Lys Phe Asn

355 360 365

Ser Leu Tyr Ser Asp Leu Thr Asn Val Met Ser Glu Val Val Tyr Ser

370 375 380

Ser Gln Tyr Asn Val Lys Asn Arg Thr His Tyr Phe Ser Arg His Tyr

385 390 395 400

Leu Pro Val Phe Ala Asn Ile Leu Asp Asp Asn Ile Tyr Thr Ile Arg

405 410 415

Asp Gly Phe Asn Leu Thr Asn Lys Gly Phe Asn Ile Glu Asn Ser Gly

420 425 430

Gln Asn Ile Glu Arg Asn Pro Ala Leu Gln Lys Leu Ser Ser Glu Ser

435 440 445

Val Val Asp Leu Phe Thr Lys Val Cys Leu Arg Leu Thr Lys

450 455 460

<210> 21

<211> 442

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/E轻链氨基酸序列

<400> 21

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro

1 5 10 15

Arg Gly Ser His Met Pro Thr Ile Asn Ser Phe Asn Tyr Asn Asp Pro

20 25 30

Val Asn Asn Arg Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln

35 40 45

Phe Tyr Lys Ser Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu

50 55 60

Arg Asn Val Ile Gly Thr Ile Pro Gln Asp Phe Leu Pro Pro Thr Ser

65 70 75 80

Leu Lys Asn Gly Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser

85 90 95

Asp Gln Glu Lys Asp Lys Phe Leu Lys Ile Val Thr Lys Ile Phe Asn

100 105 110

Arg Ile Asn Asp Asn Leu Ser Gly Arg Ile Leu Leu Glu Glu Leu Ser

115 120 125

Lys Ala Asn Pro Tyr Leu Gly Asn Asp Asn Thr Pro Asp Gly Asp Phe

130 135 140

Ile Ile Asn Asp Ala Ser Ala Val Pro Ile Gln Phe Ser Asn Gly Ser

145 150 155 160

Gln Ser Ile Leu Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp

165 170 175

Leu Phe Glu Thr Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met

180 185 190

Pro Ser Asn His Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro

195 200 205

Glu Tyr Ser Phe Arg Phe Lys Asp Asn Ser Met Asn Glu Phe Ile Gln

210 215 220

Asp Pro Ala Leu Thr Leu Met His Glu Leu Ile His Ser Leu His Gly

225 230 235 240

Leu Tyr Gly Ala Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Lys

245 250 255

Gln Asn Pro Leu Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe

260 265 270

Leu Thr Phe Gly Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Ser

275 280 285

Asn Asp Ile Tyr Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser

290 295 300

Lys Leu Ser Lys Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys

305 310 315 320

Asp Val Phe Glu Ala Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile

325 330 335

Tyr Ser Val Asn Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr

340 345 350

Ser Phe Thr Glu Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg

355 360 365

Gln Thr Tyr Ile Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu

370 375 380

Asn Asp Ser Ile Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu

385 390 395 400

Lys Val Asn Phe Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile

405 410 415

Thr Pro Ile Thr Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys

420 425 430

Lys Asn Ile Val Ser Val Lys Gly Ile Arg

435 440

<210> 22

<211> 456

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/F轻链氨基酸序列

<400> 22

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro

1 5 10 15

Arg Gly Ser His Met Pro Val Ala Ile Asn Ser Phe Asn Tyr Asn Asp

20 25 30

Pro Val Asn Asp Asp Thr Ile Leu Tyr Met Gln Ile Pro Tyr Glu Glu

35 40 45

Lys Ser Lys Lys Tyr Tyr Lys Ala Phe Glu Ile Met Arg Asn Val Trp

50 55 60

Ile Ile Pro Glu Arg Asn Thr Ile Gly Thr Asn Pro Ser Asp Phe Asp

65 70 75 80

Pro Pro Ala Ser Leu Lys Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn

85 90 95

Tyr Leu Thr Thr Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr Ile

100 105 110

Lys Leu Phe Lys Arg Ile Asn Ser Asn Pro Ala Gly Lys Val Leu Leu

115 120 125

Gln Glu Ile Ser Tyr Ala Lys Pro Tyr Leu Gly Asn Asp His Thr Pro

130 135 140

Ile Asp Glu Phe Ser Pro Val Thr Arg Thr Thr Ser Val Asn Ile Lys

145 150 155 160

Leu Ser Thr Asn Val Glu Ser Ser Met Leu Leu Asn Leu Leu Val Leu

165 170 175

Gly Ala Gly Pro Asp Ile Phe Glu Ser Cys Cys Tyr Pro Val Arg Lys

180 185 190

Leu Ile Asp Pro Asp Val Val Tyr Asp Pro Ser Asn Tyr Gly Phe Gly

195 200 205

Ser Ile Asn Ile Val Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn

210 215 220

Asp Ile Ser Gly Gly His Asn Ser Ser Thr Glu Ser Phe Ile Ala Asp

225 230 235 240

Pro Ala Ile Ser Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu

245 250 255

Tyr Gly Ala Arg Gly Val Thr Tyr Glu Glu Thr Ile Glu Val Lys Gln

260 265 270

Ala Pro Leu Met Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu

275 280 285

Thr Phe Gly Gly Gln Asp Leu Asn Ile Ile Thr Ser Ala Met Lys Glu

290 295 300

Lys Ile Tyr Asn Asn Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg

305 310 315 320

Leu Ser Glu Val Asn Ser Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr

325 330 335

Lys Asp Tyr Phe Gln Trp Lys Tyr Gly Leu Asp Lys Asn Ala Asp Gly

340 345 350

Ser Tyr Thr Val Asn Glu Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu

355 360 365

Tyr Ser Phe Thr Glu Ser Asp Leu Ala Asn Lys Phe Lys Val Lys Cys

370 375 380

Arg Asn Thr Tyr Phe Ile Lys Tyr Glu Phe Leu Lys Val Pro Asn Leu

385 390 395 400

Leu Asp Asp Asp Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn

405 410 415

Leu Ala Val Asn Asn Arg Gly Gln Ser Ile Lys Leu Asn Pro Lys Ile

420 425 430

Ile Asp Ser Ile Pro Asp Lys Gly Leu Val Glu Lys Ile Val Lys Phe

435 440 445

Cys Lys Ser Val Ile Pro Arg Lys

450 455

<210> 23

<211> 461

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/G轻链氨基酸序列

<400> 23

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro

1 5 10 15

Arg Gly Ser His Met Pro Val Asn Ile Lys Phe Asn Tyr Asn Asp Pro

20 25 30

Ile Asn Asn Asp Asp Ile Ile Met Met Glu Pro Phe Asn Asp Pro Gly

35 40 45

Pro Gly Thr Tyr Tyr Lys Ala Phe Arg Ile Ile Asp Arg Ile Trp Ile

50 55 60

Val Pro Glu Arg Phe Thr Tyr Gly Phe Gln Pro Asp Gln Phe Asn Ala

65 70 75 80

Ser Thr Gly Val Phe Ser Lys Asp Val Tyr Glu Tyr Tyr Asp Pro Thr

85 90 95

Tyr Leu Lys Thr Asp Ala Glu Lys Asp Lys Phe Leu Lys Thr Met Ile

100 105 110

Lys Leu Phe Asn Arg Ile Asn Ser Lys Pro Ser Gly Gln Arg Leu Leu

115 120 125

Asp Met Ile Val Asp Ala Ile Pro Tyr Leu Gly Asn Ala Ser Thr Pro

130 135 140

Pro Asp Lys Phe Ala Ala Asn Val Ala Asn Val Ser Ile Asn Lys Lys

145 150 155 160

Ile Ile Gln Pro Gly Ala Glu Asp Gln Ile Lys Gly Leu Met Thr Asn

165 170 175

Leu Ile Ile Phe Gly Pro Gly Pro Val Leu Ser Asp Asn Phe Thr Asp

180 185 190

Ser Met Ile Met Asn Gly His Ser Pro Ile Ser Glu Gly Phe Gly Ala

195 200 205

Arg Met Met Ile Arg Phe Cys Pro Ser Cys Leu Asn Val Phe Asn Asn

210 215 220

Val Gln Glu Asn Lys Asp Thr Ser Ile Phe Ser Arg Arg Ala Tyr Phe

225 230 235 240

Ala Asp Pro Ala Leu Thr Leu Met His Glu Leu Ile His Val Leu His

245 250 255

Gly Leu Tyr Gly Ile Lys Ile Ser Asn Leu Pro Ile Thr Pro Asn Thr

260 265 270

Lys Glu Phe Phe Met Gln His Ser Asp Pro Val Gln Ala Glu Glu Leu

275 280 285

Tyr Thr Phe Gly Gly His Asp Pro Ser Val Ile Ser Pro Ser Thr Asp

290 295 300

Met Asn Ile Tyr Asn Lys Ala Leu Gln Asn Phe Gln Asp Ile Ala Asn

305 310 315 320

Arg Leu Asn Ile Val Ser Ser Ala Gln Gly Ser Gly Ile Asp Ile Ser

325 330 335

Leu Tyr Lys Gln Ile Tyr Lys Asn Lys Tyr Asp Phe Val Glu Asp Pro

340 345 350

Asn Gly Lys Tyr Ser Val Asp Lys Asp Lys Phe Asp Lys Leu Tyr Lys

355 360 365

Ala Leu Met Phe Gly Phe Thr Glu Thr Asn Leu Ala Gly Glu Tyr Gly

370 375 380

Ile Lys Thr Arg Tyr Ser Tyr Phe Ser Glu Tyr Leu Pro Pro Ile Lys

385 390 395 400

Thr Glu Lys Leu Leu Asp Asn Thr Ile Tyr Thr Gln Asn Glu Gly Phe

405 410 415

Asn Ile Ala Ser Lys Asn Leu Lys Thr Glu Phe Asn Gly Gln Asn Lys

420 425 430

Ala Val Asn Lys Glu Ala Tyr Glu Glu Ile Ser Leu Glu His Leu Val

435 440 445

Ile Tyr Arg Ile Ala Met Cys Lys Pro Val Met Tyr Lys

450 455 460

<210> 24

<211> 1437

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/A轻链cDNA序列

<400> 24

atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60

atgatgccct ttgtcaacaa acagttcaac tacaaggacc cagttaatgg agtagacatc 120

gcatatatca agattcccaa cgctggccag atgcaacccg ttaaggcatt taaaatccat 180

aacaaaatct gggttatccc agagcgggat accttcacca accccgagga gggcgatctg 240

aaccccccgc cggaggcgaa gcaggtccca gtgagctact acgatagcac ctacctcagc 300

accgacaacg agaaggacaa ctacctcaaa ggagtcacga agttgttcga gagaatctac 360

tccacagacc tcggccgcat gcttctaacc agcattgtgc gtggcattcc cttttggggc 420

ggctctacca tcgacacaga gctgaaggtg atagacacca actgcatcaa cgtaatccag 480

cctgacggca gctaccgaag cgaggagctt aacctggtga tcatcggccc ttccgccgat 540

atcatccaat tcgagtgcaa gagcttcggc cacgaggtcc tgaacctcac ccggaacggc 600

tatggaagca cccagtacat aagattcagc cctgacttca ccttcgggtt tgaggagagc 660

ttggaggtcg acacaaaccc cctgctggga gccgggaagt tcgccactga cccagccgtg 720

actctggcac acgagctgat ccacgccggt caccgcctgt acggcatagc tataaaccca 780

aacagggtgt tcaaagtgaa caccaacgct tactatgaaa tgagcggcct ggaggtgagc 840

ttcgaggagc tgagaacgtt cgggggacat gatgctaaat ttatcgacag cctgcaggag 900

aacgagttca ggctgtacta ctacaataag ttcaaggata tagcgagcac tctgaacaag 960

gccaagtcca tcgtaggcac tactgcatcc ctccagtata tgaagaatgt gttcaaagag 1020

aaatacctgc tgagcgagga taccagcggt aagttcagcg tggataagct taagttcgac 1080

aagctgtata agatgctcac cgaaatctac accgaggata atttcgttaa gttcttcaag 1140

gtcctgaacc ggaagaccta cctgaacttc gacaaggccg tgttcaagat caacatcgtg 1200

cctaaagtga actacaccat ctacgacggg tttaacctga ggaacaccaa cctggccgct 1260

aacttcaacg ggcagaacac agagatcaac aacatgaatt tcacgaagtt gaagaacttc 1320

accggactgt ttgagttcta caaattgctg tgtgtgcgcg ggatcatcac tagcaagacc 1380

aagagccttg acaaaggcta caacaagtga ctcgagcacc accaccacca ccactga 1437

<210> 25

<211> 1413

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/B轻链cDNA序列

<400> 25

atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60

atgatgccag ttacaataaa taattttaat tataatgatc ctattgataa taataatatt 120

attatgatgg agcctccatt tgcgagaggt acggggagat attataaagc ttttaaaatc 180

acagatcgta tttggataat accggaaaga tatacttttg gatataaacc tgaggatttt 240

aataaaagtt ccggtatttt taatagagat gtttgtgaat attatgatcc agattactta 300

aatactaatg ataaaaagaa tatattttta caaacaatga tcaagttatt taatagaatc 360

aaatcaaaac cattgggtga aaagttatta gagatgatta taaatggtat accttatctt 420

ggagatagac gtgttccact cgaagagttt aacacaaaca ttgctagtgt aactgttaat 480

aaattaatca gtaatccagg agaagtggag cgaaaaaaag gtattttcgc aaatttaata 540

atatttggac ctgggccagt tttaaatgaa aatgagacta tagatatagg tatacaaaat 600

cattttgcat caagggaagg cttcgggggt ataatgcaaa tgaagttttg cccagaatat 660

gtaagcgtat ttaataatgt tcaagaaaac aaaggcgcaa gtatatttaa tagacgtgga 720

tatttttcag atccagcctt gatattaatg catgaactta tacatgtttt acatggatta 780

tatggcatta aagtagatga tttaccaatt gtaccaaatg aaaaaaaatt ttttatgcaa 840

tctacagatg ctatacaggc agaagaacta tatacatttg gaggacaaga tcccagcatc 900

ataactcctt ctacggataa aagtatctat gataaagttt tgcaaaattt tagagggata 960

gttgatagac ttaacaaggt tttagtttgc atatcagatc ctaacattaa tattaatata 1020

tataaaaata aatttaaaga taaatataaa ttcgttgaag attctgaggg aaaatatagt 1080

atagatgtag aaagttttga taaattatat aaaagcttaa tgtttggttt tacagaaact 1140

aatatagcag aaaattataa aataaaaact agagcttctt attttagtga ttccttacca 1200

ccagtaaaaa taaaaaattt attagataat gaaatctata ctatagagga agggtttaat 1260

atatctgata aagatatgga aaaagaatat agaggtcaga ataaagctat aaataaacaa 1320

gcttatgaag aaattagcaa ggagcatttg gctgtatata agatacaaat gtgtaaaagt 1380

gttaaactcg agcaccacca ccaccaccac tga 1413

<210> 26

<211> 1437

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/C轻链cDNA序列

<400> 26

atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60

atgatgccaa taacaattaa caactttaat tattcagatc ctgttgataa taaaaatatt 120

ttatatttag atactcattt aaatacacta gctaatgagc ctgaaaaagc ctttcgcatt 180

acaggaaata tatgggtaat acctgataga ttttcaagaa attctaatcc aaatttaaat 240

aaacctcctc gagttacaag ccctaaaagt ggttattatg atcctaatta tttgagtact 300

gattctgaca aagatacatt tttaaaagaa attataaagt tatttaaaag aattaattct 360

agagaaatag gagaagaatt aatatataga ctttcgacag atataccctt tcctgggaat 420

aacaatactc caattaatac ttttgatttt gatgtagatt ttaacagtgt tgatgttaaa 480

actagacaag gtaacaactg ggttaaaact ggtagcataa atcctagtgt tataataact 540

ggacctagag aaaacattat agatccagaa acttctacgt ttaaattaac taacaatact 600

tttgcggcac aagaaggatt tggtgcttta tcaataattt caatatcacc tagatttatg 660

ctaacatata gtaatgcaac taatgatgta ggagagggta gattttctaa gtctgaattt 720

tgcatggatc caatactaat tttaatgcat gaacttaatc atgcaatgca taatttatat 780

ggaatagcta taccaaatga tcaaacaatt tcatctgtaa ctagtaatat tttttattct 840

caatataatg tgaaattaga gtatgcagaa atatatgcat ttggaggtcc aactatagac 900

cttattccta aaagtgcaag gaaatatttt gaggaaaagg cattggatta ttatagatct 960

atagctaaaa gacttaatag tataactact gcaaatcctt caagctttaa taaatatata 1020

ggggaatata aacagaaact tattagaaag tatagattcg tagtagaatc ttcaggtgaa 1080

gttacagtaa atcgtaataa gtttgttgag ttatataatg aacttacaca aatatttaca 1140

gaatttaact acgctaaaat atataatgta caaaatagga aaatatatct ttcaaatgta 1200

tatactccgg ttacggcgaa tatattagac gataatgttt atgatataca aaatggattt 1260

aatataccta aaagtaattt aaatgtacta tttatgggtc aaaatttatc tcgaaatcca 1320

gcattaagaa aagtcaatcc tgaaaatatg ctttatttat ttacaaaatt ttgtcataaa 1380

gcaatagatg gtagatcatt atataataaa ctcgagcacc accaccacca ccactga 1437

<210> 27

<211> 1416

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/D轻链cDNA序列

<400> 27

atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60

atgatgacat ggccagtaaa agattttaat tatagtgatc ctgttaatga caatgatata 120

ttatatttaa gaataccaca aaataagtta attactacac ctgtaaaagc ttttatgatt 180

actcaaaata tttgggtaat accagaaaga ttttcatcag atactaatcc aagtttaagt 240

aaaccgccca gacctacttc aaagtatcaa agttattatg atcctagtta tttatctact 300

gatgaacaaa aagatacatt tttaaaaggg attataaaat tatttaaaag aattaatgaa 360

agagatatag gaaaaaaatt aataaattat ttagtagttg gttcaccttt tatgggagat 420

tcaagtacgc ctgaagatac atttgatttt acacgtcata ctactaatat tgcagttgaa 480

aagtttgaaa atggtagttg gaaagtaaca aatattataa caccaagtgt attgatattt 540

ggaccacttc ctaatatatt agactataca gcatccctta cattgcaagg acaacaatca 600

aatccatcat ttgaagggtt tggaacatta tctatactaa aagtagcacc tgaatttttg 660

ttaacattta gtgatgtaac atctaatcaa agttcagctg tattaggcaa atctatattt 720

tgtatggatc cagtaatagc tttaatgcat gagttaacac attctttgca tcaattatat 780

ggaataaata taccatctga taaaaggatt cgtccacaag ttagcgaggg atttttctct 840

caagatggac ccaacgtaca atttgaggaa ttatatacat ttggaggatt agatgttgaa 900

ataatacctc aaattgaaag atcacaatta agagaaaaag cattaggtca ctataaagat 960

atagcgaaaa gacttaataa tattaataaa actattcctt ctagttggat tagtaatata 1020

gataaatata aaaaaatatt ttctgaaaag tataattttg ataaagataa tacaggaaat 1080

tttgttgtaa atattgataa attcaatagc ttatattcag acttgactaa tgttatgtca 1140

gaagttgttt attcttcgca atataatgtt aaaaacagga ctcattattt ttcaaggcat 1200

tatctacctg tatttgcaaa tatattagat gataatattt atactataag agatggtttt 1260

aatttaacaa ataaaggttt taatatagaa aattcgggtc agaatataga aaggaatcct 1320

gcactacaaa agcttagttc agaaagtgta gtagatttat ttacaaaagt atgtttaaga 1380

ttaacaaaac tcgagcacca ccaccaccac cactga 1416

<210> 28

<211> 1356

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/E轻链cDNA序列

<400> 28

atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60

atgatgccaa caattaatag ttttaattat aatgatcctg ttaataatag aacaatttta 120

tatattaaac caggcggttg tcaacaattt tataaatcat ttaatattat gaaaaatatt 180

tggataattc cagagagaaa tgtaattggt acaattcccc aagattttct tccgcctact 240

tcattgaaaa atggagatag tagttattat gaccctaatt atttacaaag tgatcaagaa 300

aaggataaat ttttaaaaat agtcacaaaa atatttaata gaataaatga taatctttca 360

ggaaggattt tattagaaga actgtcaaaa gctaatccat atttaggaaa tgataatact 420

ccagatggtg acttcattat taatgatgca tcagcagttc caattcaatt ctcaaatggt 480

agccaaagca tactattacc taatgttatt ataatgggag cagagcctga tttatttgaa 540

actaacagtt ccaatatttc tctaagaaat aattatatgc caagcaatca cggttttgga 600

tcaatagcta tagtaacatt ctcacctgaa tattctttta gatttaaaga taatagtatg 660

aatgaattta ttcaagatcc tgctcttaca ttaatgcatg aattaataca ttcattacat 720

ggactatatg gggctaaagg gattactaca aagtatacta taacacaaaa acaaaatccc 780

ctaataacaa atataagagg tacaaatatt gaagaattct taacttttgg aggtactgat 840

ttaaacatta ttactagtgc tcagtccaat gatatctata ctaatcttct agctgattat 900

aaaaaaatag cgtctaaact tagcaaagta caagtatcta atccactact taatccttat 960

aaagatgttt ttgaagcaaa gtatggatta gataaagatg ctagcggaat ttattcggta 1020

aatataaaca aatttaatga tatttttaaa aaattataca gctttacgga atttgattta 1080

gcaactaaat ttcaagttaa atgtaggcaa acttatattg gacagtataa atacttcaaa 1140

ctttcaaact tgttaaatga ttctatttat aatatatcag aaggctataa tataaataat 1200

ttaaaggtaa attttagagg acagaatgca aatttaaatc ctagaattat tacaccaatt 1260

acaggtagag gactagtaaa aaaaatcatt agattttgta aaaatattgt ttctgtaaaa 1320

ggcataaggc tcgagcacca ccaccaccac cactga 1356

<210> 29

<211> 1398

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/F轻链cDNA序列

<400> 29

atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60

atgatgccag ttgcaataaa tagttttaat tataatgacc ctgttaatga tgatacaatt 120

ttatacatgc agataccata tgaagaaaaa agtaaaaaat attataaagc ttttgagatt 180

atgcgtaatg tttggataat tcctgagaga aatacaatag gaacgaatcc tagtgatttt 240

gatccaccgg cttcattaaa gaacggaagc agtgcttatt atgatcctaa ttatttaacc 300

actgatgctg aaaaagatag atatttaaaa acaacgataa aattatttaa gagaattaat 360

agtaatcctg cagggaaagt tttgttacaa gaaatatcat atgctaaacc atatttagga 420

aatgaccaca cgccaattga tgaattctct ccagttacta gaactacaag tgttaatata 480

