CN113677799A - 一种遗传修饰乳酸杆菌及其应用 - Google Patents

Abstract

本发明涉及通过活体递送平台将抗感染活性、免疫调节因子或生长促进生物分子直接有效递送至动物的消化道。所述活体递送平台可以是遗传修饰微生物。可以用定殖于胃肠道的乳杆菌属(Lactobacillus sp)完成递送。所述抗感染活性可以是杀菌或抑菌肽、特异性识别致病菌的抗体或其片段、或噬菌体、或来自噬菌体的特异性靶向某种致病菌的裂解肽。

Description

一种遗传修饰乳酸杆菌及其应用

本申请要求2019年2月5日提交的美国专利申请No.62/801,307的优先权，其全部内容通过引用并入本申请。

技术领域

本发明涉及一种向有需要动物的粘膜直接提供预防性或治疗性抗感染活性、免疫调节因子或促生长生物分子的活体递送平台(例如遗传修饰细菌)。

背景技术

直接饲喂微生物(DFMs，通常也称为益生菌)是定殖于动物胃肠道并为该动物提供一些有益效果的微生物。所述微生物可以是诸如来自杆菌(Bacillus)、乳酸杆菌(Lactobacillus)、乳酸乳球菌(Lactococcus)和肠球菌(Enterococcus)属的细菌种类。所述微生物也可以是酵母菌或者甚至可以是霉菌。所述微生物可以经口或经粘膜提供给动物，就鸟类而言，是提供给受精卵(即卵内)的。

DFM提供的有益活性可以是合成宿主动物健康代谢所需的维生素或其它营养分子。DFM还可以保护宿主动物免受由其他病原微生物引起的疾病、病症或临床症状。例如，DFM可以自然产生对某些种类的致病菌(例如有害的或引起疾病的细菌)具有抑制性或细胞毒性活性的因子。然而，DFM可能不能产生足量的此类因子，以减少宿主与致病菌的感染，或者这些因子可能仅影响有限的致病菌，从而使宿主易受其他致病菌的影响。

宿主动物可以使用更强或更广泛的抗生素，例如口服或肠胃外给药，但这些在宿主中持续时间有限，因此可能需要重复给药。口服还可导致抗生素降解或不能将抗生素递送到特定的最需要治疗效果的解剖部位。致病菌耐药性的形成是另一个重要的问题。

人们需要的是一种可以持续将抗感染分子直接递送到胃肠道或呼吸道(致病细菌在宿主中复制的地方)的递送系统。胃肠道和呼吸系统通常也是致病菌进入宿主的点。优选地，所述递送系统是活的遗传修饰微生物(例如细菌)，其可定殖宿主的胃肠道或呼吸道并直接递送抗生素因子，以减少致病菌的数量或阻断致病菌的进入。例如，在卵递送中，活体递送平台可能通过竞争性排除或者直接或间接抗感染效应，防止致病菌在胚胎的早期定殖。卵内递送具有绕过任何由于母体抗体干扰导致的遗传修饰微生物定殖受限的优点。优选地，活细菌递送系统合成对致病菌具有期望效果的足量抗感染因子。靶向致病菌可以是，但不限于，沙门氏菌属(Salmonella)、梭菌属(Clostridium)、弯曲菌属(Capylobacter)、梭杆菌属(Fusobacterium)、葡萄球菌属(Staphylococcus)、或链球菌属(Streptococcus)、或大肠杆菌(E.coli)、或寄生虫(例如艾美球虫属)。优选地，活体细菌系统在宿主胃肠道中持续存在一段时间。优选地，活体细菌递送系统产生广谱抗感染因子或多个抗感染因子，以靶向多种致病菌。或者，可将活体递送系统的组合给药于单个动物，其中遗传修饰细菌产生多个抗感染因子、免疫调节分子或促生长生物分子，或其任意组合。因此，一种以上的疾病状态得以预防或减少，或者多种病因的疾病和综合征可以得到有效治疗。

本发明中公开了抗感染肽，包括靶向多个细菌物种的新美杀菌素样(mercacidin-like)肽。本发明还公开了抗体，包括靶向特异性致病菌和致病分子的单链抗体。本发明还公开了靶向致病物种的噬菌体或噬菌体裂解肽。本发明还提供了一种可以产生高水平的至少一种或多种上述分子(优选作为表面展示或分泌的分子)的乳酸杆菌表达系统。

发明内容

本发明提供了一种包含遗传修饰微生物的活体递送平台。所述遗传修饰微生物包括表达盒，所述表达盒包括一个或多个：用于转录表达的启动子、编码用于分泌的信号序列的核苷酸序列、编码细胞壁锚的核苷酸序列、至少一个编码所需生物分子的异源编码区、编码用于检测的表达肽标签的核苷酸序列、以及用于终止翻译和转录的终止子。所述遗传修饰微生物可以是细菌、酵母或真菌。遗传修饰细菌优选为芽孢杆菌(Bacillus)、乳酸杆菌(Lactobacillus)、乳酸乳球菌(Lactococcus)或肠球菌(Enterococcus)。所述遗传修饰细菌也可以优选为大肠杆菌(E.coli)。所述遗传修饰细菌也可以优选为罗伊氏乳杆菌(Lactobacillus reuteri)菌株。

本发明提供了一种遗传修饰微生物中的表达盒，所述表达盒包括用于转录表达的启动子。所述转录表达的启动子包括SEQ ID NO：14、SEQ ID NO：15、SEQ ID NO：16、SEQ IDNO：17、SEQ ID NO：18、SEQ ID NO：19、SEQ ID NO：41、SEQ ID NO：42和SEQ ID NO：43的核苷酸序列。

本发明提供了一种遗传修饰微生物中的表达盒，所述表达盒包括编码用于分泌的信号序列的核苷酸序列。所述用于分泌的信号序列可以是至少20个氨基酸、至少25个氨基酸、至少30个氨基酸、至少35个氨基酸、至少40个氨基酸、至少44个氨基酸、至少50个氨基酸、至少55个氨基酸、至少60个氨基酸或至少65个氨基酸的分泌信号序列，也可以是20～65个氨基酸、20～60个氨基酸；20～55个氨基酸；20～50个氨基酸、20～45个氨基酸、20～40个氨基酸、20～35个氨基酸、20～30个氨基酸、25～65个氨基酸、25～60个氨基酸；25～55个氨基酸；25～50个氨基酸、25～45个氨基酸、25～40个氨基酸、25～35个氨基酸、25～30个氨基酸、30～65个氨基酸、30～60个氨基酸；30～55个氨基酸；30～50个氨基酸、30～45个氨基酸、30～40个氨基酸或30～35个氨基酸。分泌的信号序列可以包括氨基酸序列，所述氨基酸序列是如SEQ ID NO：20、SEQ ID NO：21、SEQ ID NO：22、SEQ ID NO：23和SEQ ID NO：40的片段，其中所述片段具有如本发明所示的长度。所述用于分泌的信号序列可以包括氨基酸序列，所述氨基酸序列是如SEQ ID NO：20、SEQ ID NO：21、SEQ ID NO：2、SEQ ID NO：23和SEQID NO：40的氨基末端片段，其中所述片段的长度如本发明所示。

本发明提供了一种遗传修饰微生物中的表达盒，所述表达盒包括编码细胞壁锚肽的核苷酸序列。所述细胞壁锚肽长度为100～250个氨基酸、100～225个氨基酸、100～200个氨基酸、100～175个氨基酸、100～150个氨基酸、125～250个氨基酸、125～225个氨基酸、125～200个氨基酸、125～175个氨基酸或125～150个氨基酸。所述细胞壁锚包含氨基酸序列，所述氨基酸序列是如SEQ ID NO：20、SEQ ID NO：21、SEQ ID NO：22、SEQ ID NO：23和SEQID NO：40的片段，其中所述片段的长度如本发明所示。所述细胞壁锚包含氨基酸序列，所述氨基酸序列是SEQ ID NO：20、SEQ ID NO：21、SEQ ID NO：22、SEQ ID NO：23和SEQ ID NO：40的羧基末端片段，其中所述片段的长度如本发明所示。

本发明提供了一种遗传修饰微生物中的表达盒，所述表达盒包括编码所需生物分子的异源编码区。所述所需生物分子可以是具有抗感染活性的生物分子、益生菌因子、免疫调节因子或促生长生物分子。所述生物分子具有抵抗致病菌或寄生虫的抗感染活性。所述寄生虫优选为艾美球虫属物种。所述致病细菌优选为沙门氏菌(Salmonella)、梭菌(Clostridium)，弯曲菌(Campylobacter)，葡萄球菌(Staphylococcus)，链球菌(Streptococcus)，梭杆菌(Fusobacterium)和大肠杆菌(E.coli)。

本发明提供了一种遗传修饰微生物中的表达盒，所述表达盒包括编码具有抗感染活性的所需生物分子的异源编码区。所述抗感染生物分子可以是杀菌肽、酶、溶素、噬菌体或抗体。所述杀菌肽为美杀菌素样分子。所述美杀菌素样分子包含本发明公开的如SEQ IDNO：2或SEQ ID NO：4所示的序列，所需生物分子可以是酶。所述酶包含本发明公开的如SEQID NO：5或SEQ ID NO：6所示的序列。所述溶素包括本发明公开的如SEQ ID NO：24、SEQ IDNO：25、SEQ ID NO：26、SEQ ID NO：27或SEQ ID NO：28所示的序列。所需生物分子可以是噬菌体或噬菌体形式的噬菌体。所述噬菌体遗传材料包括本发明公开的如SEQ ID NO：11、SEQID NO：12或SEQ ID NO：13所示的序列。所述所需生物分子来自芽孢杆菌属。所述所需生物分子为芽孢杆菌细菌素。所述芽孢杆菌抗感染分子包括本发明公开的如SEQ ID NO：44、SEQID NO：45、SEQ ID NO：46、SEQ ID NO：47或SEQ ID NO：48所示的序列。

本发明提供了一种遗传修饰微生物中的表达盒，所述表达盒包括编码所需生物分子的异源编码区，所述所需生物分子是抗体。所述抗体为单链抗体。所述单链抗体来自骆驼科。所述单链抗体可特异性识别病原微生物或致病菌产生的分子。所述单链抗体可以特异性识别细菌蛋白(例如毒素或附着分子)。所述单链抗体可以特异性识别来自产气荚膜梭菌(Clostridium perfringens)的细菌蛋白。细菌蛋白可以是由产气荚膜梭菌产生的蛋白质。细菌蛋白可以是产气荚膜梭菌α毒素或产气荚膜梭菌NetB毒素。

本发明提供了一种遗传修饰微生物中的表达盒，所述表达盒包括编码所需生物分子的异源编码区，所述所需生物分子是识别产气荚膜梭菌α毒素或产气荚膜梭菌毒素的抗体。所述单链抗体包括本发明公开的如SEQ ID NO：29、SEQ ID NO：30、SEQ ID NO：33或SEQID NO：34所示的氨基酸序列。

本发明提供了一种遗传修饰微生物中的表达盒，所述表达盒包括编码所需生物分子的异源编码区，所述所需生物分子是益生因子。所述益生因子为凝集素受体。所述凝集素受体包括本发明公开的如SEQ ID NO：9或SEQ ID NO：10所示的序列。

本发明提供了一种包含表达盒的遗传修饰微生物，所述表达盒包括转录表达的启动子和至少一个编码所需生物分子的异源编码区。所述表达盒还可任选地包括编码用于分泌的信号序列的核苷酸序列、编码细胞壁锚的核苷酸序列、编码用于检测的表达肽标签的核苷酸序列、以及用于终止翻译和转录的终止子中的一种或多种。所述表达盒包含具有本发明公开的如SEQ ID NO：14、SEQ ID NO：15、SEQ ID NO：16、SEQ ID NO：17、SEQ ID NO：18、SEQ ID NO：19、SEQ ID NO：41、SEQ ID NO：42或SEQ ID NO：43所示序列的启动子。所述至少一个编码所需生物分子的异源编码区可以编码诸如杀菌肽、酶、溶素、噬菌体和抗体等生物分子。所述至少一个编码所需生物分子的异源编码区可编码具有抗感染活性的生物分子。所述至少一个编码所需生物分子的异源编码区可编码具有本发明公开的如SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：5、SEQ ID NO：6、SEQ ID NO：24,SEQ ID NO：25、SEQ ID NO：26、SEQ ID NO：27、SEQ ID NO：28、SEQ ID NO：29、SEQ ID NO：30、SEQ ID NO：33、SEQ ID NO：34、SEQ ID NO：44、SEQ ID NO：45、SEQ ID NO：46、SEQ ID NO：47、SEQ ID NO：48、SEQ IDNO：49或SEQ ID NO：50所示序列的生物分子。

本发明提供了一种包含表达盒的遗传修饰微生物，其中所述遗传修饰细菌是芽孢杆菌、乳酸杆菌，乳酸乳球菌或肠球菌。所述遗传修饰细菌可以是罗伊氏乳杆菌菌株。

本发明提供了一种包含表达盒的遗传修饰微生物，其中所述表达盒位于质粒上。所述质粒包含本发明公开的如SEQ ID NO：8所示的序列。本发明提供一种包含表达盒的遗传修饰微生物，其中所述表达盒位于细菌染色体上。所述位于细菌染色体上的表达盒可插入转座酶基因座中。所述位于细菌染色体上的表达盒可插入尿嘧啶磷酸核糖(UPP)转移酶基因座中。所述位于细菌染色体上的表达盒可插入到pyrE基因座中。

本发明提供了一种减少动物被致病细菌定殖的方法。所述方法包括用活体递送平台处理对其有需要的动物。所述活体递送平台包括遗传修饰微生物。所述遗传修饰微生物包括表达盒，所述表达盒包括一个或多个：用于转录表达的启动子、编码用于分泌的信号序列的核苷酸序列、编码细胞壁锚的核苷酸序列、至少一个编码所需生物分子的异源编码区、编码用于检测的表达肽标签的核苷酸序列、以及用于终止翻译和转录的终止子。所述遗传修饰微生物可以是细菌、酵母或真菌。所述遗传修饰细菌优选为芽孢杆菌、乳酸杆菌、乳酸乳球菌或肠球菌。所述遗传修饰细菌也可优选为大肠杆菌细菌。所述遗传修饰细菌也可以优选为罗伊氏乳杆菌菌株。

本发明提供了一种减少动物被致病细菌定殖的方法，其中所述方法包括用遗传修饰微生物处理对其有需要的动物。所述遗传修饰微生物经修饰后含有表达盒。所述表达盒包括用于转录表达的启动子，所述启动子具有本发明公开的如SEQ ID NO：14、SEQ ID NO：15、SEQ ID NO：16、SEQ ID NO：17、SEQ ID NO：18、SEQ ID NO：19、SEQ ID NO：41、SEQ IDNO：42或SEQ ID NO：43的序列。所述表达盒还包括用于分泌的信号序列，所述信号序列的氨基酸序列如SEQ ID NO：20、SEQ ID NO：21、SEQ ID NO：22、SEQ ID NO：23和SEQ ID NO：40的片段所示，其中所述片段的长度如本发明所示。用于分泌的信号序列包括氨基酸序列，其是SEQ ID NO：20、SEQ ID NO：21、SEQ ID NO：22、SEQ ID NO：23和SEQ ID NO：40的氨基末端片段，其中所述片段的长度如本发明所示。所述表达盒优选还包括编码所需生物分子的异源编码区。所述所需生物分子可以是具有抗感染活性的生物分子、益生菌因子、免疫调节因子或促生长生物分子。所述生物分子具有对抗致病细菌或寄生虫的抗感染活性。所述寄生虫优选为艾美球虫属物种。所述致病菌优选为沙门氏菌、梭菌、弯曲菌、葡萄球菌、链球菌、梭杆菌和大肠杆菌。所述抗感染生物分子可以是杀菌肽、酶、溶素、噬菌体或抗体。所述表达盒可任选地包含编码细胞壁锚的核苷酸序列、编码用于检测的表达肽标签的核苷酸序列、和/或终止翻译和转录的终止子。

本发明提供了一种减少动物被致病细菌定殖的方法，其中所述方法包括用遗传修饰微生物处理对其有需要动物。所述动物可以是鸟、人或非人哺乳动物。所述治疗可经口、经肠胃外、经鼻或经粘膜给药。当所述动物是鸟时，可以在卵内给药治疗。

本发明提供了本发明公开的任意遗传修饰微生物在治疗中的应用。本发明提供了经遗传修饰包含了本发明所公开的任何表达盒的细菌在治疗中的应用。治疗可以是减少动物被致病菌定殖。

本发明提供了如本发明公开的任意表达盒在治疗方面的应用。本发明提供了如本发明所公开的任意表达盒在治疗中的应用。治疗可以是减少动物被致病细菌定殖。所述表达盒应包含至少一个编码所需生物分子的异源编码区。所述至少一个编码所需生物分子的异源编码区可编码诸如杀菌肽、酶、溶素、噬菌体和抗体的生物分子。所述至少一个编码所需生物分子的异源编码区可编码具有抗感染活性的生物分子。所述至少一个编码所需生物分子的异源编码区可编码本发明公开的如SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：5、SEQID NO：6、SEQ ID NO：24、SEQ ID NO：25、SEQ ID NO：26、SEQ ID NO：27、SEQ ID NO：28、SEQID NO：29、SEQ ID NO：30、SEQ ID NO：33、SEQ ID NO：34、SEQ ID NO：44、SEQ ID NO：45、SEQID NO：46、SEQ ID NO：47、SEQ ID NO：48、SEQ ID NO：49或SEQ ID NO：50所示序列的生物分子。

本发明提供了如本发明所公开的任意遗传修饰微生物在制备减少动物被治病细菌定殖的药物中的应用。本发明提供了经遗传修饰以包含如本发明所公开的任意表达盒的细菌在制备用于减少致病菌对动物的定殖的药物中的应用。

本发明提供了如本发明所公开的任意表达盒在制备减少动物被致病细菌定殖的药物中的应用。本发明提供了在制备如本发明所公开的任何表达盒的药物中的用途。所述药物可以用于减少动物被致病细菌定殖。所述表达盒应包含至少一个编码所需生物分子的异源编码区。所述至少一个编码所需生物分子的异源编码区可编码诸如杀菌肽、酶、溶素、噬菌体和抗体的生物分子。所述至少一个编码所需生物分子的异源编码区可编码具有抗感染活性的生物分子。所述至少一个编码所需生物分子的异源编码区可编码具有本发明公开的如SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：5、SEQ ID NO：6、SEQ ID NO：24、SEQ ID NO：25、SEQ ID NO：26、SEQ ID NO：27、SEQ ID NO：28、SEQ ID NO：29、SEQ ID NO：30、SEQ IDNO：33、SEQ ID NO：34、SEQ ID NO：44、SEQ ID NO：45、SEQ ID NO：46、SEQ ID NO：47、SEQ IDNO：48、SEQ ID NO：49或SEQ ID NO：50所示序列的生物分子。

本发明提供了包含如SEQ ID NO：29、SEQ ID NO：49、SEQ ID NO：30、SEQ ID NO：33、SEQ ID NO：50或SEQ ID NO：34所示氨基酸序列的抗体，所述抗体优选地结合产气荚膜梭菌产生的毒素。所述抗体优选地结合产气荚膜梭菌α毒素和产气荚膜梭菌NetB毒素。本发明提供了包含如SEQ ID NO：29、SEQ ID NO：49、SEQ ID NO：30、SEQ ID NO：33、SEQ ID NO：50或SEQ ID NO：34所示氨基酸序列的抗体。本发明提供了包含如SEQ ID NO：29、SEQ IDNO：49、SEQ ID NO：30所示氨基酸序列的抗体在治疗中应用，本发明提供了如SEQ ID NO：33、SEQ ID NO：50或SEQ ID NO：34所示氨基酸序列的抗体在减少动物被产气荚膜杆菌定殖中的应用。本发明提供了包含如SEQ ID NO：29、SEQ ID NO：49、SEQ ID NO：30、SEQ ID NO：33、SEQ ID NO：50或SEQ ID NO：34所示氨基酸序列的抗体在制备减少动物被产气荚膜杆菌定殖的药物中的应用。本发明提供了一种动物产气荚膜杆菌感染或定制的方法，所述方法包括将包含如SEQ ID NO：29、SEQ ID NO：49、SEQ ID NO：30、SEQ ID NO：33、SEQ ID NO：50或SEQ ID NO：34所示氨基酸序列的抗体施用给对其有需要的动物。

附图说明

图1.用

平板测定罗伊氏乳杆菌菌株的抗菌性或敏感性。

图2.菌株3632周围清晰的“圈”证明，罗伊氏乳杆菌菌株3632可杀死病原性细菌，但不杀死非致病性嗜酸乳杆菌(Lactobacillus acidophilus)。

图3.表达盒的染色体整合。A.代表性表达盒的自杀载体图的非限制性示例。B.可能的染色体整合位点图。

具体实施方式

本发明所述“遗传修饰微生物”是指任何使用分子生物技术改变自然状态的微生物。遗传修饰可以是缺失细菌染色体或天然存在的质粒的一部分。所述遗传修饰也可以是将人工或外源核酸引入到部分染色体中。所述引入为扰乱或干扰细菌基因的表达。所述遗传修饰也可以是引入人工质粒。遗传修饰微生物可以是细菌、病毒、酵母、霉菌或单细胞生物。

“人工核酸”或“人工质粒”是指不是天然存在的而是使用分子生物技术构建的任何核酸或质粒。核酸或质粒的部分可以自然产生，但是这些部分都处于人工关系或组织中。

本发明所述“表达盒”是经构建以引起遗传修饰微生物表达所需生物分子的人工核酸。表达盒包括转录表达的启动子、编码用于分泌的信号序列的核苷酸序列、编码细胞壁锚的核苷酸序列，至少一个编码所需生物分子的异源编码区，编码用于检测的表达肽标签的核苷酸序列，以及用于终止翻译和转录的终止子中的一个或多个。启动子指导将编码区转录成信使RNA和mRNA翻译成肽。用于分泌信号的序列或分泌信号序列指导肽定位于细胞膜外。胞外肽可以是可溶的分泌蛋白，或者它可以是细胞相关的，特别是如果表达盒含有将胞外肽连接到细菌细胞壁的细胞壁锚序列。所述表达肽标签是可以被抗体或其它结合蛋白识别的任何氨基酸序列。所述表达肽标签还可以结合无机物质，例如与镍分子结合的六组氨酸的标签。用于翻译的终止子可以是终止密码子或含有终止密码子的间隔开放阅读框。

本发明所述“异源编码区”是含有编码肽的开放阅读框核苷酸序列。编码区与相关启动子是异源的，这意味着编码区与启动子在其自然状态下的不相关联。

本发明所述“蛋白”是呈现三维结构的氨基酸序列。肽可以与蛋白质互换使用，但也可以是不具有限定三维结构的氨基酸短线性序列。

本发明所述“所需生物分子”可以是当经由活体递送平台给药时对宿主有利的任何肽。所述所需生物分子可以是具有抗感染活性的肽、益生菌因子、免疫调节因子、抗肿瘤因子或促生长因子。所述所需生物分子还可以是产生抗感染活性物质或益生因子(例如维生素)的酶。

本发明所述“抗感染活性”包括防止宿主感染致病生物的任何活性。以下分子是有抗感染活性生物分子的实例：抗菌肽；溶素或裂解酶；原噬菌体、噬菌体或病毒；酶，例如裂解致病菌合成的蛋白质的酶或使致病菌失去合成蛋白能力的酶；阻断、抑制或清除致病分子的抗体。抗感染性可具有减慢、减少或防止致病物种生长的抑菌活性。抗菌肽的非限制性实例包括例如在EP0700998中所述的美杀菌素家族成员或美杀菌素样分子。溶素的非限制性实例包括由噬菌体产生的裂解分子。溶素对于某些致病种类的细菌可能具有特异性，并且已被建议用于替代传统抗生素(V.A.Fischetti,Viruses，vol.10，no.(2018)；R.Vazquezetal.Frontier in Immunology，vol.9，2252(2018))。

本发明所述“益生因子”是由遗传修饰微生物产生的，证明有益于宿主的物质。益生因子可以是促进所述遗传修饰微生物定殖所述宿主和/或延长所述遗传修饰微生物定殖所述宿主时间周期的附着分子或凝集分子。所述遗传修饰微生物在宿主中持续时间越长，提供的有益效果越持久。

本发明所述“免疫调节因子”可以是细胞因子、淋巴因子、趋化因子、白介素、干扰素、集落刺激因子或生长因子。所述免疫调节因子可以增强免疫应答的非特异性，或者所述免疫调节因子可以增加宿主中存在的免疫细胞的数量或组织分布。所述免疫调节因子还可以减少不适当的免疫应答，例如但不限于自身免疫应答。

本发明所述“促生长生物分子”可以是生长因子、转移因子(例如铁螯合分子)、激素或促进健康代谢活性的任何其它因子。

本发明所述“抗营养因子”可以包括蛋白酶抑制剂，例如胰蛋白酶抑制剂。

本发明所述“递送”或“给药”是指向宿主提供有益活性的动作。所述递送可以是直接的或间接的。给药可以是口服、鼻腔或粘膜途径。例如但不限于，口服途径可以是通过饮用水给药，鼻途径给药可以通过喷雾或蒸气，粘膜途径给药可以通过与粘膜组织的直接接触。粘膜组织是富含粘液的膜，例如排列在鼻部、嘴部、食道、气管、肺、胃、肠道(gut)、肠(intestine)和肛门的内表面上的膜。就鸟类的而言，给药可能在卵内，即受精卵给药。卵内给药可以通过喷洒到蛋壳表面上的液体或通过外壳注入的液体。

本发明所述术语“治疗”(treating)、“治疗”(to treat)或“治疗”(treatment)包括抑制、减慢、停止、减少、改善或逆转现有症状、紊乱、状况或疾病的进展或严重性。还可以预防性地给药治疗以预防或减少临床症状、紊乱、状况或疾病的发生率、发生、风险或严重程度。本发明所述术语“减少”可应用于预防疾病的(例如预防性)治疗或治疗性治疗。

以下实验实施例说明了包含可由所公开的活体递送平台递送的乳杆菌表达盒的活体递送系统。应当理解，其它实施方式和用途对于本领域技术人员将是显而易见的，并且本发明不限于这些具体的说明性实施例或优选实施方式。这些实施例的描述标题仅是为了方便，并且不应影响对其中呈现的任何结果的解释。

实施例1.罗伊氏乳杆菌菌株的鉴定

7种罗伊氏乳杆菌菌株分离自汉诺威研究中心较老的鸟类，来自汉诺威大学家禽诊所的盲肠内容物。所有七种菌株经16S rRNA测序都被鉴定为罗伊氏乳杆菌。

虽然需氧条件下大多数菌株在MRS肉汤和琼脂中生长有限(de Man,J.D.；Rogosa,M.；and Sharpe,M.E.“A Medium for the Cultivation of Lactobacilli”.J ApplBad.23：130-135(1960))，在39℃厌氧条件下，所有菌株在MRS琼脂和MRS肉汤上均表现出良好的生长。在厌氧条件下，在血液琼脂上培养细菌菌株主要导致有限的生长。这些菌株都不能在厌氧条件下在米勒顿(Mueller Hinton)肉汤中生长。对于所有进一步的分析，细菌菌株在39℃的厌氧条件下在MRS培养基中生长。

使用

平板测试细菌分离物的抗菌敏感性(图1)。所有的菌株都抗大肠杆菌素、多西环素、恩诺沙星、红霉素、新霉素、苯唑西林、青霉素G、联磺甲氧苄啶、四环素、替米考星和泰乐菌素。除了菌株3632之外，所有菌株都耐受链霉素，除了菌株2098之外都抗泰妙菌素。另外，菌株2091、2095、2097和3630耐受头孢泊肟酯；菌株2091、2095和2097耐受头孢噻肟；菌株3630和3632耐受林可霉素。在测试浓度下，未发现菌株耐受阿莫西林、头孢噻呋、红霉素D、林可霉素-大观霉素和利福平。

实施例2.罗伊氏乳杆菌菌株的选择及抗感染活性

为了选择用于进一步工程的最佳菌株，测试了罗伊氏乳杆菌各种期望的益生菌抗感染性性质，例如生长动力学、产生过氧化氢的能力、自凝集、酶谱、在牛胆汁和胰酶存在下的存活率以及对热休克和pH变化的敏感性。还使用溶血试验对罗伊氏乳杆菌菌株进行了安全性测试。

总体来看，所有菌株在益生菌性质方面表现得非常相似，包括生长动力学和产生过氧化氢的能力，除了菌株3632显示出一些独特的性质，包括在液体培养基中的自凝集能力(与良好表征的人益生菌株罗伊氏乳杆菌ATCC23272相当)。发现这些菌株都没有在血液琼脂平板上溶血，表明这些分离物不太可能对人致病。

用

测序(扩增子表达)对罗伊氏乳杆菌菌株2091、3632和一株来自欧洲的独立分离罗伊氏乳杆菌菌株170331进行了全基因组测序。表1总结了测序、组装和注释统计信息。被测菌株之间的基因组结构和组织有所不同。

表1：基因组测序，组装和注释统计

在基因组测序数据的基础上，基于与美杀菌素保守结构域的同源性菌株3632编码属于美杀菌素家族的两个细菌素。这些细菌素似乎是菌株3632特有的。编码一种美杀菌素(美杀菌素-El)的cDNA可以是：

(SEQ ID NO：1)。该新型开放阅读框将编码美杀菌素-El的多肽：

MDKEELEKIVGNNFEEMSLQKMTEIQGMGEYQVDSTPAASAISRATIQVSRASSGKCLSWGSGAAFSAYFTHKRWC(SEQ ID NO：2)

编码第二种形式的美杀菌素(美杀菌素-E2)的另一种cDNA可以是：

(SEQ ID NO：3)。该新型开放阅读框将编码美杀菌素-E2的多肽：

MEEKELEGVIGNSFESMTVEEMTKIQGMGEYQVDSTPGYFMESAAFSALTANITRHAMHHH(SEQ IDNO：4)

罗伊氏乳杆菌菌株3632编码对该菌株抗感染活性有贡献的其它多肽。卷曲霉啶(capreomycidine)合成酶参与卷曲霉素的合成，在培养上清和颗粒化(pelleted)细胞中均有表达。

MVEIAHFGVEAWLNKWEKSATYDISQSTIASLSMHDLLNLDGNNGEEFYEMLDKQQMNYGWIEGSPEFKEEVAKLYHHVDPENILQTNGATGANILALYALINPGDHVIAEYPSYQQLYDIPKSLGADVDYWHIHEEDNWYPRIDDLKAMVKPNTKMICLNNANNPTGTVLDKEFLEQVVEIAKSVDAYVLVDEVYLPLDHPEKFAQIIDLYDKGISTNSLSKTYSVPGVRIGWTATNAEVADIFRKFRDYTMICGGVFNDQLATYVLRHRDQVLARNRKLVLGNLAIYKDWIDHEDRASVIMPQAVSTSFPKLDVPVDIHTFCENLLHDEGVLLVPGDAFDTPGHVRLGYCAPEATLKEGLKRLSKYMHQYD(SEQ ID NO：5)

大肠杆菌素V产生蛋白也在菌株3632的颗粒细胞中表达：

MILTTFIILILMGCFINGHRRGLLTMTLMLGTYIVAWIVARQGAQLIGGWLKSLLPSIGTPATFSESLLANVNSNLFFYNGIAFMIIFTIVSILCHWGIRQLNWIKRIPVVGTVDKIAGGLISFLIGYLIIYVVLLIMQLFPAGWWQMQIANSELARFMINQTPGIAHLVIDTLVQGG(SEQ ID NO：6)。本领域技术人员应认识到，由于遗传密码的冗余，多个核苷酸序列可编码上述肽。

用菌株3632和病原菌共种植来测试这些推定细菌素的功能。如图2所示，罗伊氏乳杆菌3632杀死可杀死致病细菌产气荚膜梭菌(Clostridium perfringens)、金黄色葡萄球菌(Staphylococcus aureus)、猪链球菌(Streptococcus suis)、乳房链球菌(Streptococcus uberis)、大肠杆菌(Escherichia coli，APEC)，鼠伤寒沙门氏菌(Salmonella typhimurium)，坏死梭杆菌(Fusobacterium necrophorum)和空肠弯曲菌(Capylobacter jejuni)。非致病性对照嗜酸乳杆菌未被杀死。例如3632的DFM菌株可用于控制、限制、预防或消除猪致病菌(猪链球菌)，家禽致病菌(产气荚膜梭菌、大肠杆菌、鼠伤寒沙门氏菌和空肠弯曲菌)和牛致病菌(金黄色葡萄球菌和坏死梭杆菌)的感染。诸如3632的DFM菌株可用于控制、限制、预防或消除人类中食源性致病菌的感染。

实施例3：罗伊氏乳杆菌菌株3632的其它遗传元件

为了鉴定任何含有抗菌抗性基因的潜在质粒，通过高通量测序(ACGT，Inc.)分离和鉴定了天然质粒。3632菌株含有三种天然质粒，但没有任何已知的抗菌基因。然而，其中一个质粒保持高拷贝数：

(SEQ ID NO：7)。基于BLAST分析，假定的罗伊氏乳杆菌质粒复制原点可以是：

(SEQ ID NO：8)。

对菌株3632序列的分析显示，与罗伊氏乳杆菌菌株2091、170331和ATCC 23272(人类)相比，在罗伊氏乳杆菌菌株3632中发现了两种独特的凝集素受体前体(ARP)。这些凝集素受体前体可能导致该菌株观察到的自凝集水平增加，并且这些受体将有助于遗传修饰DFM表达凝集素受体的益生潜力。ARP-1的氨基酸序列为：

MNEYNAEMAKLNQGANAPVITTNSVNQALSLKPENNATVDIEALNPRITFKRVEEGTKYAGYQIFDKNNAYVNNIDGEFLRVTYTNLKNSTYKGSKISKIVVTYSDSTPTGNRITQSGLNAVTEGANDNFLVVFEDPVRGDMHSTTVTATYQYYDANGNLIDFSGTNNAWLSVGSLNFDQGNDYQGGKNEGNPTSGISEGVKLISGAQIKQLAGSSISVHDDGWAYAGFNNYSGTGMNNGINTDNGGSGWDMDGSPNAYYGAIVFQLTGSSVSLRQGLVSWGGADIASQYNNQFLNNAWFTAGTTLPETQIKQPIRKTSETHYHYNPSVIRL(SEQ ID NO：9)。ARP 2的氨基酸序列为：

MAQKLMSANSTDKNFKMYKSKKSWVFAYSTTLALAAVAGITLSTTNVHADTTNGGDNQVNATAVTQNTTSNTVDQIAANTAQTDNTSTSINIRSLMDDLASGDDTSSSQNGQEQSQNYASSNQNSQTQQENGTTGQSTASQNGTTSDQTNSDQSDKNYYVISTRDLDKNGNVNYLTQKNYTSIKGQEVADGTVVTWPLSVSALPANRAQDLKSHVISETLDPHLEYLHYRAYLTNTDGTVTDVTNHVNLNRSGQTLIFTDDNYLLSIYNNNRYRVQNLPVIKLVTKANGNGYIIPNAFKSSYVFNDGSHDVSFTTTSNNVQIKTFNPGNSKDVEIGGNVQGDPSGTINGQVVADGSVVTWPMSVGDLPANRAQDVLSHIETDTLYNGLNYEGYHAYLPQADGSFQDVSSHINVQQNGQDLTFIADDYLIGLYNQDKSTAFKMPIIDLITSVHGTSIIAPNKFNSQLAFKDGNGQTVINNTSNQVQISTYHPTNTKDVELGGNVQGDTPNSINDKVVANGAIVTWPMASSELPANRVQDLQSRVISETLDSHLQYQGYKAWLQNADGKYTDVTSHVKLTQDGQNLTFADDEYLLNLYNSNKGTAYKLPIIDLVTKVNGAGITAPNSYTTKYVYSDGDGNTTINVTSNTVKISTFNPTTNKDVELGDNIHGDTESSIAGKLVSEGTIVTWPLSTSDLPANRAQDVVSHTAVDALEPTLQYISYTAWLPDSNGQLQDVTSHVKMTRDGQKLTFTDDDYLIGLYNQNKDIALKMPIIDLVTKATGNTKLLPNSFDSQFVYNDVDGNTIINVSSNKPTVETFDPTVHKDVELGGNNVQGDTPNSIDGKIVAQGTVVTWPMSTSDLPANRTQDVVSHSTSETLNQNLQYVGYHAYMPDANGKLQDVTSHVQLQQNGQNLVFTDDSYLINLYNQDKSIAFKMPIIDLMTKAISDSATIPNTFESQYVFNDGNGNTTFKSTSNTVQIITYKPKTTKDVELGDNIHGDTNASIAGQMITDGTVVTWPMSTSDLPANRTQDLQQHVVTDNLNDNLIFQGYTAWLPTANGLVDVTNHIELTRDGQNLTFTDDAYLLNLYNQNKDTAYKLPIIDLVTKANGNTKLIPNNFDSMFVYNDGDQQTTVNVTSNTVNISTYDPTATKDVELGDDIEGDTADTINNLMVQIGTKMTYPLTVSDLPANRADEITAHQSVDTLSDYLEYQGYKAYLPDADGKLQDITEHVNLKREGQKLSFNDDDYLINLYNNSKATKQALPVIDLVAKVTGSNDGKKVHIIPNHFDSTITTKDGKINTTSNTVVINSNDPEAVKDVELGDNVVGDTPNSVTGTTVADGTIVTWPMSVGSLGANRAQNVIKHTETENLDSGLTYLSFKAYLPDADGKMQDITEHINIQQDGQKLVFTDDDYLISLYNKDKSQRFALPVIDLVTRVNGDNKIIPNTFVSQFTFNDGKGNTITSVTSNQVNVSTFKSNPEKHVTLGTDIEGDDAENADGTVVAQGSEVTWPLSDKSPLPANRSQDVKSHTLVDKLDDNLQYNSYKAYLKGTDGKLQDVTDHIKLTRDGQNLTFIDDDYLLDLYNKDKSTAFNLPIIDLVTTVVGNDKLIPNKFDSNFVFSDGNKDTSMKTTSNEVSISTYTPVTNKDAELGDNVVGDTSDSIANETVPDGTIVTWPLSVSSLPANRSQDVFKHVIEDILDGNLTYNSFKAYLKDAAGNLQEVTDHVKLAQEGQHLTFTDDDYLINLYNSSKNKEQSLPIIDLVTTVHGDSKLIPNEFDNVFVFKDGKGQTTVKTTSNKVTIKTASLPTPTKEETDDQGNNINGNEVKAGEHVNYTLNWDLSNDKDVKATPEMIKKGFFFIDPIDSRALSVDDLSKAKVVDQNGNKVDGISFHLYNSLSEVPEFIQEQVKANNLQDKITGPFVVAQADDLQAFFDKYVKTGAKLKVTIPTIVKSGFTGEFSNTAYQFGFGKATPTNTVTNYVKPMHKPASPETPAAIAPQVISATAQPMTSDAPVTPSEKTAKLPQTGNADEGALLGLAAVSLVGSLGLAALGLKQNRNDD(SEQ ID NO：10)。本领域技术人员应认识到，由于遗传密码的冗余，多个核苷酸序列可以编码上述肽。

罗伊氏乳杆菌菌株36323632包含八个噬菌体编码位点，但其中只有三个位点似乎编码多产原噬菌体。噬菌体基因座1具有以下序列：

(SEQ ID NO：11)。噬菌体基因座3具有以下序列：

(SEQ ID NO：12)噬菌体基因座6具有以下序列：

(SEQ ID NO：13)

实施例4.罗伊氏乳杆菌启动子和转录终止子的选择

分别对罗伊氏乳杆菌3632颗粒(pellets)和培养上清液进行全蛋白组学分析，以鉴定潜在的强启动子和分泌信号。颗粒和上清液中的蛋白质按其表达水平排序，高表达和分泌蛋白质的启动子和分泌信号通过全基因组测序进行鉴定(实施例2)。所述遗传元件被选择用于表达盒中，所述表达盒可用于产生遗传修饰细菌，所述遗传修饰细菌将所需生物分子递送给对其有需要的宿主动物。表达盒将包括合适的启动子、编码所需生物分子的异源编码序列和转录终止子。异源编码序列可包括用于分泌的信号序列、细胞壁锚定序列和/或可检测的肽标签。