aaattatcaa ctaatgttga aagttcaatg ttattgaatc ttcttgtatt gggagcagga 540

cctgatatat ttgaaagttg ttgttacccc gttagaaaac taatagatcc agatgtagtt 600

tatgatccaa gtaattatgg ttttggatca attaatatcg tgacattttc acctgagtat 660

gaatatactt ttaatgatat tagtggaggg cataatagta gtacagaatc atttattgca 720

gatcctgcaa tttcactagc tcatgaattg atacatgcac tgcatggatt atacggggct 780

aggggagtta cttatgaaga gactatagaa gtaaagcaag cacctcttat gatagccgaa 840

aaacccataa ggctagaaga atttttaacc tttggaggtc aggatttaaa tattattact 900

agtgctatga aggaaaaaat atataacaat cttttagcta actatgaaaa aatagctact 960

agacttagtg aagttaatag tgctcctcct gaatatgata ttaatgaata taaagattat 1020

tttcaatgga agtatgggct agataaaaat gctgatggaa gttatactgt aaatgaaaat 1080

aaatttaatg aaatttataa aaaattatat agttttacag agagtgactt agcaaataaa 1140

tttaaagtaa aatgtagaaa tacttatttt attaaatatg aatttttaaa agttccaaat 1200

ttgttagatg atgatattta tactgtatca gaggggttta atataggtaa tttagcagta 1260

aacaatcgcg gacaaagtat aaagttaaat cctaaaatta ttgattccat tccagataaa 1320

ggtctagtag aaaagatcgt taaattttgt aagagcgtta ttcctagaaa actcgagcac 1380

caccaccacc accactga 1398

<210> 30

<211> 1416

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/G轻链cDNA序列

<400> 30

atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60

atgatgccag ttaatataaa aanctttaat tataatgacc ctattaataa tgatgacatt 120

attatgatgg aaccattcaa tgacccaggg ccaggaacat attataaagc ttttaggatt 180

atagatcgta tttggatagt accagaaagg tttacttatg gatttcaacc tgaccaattt 240

aatgccagta caggagtttt tagtaaagat gtctacgaat attacgatcc aacttattta 300

aaaaccgatg ctgaaaaaga taaattttta aaaacaatga ttaaattatt taatagaatt 360

aattcaaaac catcaggaca gagattactg gatatgatag tagatgctat accttatctt 420

ggaaatgcat ctacaccgcc cgacaaattt gcagcaaatg ttgcaaatgt atctattaat 480

aaaaaaatta tccaacctgg agctgaagat caaataaaag gtttaatgac aaatttaata 540

atatttggac caggaccagt tctaagtgat aattttactg atagtatgat tatgaatggc 600

cattccccaa tatcagaagg atttggtgca agaatgatga taagattttg tcctagttgt 660

ttaaatgtat ttaataatgt tcaggaaaat aaagatacat ctatatttag tagacgcgcg 720

tattttgcag atccagctct aacgttaatg catgaactta tacatgtgtt acatggatta 780

tatggaatta agataagtaa tttaccaatt actccaaata caaaagaatt tttcatgcaa 840

catagcgatc ctgtacaagc agaagaacta tatacattcg gaggacatga tcctagtgtt 900

ataagtcctt ctacggatat gaatatttat aataaagcgt tacaaaattt tcaagatata 960

gctaataggc ttaatattgt ttcaagtgcc caagggagtg gaattgatat ttccttatat 1020

aaacaaatat ataaaaataa atatgatttt gttgaagatc ctaatggaaa atatagtgta 1080

gataaggata agtttgataa attatataag gccttaatgt ttggctttac tgaaactaat 1140

ctagctggtg aatatggaat aaaaactagg tattcttatt ttagtgaata tttgccaccg 1200

ataaaaactg aaaaattgtt agacaataca atttatactc aaaatgaagg ctttaacata 1260

gctagtaaaa atctcaaaac ggaatttaat ggtcagaata aggcggtaaa taaagaggct 1320

tatgaagaaa tcagcctaga acatctcgtt atatatagaa tagcaatgtg caagcctgta 1380

atgtacaaac tcgagcacca ccaccaccac cactga 1416

<210> 31

<211> 461

<212> PRT

<213> 人工序列

<220>

<223> MTD-BoNT/A轻链氨基酸序列

<400> 31

Met Lys Ala Met Ile Asn Ile Asn Lys Phe Leu Asn Gln Cys Pro Phe

1 5 10 15

Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly Val Asp Ile

20 25 30

Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro Val Lys Ala

35 40 45

Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg Asp Thr Phe

50 55 60

Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu Ala Lys Gln

65 70 75 80

Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr Asp Asn Glu

85 90 95

Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu Arg Ile Tyr

100 105 110

Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val Arg Gly Ile

115 120 125

Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys Val Ile Asp

130 135 140

Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr Arg Ser Glu

145 150 155 160

Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile Ile Gln Phe

165 170 175

Glu Cys Lys Ser Phe Gly His Glu Val Leu Asn Leu Thr Arg Asn Gly

180 185 190

Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe Thr Phe Gly

195 200 205

Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu Gly Ala Gly

210 215 220

Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu Leu Ile His

225 230 235 240

Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn Arg Val Phe

245 250 255

Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu Glu Val Ser

260 265 270

Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys Phe Ile Asp

275 280 285

Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn Lys Phe Lys

290 295 300

Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val Gly Thr Thr

305 310 315 320

Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys Tyr Leu Leu

325 330 335

Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu Lys Phe Asp

340 345 350

Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp Asn Phe Val

355 360 365

Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn Phe Asp Lys

370 375 380

Ala Val Phe Lys Ile Asn Ile Val Pro Lys Val Asn Tyr Thr Ile Tyr

385 390 395 400

Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn Phe Asn Gly

405 410 415

Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu Lys Asn Phe

420 425 430

Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg Gly Ile Ile

435 440 445

Thr Ser Lys Thr Lys Ser Leu Asp Lys Gly Tyr Asn Lys

450 455 460

<210> 32

<211> 454

<212> PRT

<213> 人工序列

<220>

<223> MTD-BoNT/B轻链氨基酸序列

<400> 32

Met Lys Ala Met Ile Asn Ile Asn Lys Phe Leu Asn Gln Cys Pro Val

1 5 10 15

Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn Asn Asn Ile

20 25 30

Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg Tyr Tyr Lys

35 40 45

Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu Arg Tyr Thr

50 55 60

Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly Ile Phe Asn

65 70 75 80

Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn Thr Asn Asp

85 90 95

Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe Asn Arg Ile

100 105 110

Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile Ile Asn Gly

115 120 125

Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu Phe Asn Thr

130 135 140

Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn Pro Gly Glu

145 150 155 160

Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile Phe Gly Pro

165 170 175

Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly Ile Gln Asn

180 185 190

His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln Met Lys Phe

195 200 205

Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu Asn Lys Gly

210 215 220

Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro Ala Leu Ile

225 230 235 240

Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr Gly Ile Lys

245 250 255

Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe Phe Met Gln

260 265 270

Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe Gly Gly Gln

275 280 285

Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile Tyr Asp Lys

290 295 300

Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn Lys Val Leu

305 310 315 320

Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr Lys Asn Lys

325 330 335

Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly Lys Tyr Ser

340 345 350

Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu Met Phe Gly

355 360 365

Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys Thr Arg Ala

370 375 380

Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys Asn Leu Leu

385 390 395 400

Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile Ser Asp Lys

405 410 415

Asp Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile Asn Lys Gln

420 425 430

Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr Lys Ile Gln

435 440 445

Met Cys Lys Ser Val Lys

450

<210> 33

<211> 462

<212> PRT

<213> 人工序列

<220>

<223> MTD-BoNT/C轻链氨基酸序列

<400> 33

Met Lys Ala Met Ile Asn Ile Asn Lys Phe Leu Asn Gln Cys Pro Ile

1 5 10 15

Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro Val Asp Asn Lys Asn Ile

20 25 30

Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu Pro Glu Lys

35 40 45

Ala Phe Arg Ile Thr Gly Asn Ile Trp Val Ile Pro Asp Arg Phe Ser

50 55 60

Arg Asn Ser Asn Pro Asn Leu Asn Lys Pro Pro Arg Val Thr Ser Pro

65 70 75 80

Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp Ser Asp Lys

85 90 95

Asp Pro Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg Ile Asn Ser

100 105 110

Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ser Thr Asp Ile Pro

115 120 125

Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp Phe Asp Val

130 135 140

Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn Asn Trp Val

145 150 155 160

Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly Pro Arg Glu

165 170 175

Asn Ile Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr Asn Asn Thr

180 185 190

Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile Ser Ile Ser

195 200 205

Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asp Val Gly Glu

210 215 220

Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile Leu Ile Leu

225 230 235 240

Met His Glu Leu Asn His Ala Met His Asn Leu Tyr Gly Ile Ala Ile

245 250 255

Pro Asn Asp Gln Thr Ile Ser Ser Val Thr Ser Asn Ile Phe Tyr Ser

260 265 270

Gln Tyr Asn Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala Phe Gly Gly

275 280 285

Pro Thr Ile Asp Leu Ile Pro Lys Ser Ala Arg Lys Tyr Phe Glu Glu

290 295 300

Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu Asn Ser Ile

305 310 315 320

Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly Glu Tyr Lys

325 330 335

Gln Lys Leu Ile Arg Lys Tyr Arg Phe Val Val Glu Ser Ser Gly Glu

340 345 350

Val Thr Val Asn Arg Asn Lys Phe Val Glu Leu Tyr Asn Glu Leu Thr

355 360 365

Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn Val Gln Asn

370 375 380

Arg Lys Ile Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr Ala Asn Ile

385 390 395 400

Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn Ile Pro Lys

405 410 415

Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser Arg Asn Pro

420 425 430

Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu Phe Thr Lys

435 440 445

Phe Cys His Lys Ala Ile Asp Gly Arg Ser Leu Tyr Asn Lys

450 455 460

<210> 34

<211> 455

<212> PRT

<213> 人工序列

<220>

<223> MTD-BoNT/D轻链氨基酸序列

<400> 34

Met Lys Ala Met Ile Asn Ile Asn Lys Phe Leu Asn Gln Cys Thr Trp

1 5 10 15

Pro Val Lys Asp Phe Asn Tyr Ser Asp Pro Val Asn Asp Asn Asp Ile

20 25 30

Leu Tyr Leu Arg Ile Pro Gln Asn Lys Leu Ile Thr Thr Pro Val Lys

35 40 45

Ala Phe Met Ile Thr Gln Asn Ile Trp Val Ile Pro Glu Arg Phe Ser

50 55 60

Ser Asp Thr Asn Pro Ser Leu Ser Lys Pro Pro Arg Pro Thr Ser Lys

65 70 75 80

Tyr Gln Ser Tyr Tyr Asp Pro Ser Tyr Leu Ser Thr Asp Glu Gln Lys

85 90 95

Asp Thr Phe Leu Lys Gly Ile Ile Lys Leu Phe Lys Arg Ile Asn Glu

100 105 110

Arg Asp Ile Gly Lys Lys Leu Ile Asn Tyr Leu Val Val Gly Ser Pro

115 120 125

Phe Met Gly Asp Ser Ser Thr Pro Glu Asp Thr Phe Asp Phe Thr Arg

130 135 140

His Thr Thr Asn Ile Ala Val Glu Lys Phe Glu Asn Gly Ser Trp Lys

145 150 155 160

Val Thr Asn Ile Ile Thr Pro Ser Val Leu Ile Phe Gly Pro Leu Pro

165 170 175

Asn Ile Leu Asp Tyr Thr Ala Ser Leu Thr Leu Gln Gly Gln Gln Ser

180 185 190

Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser Ile Leu Lys Val Ala

195 200 205

Pro Glu Phe Leu Leu Thr Phe Ser Asp Val Thr Ser Asn Gln Ser Ser

210 215 220

Ala Val Leu Gly Lys Ser Ile Phe Cys Met Asp Pro Val Ile Ala Leu

225 230 235 240

Met His Glu Leu Thr His Ser Leu His Gln Leu Tyr Gly Ile Asn Ile

245 250 255

Pro Ser Asp Lys Arg Ile Arg Pro Gln Val Ser Glu Gly Phe Phe Ser

260 265 270

Gln Asp Gly Pro Asn Val Gln Phe Glu Glu Leu Tyr Thr Phe Gly Gly

275 280 285

Leu Asp Val Glu Ile Ile Pro Gln Ile Glu Arg Ser Gln Leu Arg Glu

290 295 300

Lys Ala Leu Gly His Tyr Lys Asp Ile Ala Lys Arg Leu Asn Asn Ile

305 310 315 320

Asn Lys Thr Ile Pro Ser Ser Trp Ile Ser Asn Ile Asp Lys Tyr Lys

325 330 335

Lys Ile Phe Ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn Thr Gly Asn

340 345 350

Phe Val Val Asn Ile Asp Lys Phe Asn Ser Leu Tyr Ser Asp Leu Thr

355 360 365

Asn Val Met Ser Glu Val Val Tyr Ser Ser Gln Tyr Asn Val Lys Asn

370 375 380

Arg Thr His Tyr Phe Ser Arg His Tyr Leu Pro Val Phe Ala Asn Ile

385 390 395 400

Leu Asp Asp Asn Ile Tyr Thr Ile Arg Asp Gly Phe Asn Leu Thr Asn

405 410 415

Lys Gly Phe Asn Ile Glu Asn Ser Gly Gln Asn Ile Glu Arg Asn Pro

420 425 430

Ala Leu Gln Lys Leu Ser Ser Glu Ser Val Val Asp Leu Phe Thr Lys

435 440 445

Val Cys Leu Arg Leu Thr Lys

450 455

<210> 35

<211> 435

<212> PRT

<213> 人工序列

<220>

<223> MTD-BoNT/E轻链氨基酸序列

<400> 35

Met Lys Ala Met Ile Asn Ile Asn Lys Phe Leu Asn Gln Cys Pro Thr

1 5 10 15

Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asn Arg Thr Ile Leu

20 25 30

Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser Phe Asn Ile

35 40 45

Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile Gly Thr Ile

50 55 60

Pro Gln Asp Phe Leu Pro Pro Thr Ser Leu Lys Asn Gly Asp Ser Ser

65 70 75 80

Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Gln Glu Lys Asp Lys Phe

85 90 95

Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asp Asn Leu Ser

100 105 110

Gly Arg Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro Tyr Leu Gly

115 120 125

Asn Asp Asn Thr Pro Asp Gly Asp Phe Ile Ile Asn Asp Ala Ser Ala

130 135 140

Val Pro Ile Gln Phe Ser Asn Gly Ser Gln Ser Ile Leu Leu Pro Asn

145 150 155 160

Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr Asn Ser Ser

165 170 175

Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His Gly Phe Gly

180 185 190

Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe Arg Phe Lys

195 200 205

Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu Thr Leu Met

210 215 220

His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala Lys Gly Ile

225 230 235 240

Thr Thr Lys Tyr Thr Ile Thr Gln Lys Gln Asn Pro Leu Ile Thr Asn

245 250 255

Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly Gly Thr Asp

260 265 270

Leu Asn Ile Ile Thr Ser Ala Gln Ser Asn Asp Ile Tyr Thr Asn Leu

275 280 285

Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys Val Gln Val

290 295 300

Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp Val Phe Glu Ala Lys Tyr

305 310 315 320

Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn Ile Asn Lys

325 330 335

Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu Phe Asp Leu

340 345 350

Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile Gly Gln Tyr

355 360 365

Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile Tyr Asn Ile

370 375 380

Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe Arg Gly Gln

385 390 395 400

Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr Gly Arg Gly

405 410 415

Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val Ser Val Lys

420 425 430

Gly Ile Arg

435

<210> 36

<211> 449

<212> PRT

<213> 人工序列

<220>

<223> MTD-BoNT/F轻链氨基酸序列

<400> 36

Met Lys Ala Met Ile Asn Ile Asn Lys Phe Leu Asn Gln Cys Pro Val

1 5 10 15

Ala Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Asp Thr Ile

20 25 30

Leu Tyr Met Gln Ile Pro Tyr Glu Glu Lys Ser Lys Lys Tyr Tyr Lys

35 40 45

Ala Phe Glu Ile Met Arg Asn Val Trp Ile Ile Pro Glu Arg Asn Thr

50 55 60

Ile Gly Thr Asn Pro Ser Asp Phe Asp Pro Pro Ala Ser Leu Lys Asn

65 70 75 80

Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr Asp Ala Glu

85 90 95

Lys Asp Arg Tyr Leu Lys Thr Thr Ile Lys Leu Phe Lys Arg Ile Asn

100 105 110

Ser Asn Pro Ala Gly Lys Val Leu Leu Gln Glu Ile Ser Tyr Ala Lys

115 120 125

Pro Tyr Leu Gly Asn Asp His Thr Pro Ile Asp Glu Phe Ser Pro Val

130 135 140

Thr Arg Thr Thr Ser Val Asn Ile Lys Leu Ser Thr Asn Val Glu Ser

145 150 155 160

Ser Met Leu Leu Asn Leu Leu Val Leu Gly Ala Gly Pro Asp Ile Phe

165 170 175

Glu Ser Cys Cys Tyr Pro Val Arg Lys Leu Ile Asp Pro Asp Val Val

180 185 190

Tyr Asp Pro Ser Asn Tyr Gly Phe Gly Ser Ile Asn Ile Val Thr Phe

195 200 205

Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp Ile Ser Gly Gly His Asn

210 215 220

Ser Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser Leu Ala His

225 230 235 240

Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Arg Gly Val Thr

245 250 255

Tyr Glu Glu Thr Ile Glu Val Lys Gln Ala Pro Leu Met Ile Ala Glu

260 265 270

Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly Gln Asp Leu

275 280 285

Asn Ile Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn Asn Leu Leu

290 295 300

Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Ser Glu Val Asn Ser Ala

305 310 315 320

Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe Gln Trp Lys

325 330 335

Tyr Gly Leu Asp Lys Asn Ala Asp Gly Ser Tyr Thr Val Asn Glu Asn

340 345 350

Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr Glu Ser Asp

355 360 365

Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr Phe Ile Lys

370 375 380

Tyr Glu Phe Leu Lys Val Pro Asn Leu Leu Asp Asp Asp Ile Tyr Thr

385 390 395 400

Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn Asn Arg Gly

405 410 415

Gln Ser Ile Lys Leu Asn Pro Lys Ile Ile Asp Ser Ile Pro Asp Lys

420 425 430

Gly Leu Val Glu Lys Ile Val Lys Phe Cys Lys Ser Val Ile Pro Arg

435 440 445

Lys

<210> 37

<211> 454

<212> PRT

<213> 人工序列

<220>

<223> MTD-BoNT/G轻链氨基酸序列

<400> 37

Met Lys Ala Met Ile Asn Ile Asn Lys Phe Leu Asn Gln Cys Pro Val

1 5 10 15

Asn Ile Lys Phe Asn Tyr Asn Asp Pro Ile Asn Asn Asp Asp Ile Ile

20 25 30

Met Met Glu Pro Phe Asn Asp Pro Gly Pro Gly Thr Tyr Tyr Lys Ala

35 40 45

Phe Arg Ile Ile Asp Arg Ile Trp Ile Val Pro Glu Arg Phe Thr Tyr

50 55 60

Gly Phe Gln Pro Asp Gln Phe Asn Ala Ser Thr Gly Val Phe Ser Lys

65 70 75 80

Asp Val Tyr Glu Tyr Tyr Asp Pro Thr Tyr Leu Lys Thr Asp Ala Glu

85 90 95

Lys Asp Lys Phe Leu Lys Thr Met Ile Lys Leu Phe Asn Arg Ile Asn

100 105 110

Ser Lys Pro Ser Gly Gln Arg Leu Leu Asp Met Ile Val Asp Ala Ile

115 120 125

Pro Tyr Leu Gly Asn Ala Ser Thr Pro Pro Asp Lys Phe Ala Ala Asn

130 135 140

Val Ala Asn Val Ser Ile Asn Lys Lys Ile Ile Gln Pro Gly Ala Glu

145 150 155 160

Asp Gln Ile Lys Gly Leu Met Thr Asn Leu Ile Ile Phe Gly Pro Gly

165 170 175

Pro Val Leu Ser Asp Asn Phe Thr Asp Ser Met Ile Met Asn Gly His

180 185 190

Ser Pro Ile Ser Glu Gly Phe Gly Ala Arg Met Met Ile Arg Phe Cys

195 200 205

Pro Ser Cys Leu Asn Val Phe Asn Asn Val Gln Glu Asn Lys Asp Thr

210 215 220

Ser Ile Phe Ser Arg Arg Ala Tyr Phe Ala Asp Pro Ala Leu Thr Leu

225 230 235 240

Met His Glu Leu Ile His Val Leu His Gly Leu Tyr Gly Ile Lys Ile

245 250 255

Ser Asn Leu Pro Ile Thr Pro Asn Thr Lys Glu Phe Phe Met Gln His

260 265 270

Ser Asp Pro Val Gln Ala Glu Glu Leu Tyr Thr Phe Gly Gly His Asp

275 280 285

Pro Ser Val Ile Ser Pro Ser Thr Asp Met Asn Ile Tyr Asn Lys Ala

290 295 300

Leu Gln Asn Phe Gln Asp Ile Ala Asn Arg Leu Asn Ile Val Ser Ser

305 310 315 320

Ala Gln Gly Ser Gly Ile Asp Ile Ser Leu Tyr Lys Gln Ile Tyr Lys

325 330 335

Asn Lys Tyr Asp Phe Val Glu Asp Pro Asn Gly Lys Tyr Ser Val Asp

340 345 350

Lys Asp Lys Phe Asp Lys Leu Tyr Lys Ala Leu Met Phe Gly Phe Thr

355 360 365

Glu Thr Asn Leu Ala Gly Glu Tyr Gly Ile Lys Thr Arg Tyr Ser Tyr

370 375 380

Phe Ser Glu Tyr Leu Pro Pro Ile Lys Thr Glu Lys Leu Leu Asp Asn

385 390 395 400

Thr Ile Tyr Thr Gln Asn Glu Gly Phe Asn Ile Ala Ser Lys Asn Leu

405 410 415

Lys Thr Glu Phe Asn Gly Gln Asn Lys Ala Val Asn Lys Glu Ala Tyr

420 425 430

Glu Glu Ile Ser Leu Glu His Leu Val Ile Tyr Arg Ile Ala Met Cys

435 440 445

Lys Pro Val Met Tyr Lys

450

<210> 38

<211> 461

<212> PRT

<213> 人工序列

<220>

<223> BoNT/A轻链-MTDr氨基酸序列

<400> 38

Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly

1 5 10 15

Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro

20 25 30

Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg

35 40 45

Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu

50 55 60

Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr

65 70 75 80

Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu

85 90 95

Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val

100 105 110

Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys

115 120 125

Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr

130 135 140

Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile

145 150 155 160

Ile Gln Phe Glu Cys Lys Ser Phe Gly His Glu Val Leu Asn Leu Thr

165 170 175

Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe

180 185 190

Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu

195 200 205

Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu

210 215 220

Leu Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn

225 230 235 240

Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu

245 250 255

Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys

260 265 270

Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn

275 280 285

Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val

290 295 300

Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys

305 310 315 320

Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu

325 330 335

Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp

340 345 350

Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn

355 360 365

Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Lys Val Asn Tyr

370 375 380

Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn

385 390 395 400

Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu

405 410 415

Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg

420 425 430

Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Lys Gly Tyr Asn Lys

435 440 445

Cys Gln Asn Leu Phe Lys Asn Ile Asn Ile Met Ala Lys

450 455 460

<210> 39

<211> 454

<212> PRT

<213> 人工序列

<220>

<223> BoNT/B轻链-MTDr氨基酸序列

<400> 39

Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn

1 5 10 15

Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg

20 25 30

Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu

35 40 45

Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly

50 55 60

Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn

65 70 75 80

Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe

85 90 95

Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile

100 105 110

Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu

115 120 125

Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn

130 135 140

Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile

145 150 155 160

Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly

165 170 175

Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln

180 185 190

Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu

195 200 205

Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro

210 215 220

Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr

225 230 235 240

Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe

245 250 255

Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe

260 265 270

Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile

275 280 285

Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn

290 295 300

Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr

305 310 315 320

Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly

325 330 335

Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu

340 345 350

Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys

355 360 365

Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys

370 375 380

Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile

385 390 395 400

Ser Asp Lys Asp Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile

405 410 415

Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr

420 425 430

Lys Ile Gln Met Cys Lys Ser Val Lys Cys Gln Asn Leu Phe Lys Asn

435 440 445

Ile Asn Ile Met Ala Lys

450

<210> 40

<211> 462

<212> PRT

<213> 人工序列

<220>

<223> BoNT/C轻链-MTDr氨基酸序列

<400> 40

Met Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro Val Asp Asn

1 5 10 15

Lys Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu

20 25 30

Pro Glu Lys Ala Phe Arg Ile Thr Gly Asn Ile Trp Val Ile Pro Asp

35 40 45

Arg Phe Ser Arg Asn Ser Asn Pro Asn Leu Asn Lys Pro Pro Arg Val

50 55 60

Thr Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp

65 70 75 80

Ser Asp Lys Asp Pro Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg

85 90 95

Ile Asn Ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ser Thr

100 105 110

Asp Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp

115 120 125

Phe Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn

130 135 140

Asn Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly

145 150 155 160

Pro Arg Glu Asn Ile Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr

165 170 175

Asn Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile

180 185 190

Ser Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asp

195 200 205

Val Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile

210 215 220

Leu Ile Leu Met His Glu Leu Asn His Ala Met His Asn Leu Tyr Gly

225 230 235 240

Ile Ala Ile Pro Asn Asp Gln Thr Ile Ser Ser Val Thr Ser Asn Ile

245 250 255

Phe Tyr Ser Gln Tyr Asn Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala

260 265 270

Phe Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser Ala Arg Lys Tyr

275 280 285

Phe Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu

290 295 300

Asn Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly

305 310 315 320

Glu Tyr Lys Gln Lys Leu Ile Arg Lys Tyr Arg Phe Val Val Glu Ser

325 330 335

Ser Gly Glu Val Thr Val Asn Arg Asn Lys Phe Val Glu Leu Tyr Asn

340 345 350

Glu Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn

355 360 365

Val Gln Asn Arg Lys Ile Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr

370 375 380

Ala Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn

385 390 395 400

Ile Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser

405 410 415

Arg Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu

420 425 430

Phe Thr Lys Phe Cys His Lys Ala Ile Asp Gly Arg Ser Leu Tyr Asn

435 440 445

Lys Cys Gln Asn Leu Phe Lys Asn Ile Asn Ile Met Ala Lys

450 455 460

<210> 41

<211> 455

<212> PRT

<213> 人工序列

<220>

<223> BoNT/D轻链-MTDr氨基酸序列

<400> 41

Met Thr Trp Pro Val Lys Asp Phe Asn Tyr Ser Asp Pro Val Asn Asp

1 5 10 15

Asn Asp Ile Leu Tyr Leu Arg Ile Pro Gln Asn Lys Leu Ile Thr Thr

20 25 30

Pro Val Lys Ala Phe Met Ile Thr Gln Asn Ile Trp Val Ile Pro Glu

35 40 45

Arg Phe Ser Ser Asp Thr Asn Pro Ser Leu Ser Lys Pro Pro Arg Pro

50 55 60

Thr Ser Lys Tyr Gln Ser Tyr Tyr Asp Pro Ser Tyr Leu Ser Thr Asp

65 70 75 80

Glu Gln Lys Asp Thr Phe Leu Lys Gly Ile Ile Lys Leu Phe Lys Arg

85 90 95

Ile Asn Glu Arg Asp Ile Gly Lys Lys Leu Ile Asn Tyr Leu Val Val

100 105 110

Gly Ser Pro Phe Met Gly Asp Ser Ser Thr Pro Glu Asp Thr Phe Asp

115 120 125

Phe Thr Arg His Thr Thr Asn Ile Ala Val Glu Lys Phe Glu Asn Gly

130 135 140

Ser Trp Lys Val Thr Asn Ile Ile Thr Pro Ser Val Leu Ile Phe Gly

145 150 155 160

Pro Leu Pro Asn Ile Leu Asp Tyr Thr Ala Ser Leu Thr Leu Gln Gly

165 170 175

Gln Gln Ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser Ile Leu

180 185 190

Lys Val Ala Pro Glu Phe Leu Leu Thr Phe Ser Asp Val Thr Ser Asn

195 200 205

Gln Ser Ser Ala Val Leu Gly Lys Ser Ile Phe Cys Met Asp Pro Val

210 215 220

Ile Ala Leu Met His Glu Leu Thr His Ser Leu His Gln Leu Tyr Gly

225 230 235 240

Ile Asn Ile Pro Ser Asp Lys Arg Ile Arg Pro Gln Val Ser Glu Gly

245 250 255

Phe Phe Ser Gln Asp Gly Pro Asn Val Gln Phe Glu Glu Leu Tyr Thr

260 265 270

Phe Gly Gly Leu Asp Val Glu Ile Ile Pro Gln Ile Glu Arg Ser Gln

275 280 285

Leu Arg Glu Lys Ala Leu Gly His Tyr Lys Asp Ile Ala Lys Arg Leu

290 295 300

Asn Asn Ile Asn Lys Thr Ile Pro Ser Ser Trp Ile Ser Asn Ile Asp

305 310 315 320

Lys Tyr Lys Lys Ile Phe Ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn

325 330 335

Thr Gly Asn Phe Val Val Asn Ile Asp Lys Phe Asn Ser Leu Tyr Ser

340 345 350

Asp Leu Thr Asn Val Met Ser Glu Val Val Tyr Ser Ser Gln Tyr Asn

355 360 365

Val Lys Asn Arg Thr His Tyr Phe Ser Arg His Tyr Leu Pro Val Phe

370 375 380

Ala Asn Ile Leu Asp Asp Asn Ile Tyr Thr Ile Arg Asp Gly Phe Asn

385 390 395 400

Leu Thr Asn Lys Gly Phe Asn Ile Glu Asn Ser Gly Gln Asn Ile Glu

405 410 415

Arg Asn Pro Ala Leu Gln Lys Leu Ser Ser Glu Ser Val Val Asp Leu

420 425 430

Phe Thr Lys Val Cys Leu Arg Leu Thr Lys Cys Gln Asn Leu Phe Lys

435 440 445

Asn Ile Asn Ile Met Ala Lys

450 455

<210> 42

<211> 435

<212> PRT

<213> 人工序列

<220>

<223> BoNT/E轻链-MTDr氨基酸序列

<400> 42

Met Pro Thr Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asn Arg

1 5 10 15

Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser

20 25 30

Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile

35 40 45

Gly Thr Ile Pro Gln Asp Phe Leu Pro Pro Thr Ser Leu Lys Asn Gly

50 55 60

Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Gln Glu Lys

65 70 75 80

Asp Lys Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asp

85 90 95

Asn Leu Ser Gly Arg Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro

100 105 110

Tyr Leu Gly Asn Asp Asn Thr Pro Asp Gly Asp Phe Ile Ile Asn Asp

115 120 125

Ala Ser Ala Val Pro Ile Gln Phe Ser Asn Gly Ser Gln Ser Ile Leu

130 135 140

Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr

145 150 155 160

Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His

165 170 175

Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe

180 185 190

Arg Phe Lys Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu

195 200 205

Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala

210 215 220

Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Lys Gln Asn Pro Leu

225 230 235 240

Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly

245 250 255

Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Ser Asn Asp Ile Tyr

260 265 270

Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys

275 280 285

Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp Val Phe Glu

290 295 300

Ala Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn

305 310 315 320

Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu

325 330 335

Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile

340 345 350

Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile

355 360 365

Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe

370 375 380

Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr

385 390 395 400

Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val

405 410 415

Ser Val Lys Gly Ile Arg Cys Gln Asn Leu Phe Lys Asn Ile Asn Ile

420 425 430

Met Ala Lys

435

<210> 43

<211> 449

<212> PRT

<213> 人工序列

<220>

<223> BoNT/F轻链-MTDr氨基酸序列

<400> 43

Met Pro Val Ala Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp

1 5 10 15

Asp Thr Ile Leu Tyr Met Gln Ile Pro Tyr Glu Glu Lys Ser Lys Lys

20 25 30

Tyr Tyr Lys Ala Phe Glu Ile Met Arg Asn Val Trp Ile Ile Pro Glu

35 40 45

Arg Asn Thr Ile Gly Thr Asn Pro Ser Asp Phe Asp Pro Pro Ala Ser

50 55 60

Leu Lys Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr

65 70 75 80

Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr Ile Lys Leu Phe Lys

85 90 95

Arg Ile Asn Ser Asn Pro Ala Gly Lys Val Leu Leu Gln Glu Ile Ser

100 105 110

Tyr Ala Lys Pro Tyr Leu Gly Asn Asp His Thr Pro Ile Asp Glu Phe

115 120 125

Ser Pro Val Thr Arg Thr Thr Ser Val Asn Ile Lys Leu Ser Thr Asn

130 135 140

Val Glu Ser Ser Met Leu Leu Asn Leu Leu Val Leu Gly Ala Gly Pro

145 150 155 160

Asp Ile Phe Glu Ser Cys Cys Tyr Pro Val Arg Lys Leu Ile Asp Pro

165 170 175

Asp Val Val Tyr Asp Pro Ser Asn Tyr Gly Phe Gly Ser Ile Asn Ile

180 185 190

Val Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp Ile Ser Gly

195 200 205

Gly His Asn Ser Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser

210 215 220

Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Arg

225 230 235 240

Gly Val Thr Tyr Glu Glu Thr Ile Glu Val Lys Gln Ala Pro Leu Met

245 250 255

Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly

260 265 270

Gln Asp Leu Asn Ile Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn

275 280 285

Asn Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Ser Glu Val

290 295 300

Asn Ser Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe

305 310 315 320

Gln Trp Lys Tyr Gly Leu Asp Lys Asn Ala Asp Gly Ser Tyr Thr Val

325 330 335

Asn Glu Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr

340 345 350

Glu Ser Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr

355 360 365

Phe Ile Lys Tyr Glu Phe Leu Lys Val Pro Asn Leu Leu Asp Asp Asp

370 375 380

Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn

385 390 395 400

Asn Arg Gly Gln Ser Ile Lys Leu Asn Pro Lys Ile Ile Asp Ser Ile

405 410 415

Pro Asp Lys Gly Leu Val Glu Lys Ile Val Lys Phe Cys Lys Ser Val

420 425 430

Ile Pro Arg Lys Cys Gln Asn Leu Phe Lys Asn Ile Asn Ile Met Ala

435 440 445

Lys

<210> 44

<211> 454

<212> PRT

<213> 人工序列

<220>

<223> BoNT/G轻链-MTDr氨基酸序列

<400> 44

Met Pro Val Asn Ile Lys Phe Asn Tyr Asn Asp Pro Ile Asn Asn Asp

1 5 10 15

Asp Ile Ile Met Met Glu Pro Phe Asn Asp Pro Gly Pro Gly Thr Tyr

20 25 30

Tyr Lys Ala Phe Arg Ile Ile Asp Arg Ile Trp Ile Val Pro Glu Arg

35 40 45

Phe Thr Tyr Gly Phe Gln Pro Asp Gln Phe Asn Ala Ser Thr Gly Val

50 55 60

Phe Ser Lys Asp Val Tyr Glu Tyr Tyr Asp Pro Thr Tyr Leu Lys Thr

65 70 75 80

Asp Ala Glu Lys Asp Lys Phe Leu Lys Thr Met Ile Lys Leu Phe Asn

85 90 95

Arg Ile Asn Ser Lys Pro Ser Gly Gln Arg Leu Leu Asp Met Ile Val

100 105 110

Asp Ala Ile Pro Tyr Leu Gly Asn Ala Ser Thr Pro Pro Asp Lys Phe

115 120 125

Ala Ala Asn Val Ala Asn Val Ser Ile Asn Lys Lys Ile Ile Gln Pro

130 135 140

Gly Ala Glu Asp Gln Ile Lys Gly Leu Met Thr Asn Leu Ile Ile Phe

145 150 155 160

Gly Pro Gly Pro Val Leu Ser Asp Asn Phe Thr Asp Ser Met Ile Met

165 170 175

Asn Gly His Ser Pro Ile Ser Glu Gly Phe Gly Ala Arg Met Met Ile

180 185 190

Arg Phe Cys Pro Ser Cys Leu Asn Val Phe Asn Asn Val Gln Glu Asn

195 200 205

Lys Asp Thr Ser Ile Phe Ser Arg Arg Ala Tyr Phe Ala Asp Pro Ala

210 215 220

Leu Thr Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr Gly

225 230 235 240

Ile Lys Ile Ser Asn Leu Pro Ile Thr Pro Asn Thr Lys Glu Phe Phe

245 250 255

Met Gln His Ser Asp Pro Val Gln Ala Glu Glu Leu Tyr Thr Phe Gly

260 265 270