通过分析全蛋白组学数据鉴定了六个合适的启动子。这些启动子序列为：

木酮糖-5-磷酸酮酶启动子：

(SEQ ID NO：14)；延长因子TU启动子

(SEQ ID NO：15)；甘油醛-3-磷酸脱氢酶启动子

(SEQ ID NO：16)；葡萄糖-6-磷酸脱氢酶启动子

(SEQ ID NO：17)；L-乳酸脱氢酶启动子

(SEQ ID NO：18)；和tuf启动子

(SEQ ID NO：19)。上述序列的下划线区域表示核糖体结合位点。

通过对全蛋白质组学数据的分析，还确定了四种合适的分泌信号肽。这些分泌信号序列可来自以下蛋白质：

C蛋白α-抗原前体

(SEQ ID NO：20)；阿拉伯半乳聚糖内酯-1,4-β-半乳糖苷酶前体

(SEQ ID NO：21)；染色体分离蛋白

(SEQ ID NO：22)；D-γ-谷氨酰-内消旋-二氨基嘌呤酸内肽酶Cwls前体

(SEQ ID NO：23)；和染色体分割蛋白Smc

(SEQ ID NO：40)。本领域技术人员应认识到，由于遗传密码的冗余，多个核苷酸序列可以编码上述肽。

实施例5.表达盒

根据前面实施例中提供的信息，可以设计产生高水平分泌目标生物分子的表达盒。例如，其中一个表达盒(CwlS_C2；SEQ ID NO：23)已进化为向细胞壁传递内肽酶(细胞壁水解酶)。内肽酶在细胞分裂和分离中起着重要作用。用异源编码区替换内肽酶将引起所需目标生物分子(例如靶向致病细菌的抗感染生物分子)的分泌。一个具体实施例为本发明所公开的新型美杀菌素可通过用强启动子(实施例4；SEQ ID Nos：14～19)和(可选地)强分泌信号(实施例4；SEQ ID Nos：20～23)换掉天然美杀菌素启动子而高水平表达。

表达载体是用以下部分或全部组分的不同组合来设计和合成的：罗伊氏乳杆菌复制的复制原点(例如，来自高拷贝数质粒；SEQ ID NO：8)、大肠杆菌复制的复制原点、用于选择的耐药标记、用于在乳酸杆菌中表达的强启动子、用于分泌的信号序列、编码所需生物分子的异源编码区、用于检测的表达肽标签、用于分泌的细胞壁锚、以及用于转录终止的终止子。

可选地，编码所需生物分子的异源编码区可整合到遗传修饰微生物的染色体中。表达盒(用于在乳酸杆菌中表达的强启动子、用于分泌的信号序列、编码所需生物分子的异源编码区、用于转录终止的终止子、以及任选地用于检测的表达肽标签和用于分泌的细胞壁锚)的染色体整合用自杀载体完成。自杀载体包括大肠杆菌中复制的复制原点、用于选择的耐药标记和两侧有与染色体特定区域同源核酸的表达盒。这种自杀载体的示例呈现在图3A中。

对于染色体整合，罗伊氏乳杆菌基因可以通过插入表达盒而被打断。这些染色体位点的图谱如图3B所示。可能被打断的两个基因是转座酶和尿嘧啶磷酸核糖(UPP)转移酶。染色体整合也可以在高温基因座内完成，并且高温缺失和随后的再插入可以用作整合的选择标记(Sakaguchi等,Bioscience of Microbiota，Food and Health，32：5968(2013))。修饰的pyrE基因还可用于重组细菌的生物抑制，因为重组细菌不能在缺少代谢补充剂的环境中存活。

实施例6.包含在表达盒中的酶基因

表达盒将包括编码所需生物分子的异源编码区。所述所需生物分子可以是具有抗感染活性的生物分子。所述抗感染活性可以是特异性针对特定属的致病菌，由裂解酶裂解致病细菌，例如来自噬菌体的裂解酶。

裂解酶可以包括PlyCM、靶向产气荚膜梭菌的裂解酶，其由以下序列编码：

(SEQ ID NO：24)；PlySS2，靶向猪链球菌和金黄色葡萄球菌的裂解酶，由以下序列编码：

(SEQ ID NO：25)；以及CP025C，一种靶向产气荚膜梭菌的裂解酶，由以下序列编码：

(SEQ ID NO：26)。

另外，在罗伊氏乳杆菌菌株3632中发现的三种多产原噬菌体中的两种含有假定的裂解酶。这些裂解酶可用于靶向致病菌或控制遗传修饰乳酸菌的环境扩散。这两种裂解酶是：来自原噬菌体基因座1的N-乙酰壁氨酰-L-丙氨酸酰胺酶sle1前体，其核苷酸序列为：

(SEQ ID NO：27)，以及来自原噬菌体基因座6的肽酶家族M23，其核苷酸序列为：

(SEQ ID NO：28)

实施例7.单链抗体及其活性

表达盒包括编码所需生物分子的异源编码区。所述所需生物分子可以是具有抗感染活性的生物分子。所述抗感染活性可以是抑制或中和致病菌产生的毒素。所述抑制或中和可以用单链抗体完成。乳酸杆菌被描述为针对宿主附着因子的单链抗体的表达系统(W02012/019054)。单链抗体靶向的毒素包括产气荚膜梭菌α毒素和NetB。

通过QVQ Holding BV公司(乌特勒克，荷兰)制备针对产气荚膜梭菌α毒素和NetB的骆驼科单重链(VHH)抗体。简而言之，用重组α毒素或NetB变体W262A免疫两个美洲驼幼崽。这些免疫原都不溶血。用毒素肽将免疫后的美洲驼增强两次。在初次免疫后的第44和72天，取血样并分离出用于噬菌体文库构建的RNA。对噬菌体库进行筛选，以确定其对两种毒素的结合活性。对候选抗体进行测序并在如下文所述的生物测定中进一步筛选。

α毒素对多种红细胞和培养细胞造成膜损伤。α毒素优先对真核细胞膜外小叶的两种主要成分磷脂酰胆碱(PC或卵磷脂)和鞘磷脂(SM)具有活性。N端结构域对磷脂酰胆碱具有完全活性，但缺乏鞘磷脂酶活性，且不溶血或细胞毒性。C端结构域缺乏酶活性，但N端和C端结构域之间的相互作用对于赋予该毒素鞘磷脂酶活性、溶血活性和细胞毒性至关重要。虽然α毒素是一种有效的溶血素，但只有在实验动物或梭状芽孢杆菌败血症的病例静脉注射毒素后，才能看到红细胞的溶解。

针对α毒素的VHH抗体对α毒素卵磷脂酶活性的抑制能力是通过测量其对蛋黄脂蛋白的影响来确定的。新鲜蛋黄离心(10000×g，4℃下20分钟)并在PBS中稀释1：10。在添加10％蛋黄乳剂之前，将两倍稀释梯度的VHHs(每稀释两个孔，5mM起始浓度)与恒定量的α-毒素(5μg/ml重组α-毒素或来自Sigma，P7633的3.33×10^-4U/μlα-毒素)在37℃下预孵育30分钟，以评估VHHs中和α-毒素活性的能力。作为阳性对照，使用用重组α毒素免疫的幼崽血清，从1：4的稀释开始。在37℃下培养1小时后，测定650nm处的吸光度(A₆₅₀)。α-毒素活性由浊度的增加表示，浊度的增加导致吸光度的增加。

对照血清能够中和商业和重组α毒素的卵磷脂酶活性。八倍稀释的抗血清(相当于3.12％的血清)能够完全中和重组α毒素的α毒素卵磷脂酶活性，而只有最高稀释的抗血清(相当于25％的血清)能够完全中和商业α毒素的卵磷脂酶活性。五种候选VHH抗体的抑制能力存在显著差异。VHH EAT-1F3对两种α毒素的卵磷脂酶活性均无影响。EAT-1 A2和EAT-1C8的中和能力非常相似，重组和商用α毒素的中和能力相同。在VHHs稀释32倍(0.16μM VHH)之前，保持最大抑制能力。然而，EAT-1 A2和EAT-1C8都不能完全中和卵磷脂酶活性，导致40％到50％的剩余卵磷脂酶活性。另外两种VHH，EAT-1F2和EAT-1G4显示出对重组和商业α毒素的中和能力不同。EAT-1F2对重组α毒素具有较高的中和能力，但不能完全中和商业α毒素，导致约25％的卵磷脂酶残留活性。与EAT-1F2相比，EAT-1G4可中和商业α毒素100％的卵磷脂酶活性，但中和重组α毒素的能力较弱。

通过测量针对α毒素的VHH抗体对绵羊红细胞的影响，确定其对α毒素溶血活性的中和作用。与抑制α-毒素卵磷脂酶活性类似，添加1％绵羊红细胞之前，将VHH抗体的两倍梯度稀释(每稀释两孔，5μM起始浓度)与恒定量的α-毒素(来自Sigma，P7633的6.25×10⁵U/μlα-毒素)在37℃下预孵育30分钟来评估中和溶血活性的能力。作为阳性对照，使用重组α毒素免疫的幼崽血清，从1：4稀释开始。在37℃下培养1小时后，离心培养皿以获得完整的红细胞颗粒。将上清液转移到新的96孔板上，并测定A₅₅₀。α-毒素活性由红细胞释放血红蛋白引起的吸光度增加表示。

仅使用商业α毒素确定VHH抗体对α毒素溶血活性的抑制能力，因为重组α毒素显示没有溶血活性。对照血清的16倍稀释(对应于1.56％血清)完全抑制了α毒素溶血。相反，候选VHHs中没有一个对α毒素的溶血活性有影响。因为对照血清含有多克隆抗体，而VHHs是单克隆抗体，所以测定所有的5个VHHs对α毒素的联合作用(在最高稀释度中，每个VHH的1μM，对应于总共5μM VHHs)。组合VHHs对α毒素溶血没有影响。

基于上述结果，选择EAT-1F2和EAT-1G4用于进一步表征。EAT-1F2的肽序列为：

EVQLVESGGGLVQAGGSLRLSCAGSGRTGSLYSMGWFRQAPGKEREFVAAITWRPSSTYYADSVKGRFTISRDDAKNTVYLQMNSLKPEDTAVYFCAARPRGGLSPTPQAYDYWGQGTQVTVSSAAASGSLEQKLISEEDLNGAAHHHHHHGAA(SEQ ID NO：29)，EAT-1G4的肽序列为：

EVQLVESGGGLVQPGGSLRLSCAASGSIATINDMGWFRQAPGKQRDWVATIVSDGSTAYADSVKGRFTISRDNAKNTVYLQMNSLKPEDTAVYYCSARRHYGQGTQVTVSSAAASGSLEQKLISEEDLNGAAHHHHHHGAA(SEQID NO：30)。

还可以产生EAT-1F2(SEQ ID NO：49)的R27H突变体，以提高抗体的蛋白酶抗性。

本领域技术人员应认识到，由于遗传密码的冗余，多个核苷酸序列可以编码上述肽。然而，编码EAT-1F2的示例性序列可以是：

(SEQ ID NO：31)，编码EAT-1G4的示例性序列可以是：

(SEQ ID NO：32)。

NetB是在平面磷脂双层中形成单通道的七聚β-成孔毒素。NetB活性受膜流动性和胆固醇的影响，这增强了NetB的低聚且在孔形成中起重要作用。NetB对禽类红细胞具有高溶血活性。

通过测定NetB介导的鸡肉红细胞的裂解来测定针对NetB的骆驼科VHH抗体对NetB溶血活性的中和。在添加1％鸡红细胞之前，将两倍梯度稀释的VHH抗体(每稀释两个孔，5μM起始浓度)与恒定量的NetB毒素(20μg重组NetB)在37℃下预孵育30分钟来评估中和NetB溶血活性的能力。阴性对照包括非毒性NetB变体W262a，因为该变体不显示溶血活性。使用来自用重组NetB免疫的兔子的阳性对照血清(野生型NetB)，从1∶4稀释开始。在37℃孵育1小时后，离心所述板以沉淀完整的红细胞。将上清液转移到新的96孔板上，并测定A₅₅₀。通过从红细胞中释放血红蛋白引起的吸光度增加来指示NetB活性。

对照血清能够中和NetB的溶血活性。VHH抗体ENB-1F4和ENB-1F10对NetB溶血没有影响。ENB-1B9具有中等抑制能力，而ENB-1D11和ENB-1A4能够中和NetB溶血直至4-8倍稀释(1.25μM～0.625μM VHHs)

基于上述结果，选择ENB-1A4和ENB-1D11用于进一步表征。ENB-1A4的肽序列为：

EVQLVESGGGLVQAGGSLRLSCAASGSIFSTNVMGWYRQAPGKQREFVAGITIGGTARYPDSVKGRFTISRDNTQNTVYLQMNNLKPEDTAVYYCNAVLPSDQRRWSWGQGTQVTVSSAAASGSLEQKLISEEDLNGAAHHHHHHGAA(SEQ ID NO：33)，ENB-1D11的肽序列为：

EVQLVESGGGLVQTGGSLRLSCTASGTIDMTYGLIWYRQAPGKERELVASIRRDGRTNYADSVKGRFTISIDNAKNSIHLQMNSLKPDDTARYYCNSPYHALWGQGTQVTVSSAAASGSLEQKLISEEDLNGAAHHHHHHGAA(SEQ ID NO：34)

还可以产生ENB-1D11(SEQ ID NO：50)的R56H突变体，以提高抗体的蛋白酶抗性

本领域技术人员应认识到，由于遗传密码的冗余，多个核苷酸序列可以编码上述肽。然而，编码ENB1-1A4的示例性序列可以是：

(SEQ ID NO：35)，编码ENB 1G4的示例性序列可以是：

(SEQ ID NO：36)

实施例8.多因子表达盒

表达盒可以设计和合成有以下组分的不同组合：用于表达的启动子、用于分泌的信号序列、用于分泌的细胞壁锚、至少一个编码所需生物分子的异源编码区和用于转录终止的终止子。表达盒可以包含多个异源编码区。

用于表达骆驼科抗体的一种代表性的表达盒是：

(SEQ ID NO：37)；其中核苷酸1至185表示来自Cwls内肽酶(SEQ ID NO：41)的启动子序列，核苷酸186至262编码Cwls内肽酶分泌信号(SEQ ID NO：23)，核苷酸263至407编码用于附接至细胞外多糖的LysM域，核苷酸408至431是间隔序列，核苷酸432至854编码VHHEAT-1G4(SEQ ID NO：30)，核苷酸855至890是终止于终止密码子以用于终止翻译的另一间隔序列。

用于表达骆驼科抗体的第二代表性表达盒是：

(SEQ ID NO：38)，其中核苷酸1至275表示来自染色体分离蛋白(SEQ ID NO：42)的启动子序列，核苷酸276至383编码来自染色体分离蛋白(SEQ ID NO：22)的分泌信号，核苷酸384至806编码VHH EAT-1G4(SEQ ID NO：30)，核苷酸807至1190编码来自染色体分离蛋白(SEQ ID NO：22)的细胞壁锚序列，核苷酸1191至1193是用于终止翻译的终止密码子。

用于表达骆驼科抗体的第三代表性表达盒是：

(SEQ ID NO：39)，其中核苷酸1至313表示来自染色体分割蛋白Smc(SEQ ID NO：43)的启动子序列，核苷酸314至499编码来自染色体分离蛋白Smc(SEQ ID NO：40)的分泌信号，核苷酸500至922编码VHH EAT-1G4(SEQ ID NO：30)，核苷酸923至1333编码来自染色体分离蛋白Smc(SEQ ID NO：40)的细胞壁锚定序列，核苷酸1334至1336是用于终止翻译的终止密码子。

所公开的表达盒元件的许多其它组合也是可能的。

实施例9.源自芽孢杆菌的抗感染分子

某些芽孢杆菌菌株也可以用作DFM，例如用于家禽。从鸡盲肠中收集到100多株芽孢杆菌。这些鸡是从美国印第安纳州采购的。分离的毒株经过挑选，可耐受100℃10分钟，作为颗粒稳定的产品进行开发。经16srRNA测序证实分离菌株为芽孢杆菌。对这些分离物进行了对各种家禽致病菌(如肠炎沙门氏菌(Salmonella Enteritidis)、禽致病性大肠杆菌、产气荚膜梭菌、盲肠肠球菌(Enterococcus cecorum)和空肠弯曲菌)抗菌活性的测试。不同的分离物对这些致病菌显示出不同的抗菌活性。在所有的分离物中，五个分离物对所有测试的致病菌都有细胞毒性。所有5个分离物均分离自相同研究收集的样品，表明它们可能是可重复的。选择2个分离株D24和D72用于进一步表征。

两个选择的分离株(D24和D72)经

测序确定最可能是芽孢杆菌。基因组序列分析潜在的细菌素、抗菌肽和消化酶。这些分离物包含几个可能编码抗菌肽和细菌素的完整基因座。考虑到将这些抗菌肽和细菌素转化到罗伊氏乳杆菌表达系统中的可行性，选择了五种核糖体细菌素进行进一步研究。根据序列同源性，五种芽孢杆菌细菌素被鉴定为抗菌肽LCI：

AIKPVPSPNGIFAASFELNGTTWIFKYKYYDSSKGYWVGIYESVDK(SEQ ID NO：44)；环素A：

LASTLGISTAAAKKAIDIIDAASTIASIISLIGIVTGAGAISYAIVATAKTMIKKYGKKYAAAW(SEQID NO：45)；羊毛硫肽3(植物乳杆菌素C)：

EFSGGGGAEQRGISQGNDGKLCTLTWECGLCPTHTCWC(SEQ ID NO：46)；羊毛硫肽5(地衣杀菌素(lichenicidin)A2)：SDATPMTVTPTTITIPISLAGCPTTKCASIVSPCN(SEQ ID NO：47)；和羊毛硫肽化合物6：

SEATPMTVTPTTITIPISLAGCPTTKCASIVSPCND(SEQ ID NO：47)。这些杀菌素此前已被证明能杀死各种革兰氏阴性和革兰氏阳性致病菌。

这些芽孢杆菌源的抗感染肽可整合进表达盒中并重组表达。重组芽孢杆菌源抗感染肽可单独表达或(但不限于)与实施例2中公开的其他抗感染肽或噬菌体、实施例6中公开的裂解酶或如实施例7中公开的抗体中的一个或多个共同表达。

实施例10.罗伊氏乳杆菌的体内效果

只有当菌株能够定殖于宿主胃肠道时，鉴定的菌株才能有效地起到直接饲喂微生物的作用。在不同给药途径之后评估罗伊氏乳杆菌菌株定植鸡的能力。

罗伊氏乳杆菌口服给药10天龄的雏鸡。在接种后的不同时间，处死雏鸡，并且在作物(crop)、小肠和盲肠中测定罗伊氏乳杆菌的存在。菌株3632能够对所有三种组织进行定殖，并在至少18天内保持可检测状态。

罗伊氏乳杆菌在孵化前3天通过卵接种(即，注射到蛋中)给药。在孵化后3天和7天在雏鸡的组织中发现菌株3632，而对照菌株的定植在第3天后下降。

罗伊氏乳杆菌通过将含有细菌的液体喷撒到蛋表面上而在卵内给药。同样，菌株3630和3632能够在孵化后3天和7天定殖和保持存在。通过喷雾接种如卵内注射一样有效，但需要较少的孵卵操作。

携带表达盒的重组菌株3630和3632，所述表达盒包含实施例7的抗体之一或PlyCM裂解酶通过卵内接种给药。所述重组菌株能够以与未修饰的亲本菌株相当的水平有效地定植鸡胃肠道。因此，如本发明所考虑的遗传操作不改变经修饰的菌株充当DFM的能力。

对携带罗伊氏乳杆菌菌株或修饰菌株的雏鸡进行坏死性肠炎的敏感性评估。雏鸡接种罗伊氏乳杆菌活表达系统，其中表达盒含有单链抗体或裂解酶。通过提供含有10⁸个重组罗伊氏乳杆菌的雏鸡饮用水，口服进行接种。