Gly His Asp Pro Ser Val Ile Ser Pro Ser Thr Asp Met Asn Ile Tyr

275 280 285

Asn Lys Ala Leu Gln Asn Phe Gln Asp Ile Ala Asn Arg Leu Asn Ile

290 295 300

Val Ser Ser Ala Gln Gly Ser Gly Ile Asp Ile Ser Leu Tyr Lys Gln

305 310 315 320

Ile Tyr Lys Asn Lys Tyr Asp Phe Val Glu Asp Pro Asn Gly Lys Tyr

325 330 335

Ser Val Asp Lys Asp Lys Phe Asp Lys Leu Tyr Lys Ala Leu Met Phe

340 345 350

Gly Phe Thr Glu Thr Asn Leu Ala Gly Glu Tyr Gly Ile Lys Thr Arg

355 360 365

Tyr Ser Tyr Phe Ser Glu Tyr Leu Pro Pro Ile Lys Thr Glu Lys Leu

370 375 380

Leu Asp Asn Thr Ile Tyr Thr Gln Asn Glu Gly Phe Asn Ile Ala Ser

385 390 395 400

Lys Asn Leu Lys Thr Glu Phe Asn Gly Gln Asn Lys Ala Val Asn Lys

405 410 415

Glu Ala Tyr Glu Glu Ile Ser Leu Glu His Leu Val Ile Tyr Arg Ile

420 425 430

Ala Met Cys Lys Pro Val Met Tyr Lys Cys Gln Asn Leu Phe Lys Asn

435 440 445

Ile Asn Ile Met Ala Lys

450

<210> 45

<211> 469

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的MTD-BoNT/A轻链氨基酸序列

<400> 45

Met Lys Ala Met Ile Asn Ile Asn Lys Phe Leu Asn Gln Cys Pro Phe

1 5 10 15

Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly Val Asp Ile

20 25 30

Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro Val Lys Ala

35 40 45

Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg Asp Thr Phe

50 55 60

Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu Ala Lys Gln

65 70 75 80

Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr Asp Asn Glu

85 90 95

Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu Arg Ile Tyr

100 105 110

Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val Arg Gly Ile

115 120 125

Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys Val Ile Asp

130 135 140

Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr Arg Ser Glu

145 150 155 160

Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile Ile Gln Phe

165 170 175

Glu Cys Lys Ser Phe Gly His Glu Val Leu Asn Leu Thr Arg Asn Gly

180 185 190

Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe Thr Phe Gly

195 200 205

Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu Gly Ala Gly

210 215 220

Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu Leu Ile His

225 230 235 240

Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn Arg Val Phe

245 250 255

Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu Glu Val Ser

260 265 270

Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys Phe Ile Asp

275 280 285

Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn Lys Phe Lys

290 295 300

Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val Gly Thr Thr

305 310 315 320

Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys Tyr Leu Leu

325 330 335

Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu Lys Phe Asp

340 345 350

Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp Asn Phe Val

355 360 365

Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn Phe Asp Lys

370 375 380

Ala Val Phe Lys Ile Asn Ile Val Pro Lys Val Asn Tyr Thr Ile Tyr

385 390 395 400

Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn Phe Asn Gly

405 410 415

Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu Lys Asn Phe

420 425 430

Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg Gly Ile Ile

435 440 445

Thr Ser Lys Thr Lys Ser Leu Asp Lys Gly Tyr Asn Lys Leu Glu His

450 455 460

His His His His His

465

<210> 46

<211> 462

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的MTD-BoNT/B轻链氨基酸序列

<400> 46

Met Lys Ala Met Ile Asn Ile Asn Lys Phe Leu Asn Gln Cys Pro Val

1 5 10 15

Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn Asn Asn Ile

20 25 30

Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg Tyr Tyr Lys

35 40 45

Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu Arg Tyr Thr

50 55 60

Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly Ile Phe Asn

65 70 75 80

Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn Thr Asn Asp

85 90 95

Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe Asn Arg Ile

100 105 110

Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile Ile Asn Gly

115 120 125

Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu Phe Asn Thr

130 135 140

Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn Pro Gly Glu

145 150 155 160

Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile Phe Gly Pro

165 170 175

Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly Ile Gln Asn

180 185 190

His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln Met Lys Phe

195 200 205

Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu Asn Lys Gly

210 215 220

Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro Ala Leu Ile

225 230 235 240

Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr Gly Ile Lys

245 250 255

Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe Phe Met Gln

260 265 270

Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe Gly Gly Gln

275 280 285

Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile Tyr Asp Lys

290 295 300

Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn Lys Val Leu

305 310 315 320

Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr Lys Asn Lys

325 330 335

Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly Lys Tyr Ser

340 345 350

Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu Met Phe Gly

355 360 365

Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys Thr Arg Ala

370 375 380

Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys Asn Leu Leu

385 390 395 400

Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile Ser Asp Lys

405 410 415

Asp Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile Asn Lys Gln

420 425 430

Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr Lys Ile Gln

435 440 445

Met Cys Lys Ser Val Lys Leu Glu His His His His His His

450 455 460

<210> 47

<211> 470

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的MTD-BoNT/C轻链氨基酸序列

<400> 47

Met Lys Ala Met Ile Asn Ile Asn Lys Phe Leu Asn Gln Cys Pro Ile

1 5 10 15

Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro Val Asp Asn Lys Asn Ile

20 25 30

Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu Pro Glu Lys

35 40 45

Ala Phe Arg Ile Thr Gly Asn Ile Trp Val Ile Pro Asp Arg Phe Ser

50 55 60

Arg Asn Ser Asn Pro Asn Leu Asn Lys Pro Pro Arg Val Thr Ser Pro

65 70 75 80

Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp Ser Asp Lys

85 90 95

Asp Pro Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg Ile Asn Ser

100 105 110

Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ser Thr Asp Ile Pro

115 120 125

Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp Phe Asp Val

130 135 140

Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn Asn Trp Val

145 150 155 160

Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly Pro Arg Glu

165 170 175

Asn Ile Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr Asn Asn Thr

180 185 190

Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile Ser Ile Ser

195 200 205

Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asp Val Gly Glu

210 215 220

Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile Leu Ile Leu

225 230 235 240

Met His Glu Leu Asn His Ala Met His Asn Leu Tyr Gly Ile Ala Ile

245 250 255

Pro Asn Asp Gln Thr Ile Ser Ser Val Thr Ser Asn Ile Phe Tyr Ser

260 265 270

Gln Tyr Asn Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala Phe Gly Gly

275 280 285

Pro Thr Ile Asp Leu Ile Pro Lys Ser Ala Arg Lys Tyr Phe Glu Glu

290 295 300

Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu Asn Ser Ile

305 310 315 320

Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly Glu Tyr Lys

325 330 335

Gln Lys Leu Ile Arg Lys Tyr Arg Phe Val Val Glu Ser Ser Gly Glu

340 345 350

Val Thr Val Asn Arg Asn Lys Phe Val Glu Leu Tyr Asn Glu Leu Thr

355 360 365

Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn Val Gln Asn

370 375 380

Arg Lys Ile Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr Ala Asn Ile

385 390 395 400

Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn Ile Pro Lys

405 410 415

Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser Arg Asn Pro

420 425 430

Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu Phe Thr Lys

435 440 445

Phe Cys His Lys Ala Ile Asp Gly Arg Ser Leu Tyr Asn Lys Leu Glu

450 455 460

His His His His His His

465 470

<210> 48

<211> 463

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的MTD-BoNT/D轻链氨基酸序列

<400> 48

Met Lys Ala Met Ile Asn Ile Asn Lys Phe Leu Asn Gln Cys Thr Trp

1 5 10 15

Pro Val Lys Asp Phe Asn Tyr Ser Asp Pro Val Asn Asp Asn Asp Ile

20 25 30

Leu Tyr Leu Arg Ile Pro Gln Asn Lys Leu Ile Thr Thr Pro Val Lys

35 40 45

Ala Phe Met Ile Thr Gln Asn Ile Trp Val Ile Pro Glu Arg Phe Ser

50 55 60

Ser Asp Thr Asn Pro Ser Leu Ser Lys Pro Pro Arg Pro Thr Ser Lys

65 70 75 80

Tyr Gln Ser Tyr Tyr Asp Pro Ser Tyr Leu Ser Thr Asp Glu Gln Lys

85 90 95

Asp Thr Phe Leu Lys Gly Ile Ile Lys Leu Phe Lys Arg Ile Asn Glu

100 105 110

Arg Asp Ile Gly Lys Lys Leu Ile Asn Tyr Leu Val Val Gly Ser Pro

115 120 125

Phe Met Gly Asp Ser Ser Thr Pro Glu Asp Thr Phe Asp Phe Thr Arg

130 135 140

His Thr Thr Asn Ile Ala Val Glu Lys Phe Glu Asn Gly Ser Trp Lys

145 150 155 160

Val Thr Asn Ile Ile Thr Pro Ser Val Leu Ile Phe Gly Pro Leu Pro

165 170 175

Asn Ile Leu Asp Tyr Thr Ala Ser Leu Thr Leu Gln Gly Gln Gln Ser

180 185 190

Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser Ile Leu Lys Val Ala

195 200 205

Pro Glu Phe Leu Leu Thr Phe Ser Asp Val Thr Ser Asn Gln Ser Ser

210 215 220

Ala Val Leu Gly Lys Ser Ile Phe Cys Met Asp Pro Val Ile Ala Leu

225 230 235 240

Met His Glu Leu Thr His Ser Leu His Gln Leu Tyr Gly Ile Asn Ile

245 250 255

Pro Ser Asp Lys Arg Ile Arg Pro Gln Val Ser Glu Gly Phe Phe Ser

260 265 270

Gln Asp Gly Pro Asn Val Gln Phe Glu Glu Leu Tyr Thr Phe Gly Gly

275 280 285

Leu Asp Val Glu Ile Ile Pro Gln Ile Glu Arg Ser Gln Leu Arg Glu

290 295 300

Lys Ala Leu Gly His Tyr Lys Asp Ile Ala Lys Arg Leu Asn Asn Ile

305 310 315 320

Asn Lys Thr Ile Pro Ser Ser Trp Ile Ser Asn Ile Asp Lys Tyr Lys

325 330 335

Lys Ile Phe Ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn Thr Gly Asn

340 345 350

Phe Val Val Asn Ile Asp Lys Phe Asn Ser Leu Tyr Ser Asp Leu Thr

355 360 365

Asn Val Met Ser Glu Val Val Tyr Ser Ser Gln Tyr Asn Val Lys Asn

370 375 380

Arg Thr His Tyr Phe Ser Arg His Tyr Leu Pro Val Phe Ala Asn Ile

385 390 395 400

Leu Asp Asp Asn Ile Tyr Thr Ile Arg Asp Gly Phe Asn Leu Thr Asn

405 410 415

Lys Gly Phe Asn Ile Glu Asn Ser Gly Gln Asn Ile Glu Arg Asn Pro

420 425 430

Ala Leu Gln Lys Leu Ser Ser Glu Ser Val Val Asp Leu Phe Thr Lys

435 440 445

Val Cys Leu Arg Leu Thr Lys Leu Glu His His His His His His

450 455 460

<210> 49

<211> 443

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的MTD-BoNT/E轻链氨基酸序列

<400> 49

Met Lys Ala Met Ile Asn Ile Asn Lys Phe Leu Asn Gln Cys Pro Thr

1 5 10 15

Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asn Arg Thr Ile Leu

20 25 30

Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser Phe Asn Ile

35 40 45

Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile Gly Thr Ile

50 55 60

Pro Gln Asp Phe Leu Pro Pro Thr Ser Leu Lys Asn Gly Asp Ser Ser

65 70 75 80

Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Gln Glu Lys Asp Lys Phe

85 90 95

Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asp Asn Leu Ser

100 105 110

Gly Arg Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro Tyr Leu Gly

115 120 125

Asn Asp Asn Thr Pro Asp Gly Asp Phe Ile Ile Asn Asp Ala Ser Ala

130 135 140

Val Pro Ile Gln Phe Ser Asn Gly Ser Gln Ser Ile Leu Leu Pro Asn

145 150 155 160

Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr Asn Ser Ser

165 170 175

Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His Gly Phe Gly

180 185 190

Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe Arg Phe Lys

195 200 205

Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu Thr Leu Met

210 215 220

His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala Lys Gly Ile

225 230 235 240

Thr Thr Lys Tyr Thr Ile Thr Gln Lys Gln Asn Pro Leu Ile Thr Asn

245 250 255

Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly Gly Thr Asp

260 265 270

Leu Asn Ile Ile Thr Ser Ala Gln Ser Asn Asp Ile Tyr Thr Asn Leu

275 280 285

Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys Val Gln Val

290 295 300

Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp Val Phe Glu Ala Lys Tyr

305 310 315 320

Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn Ile Asn Lys

325 330 335

Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu Phe Asp Leu

340 345 350

Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile Gly Gln Tyr

355 360 365

Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile Tyr Asn Ile

370 375 380

Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe Arg Gly Gln

385 390 395 400

Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr Gly Arg Gly

405 410 415

Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val Ser Val Lys

420 425 430

Gly Ile Arg Leu Glu His His His His His His

435 440

<210> 50

<211> 457

<212> PRT

<213> 人工序列

<220>

<223> MTD-具有六组氨酸的BoNT/F轻链氨基酸序列

<400> 50

Met Lys Ala Met Ile Asn Ile Asn Lys Phe Leu Asn Gln Cys Pro Val

1 5 10 15

Ala Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Asp Thr Ile

20 25 30

Leu Tyr Met Gln Ile Pro Tyr Glu Glu Lys Ser Lys Lys Tyr Tyr Lys

35 40 45

Ala Phe Glu Ile Met Arg Asn Val Trp Ile Ile Pro Glu Arg Asn Thr

50 55 60

Ile Gly Thr Asn Pro Ser Asp Phe Asp Pro Pro Ala Ser Leu Lys Asn

65 70 75 80

Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr Asp Ala Glu

85 90 95

Lys Asp Arg Tyr Leu Lys Thr Thr Ile Lys Leu Phe Lys Arg Ile Asn

100 105 110

Ser Asn Pro Ala Gly Lys Val Leu Leu Gln Glu Ile Ser Tyr Ala Lys

115 120 125

Pro Tyr Leu Gly Asn Asp His Thr Pro Ile Asp Glu Phe Ser Pro Val

130 135 140

Thr Arg Thr Thr Ser Val Asn Ile Lys Leu Ser Thr Asn Val Glu Ser

145 150 155 160

Ser Met Leu Leu Asn Leu Leu Val Leu Gly Ala Gly Pro Asp Ile Phe

165 170 175

Glu Ser Cys Cys Tyr Pro Val Arg Lys Leu Ile Asp Pro Asp Val Val

180 185 190

Tyr Asp Pro Ser Asn Tyr Gly Phe Gly Ser Ile Asn Ile Val Thr Phe

195 200 205

Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp Ile Ser Gly Gly His Asn

210 215 220

Ser Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser Leu Ala His

225 230 235 240

Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Arg Gly Val Thr

245 250 255

Tyr Glu Glu Thr Ile Glu Val Lys Gln Ala Pro Leu Met Ile Ala Glu

260 265 270

Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly Gln Asp Leu

275 280 285

Asn Ile Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn Asn Leu Leu

290 295 300

Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Ser Glu Val Asn Ser Ala

305 310 315 320

Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe Gln Trp Lys

325 330 335

Tyr Gly Leu Asp Lys Asn Ala Asp Gly Ser Tyr Thr Val Asn Glu Asn

340 345 350

Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr Glu Ser Asp

355 360 365

Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr Phe Ile Lys

370 375 380

Tyr Glu Phe Leu Lys Val Pro Asn Leu Leu Asp Asp Asp Ile Tyr Thr

385 390 395 400

Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn Asn Arg Gly

405 410 415

Gln Ser Ile Lys Leu Asn Pro Lys Ile Ile Asp Ser Ile Pro Asp Lys

420 425 430

Gly Leu Val Glu Lys Ile Val Lys Phe Cys Lys Ser Val Ile Pro Arg

435 440 445

Lys Leu Glu His His His His His His

450 455

<210> 51

<211> 462

<212> PRT

<213> 人工序列

<220>

<223> MTD-具有六组氨酸的BoNT/G轻链氨基酸序列

<400> 51

Met Lys Ala Met Ile Asn Ile Asn Lys Phe Leu Asn Gln Cys Pro Val

1 5 10 15

Asn Ile Lys Phe Asn Tyr Asn Asp Pro Ile Asn Asn Asp Asp Ile Ile

20 25 30

Met Met Glu Pro Phe Asn Asp Pro Gly Pro Gly Thr Tyr Tyr Lys Ala

35 40 45

Phe Arg Ile Ile Asp Arg Ile Trp Ile Val Pro Glu Arg Phe Thr Tyr

50 55 60

Gly Phe Gln Pro Asp Gln Phe Asn Ala Ser Thr Gly Val Phe Ser Lys

65 70 75 80

Asp Val Tyr Glu Tyr Tyr Asp Pro Thr Tyr Leu Lys Thr Asp Ala Glu

85 90 95

Lys Asp Lys Phe Leu Lys Thr Met Ile Lys Leu Phe Asn Arg Ile Asn

100 105 110

Ser Lys Pro Ser Gly Gln Arg Leu Leu Asp Met Ile Val Asp Ala Ile

115 120 125

Pro Tyr Leu Gly Asn Ala Ser Thr Pro Pro Asp Lys Phe Ala Ala Asn

130 135 140

Val Ala Asn Val Ser Ile Asn Lys Lys Ile Ile Gln Pro Gly Ala Glu

145 150 155 160

Asp Gln Ile Lys Gly Leu Met Thr Asn Leu Ile Ile Phe Gly Pro Gly

165 170 175

Pro Val Leu Ser Asp Asn Phe Thr Asp Ser Met Ile Met Asn Gly His

180 185 190

Ser Pro Ile Ser Glu Gly Phe Gly Ala Arg Met Met Ile Arg Phe Cys

195 200 205

Pro Ser Cys Leu Asn Val Phe Asn Asn Val Gln Glu Asn Lys Asp Thr

210 215 220

Ser Ile Phe Ser Arg Arg Ala Tyr Phe Ala Asp Pro Ala Leu Thr Leu

225 230 235 240

Met His Glu Leu Ile His Val Leu His Gly Leu Tyr Gly Ile Lys Ile

245 250 255

Ser Asn Leu Pro Ile Thr Pro Asn Thr Lys Glu Phe Phe Met Gln His

260 265 270

Ser Asp Pro Val Gln Ala Glu Glu Leu Tyr Thr Phe Gly Gly His Asp

275 280 285

Pro Ser Val Ile Ser Pro Ser Thr Asp Met Asn Ile Tyr Asn Lys Ala

290 295 300