如表2所示进行实验，其中40只一天龄的雏鸡分为10组。第1组雏鸡作为未处理和未攻毒的对照。第2组雏鸡作为未处理的攻毒对照。第3组表示给定的标准抗生素治疗的攻毒雏鸡。第2～10组在第17天和第18天的每一天各接受2×10⁸CFU的产气荚膜梭菌。第4～9组从第一天开始预防性接种罗伊氏乳杆菌。第10组仅在第18～21天接受治疗剂量的罗伊氏乳杆菌。第21天处死雏鸡，检查小肠组织是否存在病变。还记录了每个治疗组的雏鸡存活情况。

如表2所示，四种抗体可以各自一定程度改善疾病。菌株的组合可能比任何单个菌株都更有效。

表2重组罗伊氏乳杆菌产生的抗体可以治疗坏死性肠炎

^a用BMD(亚甲基双水杨酸杆菌肽)处理。

序列表

<110> 伊兰科美国公司

<120> 一种活体递送平台

<130> X21471

<150> 62/801307

<151> 2019-02-05

<160> 50

<170> PatentIn version 3.5

<210> 1

<211> 231

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 1

atggacaaag aagaattaga aaaaattgta ggtaataact ttgaggaaat gagtttacaa 60

aaaatgacag aaattcaagg tatgggtgaa taccaagtgg attcaacacc agcagcttct 120

gcgatttcac gggcaacaat tcaagtatca cgtgcatctt ctggaaaatg tctaagttgg 180

ggtagtggtg cagcatttag tgcttatttt actcataaaa gatggtgcta g 231

<210> 2

<211> 76

<212> PRT

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 2

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

1 5 10 15

Met Ser Leu Gln Lys Met Thr Glu Ile Gln Gly Met Gly Glu Tyr Gln

20 25 30

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

35 40 45

Val Ser Arg Ala Ser Ser Gly Lys Cys Leu Ser Trp Gly Ser Gly Ala

50 55 60

Ala Phe Ser Ala Tyr Phe Thr His Lys Arg Trp Cys

65 70 75

<210> 3

<211> 186

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 3

atggaagaaa aagaattaga aggtgtaata gggaattcgt ttgaaagtat gactgtagag 60

gaaatgacaa aaattcaagg tatgggtgaa tatcaagtag attcgacgcc tggatatttt 120

atggaaagtg ctgccttttc agctcttaca gccaatataa caagacatgc tatgcatcat 180

cattaa 186

<210> 4

<211> 61

<212> PRT

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 4

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

1 5 10 15

Met Thr Val Glu Glu Met Thr Lys Ile Gln Gly Met Gly Glu Tyr Gln

20 25 30

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

35 40 45

Leu Thr Ala Asn Ile Thr Arg His Ala Met His His His

50 55 60

<210> 5

<211> 373

<212> PRT

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

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Met Val Glu Ile Ala His Phe Gly Val Glu Ala Trp Leu Asn Lys Trp

1 5 10 15

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

20 25 30

Ser Met His Asp Leu Leu Asn Leu Asp Gly Asn Asn Gly Glu Glu Phe

35 40 45

Tyr Glu Met Leu Asp Lys Gln Gln Met Asn Tyr Gly Trp Ile Glu Gly

50 55 60

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

65 70 75 80

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

85 90 95

Ala Leu Tyr Ala Leu Ile Asn Pro Gly Asp His Val Ile Ala Glu Tyr

100 105 110

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

115 120 125

Val Asp Tyr Trp His Ile His Glu Glu Asp Asn Trp Tyr Pro Arg Ile

130 135 140

Asp Asp Leu Lys Ala Met Val Lys Pro Asn Thr Lys Met Ile Cys Leu

145 150 155 160

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

165 170 175

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

180 185 190

Asp Glu Val Tyr Leu Pro Leu Asp His Pro Glu Lys Phe Ala Gln Ile

195 200 205

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

210 215 220

Tyr Ser Val Pro Gly Val Arg Ile Gly Trp Thr Ala Thr Asn Ala Glu

225 230 235 240

Val Ala Asp Ile Phe Arg Lys Phe Arg Asp Tyr Thr Met Ile Cys Gly

245 250 255

Gly Val Phe Asn Asp Gln Leu Ala Thr Tyr Val Leu Arg His Arg Asp

260 265 270

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

275 280 285

Tyr Lys Asp Trp Ile Asp His Glu Asp Arg Ala Ser Val Ile Met Pro

290 295 300

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

305 310 315 320

His Thr Phe Cys Glu Asn Leu Leu His Asp Glu Gly Val Leu Leu Val

325 330 335

Pro Gly Asp Ala Phe Asp Thr Pro Gly His Val Arg Leu Gly Tyr Cys

340 345 350

Ala Pro Glu Ala Thr Leu Lys Glu Gly Leu Lys Arg Leu Ser Lys Tyr

355 360 365

Met His Gln Tyr Asp

370

<210> 6

<211> 178

<212> PRT

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 6

Met Ile Leu Thr Thr Phe Ile Ile Leu Ile Leu Met Gly Cys Phe Ile

1 5 10 15

Asn Gly His Arg Arg Gly Leu Leu Thr Met Thr Leu Met Leu Gly Thr

20 25 30

Tyr Ile Val Ala Trp Ile Val Ala Arg Gln Gly Ala Gln Leu Ile Gly

35 40 45

Gly Trp Leu Lys Ser Leu Leu Pro Ser Ile Gly Thr Pro Ala Thr Phe

50 55 60

Ser Glu Ser Leu Leu Ala Asn Val Asn Ser Asn Leu Phe Phe Tyr Asn

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Gly Ile Ala Phe Met Ile Ile Phe Thr Ile Val Ser Ile Leu Cys His

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Trp Gly Ile Arg Gln Leu Asn Trp Ile Lys Arg Ile Pro Val Val Gly

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

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Leu Ile Ile Tyr Val Val Leu Leu Ile Met Gln Leu Phe Pro Ala Gly

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Trp Trp Gln Met Gln Ile Ala Asn Ser Glu Leu Ala Arg Phe Met Ile

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Asn Gln Thr Pro Gly Ile Ala His Leu Val Ile Asp Thr Leu Val Gln