Leu Gln Asn Phe Gln Asp Ile Ala Asn Arg Leu Asn Ile Val Ser Ser

305 310 315 320

Ala Gln Gly Ser Gly Ile Asp Ile Ser Leu Tyr Lys Gln Ile Tyr Lys

325 330 335

Asn Lys Tyr Asp Phe Val Glu Asp Pro Asn Gly Lys Tyr Ser Val Asp

340 345 350

Lys Asp Lys Phe Asp Lys Leu Tyr Lys Ala Leu Met Phe Gly Phe Thr

355 360 365

Glu Thr Asn Leu Ala Gly Glu Tyr Gly Ile Lys Thr Arg Tyr Ser Tyr

370 375 380

Phe Ser Glu Tyr Leu Pro Pro Ile Lys Thr Glu Lys Leu Leu Asp Asn

385 390 395 400

Thr Ile Tyr Thr Gln Asn Glu Gly Phe Asn Ile Ala Ser Lys Asn Leu

405 410 415

Lys Thr Glu Phe Asn Gly Gln Asn Lys Ala Val Asn Lys Glu Ala Tyr

420 425 430

Glu Glu Ile Ser Leu Glu His Leu Val Ile Tyr Arg Ile Ala Met Cys

435 440 445

Lys Pro Val Met Tyr Lys Leu Glu His His His His His His

450 455 460

<210> 52

<211> 481

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/A轻链-MTDr氨基酸序列

<400> 52

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro

1 5 10 15

Arg Gly Ser His Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp

20 25 30

Pro Val Asn Gly Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly

35 40 45

Gln Met Gln Pro Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val

50 55 60

Ile Pro Glu Arg Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn

65 70 75 80

Pro Pro Pro Glu Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr

85 90 95

Tyr Leu Ser Thr Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr

100 105 110

Lys Leu Phe Glu Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu

115 120 125

Thr Ser Ile Val Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp

130 135 140

Thr Glu Leu Lys Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro

145 150 155 160

Asp Gly Ser Tyr Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro

165 170 175

Ser Ala Asp Ile Ile Gln Phe Glu Cys Lys Ser Phe Gly His Glu Val

180 185 190

Leu Asn Leu Thr Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe

195 200 205

Ser Pro Asp Phe Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr

210 215 220

Asn Pro Leu Leu Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr

225 230 235 240

Leu Ala His Glu Leu Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala

245 250 255

Ile Asn Pro Asn Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu

260 265 270

Met Ser Gly Leu Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly

275 280 285

His Asp Ala Lys Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu

290 295 300

Tyr Tyr Tyr Asn Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala

305 310 315 320

Lys Ser Ile Val Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val

325 330 335

Phe Lys Glu Lys Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser

340 345 350

Val Asp Lys Leu Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile

355 360 365

Tyr Thr Glu Asp Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys

370 375 380

Thr Tyr Leu Asn Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro

385 390 395 400

Lys Val Asn Tyr Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn

405 410 415

Leu Ala Ala Asn Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn

420 425 430

Phe Thr Lys Leu Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu

435 440 445

Leu Cys Val Arg Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Lys

450 455 460

Gly Tyr Asn Lys Cys Gln Asn Leu Phe Lys Asn Ile Asn Ile Met Ala

465 470 475 480

Lys

<210> 53

<211> 474

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/B轻链-MTDr氨基酸序列

<400> 53

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro

1 5 10 15

Arg Gly Ser His Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp

20 25 30

Pro Ile Asp Asn Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg

35 40 45

Gly Thr Gly Arg Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp

50 55 60

Ile Ile Pro Glu Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn

65 70 75 80

Lys Ser Ser Gly Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro

85 90 95

Asp Tyr Leu Asn Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met

100 105 110

Ile Lys Leu Phe Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu

115 120 125

Leu Glu Met Ile Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val

130 135 140

Pro Leu Glu Glu Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys

145 150 155 160

Leu Ile Ser Asn Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala

165 170 175

Asn Leu Ile Ile Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr

180 185 190

Ile Asp Ile Gly Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly

195 200 205

Gly Ile Met Gln Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn

210 215 220

Asn Val Gln Glu Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr

225 230 235 240

Phe Ser Asp Pro Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu

245 250 255

His Gly Leu Tyr Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn

260 265 270

Glu Lys Lys Phe Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu

275 280 285

Leu Tyr Thr Phe Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr

290 295 300

Asp Lys Ser Ile Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val

305 310 315 320

Asp Arg Leu Asn Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn

325 330 335

Ile Asn Ile Tyr Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu

340 345 350

Asp Ser Glu Gly Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu

355 360 365

Tyr Lys Ser Leu Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn

370 375 380

Tyr Lys Ile Lys Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro

385 390 395 400

Val Lys Ile Lys Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu

405 410 415

Gly Phe Asn Ile Ser Asp Lys Asp Met Glu Lys Glu Tyr Arg Gly Gln

420 425 430

Asn Lys Ala Ile Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His

435 440 445

Leu Ala Val Tyr Lys Ile Gln Met Cys Lys Ser Val Lys Cys Gln Asn

450 455 460

Leu Phe Lys Asn Ile Asn Ile Met Ala Lys

465 470

<210> 54

<211> 482

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/C轻链-MTDr氨基酸序列

<400> 54

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro

1 5 10 15

Arg Gly Ser His Met Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp

20 25 30

Pro Val Asp Asn Lys Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr

35 40 45

Leu Ala Asn Glu Pro Glu Lys Ala Phe Arg Ile Thr Gly Asn Ile Trp

50 55 60

Val Ile Pro Asp Arg Phe Ser Arg Asn Ser Asn Pro Asn Leu Asn Lys

65 70 75 80

Pro Pro Arg Val Thr Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr

85 90 95

Leu Ser Thr Asp Ser Asp Lys Asp Pro Phe Leu Lys Glu Ile Ile Lys

100 105 110

Leu Phe Lys Arg Ile Asn Ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr

115 120 125

Arg Leu Ser Thr Asp Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile

130 135 140

Asn Thr Phe Asp Phe Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr

145 150 155 160

Arg Gln Gly Asn Asn Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val

165 170 175

Ile Ile Thr Gly Pro Arg Glu Asn Ile Ile Asp Pro Glu Thr Ser Thr

180 185 190

Phe Lys Leu Thr Asn Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala

195 200 205

Leu Ser Ile Ile Ser Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn

210 215 220

Ala Thr Asn Asp Val Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys

225 230 235 240

Met Asp Pro Ile Leu Ile Leu Met His Glu Leu Asn His Ala Met His

245 250 255

Asn Leu Tyr Gly Ile Ala Ile Pro Asn Asp Gln Thr Ile Ser Ser Val

260 265 270

Thr Ser Asn Ile Phe Tyr Ser Gln Tyr Asn Val Lys Leu Glu Tyr Ala

275 280 285

Glu Ile Tyr Ala Phe Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser

290 295 300

Ala Arg Lys Tyr Phe Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile

305 310 315 320

Ala Lys Arg Leu Asn Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn

325 330 335

Lys Tyr Ile Gly Glu Tyr Lys Gln Lys Leu Ile Arg Lys Tyr Arg Phe

340 345 350

Val Val Glu Ser Ser Gly Glu Val Thr Val Asn Arg Asn Lys Phe Val

355 360 365

Glu Leu Tyr Asn Glu Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala

370 375 380

Lys Ile Tyr Asn Val Gln Asn Arg Lys Ile Tyr Leu Ser Asn Val Tyr

385 390 395 400

Thr Pro Val Thr Ala Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln

405 410 415

Asn Gly Phe Asn Ile Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly

420 425 430

Gln Asn Leu Ser Arg Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn

435 440 445

Met Leu Tyr Leu Phe Thr Lys Phe Cys His Lys Ala Ile Asp Gly Arg

450 455 460

Ser Leu Tyr Asn Lys Cys Gln Asn Leu Phe Lys Asn Ile Asn Ile Met

465 470 475 480

Ala Lys

<210> 55

<211> 475

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/D轻链-MTDr氨基酸序列

<400> 55

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro

1 5 10 15

Arg Gly Ser His Met Thr Trp Pro Val Lys Asp Phe Asn Tyr Ser Asp

20 25 30

Pro Val Asn Asp Asn Asp Ile Leu Tyr Leu Arg Ile Pro Gln Asn Lys

35 40 45

Leu Ile Thr Thr Pro Val Lys Ala Phe Met Ile Thr Gln Asn Ile Trp

50 55 60

Val Ile Pro Glu Arg Phe Ser Ser Asp Thr Asn Pro Ser Leu Ser Lys

65 70 75 80

Pro Pro Arg Pro Thr Ser Lys Tyr Gln Ser Tyr Tyr Asp Pro Ser Tyr

85 90 95

Leu Ser Thr Asp Glu Gln Lys Asp Thr Phe Leu Lys Gly Ile Ile Lys

100 105 110

Leu Phe Lys Arg Ile Asn Glu Arg Asp Ile Gly Lys Lys Leu Ile Asn

115 120 125

Tyr Leu Val Val Gly Ser Pro Phe Met Gly Asp Ser Ser Thr Pro Glu

130 135 140

Asp Thr Phe Asp Phe Thr Arg His Thr Thr Asn Ile Ala Val Glu Lys

145 150 155 160

Phe Glu Asn Gly Ser Trp Lys Val Thr Asn Ile Ile Thr Pro Ser Val

165 170 175

Leu Ile Phe Gly Pro Leu Pro Asn Ile Leu Asp Tyr Thr Ala Ser Leu

180 185 190

Thr Leu Gln Gly Gln Gln Ser Asn Pro Ser Phe Glu Gly Phe Gly Thr

195 200 205

Leu Ser Ile Leu Lys Val Ala Pro Glu Phe Leu Leu Thr Phe Ser Asp

210 215 220

Val Thr Ser Asn Gln Ser Ser Ala Val Leu Gly Lys Ser Ile Phe Cys

225 230 235 240

Met Asp Pro Val Ile Ala Leu Met His Glu Leu Thr His Ser Leu His

245 250 255

Gln Leu Tyr Gly Ile Asn Ile Pro Ser Asp Lys Arg Ile Arg Pro Gln

260 265 270

Val Ser Glu Gly Phe Phe Ser Gln Asp Gly Pro Asn Val Gln Phe Glu

275 280 285

Glu Leu Tyr Thr Phe Gly Gly Leu Asp Val Glu Ile Ile Pro Gln Ile

290 295 300

Glu Arg Ser Gln Leu Arg Glu Lys Ala Leu Gly His Tyr Lys Asp Ile

305 310 315 320

Ala Lys Arg Leu Asn Asn Ile Asn Lys Thr Ile Pro Ser Ser Trp Ile

325 330 335

Ser Asn Ile Asp Lys Tyr Lys Lys Ile Phe Ser Glu Lys Tyr Asn Phe

340 345 350

Asp Lys Asp Asn Thr Gly Asn Phe Val Val Asn Ile Asp Lys Phe Asn

355 360 365

Ser Leu Tyr Ser Asp Leu Thr Asn Val Met Ser Glu Val Val Tyr Ser

370 375 380

Ser Gln Tyr Asn Val Lys Asn Arg Thr His Tyr Phe Ser Arg His Tyr

385 390 395 400

Leu Pro Val Phe Ala Asn Ile Leu Asp Asp Asn Ile Tyr Thr Ile Arg

405 410 415

Asp Gly Phe Asn Leu Thr Asn Lys Gly Phe Asn Ile Glu Asn Ser Gly

420 425 430

Gln Asn Ile Glu Arg Asn Pro Ala Leu Gln Lys Leu Ser Ser Glu Ser

435 440 445

Val Val Asp Leu Phe Thr Lys Val Cys Leu Arg Leu Thr Lys Cys Gln

450 455 460

Asn Leu Phe Lys Asn Ile Asn Ile Met Ala Lys

465 470 475

<210> 56

<211> 455

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/E轻链-MTDr氨基酸序列

<400> 56

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro

1 5 10 15

Arg Gly Ser His Met Pro Thr Ile Asn Ser Phe Asn Tyr Asn Asp Pro

20 25 30

Val Asn Asn Arg Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln

35 40 45

Phe Tyr Lys Ser Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu

50 55 60

Arg Asn Val Ile Gly Thr Ile Pro Gln Asp Phe Leu Pro Pro Thr Ser

65 70 75 80

Leu Lys Asn Gly Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser

85 90 95

Asp Gln Glu Lys Asp Lys Phe Leu Lys Ile Val Thr Lys Ile Phe Asn

100 105 110

Arg Ile Asn Asp Asn Leu Ser Gly Arg Ile Leu Leu Glu Glu Leu Ser

115 120 125

Lys Ala Asn Pro Tyr Leu Gly Asn Asp Asn Thr Pro Asp Gly Asp Phe

130 135 140

Ile Ile Asn Asp Ala Ser Ala Val Pro Ile Gln Phe Ser Asn Gly Ser

145 150 155 160

Gln Ser Ile Leu Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp

165 170 175

Leu Phe Glu Thr Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met

180 185 190

Pro Ser Asn His Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro

195 200 205

Glu Tyr Ser Phe Arg Phe Lys Asp Asn Ser Met Asn Glu Phe Ile Gln

210 215 220

Asp Pro Ala Leu Thr Leu Met His Glu Leu Ile His Ser Leu His Gly

225 230 235 240

Leu Tyr Gly Ala Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Lys

245 250 255

Gln Asn Pro Leu Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe

260 265 270

Leu Thr Phe Gly Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Ser

275 280 285

Asn Asp Ile Tyr Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser

290 295 300

Lys Leu Ser Lys Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys

305 310 315 320

Asp Val Phe Glu Ala Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile

325 330 335

Tyr Ser Val Asn Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr

340 345 350

Ser Phe Thr Glu Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg

355 360 365

Gln Thr Tyr Ile Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu

370 375 380

Asn Asp Ser Ile Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu

385 390 395 400

Lys Val Asn Phe Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile

405 410 415

Thr Pro Ile Thr Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys

420 425 430

Lys Asn Ile Val Ser Val Lys Gly Ile Arg Cys Gln Asn Leu Phe Lys

435 440 445

Asn Ile Asn Ile Met Ala Lys

450 455

<210> 57

<211> 469

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/F轻链-MTDr氨基酸序列

<400> 57

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro

1 5 10 15

Arg Gly Ser His Met Pro Val Ala Ile Asn Ser Phe Asn Tyr Asn Asp

20 25 30

Pro Val Asn Asp Asp Thr Ile Leu Tyr Met Gln Ile Pro Tyr Glu Glu

35 40 45

Lys Ser Lys Lys Tyr Tyr Lys Ala Phe Glu Ile Met Arg Asn Val Trp

50 55 60

Ile Ile Pro Glu Arg Asn Thr Ile Gly Thr Asn Pro Ser Asp Phe Asp

65 70 75 80

Pro Pro Ala Ser Leu Lys Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn

85 90 95

Tyr Leu Thr Thr Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr Ile

100 105 110

Lys Leu Phe Lys Arg Ile Asn Ser Asn Pro Ala Gly Lys Val Leu Leu

115 120 125

Gln Glu Ile Ser Tyr Ala Lys Pro Tyr Leu Gly Asn Asp His Thr Pro

130 135 140

Ile Asp Glu Phe Ser Pro Val Thr Arg Thr Thr Ser Val Asn Ile Lys

145 150 155 160

Leu Ser Thr Asn Val Glu Ser Ser Met Leu Leu Asn Leu Leu Val Leu

165 170 175

Gly Ala Gly Pro Asp Ile Phe Glu Ser Cys Cys Tyr Pro Val Arg Lys

180 185 190

Leu Ile Asp Pro Asp Val Val Tyr Asp Pro Ser Asn Tyr Gly Phe Gly

195 200 205

Ser Ile Asn Ile Val Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn

210 215 220

Asp Ile Ser Gly Gly His Asn Ser Ser Thr Glu Ser Phe Ile Ala Asp

225 230 235 240

Pro Ala Ile Ser Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu

245 250 255

Tyr Gly Ala Arg Gly Val Thr Tyr Glu Glu Thr Ile Glu Val Lys Gln

260 265 270

Ala Pro Leu Met Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu

275 280 285

Thr Phe Gly Gly Gln Asp Leu Asn Ile Ile Thr Ser Ala Met Lys Glu

290 295 300

Lys Ile Tyr Asn Asn Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg

305 310 315 320

Leu Ser Glu Val Asn Ser Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr

325 330 335

Lys Asp Tyr Phe Gln Trp Lys Tyr Gly Leu Asp Lys Asn Ala Asp Gly

340 345 350

Ser Tyr Thr Val Asn Glu Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu

355 360 365

Tyr Ser Phe Thr Glu Ser Asp Leu Ala Asn Lys Phe Lys Val Lys Cys

370 375 380

Arg Asn Thr Tyr Phe Ile Lys Tyr Glu Phe Leu Lys Val Pro Asn Leu

385 390 395 400

Leu Asp Asp Asp Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn

405 410 415

Leu Ala Val Asn Asn Arg Gly Gln Ser Ile Lys Leu Asn Pro Lys Ile

420 425 430

Ile Asp Ser Ile Pro Asp Lys Gly Leu Val Glu Lys Ile Val Lys Phe

435 440 445

Cys Lys Ser Val Ile Pro Arg Lys Cys Gln Asn Leu Phe Lys Asn Ile

450 455 460

Asn Ile Met Ala Lys

465

<210> 58

<211> 474

<212> PRT

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/G轻链-MTDr氨基酸序列

<400> 58

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro

1 5 10 15

Arg Gly Ser His Met Pro Val Asn Ile Lys Phe Asn Tyr Asn Asp Pro

20 25 30

Ile Asn Asn Asp Asp Ile Ile Met Met Glu Pro Phe Asn Asp Pro Gly

35 40 45

Pro Gly Thr Tyr Tyr Lys Ala Phe Arg Ile Ile Asp Arg Ile Trp Ile

50 55 60

Val Pro Glu Arg Phe Thr Tyr Gly Phe Gln Pro Asp Gln Phe Asn Ala

65 70 75 80

Ser Thr Gly Val Phe Ser Lys Asp Val Tyr Glu Tyr Tyr Asp Pro Thr

85 90 95

Tyr Leu Lys Thr Asp Ala Glu Lys Asp Lys Phe Leu Lys Thr Met Ile

100 105 110

Lys Leu Phe Asn Arg Ile Asn Ser Lys Pro Ser Gly Gln Arg Leu Leu

115 120 125

Asp Met Ile Val Asp Ala Ile Pro Tyr Leu Gly Asn Ala Ser Thr Pro

130 135 140

Pro Asp Lys Phe Ala Ala Asn Val Ala Asn Val Ser Ile Asn Lys Lys

145 150 155 160

Ile Ile Gln Pro Gly Ala Glu Asp Gln Ile Lys Gly Leu Met Thr Asn

165 170 175

Leu Ile Ile Phe Gly Pro Gly Pro Val Leu Ser Asp Asn Phe Thr Asp

180 185 190

Ser Met Ile Met Asn Gly His Ser Pro Ile Ser Glu Gly Phe Gly Ala

195 200 205

Arg Met Met Ile Arg Phe Cys Pro Ser Cys Leu Asn Val Phe Asn Asn