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Gly Gly

<210> 7

<211> 4653

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 7

ttggcaatct ttcacctatc agcaaaaatc attagtcgag gaaaagggca atcagcaatt 60

gcctcggctg cttatcgttc agggaacaaa cttcacgatg aacgatacga cgaaacacaa 120

gattacacaa acaaacgttt catcgaacac tcagaaatcc aacttccaga gaatgcccca 180

gctaagtatc aagatagggc aaccctttgg aacagcgtag aaaaggcaga aaaagctaag 240

aattcgcagt tagctagaga aattgaaata gccttaccac gagagctaac accagaacaa 300

cgagttaaat tagtccacga ttacgttcag aaaacctttg tcgataaggg aatggtagcc 360

gattggtcta ttcacaaccc acaaccagat aaggataatc cagaaaagcc agcaaacccg 420

cacgctcata tcatgctcac actgcgtagc ttgcgttcta acggctcttg ggcaccaaag 480

aagacaagtc actatgaact agacgaaaac ggccagaagg tgcctgtaat cgatcctgag 540

acaggcaaac agaagttagg ggcgaaaaac caaaaaatct ggaaacgagt aattacccca 600

actaatgact ggaataaccc caaaaacgtt gaaaaatggc gggctgaatg ggcgaagact 660

tgtaataagt acttggctcc tgaccaccag attgaccacc gcagttataa acggcagggc 720

aaaaaacaaa ttccaacaat ccatgagggc tatgttgctc gtaaaatgga acgagaagcc 780

acaggaagct cagagagggc ttcttttaac caaatcgtaa aatatatcaa caacgagtta 840

aaatcgctta gaaagcaaat taggggcatt atacgcgaaa tacggaatct agaaaaagga 900

cgtgaccaag atgagaaagt tcgacttcag caagacaccc gagcaaatca aacaagaaca 960

agcagagatc aagcgctggg aggaggagtt caacgcacga ttacagacaa tcgggagcaa 1020

tccggaaacg gtagagccaa cggaactgcc gaagaagccc aacgaacaga tcctaatcaa 1080

cttcatcaac agtctttcgg agaaacagca agaagaattt tccgagattt ccggctattt 1140

gcggagcgtc aacgagaagc tcgacagcga caacgccgac ttagccaagg cttacaacat 1200

catcaagaaa cagaaaaacg acttgaagca gggcaacaac aacttactcg aacaaaacat 1260

acagctgacg aacgaaaaca acagcttaca gaacgacaaa ttcgagttag aaaagctttt 1320

agcgacgctt ctaagtccaa agtcttggaa cgatactaca ctcacttccg tcaaacaaaa 1380

actcaagcaa atcatgcaaa aggattagac taccaaggac caattctggg acattcccga 1440

ggacgctcaa ggtagttatg ctagaatgat taagccgtcc gtggcaactc ctacgtaaag 1500

gaggtggcaa catggcacat atactcctct cagcagtttt gagccttctg ggttccttga 1560

taactgcact gttcgctgat tggcttcgtc ggcgaaaata gtttttgacg gcatgaagcg 1620

caaattaaaa gccactcaac tattcgcact agttgggtgg cttttttgca acgtggcaat 1680

atacttctct cggttaaatt ataacatggt tgttccagaa gtgcattagg taggcggttt 1740

gtcccccccc atgggttttg gcttcgaaaa tcaatcgttg aagtgaccag cgaagctcag 1800

ctgatggagc cctcatattc aagaagcagg tcaacgaatg ccaattccaa catgagggcc 1860

atgacttagg ccactgatac tcgcaaatga ctgtcacacc aacggtatta gccattctga 1920

taagcgagtc taacgacttt tgacgcataa tcgaccacct accttatttt caaagcgcct 1980

cagagctaat tagagtattc ttcaaggagc ttagcgagga ttatagacgc ttggcggact 2040

tcactttcgg cttgattgac ccgtttagcg atcttgtcat catggcgagc caccttttcc 2100

aagtcatgga gattatcagc gatttggtca gtcatgacgg agtagttctt aatacccgac 2160

agcaggcttt caacgttagt attctcaagg ttttcgttga atatcatggt tattcctcct 2220

tatcctcaaa cttagtcatc ttctgtctcc tgttttagag aattttcaat catgtctaat 2280

aatgttgaca tgtcaatgtt aaatgaatca aggctaccat accgcttgat ttgagtttta 2340

acactatcca atccatattc ttcaagtaat gcttttaatt taccagtaac tttttcacct 2400

aaatgcccaa aatactgaag ccagctctta agtaattctc ggtactggtc aaatttttct 2460

ttttcagtaa gctgtttctc aacttcttct tggtcctcag aaccgttaac tactttccag 2520

aggtcagggt cagtagtacc caacttataa ccaagtaccc gatcatatgc tctggccttt 2580

tcctttggag tcaaatcagg attaaggtca atattatcta gcttcatacg ttggtcagca 2640

taacgacctt tttcaaagtc atcggcattc ttggcttcag gttgccaagt gaactcatag 2700

ccaacaacag gacgtcctct accagctcct cgaatagttt ttacggttaa gccacgaata 2760

acaggggtta gctcttcttt aattggacgt aacactcttt ttcttacgtt accctggtct 2820

ttcttgtagc tcttgggtag atctagttgc ttaaataatt cgtcactggt taacttcaac 2880

cgccctactg ttcgatattg tttaattaac actctttttc ttacgttacc ctggtctttc 2940

ttgtagctct tgggtagatc tagttgctta aataattcgt cactggttaa cttcaaccgc 3000

cctactgttc gatattgttt aatcaagcga aacatgttct tagcataagc agattgaaga 3060

ctattaaact gagctagttg aaatcttgtc caatgactta aatcattgaa tagcttctga 3120

aatagagggt taacttgaac agtcagaatc tgtttactcc tctgaatctg aaatacattc 3180

caacaagcaa attgagtaat ggtgtctcca tcatcggtgt aggcattaat ctctaatagc 3240

tttttattag ttttcatcaa atcttttaca aaattagatg tactccgttc atacttacta 3300

agactttgta gttgcgtaaa tgaaaacttt aatttctgag tacccttttg atacgactgc 3360

tgaactagtg taataaatag atttagctca ttagagttca gtcggccaag tggaatagta 3420

ttaagccgat tactatactt cacaatttca ttactcaatt ttctcacctc aaatatatta 3480

taacatttta tcgtacacaa tcaagagtac gtacttatag cacacgtaca gtattagcgc 3540

gtatattaag acaaaaattt agacaaaatg taccttataa cggacaaaat gtaccttata 3600

aaccggacaa aatgtacctt ataacggaca aaatgtacct tataaaaaac gaaactctta 3660

ctctcccaag gagtttcaaa cccctaaaga gtatttaaag aatatatata aagatattta 3720

aagaggcacc atacggaaaa tctcctttct gatttcaatt tcagagagga gattatcact 3780

aaatttcaaa attcaatttt tcgccaaaaa cttttttagt agttttcggt aactagcaaa 3840

atcaacttcg ttgattattt tgactatttc gaaattcatc aaaaactgaa tgcccgtctt 3900

tatagtgaat ttgtttagtg aagttcacta actgcttcag catattcagt tgaccacgta 3960

accattgttc actatcagca ggactaattt gttctgataa ttgctctttt tggtggttat 4020

aagccttata agcctgatat ttagcataac cttgtgggtc atctgattta gccactctag 4080

ccactttagg tacttttcta agccccttag cttgagcttg aatatcttct aaaaccgccc 4140

ctttttcgat gagacgctta tttcgttgct ttctcttttc agtattgatg cgagccattt 4200

gacgctctcg acgagcctta aacactgctt ccttttcttg aaggcgttca agttctgctt 4260

tcattttttc aatatcagcc ataacaaatc ctcctaatgt gaataatgtt ttctgaaatt 4320

caaatgttat ctagattata tgatgttaca attgatttaa ccactacgtt tggtctccct 4380

gttaagggcg cacttataca cattcaaaga atgtaattac gcctgacggc gacgtgcgct 4440

ctccgagggg acttacccag tcggggcgac ttcatcgaga ccatgtcatg gagattcagt 4500

ttcaccgaaa cggaccacta accccgtgga ggggagaagt ttcacttcta gggaccattc 4560

catggagggt cgctttttac aaaagcaacg caaaacaaaa gactagctcc caagcttacg 4620

caaggtcgtt agtccctaag gaggcagcac aga 4653

<210> 8

<211> 1026

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 8

ttagtcatct tctgtctcct gttttagaga attttcaatc atgtctaata atgttgacat 60

gtcaatgtta aatgaatcaa ggctaccata ccgcttgatt tgagttttaa cactatccaa 120

tccatattct tcaagtaatg cttttaattt accagtaact ttttcaccta aatgcccaaa 180

atactgaagc cagctcttaa gtaattctcg gtactggtca aatttttctt tttcagtaag 240

ctgtttctca acttcttctt ggtcctcaga accgttaact actttccaga ggtcagggtc 300

agtagtaccc aacttataac caagtacccg atcatatgct ctggcctttt cctttggagt 360

caaatcagga ttaaggtcaa tattatctag cttcatacgt tggtcagcat aacgaccttt 420

ttcaaagtca tcggcattct tggcttcagg ttgccaagtg aactcatagc caacaacagg 480

acgtcctcta ccagctcctc gaatagtttt tacggttaag ccacgaataa caggggttag 540

ctcttcttta attggacgta acactctttt tcttacgtta ccctggtctt tcttgtagct 600

cttgggtaga tctagttgct taaataattc gtcactggtt aacttcaacc gccctactgt 660

tcgatattgt ttaattaaca ctctttttct tacgttaccc tggtctttct tgtagctctt 720

gggtagatct agttgcttaa ataattcgtc actggttaac ttcaaccgcc ctactgttcg 780

atattgttta atcaagcgaa acatgttctt agcataagca gattgaagac tattaaactg 840

agctagttga aatcttgtcc aatgacttaa atcattgaat agcttctgaa atagagggtt 900

aacttgaaca gtcagaatct gtttactcct ctgaatctga aatacattcc aacaagcaaa 960

ttgagtaatg gtgtctccat catcggtgta ggcattaatc tctaatagct ttttattagt 1020

tttcat 1026

<210> 9

<211> 332

<212> PRT

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 9

Met Asn Glu Tyr Asn Ala Glu Met Ala Lys Leu Asn Gln Gly Ala Asn

1 5 10 15

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

20 25 30

Pro Glu Asn Asn Ala Thr Val Asp Ile Glu Ala Leu Asn Pro Arg Ile

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

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

115 120 125

Asn Phe Leu Val Val Phe Glu Asp Pro Val Arg Gly Asp Met His Ser

130 135 140

Thr Thr Val Thr Ala Thr Tyr Gln Tyr Tyr Asp Ala Asn Gly Asn Leu

145 150 155 160

Ile Asp Phe Ser Gly Thr Asn Asn Ala Trp Leu Ser Val Gly Ser Leu

165 170 175

Asn Phe Asp Gln Gly Asn Asp Tyr Gln Gly Gly Lys Asn Glu Gly Asn

180 185 190

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

195 200 205

Ile Lys Gln Leu Ala Gly Ser Ser Ile Ser Val His Asp Asp Gly Trp

210 215 220

Ala Tyr Ala Gly Phe Asn Asn Tyr Ser Gly Thr Gly Met Asn Asn Gly

225 230 235 240

Ile Asn Thr Asp Asn Gly Gly Ser Gly Trp Asp Met Asp Gly Ser Pro

245 250 255

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

260 265 270

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

275 280 285

Gln Tyr Asn Asn Gln Phe Leu Asn Asn Ala Trp Phe Thr Ala Gly Thr

290 295 300

Thr Leu Pro Glu Thr Gln Ile Lys Gln Pro Ile Arg Lys Thr Ser Glu

305 310 315 320

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

325 330

<210> 10

<211> 2059

<212> PRT

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 10

Met Ala Gln Lys Leu Met Ser Ala Asn Ser Thr Asp Lys Asn Phe Lys

1 5 10 15

Met Tyr Lys Ser Lys Lys Ser Trp Val Phe Ala Tyr Ser Thr Thr Leu

20 25 30

Ala Leu Ala Ala Val Ala Gly Ile Thr Leu Ser Thr Thr Asn Val His

35 40 45

Ala Asp Thr Thr Asn Gly Gly Asp Asn Gln Val Asn Ala Thr Ala Val

50 55 60

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

65 70 75 80

Ala Gln Thr Asp Asn Thr Ser Thr Ser Ile Asn Ile Arg Ser Leu Met

85 90 95

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

100 105 110

Glu Gln Ser Gln Asn Tyr Ala Ser Ser Asn Gln Asn Ser Gln Thr Gln

115 120 125

Gln Glu Asn Gly Thr Thr Gly Gln Ser Thr Ala Ser Gln Asn Gly Thr

130 135 140

Thr Ser Asp Gln Thr Asn Ser Asp Gln Ser Asp Lys Asn Tyr Tyr Val

145 150 155 160

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

165 170 175

Gln Lys Asn Tyr Thr Ser Ile Lys Gly Gln Glu Val Ala Asp Gly Thr

180 185 190

Val Val Thr Trp Pro Leu Ser Val Ser Ala Leu Pro Ala Asn Arg Ala

195 200 205

Gln Asp Leu Lys Ser His Val Ile Ser Glu Thr Leu Asp Pro His Leu

210 215 220

Glu Tyr Leu His Tyr Arg Ala Tyr Leu Thr Asn Thr Asp Gly Thr Val

225 230 235 240

Thr Asp Val Thr Asn His Val Asn Leu Asn Arg Ser Gly Gln Thr Leu

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

Asn Asp Gly Ser His Asp Val Ser Phe Thr Thr Thr Ser Asn Asn Val

305 310 315 320

Gln Ile Lys Thr Phe Asn Pro Gly Asn Ser Lys Asp Val Glu Ile Gly

325 330 335

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

340 345 350

Ala Asp Gly Ser Val Val Thr Trp Pro Met Ser Val Gly Asp Leu Pro

355 360 365

Ala Asn Arg Ala Gln Asp Val Leu Ser His Ile Glu Thr Asp Thr Leu

370 375 380

Tyr Asn Gly Leu Asn Tyr Glu Gly Tyr His Ala Tyr Leu Pro Gln Ala

385 390 395 400

Asp Gly Ser Phe Gln Asp Val Ser Ser His Ile Asn Val Gln Gln Asn

405 410 415

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

420 425 430

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

435 440 445

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

450 455 460

Gln Leu Ala Phe Lys Asp Gly Asn Gly Gln Thr Val Ile Asn Asn Thr

465 470 475 480

Ser Asn Gln Val Gln Ile Ser Thr Tyr His Pro Thr Asn Thr Lys Asp

485 490 495

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

500 505 510

Asp Lys Val Val Ala Asn Gly Ala Ile Val Thr Trp Pro Met Ala Ser

515 520 525

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

530 535 540

Ser Glu Thr Leu Asp Ser His Leu Gln Tyr Gln Gly Tyr Lys Ala Trp

545 550 555 560

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

565 570 575

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

580 585 590

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

595 600 605

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

610 615 620

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

625 630 635 640

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

645 650 655

Thr Asn Lys Asp Val Glu Leu Gly Asp Asn Ile His Gly Asp Thr Glu

660 665 670

Ser Ser Ile Ala Gly Lys Leu Val Ser Glu Gly Thr Ile Val Thr Trp

675 680 685

Pro Leu Ser Thr Ser Asp Leu Pro Ala Asn Arg Ala Gln Asp Val Val

690 695 700

Ser His Thr Ala Val Asp Ala Leu Glu Pro Thr Leu Gln Tyr Ile Ser

705 710 715 720

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

725 730 735

Ser His Val Lys Met Thr Arg Asp Gly Gln Lys Leu Thr Phe Thr Asp

740 745 750

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

755 760 765

Lys Met Pro Ile Ile Asp Leu Val Thr Lys Ala Thr Gly Asn Thr Lys

770 775 780

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

785 790 795 800

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

805 810 815

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

820 825 830

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

835 840 845

Thr Val Val Thr Trp Pro Met Ser Thr Ser Asp Leu Pro Ala Asn Arg

850 855 860

Thr Gln Asp Val Val Ser His Ser Thr Ser Glu Thr Leu Asn Gln Asn

865 870 875 880

Leu Gln Tyr Val Gly Tyr His Ala Tyr Met Pro Asp Ala Asn Gly Lys

885 890 895

Leu Gln Asp Val Thr Ser His Val Gln Leu Gln Gln Asn Gly Gln Asn

900 905 910

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

915 920 925

Lys Ser Ile Ala Phe Lys Met Pro Ile Ile Asp Leu Met Thr Lys Ala

930 935 940

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

945 950 955 960

Phe Asn Asp Gly Asn Gly Asn Thr Thr Phe Lys Ser Thr Ser Asn Thr

965 970 975

Val Gln Ile Ile Thr Tyr Lys Pro Lys Thr Thr Lys Asp Val Glu Leu

980 985 990

Gly Asp Asn Ile His Gly Asp Thr Asn Ala Ser Ile Ala Gly Gln Met

995 1000 1005

Ile Thr Asp Gly Thr Val Val Thr Trp Pro Met Ser Thr Ser Asp

1010 1015 1020

Leu Pro Ala Asn Arg Thr Gln Asp Leu Gln Gln His Val Val Thr

1025 1030 1035

Asp Asn Leu Asn Asp Asn Leu Ile Phe Gln Gly Tyr Thr Ala Trp

1040 1045 1050

Leu Pro Thr Ala Asn Gly Leu Val Asp Val Thr Asn His Ile Glu

1055 1060 1065

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

1070 1075 1080

Leu Leu Asn Leu Tyr Asn Gln Asn Lys Asp Thr Ala Tyr Lys Leu

1085 1090 1095

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

1100 1105 1110

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

1115 1120 1125

Gln Gln Thr Thr Val Asn Val Thr Ser Asn Thr Val Asn Ile Ser

1130 1135 1140

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

1145 1150 1155

Ile Glu Gly Asp Thr Ala Asp Thr Ile Asn Asn Leu Met Val Gln

1160 1165 1170

Ile Gly Thr Lys Met Thr Tyr Pro Leu Thr Val Ser Asp Leu Pro

1175 1180 1185

Ala Asn Arg Ala Asp Glu Ile Thr Ala His Gln Ser Val Asp Thr

1190 1195 1200

Leu Ser Asp Tyr Leu Glu Tyr Gln Gly Tyr Lys Ala Tyr Leu Pro

1205 1210 1215

Asp Ala Asp Gly Lys Leu Gln Asp Ile Thr Glu His Val Asn Leu

1220 1225 1230

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

1235 1240 1245

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

1250 1255 1260

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

1265 1270 1275

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

1280 1285 1290

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

1295 1300 1305

Ser Asn Asp Pro Glu Ala Val Lys Asp Val Glu Leu Gly Asp Asn

1310 1315 1320

Val Val Gly Asp Thr Pro Asn Ser Val Thr Gly Thr Thr Val Ala

1325 1330 1335

Asp Gly Thr Ile Val Thr Trp Pro Met Ser Val Gly Ser Leu Gly

1340 1345 1350

Ala Asn Arg Ala Gln Asn Val Ile Lys His Thr Glu Thr Glu Asn

1355 1360 1365

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

1370 1375 1380

Asp Ala Asp Gly Lys Met Gln Asp Ile Thr Glu His Ile Asn Ile

1385 1390 1395

Gln Gln Asp Gly Gln Lys Leu Val Phe Thr Asp Asp Asp Tyr Leu

1400 1405 1410

Ile Ser Leu Tyr Asn Lys Asp Lys Ser Gln Arg Phe Ala Leu Pro

1415 1420 1425

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

1430 1435 1440

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

1445 1450 1455

Asn Thr Ile Thr Ser Val Thr Ser Asn Gln Val Asn Val Ser Thr

1460 1465 1470

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

1475 1480 1485

Glu Gly Asp Asp Ala Glu Asn Ala Asp Gly Thr Val Val Ala Gln

1490 1495 1500

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

1505 1510 1515

Ala Asn Arg Ser Gln Asp Val Lys Ser His Thr Leu Val Asp Lys

1520 1525 1530

Leu Asp Asp Asn Leu Gln Tyr Asn Ser Tyr Lys Ala Tyr Leu Lys

1535 1540 1545

Gly Thr Asp Gly Lys Leu Gln Asp Val Thr Asp His Ile Lys Leu

1550 1555 1560

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

1565 1570 1575

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

1580 1585 1590

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

1595 1600 1605

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

1610 1615 1620

Asp Thr Ser Met Lys Thr Thr Ser Asn Glu Val Ser Ile Ser Thr

1625 1630 1635

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

1640 1645 1650

Val Gly Asp Thr Ser Asp Ser Ile Ala Asn Glu Thr Val Pro Asp

1655 1660 1665

Gly Thr Ile Val Thr Trp Pro Leu Ser Val Ser Ser Leu Pro Ala

1670 1675 1680

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

1685 1690 1695

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

1700 1705 1710

Ala Ala Gly Asn Leu Gln Glu Val Thr Asp His Val Lys Leu Ala

1715 1720 1725

Gln Glu Gly Gln His Leu Thr Phe Thr Asp Asp Asp Tyr Leu Ile

1730 1735 1740