210 215 220

Val Gln Glu Asn Lys Asp Thr Ser Ile Phe Ser Arg Arg Ala Tyr Phe

225 230 235 240

Ala Asp Pro Ala Leu Thr Leu Met His Glu Leu Ile His Val Leu His

245 250 255

Gly Leu Tyr Gly Ile Lys Ile Ser Asn Leu Pro Ile Thr Pro Asn Thr

260 265 270

Lys Glu Phe Phe Met Gln His Ser Asp Pro Val Gln Ala Glu Glu Leu

275 280 285

Tyr Thr Phe Gly Gly His Asp Pro Ser Val Ile Ser Pro Ser Thr Asp

290 295 300

Met Asn Ile Tyr Asn Lys Ala Leu Gln Asn Phe Gln Asp Ile Ala Asn

305 310 315 320

Arg Leu Asn Ile Val Ser Ser Ala Gln Gly Ser Gly Ile Asp Ile Ser

325 330 335

Leu Tyr Lys Gln Ile Tyr Lys Asn Lys Tyr Asp Phe Val Glu Asp Pro

340 345 350

Asn Gly Lys Tyr Ser Val Asp Lys Asp Lys Phe Asp Lys Leu Tyr Lys

355 360 365

Ala Leu Met Phe Gly Phe Thr Glu Thr Asn Leu Ala Gly Glu Tyr Gly

370 375 380

Ile Lys Thr Arg Tyr Ser Tyr Phe Ser Glu Tyr Leu Pro Pro Ile Lys

385 390 395 400

Thr Glu Lys Leu Leu Asp Asn Thr Ile Tyr Thr Gln Asn Glu Gly Phe

405 410 415

Asn Ile Ala Ser Lys Asn Leu Lys Thr Glu Phe Asn Gly Gln Asn Lys

420 425 430

Ala Val Asn Lys Glu Ala Tyr Glu Glu Ile Ser Leu Glu His Leu Val

435 440 445

Ile Tyr Arg Ile Ala Met Cys Lys Pro Val Met Tyr Lys Cys Gln Asn

450 455 460

Leu Phe Lys Asn Ile Asn Ile Met Ala Lys

465 470

<210> 59

<211> 1386

<212> DNA

<213> 人工序列

<220>

<223> MTD-BoNT/A轻链cDNA序列

<400> 59

atgaaggcca tgatcaatat taacaagttc ttaaatcaat gtccctttgt caacaaacag 60

ttcaactaca aggacccagt taatggagta gacatcgcat atatcaagat tcccaacgct 120

ggccagatgc aacccgttaa ggcatttaaa atccataaca aaatctgggt tatcccagag 180

cgggatacct tcaccaaccc cgaggagggc gatctgaacc ccccgccgga ggcgaagcag 240

gtcccagtga gctactacga tagcacctac ctcagcaccg acaacgagaa ggacaactac 300

ctcaaaggag tcacgaagtt gttcgagaga atctactcca cagacctcgg ccgcatgctt 360

ctaaccagca ttgtgcgtgg cattcccttt tggggcggct ctaccatcga cacagagctg 420

aaggtgatag acaccaactg catcaacgta atccagcctg acggcagcta ccgaagcgag 480

gagcttaacc tggtgatcat cggcccttcc gccgatatca tccaattcga gtgcaagagc 540

ttcggccacg aggtcctgaa cctcacccgg aacggctatg gaagcaccca gtacataaga 600

ttcagccctg acttcacctt cgggtttgag gagagcttgg aggtcgacac aaaccccctg 660

ctgggagccg ggaagttcgc cactgaccca gccgtgactc tggcacacga gctgatccac 720

gccggtcacc gcctgtacgg catagctata aacccaaaca gggtgttcaa agtgaacacc 780

aacgcttact atgaaatgag cggcctggag gtgagcttcg aggagctgag aacgttcggg 840

ggacatgatg ctaaatttat cgacagcctg caggagaacg agttcaggct gtactactac 900

aataagttca aggatatagc gagcactctg aacaaggcca agtccatcgt aggcactact 960

gcatccctcc agtatatgaa gaatgtgttc aaagagaaat acctgctgag cgaggatacc 1020

agcggtaagt tcagcgtgga taagcttaag ttcgacaagc tgtataagat gctcaccgaa 1080

atctacaccg aggataattt cgttaagttc ttcaaggtcc tgaaccggaa gacctacctg 1140

aacttcgaca aggccgtgtt caagatcaac atcgtgccta aagtgaacta caccatctac 1200

gacgggttta acctgaggaa caccaacctg gccgctaact tcaacgggca gaacacagag 1260

atcaacaaca tgaatttcac gaagttgaag aacttcaccg gactgtttga gttctacaaa 1320

ttgctgtgtg tgcgcgggat catcactagc aagaccaaga gccttgacaa aggctacaac 1380

aagtga 1386

<210> 60

<211> 1362

<212> DNA

<213> 人工序列

<220>

<223> MTD-BoNT/B轻链cDNA序列

<400> 60

atgaaggcca tgatcaatat taacaagttc ttaaatcaat gtccagttac aataaataat 60

tttaattata atgatcctat tgataataat aatattatta tgatggagcc tccatttgcg 120

agaggtacgg ggagatatta taaagctttt aaaatcacag atcgtatttg gataataccg 180

gaaagatata cttttggata taaacctgag gattttaata aaagttccgg tatttttaat 240

agagatgttt gtgaatatta tgatccagat tacttaaata ctaatgataa aaagaatata 300

tttttacaaa caatgatcaa gttatttaat agaatcaaat caaaaccatt gggtgaaaag 360

ttattagaga tgattataaa tggtatacct tatcttggag atagacgtgt tccactcgaa 420

gagtttaaca caaacattgc tagtgtaact gttaataaat taatcagtaa tccaggagaa 480

gtggagcgaa aaaaaggtat tttcgcaaat ttaataatat ttggacctgg gccagtttta 540

aatgaaaatg agactataga tataggtata caaaatcatt ttgcatcaag ggaaggcttc 600

gggggtataa tgcaaatgaa gttttgccca gaatatgtaa gcgtatttaa taatgttcaa 660

gaaaacaaag gcgcaagtat atttaataga cgtggatatt tttcagatcc agccttgata 720

ttaatgcatg aacttataca tgttttacat ggattatatg gcattaaagt agatgattta 780

ccaattgtac caaatgaaaa aaaatttttt atgcaatcta cagatgctat acaggcagaa 840

gaactatata catttggagg acaagatccc agcatcataa ctccttctac ggataaaagt 900

atctatgata aagttttgca aaattttaga gggatagttg atagacttaa caaggtttta 960

gtttgcatat cagatcctaa cattaatatt aatatatata aaaataaatt taaagataaa 1020

tataaattcg ttgaagattc tgagggaaaa tatagtatag atgtagaaag ttttgataaa 1080

ttatataaaa gcttaatgtt tggttttaca gaaactaata tagcagaaaa ttataaaata 1140

aaaactagag cttcttattt tagtgattcc ttaccaccag taaaaataaa aaatttatta 1200

gataatgaaa tctatactat agaggaaggg tttaatatat ctgataaaga tatggaaaaa 1260

gaatatagag gtcagaataa agctataaat aaacaagctt atgaagaaat tagcaaggag 1320

catttggctg tatataagat acaaatgtgt aaaagtgtta aa 1362

<210> 61

<211> 1386

<212> DNA

<213> 人工序列

<220>

<223> MTD-BoNT/C轻链cDNA序列

<400> 61

atgaaggcca tgatcaatat taacaagttc ttaaatcaat gtccaataac aattaacaac 60

tttaattatt cagatcctgt tgataataaa aatattttat atttagatac tcatttaaat 120

acactagcta atgagcctga aaaagccttt cgcattacag gaaatatatg ggtaatacct 180

gatagatttt caagaaattc taatccaaat ttaaataaac ctcctcgagt tacaagccct 240

aaaagtggtt attatgatcc taattatttg agtactgatt ctgacaaaga tacattttta 300

aaagaaatta taaagttatt taaaagaatt aattctagag aaataggaga agaattaata 360

tatagacttt cgacagatat accctttcct gggaataaca atactccaat taatactttt 420

gattttgatg tagattttaa cagtgttgat gttaaaacta gacaaggtaa caactgggtt 480

aaaactggta gcataaatcc tagtgttata ataactggac ctagagaaaa cattatagat 540

ccagaaactt ctacgtttaa attaactaac aatacttttg cggcacaaga aggatttggt 600

gctttatcaa taatttcaat atcacctaga tttatgctaa catatagtaa tgcaactaat 660

gatgtaggag agggtagatt ttctaagtct gaattttgca tggatccaat actaatttta 720

atgcatgaac ttaatcatgc aatgcataat ttatatggaa tagctatacc aaatgatcaa 780

acaatttcat ctgtaactag taatattttt tattctcaat ataatgtgaa attagagtat 840

gcagaaatat atgcatttgg aggtccaact atagacctta ttcctaaaag tgcaaggaaa 900

tattttgagg aaaaggcatt ggattattat agatctatag ctaaaagact taatagtata 960

actactgcaa atccttcaag ctttaataaa tatatagggg aatataaaca gaaacttatt 1020

agaaagtata gattcgtagt agaatcttca ggtgaagtta cagtaaatcg taataagttt 1080

gttgagttat ataatgaact tacacaaata tttacagaat ttaactacgc taaaatatat 1140

aatgtacaaa ataggaaaat atatctttca aatgtatata ctccggttac ggcgaatata 1200

ttagacgata atgtttatga tatacaaaat ggatttaata tacctaaaag taatttaaat 1260

gtactattta tgggtcaaaa tttatctcga aatccagcat taagaaaagt caatcctgaa 1320

aatatgcttt atttatttac aaaattttgt cataaagcaa tagatggtag atcattatat 1380

aataaa 1386

<210> 62

<211> 1365

<212> DNA

<213> 人工序列

<220>

<223> MTD-BoNT/D轻链cDNA序列

<400> 62

atgaaggcca tgatcaatat taacaagttc ttaaatcaat gtacatggcc agtaaaagat 60

tttaattata gtgatcctgt taatgacaat gatatattat atttaagaat accacaaaat 120

aagttaatta ctacacctgt aaaagctttt atgattactc aaaatatttg ggtaatacca 180

gaaagatttt catcagatac taatccaagt ttaagtaaac cgcccagacc tacttcaaag 240

tatcaaagtt attatgatcc tagttattta tctactgatg aacaaaaaga tacattttta 300

aaagggatta taaaattatt taaaagaatt aatgaaagag atataggaaa aaaattaata 360

aattatttag tagttggttc accttttatg ggagattcaa gtacgcctga agatacattt 420

gattttacac gtcatactac taatattgca gttgaaaagt ttgaaaatgg tagttggaaa 480

gtaacaaata ttataacacc aagtgtattg atatttggac cacttcctaa tatattagac 540

tatacagcat cccttacatt gcaaggacaa caatcaaatc catcatttga agggtttgga 600

acattatcta tactaaaagt agcacctgaa tttttgttaa catttagtga tgtaacatct 660

aatcaaagtt cagctgtatt aggcaaatct atattttgta tggatccagt aatagcttta 720

atgcatgagt taacacattc tttgcatcaa ttatatggaa taaatatacc atctgataaa 780

aggattcgtc cacaagttag cgagggattt ttctctcaag atggacccaa cgtacaattt 840

gaggaattat atacatttgg aggattagat gttgaaataa tacctcaaat tgaaagatca 900

caattaagag aaaaagcatt aggtcactat aaagatatag cgaaaagact taataatatt 960

aataaaacta ttccttctag ttggattagt aatatagata aatataaaaa aatattttct 1020

gaaaagtata attttgataa agataataca ggaaattttg ttgtaaatat tgataaattc 1080

aatagcttat attcagactt gactaatgtt atgtcagaag ttgtttattc ttcgcaatat 1140

aatgttaaaa acaggactca ttatttttca aggcattatc tacctgtatt tgcaaatata 1200

ttagatgata atatttatac tataagagat ggttttaatt taacaaataa aggttttaat 1260

atagaaaatt cgggtcagaa tatagaaagg aatcctgcac tacaaaagct tagttcagaa 1320

agtgtagtag atttatttac aaaagtatgt ttaagattaa caaaa 1365

<210> 63

<211> 1305

<212> DNA

<213> 人工序列

<220>

<223> MTD-BoNT/E轻链cDNA序列

<400> 63

atgaaggcca tgatcaatat taacaagttc ttaaatcaat gtccaacaat taatagtttt 60

aattataatg atcctgttaa taatagaaca attttatata ttaaaccagg cggttgtcaa 120

caattttata aatcatttaa tattatgaaa aatatttgga taattccaga gagaaatgta 180

attggtacaa ttccccaaga ttttcttccg cctacttcat tgaaaaatgg agatagtagt 240

tattatgacc ctaattattt acaaagtgat caagaaaagg ataaattttt aaaaatagtc 300

acaaaaatat ttaatagaat aaatgataat ctttcaggaa ggattttatt agaagaactg 360

tcaaaagcta atccatattt aggaaatgat aatactccag atggtgactt cattattaat 420

gatgcatcag cagttccaat tcaattctca aatggtagcc aaagcatact attacctaat 480

gttattataa tgggagcaga gcctgattta tttgaaacta acagttccaa tatttctcta 540

agaaataatt atatgccaag caatcacggt tttggatcaa tagctatagt aacattctca 600

cctgaatatt cttttagatt taaagataat agtatgaatg aatttattca agatcctgct 660

cttacattaa tgcatgaatt aatacattca ttacatggac tatatggggc taaagggatt 720

actacaaagt atactataac acaaaaacaa aatcccctaa taacaaatat aagaggtaca 780

aatattgaag aattcttaac ttttggaggt actgatttaa acattattac tagtgctcag 840

tccaatgata tctatactaa tcttctagct gattataaaa aaatagcgtc taaacttagc 900

aaagtacaag tatctaatcc actacttaat ccttataaag atgtttttga agcaaagtat 960

ggattagata aagatgctag cggaatttat tcggtaaata taaacaaatt taatgatatt 1020

tttaaaaaat tatacagctt tacggaattt gatttagcaa ctaaatttca agttaaatgt 1080

aggcaaactt atattggaca gtataaatac ttcaaacttt caaacttgtt aaatgattct 1140

atttataata tatcagaagg ctataatata aataatttaa aggtaaattt tagaggacag 1200

aatgcaaatt taaatcctag aattattaca ccaattacag gtagaggact agtaaaaaaa 1260

atcattagat tttgtaaaaa tattgtttct gtaaaaggca taagg 1305

<210> 64

<211> 1347

<212> DNA

<213> 人工序列

<220>

<223> MTD-BoNT/F轻链cDNA序列

<400> 64

atgaaggcca tgatcaatat taacaagttc ttaaatcaat gtccagttgc aataaatagt 60

tttaattata atgaccctgt taatgatgat acaattttat acatgcagat accatatgaa 120

gaaaaaagta aaaaatatta taaagctttt gagattatgc gtaatgtttg gataattcct 180

gagagaaata caataggaac gaatcctagt gattttgatc caccggcttc attaaagaac 240

ggaagcagtg cttattatga tcctaattat ttaaccactg atgctgaaaa agatagatat 300

ttaaaaacaa cgataaaatt atttaagaga attaatagta atcctgcagg gaaagttttg 360

ttacaagaaa tatcatatgc taaaccatat ttaggaaatg accacacgcc aattgatgaa 420

ttctctccag ttactagaac tacaagtgtt aatataaaat tatcaactaa tgttgaaagt 480

tcaatgttat tgaatcttct tgtattggga gcaggacctg atatatttga aagttgttgt 540

taccccgtta gaaaactaat agatccagat gtagtttatg atccaagtaa ttatggtttt 600

ggatcaatta atatcgtgac attttcacct gagtatgaat atacttttaa tgatattagt 660

ggagggcata atagtagtac agaatcattt attgcagatc ctgcaatttc actagctcat 720

gaattgatac atgcactgca tggattatac ggggctaggg gagttactta tgaagagact 780

atagaagtaa agcaagcacc tcttatgata gccgaaaaac ccataaggct agaagaattt 840

ttaacctttg gaggtcagga tttaaatatt attactagtg ctatgaagga aaaaatatat 900

aacaatcttt tagctaacta tgaaaaaata gctactagac ttagtgaagt taatagtgct 960

cctcctgaat atgatattaa tgaatataaa gattattttc aatggaagta tgggctagat 1020

aaaaatgctg atggaagtta tactgtaaat gaaaataaat ttaatgaaat ttataaaaaa 1080

ttatatagtt ttacagagag tgacttagca aataaattta aagtaaaatg tagaaatact 1140

tattttatta aatatgaatt tttaaaagtt ccaaatttgt tagatgatga tatttatact 1200

gtatcagagg ggtttaatat aggtaattta gcagtaaaca atcgcggaca aagtataaag 1260

ttaaatccta aaattattga ttccattcca gataaaggtc tagtagaaaa gatcgttaaa 1320

ttttgtaaga gcgttattcc tagaaaa 1347

<210> 65

<211> 1365

<212> DNA

<213> 人工序列

<220>

<223> MTD-BoNT/G轻链cDNA序列

<400> 65

atgaaggcca tgatcaatat taacaagttc ttaaatcaat gtccagttaa tataaaaanc 60

tttaattata atgaccctat taataatgat gacattatta tgatggaacc attcaatgac 120

ccagggccag gaacatatta taaagctttt aggattatag atcgtatttg gatagtacca 180

gaaaggttta cttatggatt tcaacctgac caatttaatg ccagtacagg agtttttagt 240

aaagatgtct acgaatatta cgatccaact tatttaaaaa ccgatgctga aaaagataaa 300

tttttaaaaa caatgattaa attatttaat agaattaatt caaaaccatc aggacagaga 360

ttactggata tgatagtaga tgctatacct tatcttggaa atgcatctac accgcccgac 420

aaatttgcag caaatgttgc aaatgtatct attaataaaa aaattatcca acctggagct 480

gaagatcaaa taaaaggttt aatgacaaat ttaataatat ttggaccagg accagttcta 540

agtgataatt ttactgatag tatgattatg aatggccatt ccccaatatc agaaggattt 600

ggtgcaagaa tgatgataag attttgtcct agttgtttaa atgtatttaa taatgttcag 660

gaaaataaag atacatctat atttagtaga cgcgcgtatt ttgcagatcc agctctaacg 720

ttaatgcatg aacttataca tgtgttacat ggattatatg gaattaagat aagtaattta 780

ccaattactc caaatacaaa agaatttttc atgcaacata gcgatcctgt acaagcagaa 840

gaactatata cattcggagg acatgatcct agtgttataa gtccttctac ggatatgaat 900

atttataata aagcgttaca aaattttcaa gatatagcta ataggcttaa tattgtttca 960

agtgcccaag ggagtggaat tgatatttcc ttatataaac aaatatataa aaataaatat 1020

gattttgttg aagatcctaa tggaaaatat agtgtagata aggataagtt tgataaatta 1080

tataaggcct taatgtttgg ctttactgaa actaatctag ctggtgaata tggaataaaa 1140

actaggtatt cttattttag tgaatatttg ccaccgataa aaactgaaaa attgttagac 1200

aatacaattt atactcaaaa tgaaggcttt aacatagcta gtaaaaatct caaaacggaa 1260

tttaatggtc agaataaggc ggtaaataaa gaggcttatg aagaaatcag cctagaacat 1320

ctcgttatat atagaatagc aatgtgcaag cctgtaatgt acaaa 1365

<210> 66

<211> 1386

<212> DNA

<213> 人工序列

<220>

<223> BoNT/A轻链-MTDr cDNA序列

<400> 66

atgccctttg tcaacaaaca gttcaactac aaggacccag ttaatggagt agacatcgca 60

tatatcaaga ttcccaacgc tggccagatg caacccgtta aggcatttaa aatccataac 120

aaaatctggg ttatcccaga gcgggatacc ttcaccaacc ccgaggaggg cgatctgaac 180

cccccgccgg aggcgaagca ggtcccagtg agctactacg atagcaccta cctcagcacc 240

gacaacgaga aggacaacta cctcaaagga gtcacgaagt tgttcgagag aatctactcc 300

acagacctcg gccgcatgct tctaaccagc attgtgcgtg gcattccctt ttggggcggc 360

tctaccatcg acacagagct gaaggtgata gacaccaact gcatcaacgt aatccagcct 420

gacggcagct accgaagcga ggagcttaac ctggtgatca tcggcccttc cgccgatatc 480

atccaattcg agtgcaagag cttcggccac gaggtcctga acctcacccg gaacggctat 540

ggaagcaccc agtacataag attcagccct gacttcacct tcgggtttga ggagagcttg 600

gaggtcgaca caaaccccct gctgggagcc gggaagttcg ccactgaccc agccgtgact 660

ctggcacacg agctgatcca cgccggtcac cgcctgtacg gcatagctat aaacccaaac 720

agggtgttca aagtgaacac caacgcttac tatgaaatga gcggcctgga ggtgagcttc 780

gaggagctga gaacgttcgg gggacatgat gctaaattta tcgacagcct gcaggagaac 840

gagttcaggc tgtactacta caataagttc aaggatatag cgagcactct gaacaaggcc 900

aagtccatcg taggcactac tgcatccctc cagtatatga agaatgtgtt caaagagaaa 960

tacctgctga gcgaggatac cagcggtaag ttcagcgtgg ataagcttaa gttcgacaag 1020

ctgtataaga tgctcaccga aatctacacc gaggataatt tcgttaagtt cttcaaggtc 1080

ctgaaccgga agacctacct gaacttcgac aaggccgtgt tcaagatcaa catcgtgcct 1140

aaagtgaact acaccatcta cgacgggttt aacctgagga acaccaacct ggccgctaac 1200

ttcaacgggc agaacacaga gatcaacaac atgaatttca cgaagttgaa gaacttcacc 1260

ggactgtttg agttctacaa attgctgtgt gtgcgcggga tcatcactag caagaccaag 1320

agccttgaca aaggctacaa caagtgatgt caaaatttat tcaagaacat taatatcatg 1380

gccaag 1386

<210> 67

<211> 1362

<212> DNA

<213> 人工序列

<220>

<223> BoNT/B轻链-MTDr cDNA序列

<400> 67

atgccagtta caataaataa ttttaattat aatgatccta ttgataataa taatattatt 60

atgatggagc ctccatttgc gagaggtacg gggagatatt ataaagcttt taaaatcaca 120

gatcgtattt ggataatacc ggaaagatat acttttggat ataaacctga ggattttaat 180

aaaagttccg gtatttttaa tagagatgtt tgtgaatatt atgatccaga ttacttaaat 240

actaatgata aaaagaatat atttttacaa acaatgatca agttatttaa tagaatcaaa 300

tcaaaaccat tgggtgaaaa gttattagag atgattataa atggtatacc ttatcttgga 360

gatagacgtg ttccactcga agagtttaac acaaacattg ctagtgtaac tgttaataaa 420

ttaatcagta atccaggaga agtggagcga aaaaaaggta ttttcgcaaa tttaataata 480

tttggacctg ggccagtttt aaatgaaaat gagactatag atataggtat acaaaatcat 540

tttgcatcaa gggaaggctt cgggggtata atgcaaatga agttttgccc agaatatgta 600

agcgtattta ataatgttca agaaaacaaa ggcgcaagta tatttaatag acgtggatat 660

ttttcagatc cagccttgat attaatgcat gaacttatac atgttttaca tggattatat 720

ggcattaaag tagatgattt accaattgta ccaaatgaaa aaaaattttt tatgcaatct 780

acagatgcta tacaggcaga agaactatat acatttggag gacaagatcc cagcatcata 840

actccttcta cggataaaag tatctatgat aaagttttgc aaaattttag agggatagtt 900

gatagactta acaaggtttt agtttgcata tcagatccta acattaatat taatatatat 960

aaaaataaat ttaaagataa atataaattc gttgaagatt ctgagggaaa atatagtata 1020

gatgtagaaa gttttgataa attatataaa agcttaatgt ttggttttac agaaactaat 1080

atagcagaaa attataaaat aaaaactaga gcttcttatt ttagtgattc cttaccacca 1140

gtaaaaataa aaaatttatt agataatgaa atctatacta tagaggaagg gtttaatata 1200

tctgataaag atatggaaaa agaatataga ggtcagaata aagctataaa taaacaagct 1260

tatgaagaaa ttagcaagga gcatttggct gtatataaga tacaaatgtg taaaagtgtt 1320

aaatgtcaaa atttattcaa gaacattaat atcatggcca ag 1362

<210> 68

<211> 1386

<212> DNA

<213> 人工序列

<220>

<223> BoNT/C轻链-MTDr cDNA序列

<400> 68

atgccaataa caattaacaa ctttaattat tcagatcctg ttgataataa aaatatttta 60

tatttagata ctcatttaaa tacactagct aatgagcctg aaaaagcctt tcgcattaca 120

ggaaatatat gggtaatacc tgatagattt tcaagaaatt ctaatccaaa tttaaataaa 180

cctcctcgag ttacaagccc taaaagtggt tattatgatc ctaattattt gagtactgat 240

tctgacaaag atacattttt aaaagaaatt ataaagttat ttaaaagaat taattctaga 300