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

1745 1750 1755

Ile Asp Leu Val Thr Thr Val His Gly Asp Ser Lys Leu Ile Pro

1760 1765 1770

Asn Glu Phe Asp Asn Val Phe Val Phe Lys Asp Gly Lys Gly Gln

1775 1780 1785

Thr Thr Val Lys Thr Thr Ser Asn Lys Val Thr Ile Lys Thr Ala

1790 1795 1800

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

1805 1810 1815

Asn Ile Asn Gly Asn Glu Val Lys Ala Gly Glu His Val Asn Tyr

1820 1825 1830

Thr Leu Asn Trp Asp Leu Ser Asn Asp Lys Asp Val Lys Ala Thr

1835 1840 1845

Pro Glu Met Ile Lys Lys Gly Phe Phe Phe Ile Asp Pro Ile Asp

1850 1855 1860

Ser Arg Ala Leu Ser Val Asp Asp Leu Ser Lys Ala Lys Val Val

1865 1870 1875

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

1880 1885 1890

Asn Ser Leu Ser Glu Val Pro Glu Phe Ile Gln Glu Gln Val Lys

1895 1900 1905

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

1910 1915 1920

Gln Ala Asp Asp Leu Gln Ala Phe Phe Asp Lys Tyr Val Lys Thr

1925 1930 1935

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

1940 1945 1950

Phe Thr Gly Glu Phe Ser Asn Thr Ala Tyr Gln Phe Gly Phe Gly

1955 1960 1965

Lys Ala Thr Pro Thr Asn Thr Val Thr Asn Tyr Val Lys Pro Met

1970 1975 1980

His Lys Pro Ala Ser Pro Glu Thr Pro Ala Ala Ile Ala Pro Gln

1985 1990 1995

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

2000 2005 2010

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

2015 2020 2025

Asp Glu Gly Ala Leu Leu Gly Leu Ala Ala Val Ser Leu Val Gly

2030 2035 2040

Ser Leu Gly Leu Ala Ala Leu Gly Leu Lys Gln Asn Arg Asn Asp

2045 2050 2055

Asp

<210> 11

<211> 44932

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 11

ctattcaaat ttgaaatttt gaacaacttt atcaaggtct tcttgttcaa tccctagata 60

gtggcgcgta tttcgttcag aggagtgatt aaataactgc gaaatgattt caacgttaac 120

acctttctta tagagttgtc gaccaaaagt ttttctaaaa gagtgtgttc caattttagc 180

aacgatttta ttattttccg tttttctagc cattcgttgc aacatttcgt aaaagccatg 240

tactgaaaaa tgaccttctt gtttcccagg gaaaagataa tcattctcgt ctttataatt 300

caagtcatta agataatcaa taatttcggc aagacaatta ttccaaaaga gcgttttagc 360

cttacctgtt ttttgctcaa taacacgggt tttagtttta tttagaacat gaccaaccct 420

taacgttaca acgtctgatg ctcgcaatcc gttattaagg gcaattgcaa ttagtaatag 480

gtttcggtta accaattcag gacgcgaatg cagcttaatt gtctcacgta actgatcata 540

ttcttcgaga gtcaagaatt taacattttc tacccatatt ttccgtttac ccttgatttg 600

accattagtt ttatttttgg gaatataaac tcgtttttta gttgacataa tatttcttcc 660

tttgttaatt tgcgaggtaa tcaactaaag cagggacgat ttcctctcct tctttccgcc 720

cagtttcata aagtgctcga attttcttaa tatttccttc aatccgctta attttaattg 780

gtttagaggg agcaatatta aagatctttc cttcacaaga caaacgatta atttcagcag 840

tctgttggtt gtagagaagg ggacgatcaa tccccgcttt agcaaattcg ggataatctt 900

tatataccat gtcgtacaac ttctttacag cagaactaga tggcttcttt cggtagttaa 960

tatccctggt ccgtacaaca acaatcttgt catagccttg tcgttgtgca atatcaaatg 1020

gaattgaatc tgtgatccca ccatcaaggc aaggcccttg agaagtttcc tgtggatctg 1080

aaagaaatgg cattgatgag gaagctttga gggcattggt tagctcctca ccaacaggat 1140

ccgtaaataa aactgttttt cctgttttca cagaagtagc aactgctgtg aaatgagatg 1200

ctgaccgtcg atatgcagct tcattaaaat tctgccaaga atatccgtga tcttcaaaaa 1260

gataatcaag attaattatt tgctttttaa atacgcgagc catcgagata tattggcgat 1320

cgtgccgatg atcaatatta atattggcag cgcgtccgta ttgcttagag acgaaattaa 1380

ctccacaaag tgaaccagca gaaacgccaa ttacacttcg aaattcaata tggtgagcga 1440

gaaatgtatc aacaattccg gcagtatatt gtccacggaa tgcgccacct tctaaaacta 1500

atgctgcatt atataacata aaatgaaata actccttttt gtccgccaaa aatagcaaaa 1560

atgacctaat ttttagtgag acactttttc aataaaatta tacaacagtc tcacacgaaa 1620

atataggatt ttaaaagctt gttatatcaa cgtttattct cggttaccgt aagactcttt 1680

tatcaaaaga gtcttactga ttttgaaata acgcaaaagt aggggcaaaa acggattttg 1740

cctctcagac gaactgaaat tcattttgat gagtagtagg tcgttacaat tcgtctcaga 1800

agaaaaacgc atattgatat caaaaaatga gtaaatagac agtaaatatt taaatatgaa 1860

ctttttatga agaaactatt gataatttaa gataatcata ataaaatagt gcacagttaa 1920

ttgttaaacg tttagcaaaa aaggtaaaga aacggggtta tttctatgct agaacgcaag 1980

gaacataaga aaatgtataa aagtggtaaa aattgggcag ttgttacact ttctacggct 2040

gcactggtgt ttggtgtaac aactgtaaac gcatccgcag acacgaatac tgaaaacaat 2100

gattcttcta ctgtacaggt tacaacaggg gataatgata ttgctgttaa aagtgtgaca 2160

cttggtagtg gtcaggttag tgcagctagt gatgcgacta aaaattctgc taatgcaaat 2220

agtgcttctt ctgccgctaa tacacaaaat tctaacagtc aagtagcaag ttctgctgca 2280

acagcttcat ctacaagttc agcatcttca tcaactaaca cagatagtaa agcagctaca 2340

gaaaatacta atgtagccaa agaggatgat acacaaaaag ctgcacccgc taacgaatcc 2400

tctgaagcca aaaaagaacc agctgtaaac actaatgatt cttcagctgc caaaaacgat 2460

gatcaacaat ccagtaaaaa gaatactgcc gctaagttaa acaaggatgc tgaaaacgtt 2520

gtaaaaaagg cgtggattga tcctaatagt ttaactgatg accagattaa agcattacat 2580

taaataagat gaacttctcg aaagctgcaa agtctggtac acaaatgact tataatgacc 2640

ttaaaaagat tggtgaggct ttagttgatc aagatcctaa atacgctatt ccttatttta 2700

atgcaagtca aataaagaat atgcccgctg catatactag ggatgctgaa acaggtgaat 2760

atgctgattt agatatttgg gattcatggc cagttcaaga tccagtgact ggttatgttt 2820

ctaattggaa tggctatcaa cttgtgatcg ctatgatggg acggcctcat cacgaagata 2880

atcatattta tttactttat aataagtatg gtgataatga tttttcacac tggcgaaatg 2940

caggatctat atttggatat aatgaatctc cactaacaca agaatggtcc gggtctgcta 3000

ttgttaattc tgacaactct attcaactct tttatactat caatgataca aataatgcta 3060

ttaaccatca aaaattagca agtgctacta tgtacttaac agccgacaat gatggtgtcc 3120

atattaataa tgtagagaat aatcatgtgg tatttgcagg tgatggttat cattaccaaa 3180

cttatgatca atggaaagct gcaaatagtt ttgctgacaa ctacactttg cgggatggac 3240

atgttgtaca aatgccaaat ggtgatcggt atttagtatt tgaaggaaat actggaactg 3300

aaaattatca aggtgaagat caattatata attggtcaaa ttatggtggt aacgatcgct 3360

ttaatattga aagcttgttt catcttttaa gtagtgatgt tgattataaa aaagctatct 3420

ttgctaacgg ggcacttgga attattaaac taacgaatga tgaaaagaac ccgcaagtag 3480

aagaagtata tactccatta gttacatcaa atatggttag tgatgaactt gaacgtccta 3540

atgttgttaa acttggagat aagtattatc tattctcggc tacacgttta agcagaggaa 3600

ctaatatcga tactcttaat aaagctaata aagttgtcgg agataatgtt gcaatgattg 3660

gttacgtagc cgatagtctt acaggtccat ataaaccatt aaatggttca ggggttgtag 3720

tgacagcttc tgttcctgcc aattggcgta ccgctactta ttcttactat gctgttccag 3780

tagaaggaaa agaaaatcaa ttactcatta cttcgtatat gacaaataga ggcgaagttg 3840

ctggaaaagg tatgaattca acatgggcac caagttttat tgtgcaaatt aatcctgatg 3900

atacaactat ggtattagct aaagtaacta accaagggga ttggatttgg gacgaatcaa 3960

gtaacaataa taatatgtta ggaaacattc aaacagctgc tttacctggg gaatttggta 4020

aaccaattga ttgggattta attggtggtt atggattaaa accgcatgat cctgctacac 4080

caaatgatcc tgaaacgcca actacaccag aaacccctga gacacctaat actcccgaaa 4140

caccaaagac tcctgaaaat cctgggacac ctcaaactcc cgatacacct aatactccgg 4200

aagttccttt aactccagaa acgcctaagc aacctgaaac ccaaactaat aatcgtttac 4260

cacaaactgg aaataacgcc aataaagcca tgattggcct aggtatggga acattgctta 4320

gtatgtttgg tcttgcagga attaacaaac gtcgagttaa ctaaatactt taaaataaaa 4380

ccgctaagcc ttaaattcag cttaacggtt ttttattttg aaagttttta ttatcgaaaa 4440

aaacaaatcc tcgttaatcc tttaatgcaa ttgttgtaaa accttgcgac agtaataaca 4500

gtggatttgc ccatctttgt cagctaactt ccgtgcatgc attgcagaaa aagtatagtg 4560

ctcatgacaa aatggacaaa catattgctt tttcccaaat agtgaggtaa taaagcccaa 4620

aattttctcc tccataaaaa aattatacac cccttaacat tatagcgaag attatcttta 4680

tagaaaaggc ttttattcat ttgttttttt atatgaagtt tcactaattt ctttaagagc 4740

atcacgctgc tgtttctggt caacgtgagt atataaatcg gtagcagatg ttcctttttg 4800

tcccaactgt tgggccacta atacttgatc tttcgttacc tcatacattt cagaagcaag 4860

ggtgtgccgt aatttatggg gagtaagagg atggccaaat gcggttgaat actttttgac 4920

cattttttca attgcattag ctgtcatccg tcgagtttgc ttgtgataaa cggttaggaa 4980

gaaagcagta tcctttttta aagcgtggta tcgttgtgca cgaatcgctt ggtaagtttg 5040

gatataagta atagcccaag gagcaattgg taccgaatcc ctctggccac cttttcttgt 5100

tacatctagc agcgattgtt ttaagtttaa atcaccaagg tttacatttg ccgcctcgga 5160

tacccggacg cctgttccta aaataagagc aataattgca atatctcgtt ccttattgac 5220

cttataagag gggagagctc ttttatcaca tttattaggg tattcttttt caatgaaagt 5280

aataaaatca aacttcattt gtccccggta catgtgtgaa gccagcgtat gagcccgata 5340

atttaatgtt tttgtatcat taagggagtc aattttcagc attacattac ggtcaaaata 5400

agattcacca ttattgttat ctgcggtaac tgttaaaaat ttatacaatg accttaatgc 5460

attaatagat cgattgattg aagtaggaga atttaaccgt ccttgtgcat tggtagtatg 5520

ctgcaaataa tcgatataca acattacatc gctacgccgt agattcgcca gtgtatcaat 5580

tggtaagtcc ttattagaat taacagagac gagtccactt gaccgtaacc aatcaaagaa 5640

tcgacggatt tcagttaagt actgataggt tgttgtgact gcatggttgg tccctaaata 5700

atactcgtta acataatcag gaaggttctg aagctcctct tgaattagct ttaaatattt 5760

atctgcttcc atttttatat cctccttgat actaaaaaga gtcgcttgaa agaaaacttt 5820

caacgactct ctaaacctat cgggaaaaca ggattcgaac ctgcgacccc ctggtcccaa 5880

accaggtgct ctaccaagct gagctatttc ccgttaataa cgaacaaata ttattctacc 5940

aattctcaat taattagtaa aggaatattt taatttaaag tgattaatag ttaatgatta 6000

aatgacataa tagtaaaatt cctattaatt gcaaaaaaat catgattaac cgaaatcctt 6060

ttctagcatc taatgaagat gataaataat tttattttcg actagttata aaagatacct 6120

ttccgaaaat aatttgacat tcaaaatact tttgaatata atttgattat cgaatatttt 6180

gatactcgaa atattttccg aaggcaggtg aatctttttg gcaataatga atgcgcagga 6240

gattatggaa ttaattccta atcgatatcc gatttgctat atcgattatg tagataaatt 6300

agttcctgaa gaaaagatta ccgcaacaaa aaatgtcaca attaatgaat cattttttcg 6360

cggtcatttt cctaataatc ctgttatgcc tggagtttta attatcgaaa cattggccca 6420

agctgcttca attttgatat taaaatcacc gcatttttat aagaaaacag cttatcttgg 6480

cgcaattcat aaagcaaggt ttcgacaaat ggtccgtcct ggtgatgtat taaaactaaa 6540

cgttgttatg aaaaaagttc gatcatcaat ggggattgta gaaacacaag cgcttgtgaa 6600

cggcaaatta gcttgtagtg cggagcttgt ctttatcgtt gctgaacgag aagaaaagat 6660

ttagcacggt gaatcattat catttataat atattttgat aatcaaatta ttcgggctat 6720

agaataaaat tgaagggaag aaatatatta tgagtaaaga taatgattac gaaaaaatta 6780

ataaaggatt aatcaaagtt tattccggaa ttctatggat tgaagaaaat gaattgcgga 6840

aaagtacatt caatgattta acaattaaag agatgcatgc aattgatgca attacgatgt 6900

ataaccatca aacaatttct caagtagcag aaaagcttca tctaactcca ggaacaatga 6960

cttcaatggc tgatcgttta attcgaaaag gatatgtgga aagaattcgt gataaagatg 7020

atcgccgcat tgttcggtta tgtttaacca aaagaggccg ggtactatat cgtgcgcacc 7080

gggcctttca taacatgatg gttgaacgtt ttcttaaagg aatggatgat gaagaaatga 7140

aggttgttaa aaaagccttg caaaacttag aagattttgt ggatgagcat gcttagagat 7200

aaggattgac aaacattgca aaatttaaga attactagta ctgcgagtta tcatccaccc 7260

cttaaaatta ctaaccagca attatcaact attatgaata cttcagatga gtggattaag 7320

acgcggactg gaattcatca acgttatatc agcaacactg aaaatacttc agacttagct 7380

gtcaatgttg gtacccagtt attgactaat gccaatttaa aggcaactga acttgattta 7440

atcattattg caacgatgtc tcctgatgcc tatactcctt caactgctgc tattgttcaa 7500

ggaagattag gtgcgaaaaa tgcaattgcg tttgatatct cagcagcttg tactggtttt 7560

atctatgcca ttaatacagc tgaattaatg ttgaaaagct ctcattggaa aaatgcaatg 7620

gtaattggtg cagaagtatt atcaaaactg attgattggc aagatcgaag tactgccgta 7680

ttgttcggcg atgggctggc ggagtgttac ttcaaaagac aactacaaca acccctttaa 7740

ttctcggccg tgatctccat acatttggtg acttaggaga taaaattatt gccggaaaaa 7800

caacgcctaa aactggcttc cctaaacaac taacatccct ttcaccattt acgatggccg 7860

gccgtgacgt ataccgcttt gccactcatg aagtaccacg atcaatcact tctgccgttc 7920

aacaagctaa tttgaaatta gacgatatcg attatttttt attacatcaa gcaaatgaac 7980

ggataattac ccaaattgca aagagactgg cgcaaccaat tacaaagttt ccaatgaata 8040

ttagtgaata tgggaataca gctgccgcta gtgaaccaat tttattgact caagctattg 8100

ctcatgaatt gattaaacca ggtaacatta ttgcaatgag tggctttggt ggcgggttaa 8160

gtacaggaac aataatttta aactattaat agagaaagaa gaatggatat gactaaagag 8220

gaaattttta atactgtaaa aactattact gttgatgaat tagatgttga cgaaaatcgt 8280

gtaacaatgg atgcacgaat caaagatgat cttgatgctg atagtcttga tgtttttgaa 8340

attatgaatg agcttgaaga taagtttgag attgaattag atgccgatga aggaatcgag 8400

actattagtg acgttgttga tttcgtaaag aaacagttgg atgaaaaata atgtactacg 8460

gaatattatt tagcggtcaa ggtgcacagc gatcgggaat gggagtcgag cttatggccg 8520

actccctttt ttcaaggatt gttagtcagg caagcgctgt ttgtgaactt gatttgctca 8580

aaattatgaa aaacgagcat aaggagttga ataaaacagc atatgttcaa ccagcaattg 8640

taacagttag ttatggaatc taccgaatgt taaaacggga tttacctcaa ctaccaatta 8700

aagggatgat tggcctatct ttaggcgagt acgctgcttt aattgctagt aatgcacttt 8760

catttgaaga aggaattaaa ttagttgcgg accgtgctca ctttatgcaa caagacgcgg 8820

atagagaaat aagtacatta gcagctgtcc ttgatcctca acttcaagag ataaaagaac 8880

taatcaccgc tcaacaagaa aatggtcagc gagtttatat tgctaactat aattcaccac 8940

gacaaatcgt agtgggcggg gcattaaacg ctttaaaggc tacccttaaa aagattgaag 9000

aggacaagct tgctaaaaga acgatcctgc tcaaagttaa tggtgcattt catactccct 9060

tctttaacgg tgcacgtcaa cagatgcata accggttaca agcagtcgac ttccatgagc 9120

cacagattga agttattagc aatactacta acagcttgtt tcattgtgag gatcttccag 9180

gaattcttga aaaacaatta gctgttccaa cacactttgg agctaatgtt aaggaattag 9240

tcaagcacgc gaaaattgac acaatattgg aaattggtcc tggaaagacg ttatctcgct 9300

tcgctcacca agttgaccag cacttaaata cccaacacat tgaaaatctt gctgattatg 9360

aaaaatttat aaaggagcaa aaagatggaa ctgacagata aagtcgtttt tataagcgga 9420

agcacacgcg ggattggggc ggctactgca ttagagtttg ctaaggctgg tagtcggcta 9480

atcctcaatg ggcggcagga taacttacca aaaacgttta aagaaaagct agatctacta 9540

ggggcggatt atcactatct taagggcgat attgcaaatg aagaatcagt tagtgaatta 9600

gcagcagcgg cttggcaaat atacgagaag atcgacattc ttatcaataa cgcgggaatc 9660

acgaatgata agttaatgat gggaatgaaa gcgagcgatt ttgaccaggt catcaatgtt 9720

aatttacgcg gaacatttat gttaacgcaa cctattttta agaagatgct caaaaaaaga 9780

gtcggttgca ttatcaacct tgctagtatt gtgggtctcc atggtaatac gggacaagct 9840

aattatgcgg caagcaaggc aggtatcatc ggccttacta aatctattgc ccaagaagga 9900

gcacgccgtg gaattcgttg caatgcgatt gctcccggaa tgattactag tgatatgact 9960

gaaaaattat ctgagcgagt aaaagaacaa attctcagtc gcatccccct caaccgctta 10020

ggacagtcag aagaagttgc taagaccgca aaatttttag cagaaaacga ttatttgact 10080

ggtcaaacca ttgtagttga cggtggcatg acaatttagg aggaactaaa tgacaagagt 10140

tgtaataaca ggaatgggtg ctgttgctcc taatggtaac ggtattcaag aatttataag 10200

taatagtttt gcaggcaaag ttggaattaa agcgatcaag aaatttgatg ccaagtcgac 10260

aggaattacc gtggcaggtg aaattgacga ttttgaccct aatgatgtca ttggaaggaa 10320

agccgcgcgc cgaatggatc tttattctca atacgcctta caaagtgcga ttgaagcaat 10380

ggaaatggcg gagattaacg aaacaaatac caagccagtc gacatgggtg ttatctatgg 10440

atctggaatt ggcggtttga caactattca agagcaaatt atcaaaatgc atgataaggg 10500

tcctagacgg gtatcaccaa tgtttgttcc aatgtcaatt gctaacatgg cagccggtaa 10560

tatttccatt cactttaatg cgcaaaatat ttgtacatcg attgtgactg cttgtgccac 10620

tggaactaat gcaattggtg aagcctttcg tcaagttaaa gcaggtcgcg ctaaagtaat 10680

gatcgctggt ggatcagagg cttcggtgaa tgagatcggg attgctggtt ttgcggcatt 10740

aacagcatta tcacaagcaa ctgatccgct taaagcttct ttgccatttg ataaggcacg 10800

tcaaggattt gttttgggtg aaggcggtgc aacacttgtt ttagaggatc ttgagcatgc 10860

gcaaaaacgc agtgctaaca ttcttggtga gattgtcggt tatggtgcta cctcggacgc 10920

ttaccatatt acatcccctg atccaactgg tgcaggggcg gcaagagcaa tggaactggc 10980

aattaaagaa gctggaatta gtcctagcga aatttcctat attaatgccc acggaaccgc 11040

tactcatgct aatgatgaag gcgaatccaa agcaatcaat caggtatttg gttccgatag 11100

taatgttcgc gttagttcaa cgaagggaat gaccggccat ttgcttgggg ctgcgggcgc 11160

aattgaggca gtcttaacag tagccgcttt acaaaaggga caattaccgt tgaatatagg 11220

ttgctttaat caagatccaa aatgctcggt taaccttgtg acggcagaaa ataggaacgc 11280

ctcaaccgcc cgttacgcaa taagcaattc ttttggtttc ggtggtcata atgctgtttt 11340

agcctttaag aaatgggagt gatctatctt ggaatttaaa gaaattcaaa cattaatgca 11400

aaattttgaa gattctgata ttcgtgaatt agaaataaat caagattcct ttcagctcta 11460

tttaagcaaa aataagcaaa cccacaagca tgaaaatctt ataacaaccg aaaaaacaga 11520

gcaaacgact tcagctaaga aaaaagcaaa cgaacaacta actttacctt cgcaaaatat 11580

aactgcgccc ctagtcggaa cagtctatct ccaaccaacc cccgatgcag atccctatgt 11640

taaaagtggc gaccacgtaa aaaaggggga tgttgtttgt gtgattgaag caatgaaaat 11700

gatgacagag ataaaaagcc cttttaacgg aaccattact tcaatttgtg taagcaatga 11760

agaattagtt gaagtagaac aaccgctttt ctcagttcag gaggataaag acaatgccta 11820

ataaaacttt agatataact gaaattcaaa aaatccttcc gcatcgttac ccgatgttgc 11880

taattgacca agttgatgaa ttaatccctg gtaagaaggc cattacacac cgtaatgtca 11940

cgattaatga agaggttttt aatggtcatt tccccgaaaa tccagtttta ccaggagcat 12000

tgattgttga atcattggcg caaacaggtg ccgtcgctct cttatctcaa gaagagttcc 12060

aagggaaaac agcctatttt ggtggaattc ggtcagcaga atttcgtaag gtagtccgtc 12120

cgggcgatac gttaaggtta gaagtcagac tagaaaaagt tcacaaaaat attggaattg 12180

gtaaaggcat tgcaacggtc gatggcaaaa aagcctgtac agccgaatta acttttatga 12240

ttgggtaggt ggtttaatgt tttccaaagt actagtggct aatcgtggtg aaattgctgt 12300

caggataata cggtcattac gggagctagg aattaagaca gtagctatct attcaactgc 12360

agaccgcgaa agtcttcatg ttcaactagc ggatgaagct gtatgcgttg gaactgcccg 12420

ggcccaagat tcatatttga atgcgaaaaa cattttagaa gctgctcttg gtacaggtgc 12480

ccaggcaatc catcctggct ttggctttct atcagaaaat gcggaatttg cgacaatgtg 12540

tgaagaatgc ggaattacgt tcatcggtcc ccaagcctca gtgattgact taatgggaaa 12600

taaggagcat gcacgggagc aaatgaaaaa atcaggggtg cctgtgattc ctggaagcga 12660

tgactatatt accaatgtta atgacgctgt tgaggtcgca aacaagattg ggtatccaat 12720

tttgttaaaa gcagctgctg gtggtggcgg taaagggatc cgacgaatta acgatcataa 12780

ccagatgcgg caaatattta gcgaggccca aaacgaagcc cgactttcgt ttaatgatga 12840

ccgaatgtac cttgaaaaga ttatggagaa tgttaaacac attgaggtcc aagtatttcg 12900

tgataatttt ggcaatgccg ttttctttcc tgaacgtgac tgctcgattc aacggaataa 12960

acaaaaattg attgaagaaa gtccttgtgt cctagtaaat gaacaagagc gaaaaacgct 13020

aggacaaatt gcaatgcgag ccattaatgc gattaactac cataatacgg ggacaataga 13080

atttctaatg gacaaggacc atcactttta ctttatggaa atgaacactc gtatccaggt 13140

tgaacataca gtgacggaga tggtaactgg gatcgactta gtgaaggcac aggttattgt 13200

cgctgcgaat gaaccacttc cctttaccca acaggatatt caggttcatg gacatgcaat 13260

tgagtgtcgg ataaatgctg aaaatcctaa acaaaacttt atgccagtga ctgggacgat 13320

taattactta tatcttccag tcggtaattt ggggatgcgc attgacactg ctatttatcc 13380

tggcagtaag atcactcctt attatgattc aatgatcgct aaggtaattg cccttggtca 13440

agatcgccaa gaagctattg aaaaaatcaa acgactttta aatgaaatgg taattacggg 13500

cgtaacgacc aatcaaaatt tccatttagc aatcctaaac aatcctaaat ttttggcagg 13560

aacagcttca acaacgtttc ttgaagactt cttcttgcca caatggaaaa aggagctgac 13620

agcgtgaaat tatatgatca aaataatact ttaagtgaac ggcacatcaa agcagataaa 13680

aatgctgatg aaagggtccc agatcaaatg tggttaaggt gtccacattg tcatcaatta 13740

ctattcgcca agcagttaac acaatatgct gtttgtccca attgtgacta tggattacga 13800

atacctgccc gccatcgact ctcgtggtta gtagattcat ttaaagaatt cgataaggat 13860

ctccagacaa agaatccgtt acattttcct ggataccaag aaaaaatcag caaacttcaa 13920

agacaaacta agctgaatga ttcagtccta actggtgaag cttcaattaa tgatcagcta 13980

ttttcactag gtattatgga tccaacattt attatgggat ccctcggaac tgttaccggt 14040

gaaaagataa cgcgcttatt tgaatatgca actatccatc gtcaagcagt agtactattc 14100

actgcttcag gcggggcacg gatgcaagaa ggaattatgt cgctaatgca gatggcgaag 14160

atttcacaag caataaatga gcatgctgct gccggccttt tatacattgt gatcttaacc 14220

gatccaacaa ctggtggagt aacagctagt ttcgcaatgg acggagatat tattctcgct 14280

gagccccatg cacttgttgg ctttgcaggc cgtcgagtta ttgaacagac gattcatcag 14340

caaattcctg ttgatctcca atcagctgaa aacatcctgc atcatggatt tattgatcga 14400

attgtaaaac gtcaagatga aaaaaagctg cttgaatggc tattaaaaac agggagcgtt 14460

gctaatgaat gaacaattat cagcaagcga gattgttaaa cgtgctcgca atgacaataa 14520

aattacgggg atggagatta ttcaaaatgt tttcccagat tttgttgagt tgcacggcga 14580

ccgggcaggc ggggatgatc ctgcaatcgt tggtggaatc gctactttcc atcagcaagc 14640

agttaccgtc attaccactg atcgaggaaa aacaacagaa gaaaaaatca taaagcattt 14700

tggctcacca atgcctagtg gttatcgcaa ggcactccgc ttaattaagc aagcagttaa 14760

atttaagcga cctgtattct gttttgttaa taccgcagga gcatttccta gcaaggaagc 14820

cgaagaaaat gggcaaggaa gtgcgattgc ccaaaacatt ttacaaataa gtcagcttgc 14880

cattccaatt atcacgatta tttatggtga aggaggtagt gggggagcct tagcattggc 14940

atgtggagat gaagtatgga tgttagaaaa cagtacttat tctattttat ctcctgaagg 15000

gtttgcctcc atcatgtgga aagatagtac gaaagcagat aaagcggcag aattaatgca 15060

aatggtgccg caagctttat taaaacaagg gattatcgaa ggaattattc cagaaagcga 15120

agagcatcgt aaaacttgca aaaatatcga gcaggtttta ctaaagcgat taaacaagct 15180

gcaagaatta ccgccaaacc aacttctagc aaacagaaaa aaacgttatc gaaagtttta 15240

aggaggataa tatgggaaat atattaacag gaaaaaagat cgttgttatg ggagtagcta 15300

ataagcgttc tattgcatgg ggatgtgcac aaatgatggc tgaacaaggt gcccaagtta 15360

tctatactta tcagaattcc agaatgaaaa aaagcttaca acggttagta gatgatgaag 15420

atcaattaat tgaatgtgat gtagcagatg atgaaagtat tgaccaagcc tttacgatta 15480

tcaaggaacg ttttacaaaa gtagatggaa ttgtacacgc aattgctttt gcaaaaagag 15540

aagaattagc tggttcaatc cttggtgcta gtcgcaaagg ttatgcaatt gcgcaagata 15600

tttcgtctta ctcccttatt gctgtcgcta aggttgctaa tgagttaaat ctattaaata 15660

atcctgcaag tattgttacc ttaacttatt ttggctcaga acgtgctatc cctaactata 15720

atgtaatggg aattgctaaa gctgcccttg aagctagtgt tcgctattta gcacgggatt 15780

taggacaaaa acgaatccgt gttaacgcaa tctctgctgg cgcggttaag acattagcag 15840

ttacaggtat taaaggtcat gatgaacttt taaagatgtc ccaagcaaga actgttgacg 15900

gagaagatgt gactattagc gaaattggga atgtgtgtgc attcttaatg agcgatttat 15960

caactggagt tactggcgac accatttatg ttgataaggg ggtacatttg atttaaaaat 16020

ataaatttta aagactgaga aatgagtttt tttcagtctt tttgctgtct tttagaagga 16080

aaacataaca atagcaagaa ctttttttaa ttttttctaa aaagttcttg ctattttata 16140

gctcagttgg tattattaat aacgttgtga aagacgagtt aataattttg agaaaaattg 16200

tttaatatat ttaaaaatgg cccgttagtc aagtggttaa gacaccagcc tttcacgctg 16260

gtatcgtggg ttcaaatccc gcacgggtca cttttgcgga agtagttcag tggtagaaca 16320

tcaccttgcc atggtggggg tcgcgggttc gaatcccgtc ttccgcttag ccagttctat 16380

tatgccgggg tggcggaatt ggcagacgca caggacttaa aatcctgcgg ttagtgataa 16440

ccgtaccggt tcgatcccgg tcctcggcac tatttcggga aatagctcag cttggtagag 16500

cacctggttt gggaccaggg ggtcgcaggt tcgaatcctg ttttcccgat ctggctaaac 16560

agctaagagg taatcttctt agctgttttt ttggatatct gtcaactggt atggttgact 16620

aaaaattttt ttagtgcgcc cggcatgggt attagctagg tggtgaaagt ccgctatggg 16680

ccgtagtagt cggaaccatg agctgaggac aagggtgtcc accgtgaggt ggaatctgaa 16740

ggaagtctaa ggcaaagtac tgcatcgatg aacaagaagt agctataagg ctgaaattaa 16800

ctggataagg ctgctagaca agttgaagtc caatactact cgaagttggt ctcagtaaag 16860

ctaacgatga catggtacga aagctaatat tcttacccgg ggagatctgg cctacacgtt 16920

tccgacaagc agtactaacg cagccattaa ttagtttttt gaaataaaat aaactttttt 16980

gaaaaaagta ttgcttttta taggataact tgataatata atattcgtcg ctgatatgcg 17040

ggtatagttc agtggtaaaa ccacagcctt ccaagctgtt gtcgcgagtt cgattctcgt 17100

tacccgcttt taattaattt aatatggccc gttagtcaag tggttaagac accagccttt 17160

cacgctggta tcgtgggttc aaatcccgca cgggtcattt ttggaggatt agctcagttg 17220

ggagagcatc tgccttacaa gcagggggtc acaggttcga gccctgtatc ctccattgaa 17280

caattttaat agttgttcaa cattttaata taattgggct atagccaagc ggtaaggcaa 17340

cggtttttgg taccgtcatg cgctggttcg aatccagcta gcccaattag tctaaaacag 17400

tgagtaaaaa ttactcgctg ttttttatta gaaaaaatag gcaatctaat aacgtaggct 17460

aaaagcgaac atcattggat tgcccaataa ttagcataat ttattcacgt cgtaacataa 17520

tatgccagac aaagaagccg actccaataa ttaacgccaa ccatgcccat agttgcaatg 17580

gaagatagat agtacacgtt aacaagccta aaacccaact gattagcatc atcgataatc 17640

cagcaatatc gccacctaaa ccatcaatac cacaagtagc gatatagacg attcctgatg 17700

ccagatataa tagcgaaaca attaattcgg ctgttccggc ataaatagga attgcaggat 17760

taggattccc aaaacctcct caagtcttat tctacccgga tgattagaat ttataaagaa 17820

ttacagtttt tctatttaca aatagagcag ggggaacaaa aaatcaaacc agaaattatc 17880

ttctgatttg attggatctt atttagttgt atcttttact tttttcaagg cttctaaaaa 17940

tacttcataa ggttgtgcac cagtgattga atacttatta ttaatcacaa atagcggtgc 18000

acttggcatt ccaatcataa atgcgcgccg ttcatttttt cgaacttcat cttcatactg 18060

attagattca agaacttttt taacatcagc aacaggtagt ccgatttcat tcattgcaac 18120

agttagagct tcatagtccg cgattgattc attatcatta aaataaagtt gataaagacg 18180

tttaacagcc ttattaagta atgcttggtc atttaaactc tcaatgtatt ttattaagcg 18240

atgagcagct aatgtattaa caggaatcgc attttccatc ttaattggta aatcaatatc 18300

ggcagcaagt tgatcaattt tatgtatttg tgtaacagct tcttgcttag ttagttgatg 18360

agtttttgca taatattctg tcattgataa atcagttgtt gttggcaacg ttggatctag 18420

ttgaaatgac ataaatttta atggtgtctg atcggcaatc tttaattctt taagcgcccg 18480

ttgtaattgt gtaattccca tatagcaata tgggcacgca atatcagccc agtattgaat 18540

ttccatagat gatcgctcct tataaatata tattaattta atctaatcat aaaataagcg 18600

aagttgctag taaatacgta aagagggaac gatttatttt aaaattaaca tgtcattaaa 18660

attatagttt taatgcgact gaatttaaaa ttccccccag aatatcaatt tttagctttc 18720

tcaaaagaat ttatttaatg cttattttta actttaataa aactttttct aaacaaaact 18780

aatacttttg atttatgttt taaaagattt atgtaatact attgatgaag tctatgtcaa 18840

aagtatttta aaaggagttt ttatcatgaa acatacgctt aaagttgatc aagtacgtga 18900

cggtttatgg ctagattcag atattacgta tacgcaagtt cctggatggc ttggtaatac 18960

aacgcgagat ttgaagcttt cagtcattcg gcattttcaa actaatgatg atacacgtta 19020

cccagtaatt ttttggtttg ctggtggcgg ctggatggat actgatcaca atgttcatct 19080

gccaaatttg gttgattttg ctcggcatgg ttatattgtt gttggtgtcg aatatcgtga 19140

tagcaataaa gttcagtttc ctgggcaatt ggaagatgct aaggctgcta ttcgttatat 19200

aagagctaat gccaagcgct ttcaagctga tcctaatcgg tttattgcga tgggagaatc 19260

agctggtggt catatggcaa gtatgttagg tgttactaat ggtcttaacc aattcgacaa 19320

aggtgctaat ttagattact ccagtgatgt tcaagtagcg gttccttttt atggtgtagt 19380

tgatccctta accgctaaaa caggaagtgc atcaaacgat tttgattttg tttaccgtaa 19440

cttgcttggc gctgaacctg aagctgcaaa tcccctcacg tatgtaaatt ctacttctac 19500

gccctttctt atctttcatg gtacggagga tgtcgttgtt ccaatcaaag atagtgaaaa 19560

gctttatgat gtattagttg aaaacaacgt tcctactgaa ttatacgaaa ttgaaggtgc 19620

aagtcacatg gatgtaaaat tccttcaacc acaggtattt aaaattgtga tggacttttt 19680

agataagtat ttaacccggc catagatcat ttcttgcctt tactactaaa aagcaaacta 19740

ataacaatcc aactttcaca ttattgacaa aatcaaaaag caccatgaaa ttactttttc 19800

acggtgcaac tattgttaat atttatctac ttggcttcca tcaatggaga aatagacaac 19860

ttgatttgca aacctaatcc agcagcatat ttatttaatg taggcagcgt agggattgaa 19920

tctaaattct cgattttagc caattgaggt tgtgtcatac cgattttttt tgcaaattct 19980

gtttgcgaaa tgccctgttt aattctttgt acttctaaaa aagacaaggt atcaacaatc 20040

gataattctt cttttgttgc agccgtctga tgcttatcta tatcttccca ttttctcatt 20100

atttaccctt cctttcgtac caatcgtcaa gtagcgacaa ggctctttct atctgacgag 20160

gatctgtttc atcctttttc ttagcataat gatttaataa tacaaaatta tttttcttcc 20220

atactccata aaatactctt tctggcatag gtcgtaactc ccatagttga tgcctataac 20280

cctttaattt ttttgcttgt ggtgtatgca acacaggacc aagagcttgt aacatcttaa 20340

tttgatggcg catttttaaa taaattgctt tatcttgctt ttgtttactt tgcgaaattt 20400

tctcgaaata atcaccaatt tcactatttc catttttatc ttcgtaaaaa actacttcat 20460

acatgcacta tttcctttat tatagcttat ttattactaa ttatagcaag attgctatag 20520

ctttactact attaaaatat attccataat caaaaatttt tctattgaaa gcacccgcac 20580

actcaattaa atcaaataag tttacggcaa tgattatttt tggtgcacaa aaattgccga 20640

tgttccccag ctttacaaac tttcaccatt tcataccatt ttaactaaac aaaagctccg 20700

gtaaaatgcc gttatatcag cattttatcg gagcttcttt tttatacaat cttaaaaaaa 20760

tgcgtccccc gagcatagaa gattgttgat atatcaacgt tttgaaggag ttagtgtgcc 20820

aaacgtgtgc gaacttaact aaaataaaaa agagcgccta tcagggcgcc ccacgttata 20880

ccggtacgga tcaagtaacg aactgaatcc aatttgcgtg ccgatacttc tcatctgata 20940

tttaaattat aacataatta ttttatgata caataattaa tgcgtgtcgg gagtaacgac 21000

ccaaactatc acgcaaaagg ggtgaattcc ccgtgacatt tatacttctc attctgatcg 21060

tgccgaacgg gctcgtcaag caactgatgc attatgcaaa ggttgtaatc aaaaaggctc 21120

tcaaacatct aattgatgaa ttacttggtt aattctgata ggaaatgtgc gagatgctat 21180

ttcagcggta gtgtttgcta tcgctgtttt ttattatact ttaagaatat agtcatggta 21240

tttttttacc ttagtaacta ttcagtttaa tgttactcta ttaaattttg aattccttta 21300

catccttaac ctctatcata ttacaatctt tatcttcaac gtagacgaat tctgacatat 21360

catcaatatc attgaaatct ttattagctg cgagttctgc agcatcctta aaaaaattag 21420

gccttttttc ttctattttg tccatcggta ctaaacattt taatatgtaa gttttgaaat 21480

gctcttgttc ccatcttgtt tgaagaggcc cttttaatgg cctacttcta atattatcta 21540

attcatatat ggcactatca aggtacctta aaatttcagg acacttataa atgcaagaat 21600

aatcaaaatg atatggtata aaaaagccac taattccgtt atcattttcc aaacgtgctg 21660

caatatcatt ccaacttcca ccagattcaa tcaggtatga gtgcccatta tatgaaataa 21720

aaggattatt atttttacag tagaaacgaa tattattttt ttctaaaaat gatcctaaca 21780

aggtactaga atctgttata actttgctaa gtggttttaa tccatcatat aaatagttcc 21840

tttgctgtga agaacaagtt gtaaagtggt aaaaatatgc attttcaatt tcatcttttt 21900

tatatagttc taatgatctt tcagccattt catgagtcga taaatcaaat atcttgtatt 21960

ttttcttttc agacaccatg gttttctcct ttttatgata tatctaattt agattactat 22020

atgacaaaac accctagcaa ccattaatcg gctactaggg tgtttattac tataattttt 22080

cacttaaatc tgaattgttt gaccaatgaa aatcatatta ggatttgcga tgccattctt 22140

ttgagctaat gcttgccagg tcttgccaaa cttagccgca attccgctta gtgtgtcccc 22200

tgcttggaca gtataagcat tggattgtcc gttgccggct aagtagagtt tttgcccaac 22260

gtagatcaca ttcgggttag taatgtgatt acggcttacg aggtctgaga cagtggtgcc 22320

gaatttggtg gcaattccgg ataacgtatc gcctgattga acaaagtaag tattttcgtt 22380

tgatactttt ccggtgactt tgaggacttg gccgacattg atctggtttg gattaccaat 22440

accattgatt gctgctaggt tctgataagt agtcccgtat ttttctgcaa ttccactcaa 22500

tgtatcgccc ggctgaacaa tgtaggttcc agtagctggg tgaccaatat gttgaaccgg 22560

ctgtggtgct ggaataacag cttgcggaac attgcctgat actaaaccag tcgtaaaggc 22620

accgtcaaag tcataactag tatcaattcc catgatgcta tgatcagaat attgccaagc 22680

acttgcatta tcaatcccca gttcagtcac accatagcca gcaatccaaa tcttccggga 22740

atcaaagcca tgtgaattaa gaataccgcc agtgaagaag gacttcatgg agtaaatccc 22800

agtattcttg taaccaagag cttctacttc ttgaagaaag gctaaggata ctgattgata 22860

atctgcagtt gaatgaactt ccgcatcatc aatcatcaaa gtatcgtcat acataccaaa 22920

ttgcttagcg attttaacga agaatcgagc ttcattttgt gcatcagtca ctgatgtata 22980

acgagcaaag tggtaacagg aaacgcgcaa gccaaccgct aaggcattac gaatttgagc 23040

ggccgcacgt ggattaacat aagccgaacc atcttcggac ccttccgtta acttaacaac 23100

gacccctaat gcaccctgag ctttagcagc ttggaaaaag gcaacagtat ctggttgata 23160

acttgaaaca tcgatgaatt gattacgcat attattcttc ccctttcatt ggatcgattg 23220

ctggtgcttg gccggctgga atagtagcgg gtttagttgg ttgtgatgtc ggatcaacgg 23280

ttggtgtcaa tgcagacttt tcataagctg attgcacaag cgtttcaata gcgttaaggt 23340

caatattctt aataccctgc ttttgaagtg cttgttgaac aatacctgtt gcttgatgaa 23400

atttctcatg accagccatg tctttgccaa ctaaggaagc aacggcatta tctgcaagtc 23460

gttcagcaca agaccaagca gcacgagaag tttctgtttt ggcatgagtg actttagcgt 23520

gaataaatgt ctggctttgc tttaaggcaa aataaaagac cgttgagata acggccgtca 23580

tgatataggt tggaattgca ttaatgattg tgctcatgtt gtttctcttc tttctctaca 23640

taatctttgc tgagcttgta aatatgattc tttacccaag atggtaatgg tagtcccatt 23700

tggccccagt tttcgataat cgaaactagg taaaatagga aaaagaaaaa gacaagcgag 23760

tcgcctgcct gtcctaatcc gcataattct aacattggat aaacggttaa agtcaaaata 23820

ataactgcag catgtttcaa aagtccaccc gttcctttac tagaagttgt tcggtgcgta 23880

atgatacttt tgaaaaaacc tgtcacaata tcaatcacaa tcgcccaaac taaccattca 23940

atgagcacgt tgtcaatcat gctggcaaaa tattgaatgt atagaacgtg ttgtggcggt 24000

tgtgtcagta gtaagtgcat ttatagtggt cacctcctta taagggtaaa ataaaaacgc 24060

cctgtagagg acgtttaaaa ttatttaatt tttattaaaa gattcccaat ttccatcttc 24120

attattattg attttaatta ttaagtcgtt aagtttatta tcaatatcat tagctagctt 24180

atacgcactt gaatcacctc gtactctagt aatttcatat atttttcctt gacgaagttc 24240

ttcagtcaaa gatttaatat tatctatttc gttagaatat ttctgtttga gattcacttt 24300

atccccatct ccttaccgct aaaataaaag tgttatatta cgctgttact aaatatggaa 24360

tattattatc agataaccat tgaactgcat tattattaat ttttacaaca tcatctaaaa 24420

ttttatctaa attaaatgaa taaccaaata taaaaaattc atgcttatga ggatcgtata 24480

ctcttaaagt ttcatctttt gtaggagtga acactttgtc tccaactttt atagtttgat 24540

acaaattacc aattacttgt ttttcaacca ttatgtgctg acgtcggtta aatttaataa 24600

gtccttcatc aatatgttct aaactattcc tgaggcctct taaattttca ttttcgaccc 24660

tattaatata gaaatcatca gttacattca aaatttccct tacctttttt cgatcattac 24720

tatttttaga gttagaaccc catagaagct tagctatatc tgacgtatac aatgcaatat 24780

tttgtaaatt ataccaaatc atattatctt tataatattt ttcatgtttg gtattatata 24840

tatttttaat agcaataata gtaaaattac ataacagata agtttcacta attgcatact 24900

cctcgtcttt actattcata gtataatgtt gcataatatt taccattccc cttattaatt 24960

gtttagtaat attatcgtat cagtaattga aataattatg atattattat gaaaaattga 25020

agaaaaacca aaggctaata attaatttac aaaagtatgg aatcaaaatt ttaataaaat 25080

gccgcccata ataaaagccc cgctcgtttg agtgaggctt atttatgtat tgtgtatttc 25140

taaggcgact ataagtaatt accgttaagc taagaattca tttaattatt agccattttc 25200

tttagctttt ctttatcttc aatctgatac aagcgaatct tagtaattaa tccctgatac 25260

gcttttactt cattgggaaa attattagca ggagtaaatt tatcatcttc ttgggacaat 25320

tcagaaacat agaatacatt tcgataataa cctatataaa gtttatggtt cagtttccta 25380

tataatttat ttttttcaat aatttttgct tctttaggag tataaccatc ttgttttcgg 25440

tcaaaatagg cttcccattt aatgcactgt tcctgtgaat taatccgatt tgtttttaaa 25500

ccaaatgctg ctaatggata cccacctaat ttgggattgt tacacaattc attatcatat 25560

tcgtaagcag tattaacaat atcttctaat gattttgaaa tattcaagat aaccacctct 25620

attttataga gtatcactaa tttagactac taaacaataa cttaaataat tttattgtca 25680

tgatttatgc tccttgaatt tcgtaattct gaccggttac cgtcttatat tcatctttag 25740

tgatcgctcc gcatcgtaca taaactttgt agaattctaa atcatggtta ccccagtcat 25800

tccaaaacat ttgaagcatt tgtaattgtg tcatcattat gccgttacct ccttagattt 25860

ttcgactgcc tgttgctgat taacagtcat aaacatttgt tgtaattgct tgatttgagt 25920

tgcttgttga gcattagtag cctgcaattt tgcatttgat tggttctgtt gcataatcat 25980

tgcttgcaaa atagtgattt gttgattggt attcccgttt gaatgcgaaa taatcgtcac 26040

aggaatttga ctatcattca taataaagct ctccttagta tatttttgtt tgataaaaca 26100

ctgttacagt tacccctgta ttttgtatag ctgagttatc aatgtttcta acgattaatc 26160

catcgccgtt tccagcttct gaaacactca attgccagcg gtcgccttct gcaacatagt 26220