gaaataggag aagaattaat atatagactt tcgacagata taccctttcc tgggaataac 360

aatactccaa ttaatacttt tgattttgat gtagatttta acagtgttga tgttaaaact 420

agacaaggta acaactgggt taaaactggt agcataaatc ctagtgttat aataactgga 480

cctagagaaa acattataga tccagaaact tctacgttta aattaactaa caatactttt 540

gcggcacaag aaggatttgg tgctttatca ataatttcaa tatcacctag atttatgcta 600

acatatagta atgcaactaa tgatgtagga gagggtagat tttctaagtc tgaattttgc 660

atggatccaa tactaatttt aatgcatgaa cttaatcatg caatgcataa tttatatgga 720

atagctatac caaatgatca aacaatttca tctgtaacta gtaatatttt ttattctcaa 780

tataatgtga aattagagta tgcagaaata tatgcatttg gaggtccaac tatagacctt 840

attcctaaaa gtgcaaggaa atattttgag gaaaaggcat tggattatta tagatctata 900

gctaaaagac ttaatagtat aactactgca aatccttcaa gctttaataa atatataggg 960

gaatataaac agaaacttat tagaaagtat agattcgtag tagaatcttc aggtgaagtt 1020

acagtaaatc gtaataagtt tgttgagtta tataatgaac ttacacaaat atttacagaa 1080

tttaactacg ctaaaatata taatgtacaa aataggaaaa tatatctttc aaatgtatat 1140

actccggtta cggcgaatat attagacgat aatgtttatg atatacaaaa tggatttaat 1200

atacctaaaa gtaatttaaa tgtactattt atgggtcaaa atttatctcg aaatccagca 1260

ttaagaaaag tcaatcctga aaatatgctt tatttattta caaaattttg tcataaagca 1320

atagatggta gatcattata taataaatgt caaaatttat tcaagaacat taatatcatg 1380

gccaag 1386

<210> 69

<211> 1365

<212> DNA

<213> 人工序列

<220>

<223> BoNT/D轻链-MTDr cDNA序列

<400> 69

atgacatggc cagtaaaaga ttttaattat agtgatcctg ttaatgacaa tgatatatta 60

tatttaagaa taccacaaaa taagttaatt actacacctg taaaagcttt tatgattact 120

caaaatattt gggtaatacc agaaagattt tcatcagata ctaatccaag tttaagtaaa 180

ccgcccagac ctacttcaaa gtatcaaagt tattatgatc ctagttattt atctactgat 240

gaacaaaaag atacattttt aaaagggatt ataaaattat ttaaaagaat taatgaaaga 300

gatataggaa aaaaattaat aaattattta gtagttggtt caccttttat gggagattca 360

agtacgcctg aagatacatt tgattttaca cgtcatacta ctaatattgc agttgaaaag 420

tttgaaaatg gtagttggaa agtaacaaat attataacac caagtgtatt gatatttgga 480

ccacttccta atatattaga ctatacagca tcccttacat tgcaaggaca acaatcaaat 540

ccatcatttg aagggtttgg aacattatct atactaaaag tagcacctga atttttgtta 600

acatttagtg atgtaacatc taatcaaagt tcagctgtat taggcaaatc tatattttgt 660

atggatccag taatagcttt aatgcatgag ttaacacatt ctttgcatca attatatgga 720

ataaatatac catctgataa aaggattcgt ccacaagtta gcgagggatt tttctctcaa 780

gatggaccca acgtacaatt tgaggaatta tatacatttg gaggattaga tgttgaaata 840

atacctcaaa ttgaaagatc acaattaaga gaaaaagcat taggtcacta taaagatata 900

gcgaaaagac ttaataatat taataaaact attccttcta gttggattag taatatagat 960

aaatataaaa aaatattttc tgaaaagtat aattttgata aagataatac aggaaatttt 1020

gttgtaaata ttgataaatt caatagctta tattcagact tgactaatgt tatgtcagaa 1080

gttgtttatt cttcgcaata taatgttaaa aacaggactc attatttttc aaggcattat 1140

ctacctgtat ttgcaaatat attagatgat aatatttata ctataagaga tggttttaat 1200

ttaacaaata aaggttttaa tatagaaaat tcgggtcaga atatagaaag gaatcctgca 1260

ctacaaaagc ttagttcaga aagtgtagta gatttattta caaaagtatg tttaagatta 1320

acaaaatgtc aaaatttatt caagaacatt aatatcatgg ccaag 1365

<210> 70

<211> 1305

<212> DNA

<213> 人工序列

<220>

<223> BoNT/E轻链-MTDr cDNA序列

<400> 70

atgccaacaa ttaatagttt taattataat gatcctgtta ataatagaac aattttatat 60

attaaaccag gcggttgtca acaattttat aaatcattta atattatgaa aaatatttgg 120

ataattccag agagaaatgt aattggtaca attccccaag attttcttcc gcctacttca 180

ttgaaaaatg gagatagtag ttattatgac cctaattatt tacaaagtga tcaagaaaag 240

gataaatttt taaaaatagt cacaaaaata tttaatagaa taaatgataa tctttcagga 300

aggattttat tagaagaact gtcaaaagct aatccatatt taggaaatga taatactcca 360

gatggtgact tcattattaa tgatgcatca gcagttccaa ttcaattctc aaatggtagc 420

caaagcatac tattacctaa tgttattata atgggagcag agcctgattt atttgaaact 480

aacagttcca atatttctct aagaaataat tatatgccaa gcaatcacgg ttttggatca 540

atagctatag taacattctc acctgaatat tcttttagat ttaaagataa tagtatgaat 600

gaatttattc aagatcctgc tcttacatta atgcatgaat taatacattc attacatgga 660

ctatatgggg ctaaagggat tactacaaag tatactataa cacaaaaaca aaatccccta 720

ataacaaata taagaggtac aaatattgaa gaattcttaa cttttggagg tactgattta 780

aacattatta ctagtgctca gtccaatgat atctatacta atcttctagc tgattataaa 840

aaaatagcgt ctaaacttag caaagtacaa gtatctaatc cactacttaa tccttataaa 900

gatgtttttg aagcaaagta tggattagat aaagatgcta gcggaattta ttcggtaaat 960

ataaacaaat ttaatgatat ttttaaaaaa ttatacagct ttacggaatt tgatttagca 1020

actaaatttc aagttaaatg taggcaaact tatattggac agtataaata cttcaaactt 1080

tcaaacttgt taaatgattc tatttataat atatcagaag gctataatat aaataattta 1140

aaggtaaatt ttagaggaca gaatgcaaat ttaaatccta gaattattac accaattaca 1200

ggtagaggac tagtaaaaaa aatcattaga ttttgtaaaa atattgtttc tgtaaaaggc 1260

ataaggtgtc aaaatttatt caagaacatt aatatcatgg ccaag 1305

<210> 71

<211> 1347

<212> DNA

<213> 人工序列

<220>

<223> BoNT/F轻链-MTDr cDNA序列

<400> 71

atgccagttg caataaatag ttttaattat aatgaccctg ttaatgatga tacaatttta 60

tacatgcaga taccatatga agaaaaaagt aaaaaatatt ataaagcttt tgagattatg 120

cgtaatgttt ggataattcc tgagagaaat acaataggaa cgaatcctag tgattttgat 180

ccaccggctt cattaaagaa cggaagcagt gcttattatg atcctaatta tttaaccact 240

gatgctgaaa aagatagata tttaaaaaca acgataaaat tatttaagag aattaatagt 300

aatcctgcag ggaaagtttt gttacaagaa atatcatatg ctaaaccata tttaggaaat 360

gaccacacgc caattgatga attctctcca gttactagaa ctacaagtgt taatataaaa 420

ttatcaacta atgttgaaag ttcaatgtta ttgaatcttc ttgtattggg agcaggacct 480

gatatatttg aaagttgttg ttaccccgtt agaaaactaa tagatccaga tgtagtttat 540

gatccaagta attatggttt tggatcaatt aatatcgtga cattttcacc tgagtatgaa 600

tatactttta atgatattag tggagggcat aatagtagta cagaatcatt tattgcagat 660

cctgcaattt cactagctca tgaattgata catgcactgc atggattata cggggctagg 720

ggagttactt atgaagagac tatagaagta aagcaagcac ctcttatgat agccgaaaaa 780

cccataaggc tagaagaatt tttaaccttt ggaggtcagg atttaaatat tattactagt 840

gctatgaagg aaaaaatata taacaatctt ttagctaact atgaaaaaat agctactaga 900

cttagtgaag ttaatagtgc tcctcctgaa tatgatatta atgaatataa agattatttt 960

caatggaagt atgggctaga taaaaatgct gatggaagtt atactgtaaa tgaaaataaa 1020

tttaatgaaa tttataaaaa attatatagt tttacagaga gtgacttagc aaataaattt 1080

aaagtaaaat gtagaaatac ttattttatt aaatatgaat ttttaaaagt tccaaatttg 1140

ttagatgatg atatttatac tgtatcagag gggtttaata taggtaattt agcagtaaac 1200

aatcgcggac aaagtataaa gttaaatcct aaaattattg attccattcc agataaaggt 1260

ctagtagaaa agatcgttaa attttgtaag agcgttattc ctagaaaatg tcaaaattta 1320

ttcaagaaca ttaatatcat ggccaag 1347

<210> 72

<211> 1365

<212> DNA

<213> 人工序列

<220>

<223> BoNT/G轻链-MTDr cDNA序列

<400> 72

atgccagtta atataaaaan ctttaattat aatgacccta ttaataatga tgacattatt 60

atgatggaac cattcaatga cccagggcca ggaacatatt ataaagcttt taggattata 120

gatcgtattt ggatagtacc agaaaggttt acttatggat ttcaacctga ccaatttaat 180

gccagtacag gagtttttag taaagatgtc tacgaatatt acgatccaac ttatttaaaa 240

accgatgctg aaaaagataa atttttaaaa acaatgatta aattatttaa tagaattaat 300

tcaaaaccat caggacagag attactggat atgatagtag atgctatacc ttatcttgga 360

aatgcatcta caccgcccga caaatttgca gcaaatgttg caaatgtatc tattaataaa 420

aaaattatcc aacctggagc tgaagatcaa ataaaaggtt taatgacaaa tttaataata 480

tttggaccag gaccagttct aagtgataat tttactgata gtatgattat gaatggccat 540

tccccaatat cagaaggatt tggtgcaaga atgatgataa gattttgtcc tagttgttta 600

aatgtattta ataatgttca ggaaaataaa gatacatcta tatttagtag acgcgcgtat 660

tttgcagatc cagctctaac gttaatgcat gaacttatac atgtgttaca tggattatat 720

ggaattaaga taagtaattt accaattact ccaaatacaa aagaattttt catgcaacat 780

agcgatcctg tacaagcaga agaactatat acattcggag gacatgatcc tagtgttata 840

agtccttcta cggatatgaa tatttataat aaagcgttac aaaattttca agatatagct 900

aataggctta atattgtttc aagtgcccaa gggagtggaa ttgatatttc cttatataaa 960

caaatatata aaaataaata tgattttgtt gaagatccta atggaaaata tagtgtagat 1020

aaggataagt ttgataaatt atataaggcc ttaatgtttg gctttactga aactaatcta 1080

gctggtgaat atggaataaa aactaggtat tcttatttta gtgaatattt gccaccgata 1140

aaaactgaaa aattgttaga caatacaatt tatactcaaa atgaaggctt taacatagct 1200

agtaaaaatc tcaaaacgga atttaatggt cagaataagg cggtaaataa agaggcttat 1260

gaagaaatca gcctagaaca tctcgttata tatagaatag caatgtgcaa gcctgtaatg 1320

tacaaatgtc aaaatttatt caagaacatt aatatcatgg ccaag 1365

<210> 73

<211> 1413

<212> DNA

<213> 人工序列

<220>

<223> MTD-具有六组氨酸的BoNT/A轻链cDNA序列

<400> 73

atgaaggcca tgatcaatat taacaagttc ttaaatcaat gtccctttgt caacaaacag 60

ttcaactaca aggacccagt taatggagta gacatcgcat atatcaagat tcccaacgct 120

ggccagatgc aacccgttaa ggcatttaaa atccataaca aaatctgggt tatcccagag 180

cgggatacct tcaccaaccc cgaggagggc gatctgaacc ccccgccgga ggcgaagcag 240

gtcccagtga gctactacga tagcacctac ctcagcaccg acaacgagaa ggacaactac 300

ctcaaaggag tcacgaagtt gttcgagaga atctactcca cagacctcgg ccgcatgctt 360

ctaaccagca ttgtgcgtgg cattcccttt tggggcggct ctaccatcga cacagagctg 420

aaggtgatag acaccaactg catcaacgta atccagcctg acggcagcta ccgaagcgag 480

gagcttaacc tggtgatcat cggcccttcc gccgatatca tccaattcga gtgcaagagc 540

ttcggccacg aggtcctgaa cctcacccgg aacggctatg gaagcaccca gtacataaga 600

ttcagccctg acttcacctt cgggtttgag gagagcttgg aggtcgacac aaaccccctg 660

ctgggagccg ggaagttcgc cactgaccca gccgtgactc tggcacacga gctgatccac 720

gccggtcacc gcctgtacgg catagctata aacccaaaca gggtgttcaa agtgaacacc 780

aacgcttact atgaaatgag cggcctggag gtgagcttcg aggagctgag aacgttcggg 840

ggacatgatg ctaaatttat cgacagcctg caggagaacg agttcaggct gtactactac 900

aataagttca aggatatagc gagcactctg aacaaggcca agtccatcgt aggcactact 960

gcatccctcc agtatatgaa gaatgtgttc aaagagaaat acctgctgag cgaggatacc 1020

agcggtaagt tcagcgtgga taagcttaag ttcgacaagc tgtataagat gctcaccgaa 1080

atctacaccg aggataattt cgttaagttc ttcaaggtcc tgaaccggaa gacctacctg 1140

aacttcgaca aggccgtgtt caagatcaac atcgtgccta aagtgaacta caccatctac 1200

gacgggttta acctgaggaa caccaacctg gccgctaact tcaacgggca gaacacagag 1260

atcaacaaca tgaatttcac gaagttgaag aacttcaccg gactgtttga gttctacaaa 1320

ttgctgtgtg tgcgcgggat catcactagc aagaccaaga gccttgacaa aggctacaac 1380

aagtgactcg agcaccacca ccaccaccac tga 1413

<210> 74

<211> 1389

<212> DNA

<213> 人工序列

<220>

<223> MTD-具有六组氨酸的BoNT/B轻链cDNA序列

<400> 74

atgaaggcca tgatcaatat taacaagttc ttaaatcaat gtccagttac aataaataat 60

tttaattata atgatcctat tgataataat aatattatta tgatggagcc tccatttgcg 120

agaggtacgg ggagatatta taaagctttt aaaatcacag atcgtatttg gataataccg 180

gaaagatata cttttggata taaacctgag gattttaata aaagttccgg tatttttaat 240

agagatgttt gtgaatatta tgatccagat tacttaaata ctaatgataa aaagaatata 300

tttttacaaa caatgatcaa gttatttaat agaatcaaat caaaaccatt gggtgaaaag 360

ttattagaga tgattataaa tggtatacct tatcttggag atagacgtgt tccactcgaa 420

gagtttaaca caaacattgc tagtgtaact gttaataaat taatcagtaa tccaggagaa 480

gtggagcgaa aaaaaggtat tttcgcaaat ttaataatat ttggacctgg gccagtttta 540

aatgaaaatg agactataga tataggtata caaaatcatt ttgcatcaag ggaaggcttc 600

gggggtataa tgcaaatgaa gttttgccca gaatatgtaa gcgtatttaa taatgttcaa 660

gaaaacaaag gcgcaagtat atttaataga cgtggatatt tttcagatcc agccttgata 720

ttaatgcatg aacttataca tgttttacat ggattatatg gcattaaagt agatgattta 780

ccaattgtac caaatgaaaa aaaatttttt atgcaatcta cagatgctat acaggcagaa 840

gaactatata catttggagg acaagatccc agcatcataa ctccttctac ggataaaagt 900

atctatgata aagttttgca aaattttaga gggatagttg atagacttaa caaggtttta 960

gtttgcatat cagatcctaa cattaatatt aatatatata aaaataaatt taaagataaa 1020

tataaattcg ttgaagattc tgagggaaaa tatagtatag atgtagaaag ttttgataaa 1080

ttatataaaa gcttaatgtt tggttttaca gaaactaata tagcagaaaa ttataaaata 1140

aaaactagag cttcttattt tagtgattcc ttaccaccag taaaaataaa aaatttatta 1200

gataatgaaa tctatactat agaggaaggg tttaatatat ctgataaaga tatggaaaaa 1260

gaatatagag gtcagaataa agctataaat aaacaagctt atgaagaaat tagcaaggag 1320

catttggctg tatataagat acaaatgtgt aaaagtgtta aactcgagca ccaccaccac 1380

caccactga 1389

<210> 75

<211> 1413

<212> DNA

<213> 人工序列

<220>

<223> MTD-具有六组氨酸的BoNT/C轻链cDNA序列

<400> 75

atgaaggcca tgatcaatat taacaagttc ttaaatcaat gtccaataac aattaacaac 60

tttaattatt cagatcctgt tgataataaa aatattttat atttagatac tcatttaaat 120

acactagcta atgagcctga aaaagccttt cgcattacag gaaatatatg ggtaatacct 180

gatagatttt caagaaattc taatccaaat ttaaataaac ctcctcgagt tacaagccct 240

aaaagtggtt attatgatcc taattatttg agtactgatt ctgacaaaga tacattttta 300

aaagaaatta taaagttatt taaaagaatt aattctagag aaataggaga agaattaata 360

tatagacttt cgacagatat accctttcct gggaataaca atactccaat taatactttt 420

gattttgatg tagattttaa cagtgttgat gttaaaacta gacaaggtaa caactgggtt 480

aaaactggta gcataaatcc tagtgttata ataactggac ctagagaaaa cattatagat 540

ccagaaactt ctacgtttaa attaactaac aatacttttg cggcacaaga aggatttggt 600

gctttatcaa taatttcaat atcacctaga tttatgctaa catatagtaa tgcaactaat 660

gatgtaggag agggtagatt ttctaagtct gaattttgca tggatccaat actaatttta 720

atgcatgaac ttaatcatgc aatgcataat ttatatggaa tagctatacc aaatgatcaa 780

acaatttcat ctgtaactag taatattttt tattctcaat ataatgtgaa attagagtat 840

gcagaaatat atgcatttgg aggtccaact atagacctta ttcctaaaag tgcaaggaaa 900

tattttgagg aaaaggcatt ggattattat agatctatag ctaaaagact taatagtata 960

actactgcaa atccttcaag ctttaataaa tatatagggg aatataaaca gaaacttatt 1020

agaaagtata gattcgtagt agaatcttca ggtgaagtta cagtaaatcg taataagttt 1080

gttgagttat ataatgaact tacacaaata tttacagaat ttaactacgc taaaatatat 1140

aatgtacaaa ataggaaaat atatctttca aatgtatata ctccggttac ggcgaatata 1200

ttagacgata atgtttatga tatacaaaat ggatttaata tacctaaaag taatttaaat 1260

gtactattta tgggtcaaaa tttatctcga aatccagcat taagaaaagt caatcctgaa 1320

aatatgcttt atttatttac aaaattttgt cataaagcaa tagatggtag atcattatat 1380

aataaactcg agcaccacca ccaccaccac tga 1413

<210> 76

<211> 1392

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的MTD-BoNT/D轻链cDNA序列

<400> 76

atgaaggcca tgatcaatat taacaagttc ttaaatcaat gtacatggcc agtaaaagat 60

tttaattata gtgatcctgt taatgacaat gatatattat atttaagaat accacaaaat 120

aagttaatta ctacacctgt aaaagctttt atgattactc aaaatatttg ggtaatacca 180

gaaagatttt catcagatac taatccaagt ttaagtaaac cgcccagacc tacttcaaag 240

tatcaaagtt attatgatcc tagttattta tctactgatg aacaaaaaga tacattttta 300

aaagggatta taaaattatt taaaagaatt aatgaaagag atataggaaa aaaattaata 360

aattatttag tagttggttc accttttatg ggagattcaa gtacgcctga agatacattt 420

gattttacac gtcatactac taatattgca gttgaaaagt ttgaaaatgg tagttggaaa 480

gtaacaaata ttataacacc aagtgtattg atatttggac cacttcctaa tatattagac 540

tatacagcat cccttacatt gcaaggacaa caatcaaatc catcatttga agggtttgga 600

acattatcta tactaaaagt agcacctgaa tttttgttaa catttagtga tgtaacatct 660

aatcaaagtt cagctgtatt aggcaaatct atattttgta tggatccagt aatagcttta 720

atgcatgagt taacacattc tttgcatcaa ttatatggaa taaatatacc atctgataaa 780

aggattcgtc cacaagttag cgagggattt ttctctcaag atggacccaa cgtacaattt 840

gaggaattat atacatttgg aggattagat gttgaaataa tacctcaaat tgaaagatca 900

caattaagag aaaaagcatt aggtcactat aaagatatag cgaaaagact taataatatt 960

aataaaacta ttccttctag ttggattagt aatatagata aatataaaaa aatattttct 1020

gaaaagtata attttgataa agataataca ggaaattttg ttgtaaatat tgataaattc 1080

aatagcttat attcagactt gactaatgtt atgtcagaag ttgtttattc ttcgcaatat 1140

aatgttaaaa acaggactca ttatttttca aggcattatc tacctgtatt tgcaaatata 1200

ttagatgata atatttatac tataagagat ggttttaatt taacaaataa aggttttaat 1260

atagaaaatt cgggtcagaa tatagaaagg aatcctgcac tacaaaagct tagttcagaa 1320

agtgtagtag atttatttac aaaagtatgt ttaagattaa caaaactcga gcaccaccac 1380

caccaccact ga 1392

<210> 77

<211> 1332

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的MTD-BoNT/E轻链cDNA序列

<400> 77

atgaaggcca tgatcaatat taacaagttc ttaaatcaat gtccaacaat taatagtttt 60

aattataatg atcctgttaa taatagaaca attttatata ttaaaccagg cggttgtcaa 120

caattttata aatcatttaa tattatgaaa aatatttgga taattccaga gagaaatgta 180

attggtacaa ttccccaaga ttttcttccg cctacttcat tgaaaaatgg agatagtagt 240

tattatgacc ctaattattt acaaagtgat caagaaaagg ataaattttt aaaaatagtc 300

acaaaaatat ttaatagaat aaatgataat ctttcaggaa ggattttatt agaagaactg 360

tcaaaagcta atccatattt aggaaatgat aatactccag atggtgactt cattattaat 420

gatgcatcag cagttccaat tcaattctca aatggtagcc aaagcatact attacctaat 480

gttattataa tgggagcaga gcctgattta tttgaaacta acagttccaa tatttctcta 540

agaaataatt atatgccaag caatcacggt tttggatcaa tagctatagt aacattctca 600

cctgaatatt cttttagatt taaagataat agtatgaatg aatttattca agatcctgct 660

cttacattaa tgcatgaatt aatacattca ttacatggac tatatggggc taaagggatt 720

actacaaagt atactataac acaaaaacaa aatcccctaa taacaaatat aagaggtaca 780

aatattgaag aattcttaac ttttggaggt actgatttaa acattattac tagtgctcag 840

tccaatgata tctatactaa tcttctagct gattataaaa aaatagcgtc taaacttagc 900

aaagtacaag tatctaatcc actacttaat ccttataaag atgtttttga agcaaagtat 960

ggattagata aagatgctag cggaatttat tcggtaaata taaacaaatt taatgatatt 1020

tttaaaaaat tatacagctt tacggaattt gatttagcaa ctaaatttca agttaaatgt 1080

aggcaaactt atattggaca gtataaatac ttcaaacttt caaacttgtt aaatgattct 1140

atttataata tatcagaagg ctataatata aataatttaa aggtaaattt tagaggacag 1200

aatgcaaatt taaatcctag aattattaca ccaattacag gtagaggact agtaaaaaaa 1260

atcattagat tttgtaaaaa tattgtttct gtaaaaggca taaggctcga gcaccaccac 1320

caccaccact ga 1332

<210> 78

<211> 1374

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的MTD-BoNT/F轻链cDNA序列

<400> 78

atgaaggcca tgatcaatat taacaagttc ttaaatcaat gtccagttgc aataaatagt 60

tttaattata atgaccctgt taatgatgat acaattttat acatgcagat accatatgaa 120

gaaaaaagta aaaaatatta taaagctttt gagattatgc gtaatgtttg gataattcct 180

gagagaaata caataggaac gaatcctagt gattttgatc caccggcttc attaaagaac 240

ggaagcagtg cttattatga tcctaattat ttaaccactg atgctgaaaa agatagatat 300

ttaaaaacaa cgataaaatt atttaagaga attaatagta atcctgcagg gaaagttttg 360

ttacaagaaa tatcatatgc taaaccatat ttaggaaatg accacacgcc aattgatgaa 420

ttctctccag ttactagaac tacaagtgtt aatataaaat tatcaactaa tgttgaaagt 480

tcaatgttat tgaatcttct tgtattggga gcaggacctg atatatttga aagttgttgt 540

taccccgtta gaaaactaat agatccagat gtagtttatg atccaagtaa ttatggtttt 600

ggatcaatta atatcgtgac attttcacct gagtatgaat atacttttaa tgatattagt 660

ggagggcata atagtagtac agaatcattt attgcagatc ctgcaatttc actagctcat 720

gaattgatac atgcactgca tggattatac ggggctaggg gagttactta tgaagagact 780

atagaagtaa agcaagcacc tcttatgata gccgaaaaac ccataaggct agaagaattt 840

ttaacctttg gaggtcagga tttaaatatt attactagtg ctatgaagga aaaaatatat 900

aacaatcttt tagctaacta tgaaaaaata gctactagac ttagtgaagt taatagtgct 960

cctcctgaat atgatattaa tgaatataaa gattattttc aatggaagta tgggctagat 1020

aaaaatgctg atggaagtta tactgtaaat gaaaataaat ttaatgaaat ttataaaaaa 1080

ttatatagtt ttacagagag tgacttagca aataaattta aagtaaaatg tagaaatact 1140

tattttatta aatatgaatt tttaaaagtt ccaaatttgt tagatgatga tatttatact 1200

gtatcagagg ggtttaatat aggtaattta gcagtaaaca atcgcggaca aagtataaag 1260

ttaaatccta aaattattga ttccattcca gataaaggtc tagtagaaaa gatcgttaaa 1320

ttttgtaaga gcgttattcc tagaaaactc gagcaccacc accaccacca ctga 1374

<210> 79

<211> 1392

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的MTD-BoNT/G轻链cDNA序列

<400> 79

atgaaggcca tgatcaatat taacaagttc ttaaatcaat gtccagttaa tataaaaanc 60

tttaattata atgaccctat taataatgat gacattatta tgatggaacc attcaatgac 120

ccagggccag gaacatatta taaagctttt aggattatag atcgtatttg gatagtacca 180

gaaaggttta cttatggatt tcaacctgac caatttaatg ccagtacagg agtttttagt 240

aaagatgtct acgaatatta cgatccaact tatttaaaaa ccgatgctga aaaagataaa 300

tttttaaaaa caatgattaa attatttaat agaattaatt caaaaccatc aggacagaga 360

ttactggata tgatagtaga tgctatacct tatcttggaa atgcatctac accgcccgac 420

aaatttgcag caaatgttgc aaatgtatct attaataaaa aaattatcca acctggagct 480

gaagatcaaa taaaaggttt aatgacaaat ttaataatat ttggaccagg accagttcta 540

agtgataatt ttactgatag tatgattatg aatggccatt ccccaatatc agaaggattt 600

ggtgcaagaa tgatgataag attttgtcct agttgtttaa atgtatttaa taatgttcag 660

gaaaataaag atacatctat atttagtaga cgcgcgtatt ttgcagatcc agctctaacg 720

ttaatgcatg aacttataca tgtgttacat ggattatatg gaattaagat aagtaattta 780

ccaattactc caaatacaaa agaatttttc atgcaacata gcgatcctgt acaagcagaa 840

gaactatata cattcggagg acatgatcct agtgttataa gtccttctac ggatatgaat 900

atttataata aagcgttaca aaattttcaa gatatagcta ataggcttaa tattgtttca 960

agtgcccaag ggagtggaat tgatatttcc ttatataaac aaatatataa aaataaatat 1020

gattttgttg aagatcctaa tggaaaatat agtgtagata aggataagtt tgataaatta 1080

tataaggcct taatgtttgg ctttactgaa actaatctag ctggtgaata tggaataaaa 1140

actaggtatt cttattttag tgaatatttg ccaccgataa aaactgaaaa attgttagac 1200

aatacaattt atactcaaaa tgaaggcttt aacatagcta gtaaaaatct caaaacggaa 1260

tttaatggtc agaataaggc ggtaaataaa gaggcttatg aagaaatcag cctagaacat 1320

ctcgttatat atagaatagc aatgtgcaag cctgtaatgt acaaactcga gcaccaccac 1380

caccaccact ga 1392

<210> 80

<211> 1446

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/A轻链-MTDr cDNA序列

<400> 80

atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60

atgccctttg tcaacaaaca gttcaactac aaggacccag ttaatggagt agacatcgca 120

tatatcaaga ttcccaacgc tggccagatg caacccgtta aggcatttaa aatccataac 180

aaaatctggg ttatcccaga gcgggatacc ttcaccaacc ccgaggaggg cgatctgaac 240

cccccgccgg aggcgaagca ggtcccagtg agctactacg atagcaccta cctcagcacc 300

gacaacgaga aggacaacta cctcaaagga gtcacgaagt tgttcgagag aatctactcc 360

acagacctcg gccgcatgct tctaaccagc attgtgcgtg gcattccctt ttggggcggc 420

tctaccatcg acacagagct gaaggtgata gacaccaact gcatcaacgt aatccagcct 480

gacggcagct accgaagcga ggagcttaac ctggtgatca tcggcccttc cgccgatatc 540

atccaattcg agtgcaagag cttcggccac gaggtcctga acctcacccg gaacggctat 600

ggaagcaccc agtacataag attcagccct gacttcacct tcgggtttga ggagagcttg 660

gaggtcgaca caaaccccct gctgggagcc gggaagttcg ccactgaccc agccgtgact 720

ctggcacacg agctgatcca cgccggtcac cgcctgtacg gcatagctat aaacccaaac 780

agggtgttca aagtgaacac caacgcttac tatgaaatga gcggcctgga ggtgagcttc 840

gaggagctga gaacgttcgg gggacatgat gctaaattta tcgacagcct gcaggagaac 900

gagttcaggc tgtactacta caataagttc aaggatatag cgagcactct gaacaaggcc 960

aagtccatcg taggcactac tgcatccctc cagtatatga agaatgtgtt caaagagaaa 1020

tacctgctga gcgaggatac cagcggtaag ttcagcgtgg ataagcttaa gttcgacaag 1080

ctgtataaga tgctcaccga aatctacacc gaggataatt tcgttaagtt cttcaaggtc 1140

ctgaaccgga agacctacct gaacttcgac aaggccgtgt tcaagatcaa catcgtgcct 1200

aaagtgaact acaccatcta cgacgggttt aacctgagga acaccaacct ggccgctaac 1260

ttcaacgggc agaacacaga gatcaacaac atgaatttca cgaagttgaa gaacttcacc 1320

ggactgtttg agttctacaa attgctgtgt gtgcgcggga tcatcactag caagaccaag 1380

agccttgaca aaggctacaa caagtgatgt caaaatttat tcaagaacat taatatcatg 1440

gccaag 1446

<210> 81

<211> 1422

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/B轻链-MTDr cDNA序列

<400> 81

atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60

atgccagtta caataaataa ttttaattat aatgatccta ttgataataa taatattatt 120

atgatggagc ctccatttgc gagaggtacg gggagatatt ataaagcttt taaaatcaca 180

gatcgtattt ggataatacc ggaaagatat acttttggat ataaacctga ggattttaat 240

aaaagttccg gtatttttaa tagagatgtt tgtgaatatt atgatccaga ttacttaaat 300

actaatgata aaaagaatat atttttacaa acaatgatca agttatttaa tagaatcaaa 360

tcaaaaccat tgggtgaaaa gttattagag atgattataa atggtatacc ttatcttgga 420

gatagacgtg ttccactcga agagtttaac acaaacattg ctagtgtaac tgttaataaa 480

ttaatcagta atccaggaga agtggagcga aaaaaaggta ttttcgcaaa tttaataata 540

tttggacctg ggccagtttt aaatgaaaat gagactatag atataggtat acaaaatcat 600

tttgcatcaa gggaaggctt cgggggtata atgcaaatga agttttgccc agaatatgta 660

agcgtattta ataatgttca agaaaacaaa ggcgcaagta tatttaatag acgtggatat 720

ttttcagatc cagccttgat attaatgcat gaacttatac atgttttaca tggattatat 780

ggcattaaag tagatgattt accaattgta ccaaatgaaa aaaaattttt tatgcaatct 840

acagatgcta tacaggcaga agaactatat acatttggag gacaagatcc cagcatcata 900

actccttcta cggataaaag tatctatgat aaagttttgc aaaattttag agggatagtt 960

gatagactta acaaggtttt agtttgcata tcagatccta acattaatat taatatatat 1020

aaaaataaat ttaaagataa atataaattc gttgaagatt ctgagggaaa atatagtata 1080

gatgtagaaa gttttgataa attatataaa agcttaatgt ttggttttac agaaactaat 1140

atagcagaaa attataaaat aaaaactaga gcttcttatt ttagtgattc cttaccacca 1200

gtaaaaataa aaaatttatt agataatgaa atctatacta tagaggaagg gtttaatata 1260

tctgataaag atatggaaaa agaatataga ggtcagaata aagctataaa taaacaagct 1320

tatgaagaaa ttagcaagga gcatttggct gtatataaga tacaaatgtg taaaagtgtt 1380

aaatgtcaaa atttattcaa gaacattaat atcatggcca ag 1422

<210> 82

<211> 1446

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/C轻链-MTDr cDNA序列

<400> 82

atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60

atgccaataa caattaacaa ctttaattat tcagatcctg ttgataataa aaatatttta 120

tatttagata ctcatttaaa tacactagct aatgagcctg aaaaagcctt tcgcattaca 180

ggaaatatat gggtaatacc tgatagattt tcaagaaatt ctaatccaaa tttaaataaa 240

cctcctcgag ttacaagccc taaaagtggt tattatgatc ctaattattt gagtactgat 300

tctgacaaag atacattttt aaaagaaatt ataaagttat ttaaaagaat taattctaga 360

gaaataggag aagaattaat atatagactt tcgacagata taccctttcc tgggaataac 420

aatactccaa ttaatacttt tgattttgat gtagatttta acagtgttga tgttaaaact 480

agacaaggta acaactgggt taaaactggt agcataaatc ctagtgttat aataactgga 540

cctagagaaa acattataga tccagaaact tctacgttta aattaactaa caatactttt 600

gcggcacaag aaggatttgg tgctttatca ataatttcaa tatcacctag atttatgcta 660

acatatagta atgcaactaa tgatgtagga gagggtagat tttctaagtc tgaattttgc 720

atggatccaa tactaatttt aatgcatgaa cttaatcatg caatgcataa tttatatgga 780

atagctatac caaatgatca aacaatttca tctgtaacta gtaatatttt ttattctcaa 840

tataatgtga aattagagta tgcagaaata tatgcatttg gaggtccaac tatagacctt 900

attcctaaaa gtgcaaggaa atattttgag gaaaaggcat tggattatta tagatctata 960

gctaaaagac ttaatagtat aactactgca aatccttcaa gctttaataa atatataggg 1020

gaatataaac agaaacttat tagaaagtat agattcgtag tagaatcttc aggtgaagtt 1080

acagtaaatc gtaataagtt tgttgagtta tataatgaac ttacacaaat atttacagaa 1140

tttaactacg ctaaaatata taatgtacaa aataggaaaa tatatctttc aaatgtatat 1200

actccggtta cggcgaatat attagacgat aatgtttatg atatacaaaa tggatttaat 1260

atacctaaaa gtaatttaaa tgtactattt atgggtcaaa atttatctcg aaatccagca 1320

ttaagaaaag tcaatcctga aaatatgctt tatttattta caaaattttg tcataaagca 1380

atagatggta gatcattata taataaatgt caaaatttat tcaagaacat taatatcatg 1440

gccaag 1446

<210> 83

<211> 1425

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/D轻链-MTDr cDNA序列

<400> 83

atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60

atgacatggc cagtaaaaga ttttaattat agtgatcctg ttaatgacaa tgatatatta 120

tatttaagaa taccacaaaa taagttaatt actacacctg taaaagcttt tatgattact 180

caaaatattt gggtaatacc agaaagattt tcatcagata ctaatccaag tttaagtaaa 240

ccgcccagac ctacttcaaa gtatcaaagt tattatgatc ctagttattt atctactgat 300

gaacaaaaag atacattttt aaaagggatt ataaaattat ttaaaagaat taatgaaaga 360

gatataggaa aaaaattaat aaattattta gtagttggtt caccttttat gggagattca 420

agtacgcctg aagatacatt tgattttaca cgtcatacta ctaatattgc agttgaaaag 480

tttgaaaatg gtagttggaa agtaacaaat attataacac caagtgtatt gatatttgga 540

ccacttccta atatattaga ctatacagca tcccttacat tgcaaggaca acaatcaaat 600

ccatcatttg aagggtttgg aacattatct atactaaaag tagcacctga atttttgtta 660

acatttagtg atgtaacatc taatcaaagt tcagctgtat taggcaaatc tatattttgt 720

atggatccag taatagcttt aatgcatgag ttaacacatt ctttgcatca attatatgga 780

ataaatatac catctgataa aaggattcgt ccacaagtta gcgagggatt tttctctcaa 840

gatggaccca acgtacaatt tgaggaatta tatacatttg gaggattaga tgttgaaata 900

atacctcaaa ttgaaagatc acaattaaga gaaaaagcat taggtcacta taaagatata 960

gcgaaaagac ttaataatat taataaaact attccttcta gttggattag taatatagat 1020

aaatataaaa aaatattttc tgaaaagtat aattttgata aagataatac aggaaatttt 1080

gttgtaaata ttgataaatt caatagctta tattcagact tgactaatgt tatgtcagaa 1140

gttgtttatt cttcgcaata taatgttaaa aacaggactc attatttttc aaggcattat 1200

ctacctgtat ttgcaaatat attagatgat aatatttata ctataagaga tggttttaat 1260

ttaacaaata aaggttttaa tatagaaaat tcgggtcaga atatagaaag gaatcctgca 1320

ctacaaaagc ttagttcaga aagtgtagta gatttattta caaaagtatg tttaagatta 1380

acaaaatgtc aaaatttatt caagaacatt aatatcatgg ccaag 1425

<210> 84

<211> 1365

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/E轻链-MTDr cDNA序列

<400> 84

atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60

atgccaacaa ttaatagttt taattataat gatcctgtta ataatagaac aattttatat 120

attaaaccag gcggttgtca acaattttat aaatcattta atattatgaa aaatatttgg 180

ataattccag agagaaatgt aattggtaca attccccaag attttcttcc gcctacttca 240

ttgaaaaatg gagatagtag ttattatgac cctaattatt tacaaagtga tcaagaaaag 300

gataaatttt taaaaatagt cacaaaaata tttaatagaa taaatgataa tctttcagga 360

aggattttat tagaagaact gtcaaaagct aatccatatt taggaaatga taatactcca 420

gatggtgact tcattattaa tgatgcatca gcagttccaa ttcaattctc aaatggtagc 480

caaagcatac tattacctaa tgttattata atgggagcag agcctgattt atttgaaact 540

aacagttcca atatttctct aagaaataat tatatgccaa gcaatcacgg ttttggatca 600

atagctatag taacattctc acctgaatat tcttttagat ttaaagataa tagtatgaat 660

gaatttattc aagatcctgc tcttacatta atgcatgaat taatacattc attacatgga 720

ctatatgggg ctaaagggat tactacaaag tatactataa cacaaaaaca aaatccccta 780

ataacaaata taagaggtac aaatattgaa gaattcttaa cttttggagg tactgattta 840

aacattatta ctagtgctca gtccaatgat atctatacta atcttctagc tgattataaa 900

aaaatagcgt ctaaacttag caaagtacaa gtatctaatc cactacttaa tccttataaa 960

gatgtttttg aagcaaagta tggattagat aaagatgcta gcggaattta ttcggtaaat 1020

ataaacaaat ttaatgatat ttttaaaaaa ttatacagct ttacggaatt tgatttagca 1080

actaaatttc aagttaaatg taggcaaact tatattggac agtataaata cttcaaactt 1140

tcaaacttgt taaatgattc tatttataat atatcagaag gctataatat aaataattta 1200

aaggtaaatt ttagaggaca gaatgcaaat ttaaatccta gaattattac accaattaca 1260

ggtagaggac tagtaaaaaa aatcattaga ttttgtaaaa atattgtttc tgtaaaaggc 1320

ataaggtgtc aaaatttatt caagaacatt aatatcatgg ccaag 1365

<210> 85

<211> 1407

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/F轻链-MTDr cDNA序列

<400> 85

atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60

atgccagttg caataaatag ttttaattat aatgaccctg ttaatgatga tacaatttta 120

tacatgcaga taccatatga agaaaaaagt aaaaaatatt ataaagcttt tgagattatg 180

cgtaatgttt ggataattcc tgagagaaat acaataggaa cgaatcctag tgattttgat 240

ccaccggctt cattaaagaa cggaagcagt gcttattatg atcctaatta tttaaccact 300

gatgctgaaa aagatagata tttaaaaaca acgataaaat tatttaagag aattaatagt 360

aatcctgcag ggaaagtttt gttacaagaa atatcatatg ctaaaccata tttaggaaat 420

gaccacacgc caattgatga attctctcca gttactagaa ctacaagtgt taatataaaa 480

ttatcaacta atgttgaaag ttcaatgtta ttgaatcttc ttgtattggg agcaggacct 540

gatatatttg aaagttgttg ttaccccgtt agaaaactaa tagatccaga tgtagtttat 600

gatccaagta attatggttt tggatcaatt aatatcgtga cattttcacc tgagtatgaa 660

tatactttta atgatattag tggagggcat aatagtagta cagaatcatt tattgcagat 720

cctgcaattt cactagctca tgaattgata catgcactgc atggattata cggggctagg 780

ggagttactt atgaagagac tatagaagta aagcaagcac ctcttatgat agccgaaaaa 840

cccataaggc tagaagaatt tttaaccttt ggaggtcagg atttaaatat tattactagt 900

gctatgaagg aaaaaatata taacaatctt ttagctaact atgaaaaaat agctactaga 960

cttagtgaag ttaatagtgc tcctcctgaa tatgatatta atgaatataa agattatttt 1020

caatggaagt atgggctaga taaaaatgct gatggaagtt atactgtaaa tgaaaataaa 1080

tttaatgaaa tttataaaaa attatatagt tttacagaga gtgacttagc aaataaattt 1140

aaagtaaaat gtagaaatac ttattttatt aaatatgaat ttttaaaagt tccaaatttg 1200

ttagatgatg atatttatac tgtatcagag gggtttaata taggtaattt agcagtaaac 1260

aatcgcggac aaagtataaa gttaaatcct aaaattattg attccattcc agataaaggt 1320

ctagtagaaa agatcgttaa attttgtaag agcgttattc ctagaaaatg tcaaaattta 1380

ttcaagaaca ttaatatcat ggccaag 1407

<210> 86

<211> 1425

<212> DNA

<213> 人工序列

<220>

<223> 具有六组氨酸的BoNT/G轻链-MTDr cDNA序列

<400> 86

atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60

atgccagtta atataaaaan ctttaattat aatgacccta ttaataatga tgacattatt 120

atgatggaac cattcaatga cccagggcca ggaacatatt ataaagcttt taggattata 180

gatcgtattt ggatagtacc agaaaggttt acttatggat ttcaacctga ccaatttaat 240

gccagtacag gagtttttag taaagatgtc tacgaatatt acgatccaac ttatttaaaa 300

accgatgctg aaaaagataa atttttaaaa acaatgatta aattatttaa tagaattaat 360

tcaaaaccat caggacagag attactggat atgatagtag atgctatacc ttatcttgga 420

aatgcatcta caccgcccga caaatttgca gcaaatgttg caaatgtatc tattaataaa 480

aaaattatcc aacctggagc tgaagatcaa ataaaaggtt taatgacaaa tttaataata 540

tttggaccag gaccagttct aagtgataat tttactgata gtatgattat gaatggccat 600

tccccaatat cagaaggatt tggtgcaaga atgatgataa gattttgtcc tagttgttta 660

aatgtattta ataatgttca ggaaaataaa gatacatcta tatttagtag acgcgcgtat 720

tttgcagatc cagctctaac gttaatgcat gaacttatac atgtgttaca tggattatat 780

ggaattaaga taagtaattt accaattact ccaaatacaa aagaattttt catgcaacat 840

agcgatcctg tacaagcaga agaactatat acattcggag gacatgatcc tagtgttata 900

agtccttcta cggatatgaa tatttataat aaagcgttac aaaattttca agatatagct 960

aataggctta atattgtttc aagtgcccaa gggagtggaa ttgatatttc cttatataaa 1020

caaatatata aaaataaata tgattttgtt gaagatccta atggaaaata tagtgtagat 1080

aaggataagt ttgataaatt atataaggcc ttaatgtttg gctttactga aactaatcta 1140

gctggtgaat atggaataaa aactaggtat tcttatttta gtgaatattt gccaccgata 1200

aaaactgaaa aattgttaga caatacaatt tatactcaaa atgaaggctt taacatagct 1260

agtaaaaatc tcaaaacgga atttaatggt cagaataagg cggtaaataa agaggcttat 1320

gaagaaatca gcctagaaca tctcgttata tatagaatag caatgtgcaa gcctgtaatg 1380

tacaaatgtc aaaatttatt caagaacatt aatatcatgg ccaag 1425

Claims (16)

1.一种包含肉毒杆菌毒素重组蛋白作为活性成分的用于止痛的药物组合物，

其中在所述肉毒杆菌毒素重组蛋白中，由SEQ ID NO:1的氨基酸序列组成的细胞穿透性肽与肉毒杆菌毒素轻链的一端或两端融合。

2.根据权利要求1所述的药物组合物，其中所述肉毒杆菌毒素重组蛋白由一个或多个氨基酸序列组成，所述氨基酸序列选自由SEQ ID NO:31至SEQ ID NO:58组成的群组。

3.根据权利要求1所述的药物组合物，其中所述肉毒杆菌毒素轻链由一个或多个氨基酸序列组成，所述氨基酸序列选自由SEQ ID NO:3至SEQ ID NO:9组成的群组。

4.根据权利要求1所述的药物组合物，其中所述肉毒杆菌毒素轻链在一端还包括六组氨酸标签。

5.根据权利要求1所述的药物组合物，其中所述肉毒杆菌毒素轻链选自由肉毒杆菌毒素血清型A、B、C、D、E、F和G组成的群组。

6.根据权利要求1所述的药物组合物，其中所述细胞穿透性肽与所述肉毒杆菌毒素轻链的羧基末端、氨基末端或两者融合。

7.根据权利要求1所述的药物组合物，其中所述融合通过肽键或共价键实现。

8.根据权利要求1所述的药物组合物，其中所述肉毒杆菌毒素重组蛋白缓解肌肉紧张。

9.根据权利要求1所述的药物组合物，其中所述疼痛选自由以下组成的群组：机械性疼痛、热性疼痛、慢性疼痛、急性疼痛、过敏性疼痛、体源性疼痛、神经性疼痛、心理性疼痛、热量诱导的疼痛、躯体性疼痛、伤害性感觉、过敏、风湿性疼痛、头痛、背部疼痛、骨盆疼痛、肌筋膜疼痛、血管疼痛、偏头痛、创伤相关疼痛、炎性疼痛、关节炎疼痛、糖尿病性疼痛、癌症衍生性疼痛、内脏疼痛、纤维肌痛、术后疼痛、幻肢痛、三叉神经痛、带状疱疹后神经痛、疼痛性糖尿病神经病变、疼痛性糖尿病周围神经病变、糖尿病多发性神经病、坐骨神经痛、神经根病、脊髓神经根病、肌痛和腰部疼痛。

10.根据权利要求1所述的药物组合物，其中所述药物组合物用于透皮给药。

11.一种包含肉毒杆菌毒素重组蛋白作为活性成分的用于止痛的准药物组合物，

其中在所述肉毒杆菌毒素重组蛋白中，由SEQ ID NO:1的氨基酸序列组成的细胞穿透性肽与肉毒杆菌毒素轻链的一端或两端融合。

12.一种包含肉毒杆菌毒素重组蛋白作为活性成分的用于止痛的皮肤外用的组合物，

其中在所述肉毒杆菌毒素重组蛋白中，由SEQ ID NO:1的氨基酸序列组成的细胞穿透性肽与肉毒杆菌毒素轻链的一端或两端融合。

13.一种包含肉毒杆菌毒素重组蛋白作为活性成分的用于止痛的化妆品组合物，

其中在所述肉毒杆菌毒素重组蛋白中，由SEQ ID NO:1的氨基酸序列组成的细胞穿透性肽与肉毒杆菌毒素轻链的一端或两端融合。

14.一种缓解疼痛的方法，其包括将包含肉毒杆菌毒素重组蛋白作为活性成分的组合物给药至有需要的受试者，其中在所述肉毒杆菌毒素重组蛋白中，由SEQ ID NO:1的氨基酸序列组成的细胞穿透性肽与肉毒杆菌毒素轻链的一端或两端融合。

15.包含肉毒杆菌毒素重组蛋白作为活性成分的组合物用于止痛的用途，其中在所述肉毒杆菌毒素重建蛋白中，由SEQ ID NO:1的氨基酸序列组成的细胞穿透性肽与肉毒杆菌毒素轻链的一端或两端融合。

16.肉毒杆菌毒素重组蛋白用于制备止痛药物的用途，其中在所述肉毒杆菌毒素重组蛋白中，由SEQ ID NO:1的氨基酸序列组成的细胞穿透性肽与肉毒杆菌毒素轻链的一端或两端融合。