taagtatttt cccagtaact ccaagttctt tattgatagc atccattgga agtgtcttga 26280

cctcttgcga attaagttga tcaatagcta taaatttagt aatcgtcttt aaattagaat 26340

tgtaatctgt ttgatgatct tgcaaactca ttcctagcga tgtattgcct aaactaatga 26400

gcgaagttgg cgttttatca gcataaattc tcccagcttg aaaatgggct gagttttgat 26460

cgtaaaattt aactccagta ttaaaagcct ttccacctaa cttaaagtga tccacatcta 26520

gatatgttcc agtcgtacta ggcccaatat atagcccatt catgctttct ctttcagctc 26580

ctagcataat ctcgtttgct gtaaactcag cgttagtttg cagagtgagc aactgatcgc 26640

attttccccc aagtgcaagg taatcaatgt ggccctttga tagcataatt tttagattag 26700

cttgtccact tttatttgta acaaaatggc tttcccaatg tttgatagtt agatcgtaag 26760

ttgaatcaat cactgatcca atagcattgt ctgctatatc aaacgtttcc tcccacacaa 26820

aagaatattc accgaaaccg gattgttgcg ataccgaatg ctgatacaat gtttgatagc 26880

aattatagtt ccaaagtttg attttgatcc caatattatg tgtgttatct gcatctgtac 26940

cagttacgtt taatatacat cctctgaaat aatacccttt aatcgaaata ttactatgaa 27000

acatggtatc aaaagaaacc ccatacttag ctgcacctcg ctctagcata atttgaatat 27060

cattttgatt tccatttcta aatttgaatg caatttgatc cacgtttagt gctggtgtta 27120

actttaacct caacgaaata tgagactgct tcaagtcaat agctaatgga ctttcaatta 27180

gataacatcc attcccatga acacttttat tattatcaac tgcataggtt agaatgcttt 27240

gaaggatttg gtaatcatca tttttaccat ctccatgtgc tccccatgtt tcaggtataa 27300

tttcattttc aataagttcc gctaatagtc ctgatgctaa ttgttctgca tgtttagttg 27360

ttgtatctgt gacataaaaa agccctcctc ctccatcatt tggttcataa tagccaagag 27420

ttcttgcgca catatctttg cttaaagcac tatcagcttt catggcagct acatttggat 27480

acacatgtgc atagttaatc gcaattgcct tcatcatttc ctgcttaaac gcctctgctt 27540

ccgcctgggt aaacaaattg tcttgcttaa tcttagcgtc tagtgcatct aaaccatttt 27600

gaacagtaac gccaagagct ttgagcgtgt tcattgattg ggtaaactta tcaataaagt 27660

cactggtcat cttggtgagc ttgtcggtac tgtcttggta cttggcttcg atttcgttaa 27720

ttaactcttc gacgggtgaa atatatgttc gtggtaccaa gccatcaatg accttatcag 27780

caaggacatc gaggtcgaat tccaaggtag tgatagagtt accgtctttg aggatgcgaa 27840

aaaaggcttg gcgataggaa ccagcgactg taaaggcatg gccgggcatg tcgaaacgga 27900

aacgtccggc ggttgggtca agggcaacgt agcctttatc atcaattact cggaagtcgc 27960

cggcggaatt cttaggaagt aatccttcaa accagacgtt acagccagtt aaatctagtg 28020

gtgtcccgtc ttcgttctta atattgacaa aaacttgacg catacttcgc tcatactgac 28080

gcgcttggac ccagttagta ttactgccat caaagttaac cttgaagtct tgtacattgt 28140

caacatgcgg acgccggtct tgaccgatca cataagttaa agtttgtgac attaaatcat 28200

tcctttctct ttcaaaatct gcatgacagc ttgttcaacc gtttcatagt cggtccctaa 28260

agccacatgc tcaattttct ttttccacgc ttcttgagcc tcttttagtt catctcgcgt 28320

ggctttttct tgttcgagtt ggtcgagacg agtattaatc tcctcgagta gcctaatcaa 28380

ttctttatac tgcctgtcac ggagattatc gtctttgtta tcctcatttt gaattatcgt 28440

taaattattt attaattgct ggcggaattt tcttccactc tgcaaatcta gattagtttc 28500

taaatccaaa tgaccatccc cctcctcact tcaaactact ccaatccgtc actctgtaac 28560

ctagatttac ctgggcccgt gatacgtatg ctttatcaga tccggaaaca ttaaaaacca 28620

gccttccttt agtagtgccg gctggaattt taatgttaac tgtgccggcg gattgccagg 28680

cgccattatt gacgatatag acaatattcg atttgccggc agaactacca tcgctcttca 28740

agaactcgac ataagtagta gcggaacttg aattatcggt cgcatttaac ttagccatga 28800

gacgacttga gatggaggag acaccgttga ggttaaattc attgcttctg agagtgcctt 28860

gctctacttc aattactgga ctgtcattat atttttcgct ggaaacagtt ccgccagtca 28920

tcatagttac caaattatgc tggccatttt gcatccagta aatattctgg tctactgctt 28980

ccatattctt atccgtctct ttcttatcat tcacgacttg cgttaatgac ttggattgct 29040

gatccatgct atttgctaat ccactaatcc ggagtcgttg ttcttcaaga gcttcttgta 29100

gacgtttagc tttctgattt tgataatcta aaatagttgt ttttgaatta tttaacgtta 29160

tttgattatc ttgtccctta gcttccggat atatcgtata accgacagtc ttataactac 29220

ctgaatattt atctcgcgca agtactctta aaatttcacc agcaacaggt acaaatgaat 29280

ttccctgaag cgtgacttca atatttaaat ctggatcagg tttaagagta gttaaggcat 29340

atttacgcat tgcatccgca tcttgaaatc tttcatcaac aagatcaggt ccaggatgtt 29400

ctccccattc gtcaattgac ttttgatcaa ctaccataaa cggttgaaaa taatactctt 29460

cctgtgattc tgtggttgta tcatttccga cagtatcatt ggtttcttta ccaccgatac 29520

catcacgaat aacttgtaag ggatcaagcc aggttccatc atttgaccag gcacttttca 29580

atgcagtcat taaattcttt tgcttagtaa ttccaatatg aagatggcta gtatcacggt 29640

ggccaataac atcccccgtc ttaatttctt ggccaacttg aacagtaata tctcctcgat 29700

tcaaaaatgc ttcttggtag caaattaaat aatcattcga cacaatagtt atatagtttt 29760

ccagtccagc aatgtatcca atatcctgga ctttcccacc atgaatcgca tgaacatcgc 29820

gtccaggatg atcgacagaa ccaaaatcaa ggccatcgtg aaaaccatta gttcgcccca 29880

ctccatcttg agggtgtgtt ccaaaagttt gccccaatga aaaatgacct tctccaacat 29940

ctgggaaagg ccatccccac ggattaggtg gtgaaatgat caatttatct ttcgtgattg 30000

gggcaccatg aggactccaa ccagtaacac cattaatttg tcctagtgca ttaggaatat 30060

tgaagaacgc aattaactgg tcaaaaccct ttaagatatt tgtatagggc tcccgacaat 30120

aagtattaaa agttcctcgc ttaaattgaa gtaacccaag cgccggacca gaaccatcgc 30180

catcgggatc agttcctggt tgcggaatcg tctcgtttcc acctgattcc aaatgaattt 30240

gtgcccgtaa aacattaagc tgctgagcat tcggttttat tccataaaag ctagcagcat 30300

actgaataac tggtgtccaa tcaccgttga ctggctcagt tggtccatta gcattagttc 30360

cacctgtccc ggtataaaca gtgtgctgaa cctcatgctt accaccaaca caacgaatca 30420

tattaataat tgattgactg tcccgagtag ttttaactga ctttgcatcc cttggaaaat 30480

caatcactcg accattatcc ttatagaact catcttctga ataaactcgt atcttcttgt 30540

tatccggata tataactgca cttggccata actcagtaat cttactaagc atctctttac 30600

cactgccgct atcgaactta gaagttgact gcttatcaaa attaccatga acttcatagc 30660

taaagcctag cttatttcca tcaatccaag cctttagtaa gtcctcaact ccataagact 30720

gatccttact agcattacta tcctgattgt tatttaaggt aaggtcactt cgtttataaa 30780

ttcgactaat ctcattatag acatgccaag cagtgatatt tttcgtatta gtatcaaagt 30840

tttcaacaca attttttaca atatattcct gtccctcaag cacaatcgaa ctttcaacat 30900

ctaatggatc atacaatgct gaatcgtaat caaatactga aaaagtcaat tgaaaggtcg 30960

aattcttaga ccactcgctt tgcattgttg gccagagaat aatgtctccg attggctcct 31020

tagttgtctc tttatgaggt gtcatcaata ctaaactaat tgtcctcacc tcctaaccaa 31080

gatacataaa tgggaaactg aatgtaattt ccaaatcatc tgcaccagta acttcaaagt 31140

cattctttcc aggcgctaaa gtaatatacc cataatcggt attggcactg tctgggttat 31200

tattcttaaa tgttcgccgt ccctttagca agagagtatc gtttccagat agattttgat 31260

tgtacgtcca agaagtatta gtagttgtat tctttactgt gaattttcca ccattatgct 31320

taatagtaat ttttagatca tgacgctgtt caacaggatc gattgtaata tcactatcgt 31380

taaggataga aaacttattc tggcctttga aatggtatga aggagtttcc tcatccatgt 31440

tcatattcaa gtctaagaaa tcctcatctt tcatctcatc cgtatgcaat ttactaaatc 31500

gcattccgct tggattttca aaaggcactt caaacgtaca atagttaact tcttctggtt 31560

cgcttttaat ctcaaaggaa ctagcccgac agtaacggac aatatccggc tcaactcctg 31620

ttcttaacct aaagattcct ttttgtgcaa aaacgcgata aatctcatgc tttgccatct 31680

taaaatcttt tcgatcaaag aactgcaata agaacttagc ggtgatcgtc gtttggccat 31740

aacgggaata actccaaatt tgaccatctt gttgggaatc atcacgataa ttattaataa 31800

cactaggcga ttcagttaag ccaagaaaag ttaagtggtc agtaatatct gtgctggcca 31860

cttctttttg atcatcaatt ttaatgtaaa gaatgtttac agcgattacg accacctcct 31920

aaagatgttg attatcaaat aatgcttgat cagttccttc aatcccataa aactggttct 31980

tgtcaaaggc gccggcttta attgcagaaa gttgagctgc atttaagccc agcattgtcc 32040

caaagcggtt gagcatctgg tccatccgat ctaacatctc tttttgcatt ccattattac 32100

tttggccacc gttattataa ttatttacaa cttgggtgct ttgcatattg ggcttaaagt 32160

tagttaatac cgaaacagca tcatcagtac cattagcata ttgtggtaag tgcttaaaca 32220

tcttagcagt atcgctagct ttaacaactt ttgttcctcg tggtgctgga aagactacat 32280

tacgcccata aggaataaat ggtgtttcac ctgggaattg aactaactca cgaaagacgg 32340

ggccaggttg atcattaact tccatcaatc cacctttatg gtaattagtt ccctgagcat 32400

gctttgattt atgaaaaata acattaactg ttttctcaat agtattaggt atactcataa 32460

aattatggat tgcatcaagt gctgttttaa ttggcccaga agctttatca ttagcttttg 32520

caatttttgc tggtcccgta tttgatccag caaaagcatt tacaccattg cgagcactat 32580

acattggtcc actagcatta tcaacggcta aagcattttt aggagcgcca ggatttgaac 32640

tagcaaaaac atcgacacca ccagtagcat ttctcatcgg accactagca ttgtcaattg 32700

ccagtgcatt ctttggttct cccggattag catgacgcca agattgaagg gcatcgttag 32760

cagcttttag atttccacta gctttatcat cagcaatgat ttgttttgca gcactaaccg 32820

gcatcccctt ccacagccca taatcagtga tcagttgctg tagttcgggt ccacccttag 32880

catttacgat tgcttgctgt tgcttaggac taagttgatt ccagcgacca attttattga 32940

gcgcatccat taatcgacca gcaccctgag tagtaaccaa agcacgttgt tgcttaaacg 33000

aaagactatt ccaaacacca gcctgcacta atgcatcaac taattgtgga gttcctttag 33060

cgtcaataat cgcttcctgt tgttttaagg ttaaattatt ccactgtccg gaactttgca 33120

aggcttcata aattggctta gttgccttat tagtaattac tgcttcctgt tgttttaatg 33180

acaaggaatt ccatgcaccg gattcaagca gaatgtcggc catttcttgc ttacctttag 33240

ccttaacaat tgcttctttg gtcttaagat caagattatt ccattctccc gacttctcta 33300

aagcttgaac aattgtttca ctaaagccgt ctttcaacca agctttctgc tctttccaac 33360

tcatgctatc ccatttacca ttttcgatta atgcagctgc caccatttgt tgagcattgg 33420

tacttaactt accttccttt ttcagaagtt taatttggtt ccattgatct ttagaattaa 33480

cagccttatt aacttcctcc tgggcattag tccgaacttt accagttttg ggatcgaaaa 33540

ctaagtcatt ccacatatca gcggcttttt tagcttttcc acttaattta gtagtattta 33600

ccgctaagct ttgtaggttt gtttcggcag ctttggattg ccgcttaatc tcatcagtgc 33660

catctttata actaagtccc atacttttta ggtcattctt aattaattta gtactttcac 33720

cattagcacg ggccaggcga atatattcgg cagctgcttt attcgtgtaa tctgttaatg 33780

ctttcttatt agcacgcata ccgaccttat acagttcaga atcttttccg tagcgtttag 33840

caagcattga agactgcttc ttgtattggt tttccatttc agcagtttca ctggttatat 33900

ctcccatttt tgttgagcgt tgttgcatag acatatgctt aatatcgtta ttaaggacag 33960

ccattgcttt agcacgctta cttccactta atttaagaat tttcatctca tcgttaagca 34020

tttcctgctg gttgttcttt aacatgactc gctgagtatc atttaattgt gaaacttttt 34080

tattgtggtc agatagaatt gtttgaacac gctgattggc attatctgca tcattcacat 34140

atgaattata ttgttttttc tccttttcta actcatcgtc aatagcttta gatatttctg 34200

gagacagacc tttttctgcc tttttggcat tagcaaaatg ttgttttgca ctattttcaa 34260

tattagtaaa ctcacggtca aaatcagcag ctaatttctt tgttgaagtt gaggcagcaa 34320

catccatatc tgacagagtg ttctgaattc cttgagaagt cgactgcatt tttgataaag 34380

ccttatcagc agctgcacca acatcagatc cccaacgatt cgttcgttgt gatgattcga 34440

cagctttctt accccaaagc tcccaaacag taattccacc ggcaatggct aaagtagcaa 34500

ctccagcaac agcggcaact gttcctaaag atactccagc cgcagcagta gcagcacctg 34560

cttcacctgc agctacaccg gtaccttgta aagcgacagt cgcagcacca acaccgttag 34620

cagcttgaac cgcccgacca cctgcaccag tcatggttgt tccaaacttt gcagcttgaa 34680

atgttgattt tgaaaaagct gagccaataa cgtctaaccc actggctcct agtttcattg 34740

cagtttgtac acggccgatt ccggaagcaa tttttccaaa ggcaagaact gtcttcccag 34800

caccactagt cagtttaccc aatgctagga acaacgggcc tgcaccagca gtgaacaggg 34860

tagttgttac aattgccttc tggacagctg gtgataaatc accaaagcca tgtgcaagtc 34920

tggaaaggcc ttgaaccatt ggaataatag atggtaagac gtactttgcc atatccattc 34980

cagcgttagt taaagattcc ttgaagattg ctaattgtgc tttaggagat tgaaggtttt 35040

tctgtgacaa gtcgccgata taatcccgct tggcagagtt ttgaacttct ttattaagtt 35100

cacgtaaccg attagcgttt tcggttaaaa ttgccccagc ttgttgtcca gttgttccaa 35160

ataacgcatg aaaaatatca ttcttttggt gaccagataa gcctttcata tggtcattta 35220

acgtcttaaa aatagacgac attgatttta actttccact cttagtaaga aaatctttag 35280

tacttaaatt aatacttgct aaagcttttt gaccattagc ggttggcgta attagtgaat 35340

tgataacctt tcgtagaccg gtaccagctt tgtctgcttc tagaccatta ttagacaaga 35400

tccccattgc actagcagtt tctgagagac taaaacctgc ttgatgagca gttgagccaa 35460

catatgacat tcccacacca agtgattgaa aatcagttga tgtggcatcg gcagcatagg 35520

ctaactcatt taatgtttta gtagatctac gctgcatgac tgtagcattc ttgattggac 35580

gtcctgcttt atcagtagcc agtccaaaag attccattgt ctgcgaagca actttgatta 35640

catcattaaa gtcatcccca gtggccacag atgctttaag ttcgttccgc attactccaa 35700

ttgctgcttt agatgtataa ccacgcttta ctagatcttg atacccggca gcaatctttt 35760

gctgactaac accgtactcg tcagaatact ttcgggcatc tgcagtcatc gttttatatg 35820

cggcattcgt ttctgcagct gattctccgg aagttcgaat aacgttcttg acctttgtca 35880

tttgatcttg gaagtcaacg agtttcttag cggaatatgt taaccctgcc gcaattggag 35940

ctgttaaata agtcgacatc cctcgaccga cacctgatat tttagaacca acattggtgg 36000

ccacatttcc aaaatgttga gtgcgattag caagttgtgt ccacttattc gactgcaatt 36060

caatatcacg attaagagct tgcattcgtc cacgtaattg ttcaatttgc gcagacgttt 36120

tattatattg ggttgctgcc ttagttcgtg actgctcgct cctggtagta tcatccattg 36180

ctttcttagc attctggagt tgcgcattat aattgcgcat ttgttgactc atagtcgcat 36240

aagcagcacg catattatta atacttccac cggatgcttt cagtgccgct tcttgcgcac 36300

gcaaagcatt ggctgttgat ttaatttgcg cttttaaaac accattagcc gctttaaacg 36360

gatcaatatc caaactaaca gttgctgcca aatgtcctaa tgattgggtc attatttaac 36420

ctcctttcct agaataagaa tgggaatgct ttatcaatgg ttgtctgctt ttcttcaaag 36480

acgtaattca ttaatttcaa atctgaaagt gttagcttac tgacctcatt ccatttatat 36540

ccatcgcgca ttttattttt aatgaaatct gtgagactct tgattgaatc atcaatcatc 36600

tcaacagtta tttttttggc ttttcgttat cctttttctc ttctaagtca tcttcactta 36660

gaggagaacc gagagtttca ttaattgcat taacaatgct aattaaccct tctatagccg 36720

taactcctga aagaacatcc tttttagtca atccattctt ccaaaaatta gcagcaaact 36780

cagcccgtaa acttaataat cgttcattat ctttatctgt tggcaatttt ttgggatcag 36840

agtacatcac tatttcttga cgttgaacct ttaatgcatc taatagattt tcaagcattg 36900

gggcttcttt tcgttcatag attgtttctt tgccatctgt ttttactttt aatttatatg 36960

gcatttccat atcctcctca tcgtctcact taactcgtct ctgtctgatt aattagttac 37020

cctttatcaa caacagtttc tggtttagta tcttttgtat tggttggatc agttactgga 37080

gcttcttccc caaagaccat tgcatggaac ttagcaaaat caaatccttc attatcttcc 37140

cgaccgatta aaaggatggt tccagtatcc gcatccccac gcggaacgaa attaccttca 37200

atttcatctg cttctgggtc tggagcaccg tcctgagtct tgattgaaat acctggtaag 37260

gagaacatcc ccttggttaa accaacccaa caatgcttcc cgtttgaaag ctttgtccgg 37320

aacatagtag caacataatt tggtacaaga ttcttagtat aaacttctgt accgttctga 37380

atatcaattc catacaagtc tttcttcatg atggaatcaa tatcataaag gttgatggtt 37440

tccttagctt cggtaattcc accagaaaga accaagtaag ggccgtcatc ggctgctaac 37500

gtctttaatt cgttagtcag ctccagctta acttcactta acccgctcat ctttcgggta 37560

tctttaatct tttcgttctc gtctagcacc ccatattcaa aattagaggc cccaaactta 37620

gcaacttttg catttggtgt tcccattaaa tatcattcct ttcttcaaac ccttcaaagt 37680

tagctgtgac cattatgcaa ttatccaagt ccggatccgg ataagaattc ttgtagtaac 37740

gttcaaagcc atgactgtgg agggtttcat atattcgttc ttgaatatcc attagccggt 37800

catcattctc ttcacgaatc caaaaatcaa cttgtacacg tgggtattca aaaaagcgag 37860

cattgtcact gtaaattgca tcatccccag gtaggggggt gatccgaatc cacggtgctg 37920

aactcgcttt aataaatgga tcatcaggcg tactagtaaa aattggaaca tatgataatt 37980

tatcttgtcg caactcatcc atcatcacga ctaggcactc atcattcgtt agataatcgg 38040

ctacttgcat ctctggagtt ttcatacttt caagtcctca ataaatttag ccaatatttt 38100

cttcttagca gcatttcttg aatcttcaac aaagtgttgt ggatcttgct tagcagttcc 38160

agcatcgggg aagtgagcaa tccatccttt atctttgtca taaccaacat cgacagaata 38220

atctccaccg ttagtcttga gattaccgtg tttagtatga tcttttaaag gagtcttgcc 38280

tgaatgatca gagcctttag aagccacagg cgtccatttt cttaattctt cttgaaagac 38340

tttagcacca tcacgggttg cttttcgtgc cttcttttct tccgtctttt caagtttagt 38400

taagttagca atcagctctg cttcaccggt cactgccatt ttggacaacc tcctgggcgg 38460

taatttttgt tagatctcgc tttgcataat caggatccat tcccgtgatt tcataccatt 38520

ttcctcgcca gttaattaac caattcgatt gaatttcctt acgagttttg aaagcaatta 38580

aaaacgtggg tgattctttt ctgaacccca cgtttgaact gttttggaca aattcacgga 38640

ctgacagctt aggaacttct gcccacacgg taaattcctt aactttctga tccttgatcg 38700

gtcgatgagt ttccggattg atccccattt ggactgagta aaatgttatc cgttccgtca 38760

tgttccgtag tttcatcggt attcacctca ctacttattt gattaataag gccatcaatt 38820

cctgatgaaa gaactggtcg gtaactatct gcagtaattc ctcgctcgta gaagtcttct 38880

ttcacctgct tcataagtgc aattttgaat cgcggttcgt cagagtagtc agcaggctta 38940

ctctcccatt taattgccct tgcaatcatc aatgcggcag catcaacaat tattttaaga 39000

atttcatcat caaagtcctg gtcaattttg cagtagtttt ttagattggc aaagaattgt 39060

tcatcactcg agaaagtgtg gtctgtttcc atcctactca cctagctttg ctaacagttg 39120

atccttagta tcggttgaag tataggcaat tccatgatca tctaagtact ttttgattgt 39180

gtctaccgta tcagcactag tcggtttaac gtccgccccg ttgtcggacg gcgttatttt 39240

gacgctgctg atggaccagt aacaaagtaa ccagcgtttt catcggcctt cttcacatct 39300

aaacgaagaa ccgcttggag atattgacca taaatgtcat tatcaaccca ccgtacctgg 39360

agatcttttc gatttgctaa aataattgca cggtatggat caccaacaaa ggcatgagct 39420

tcaccctttg ctccaaataa gtcatcttca attacagcaa cgttgattcc agagaccgct 39480

ttaccagacg gactagaaat actatcttga agtaagtaac gtccattctt atctttcaat 39540

gtatcaagcc agttgtagaa gctttggctg gcaataatca tcttattgta agccacatct 39600

aaatcgacat tccaaatttt ctttaggtca tcgattgcat tagagccatc aacggtctta 39660

gctgaaaatc ccttaaaaat tgtagcaata gccgcattct ttgtatttaa ggattgttct 39720

tttgcgtttc gagcaactaa tccagttaaa tcaattgctg aatcatcaat cgattcttgt 39780

gaaataggaa ttgctccacg ataggtctca atttgccatg ctaccttaag aaattcaggt 39840

ttttgaagat caggattctt tgcaagttct tcaacagtat gcatttgagc agttgccttt 39900

ttaagaattg gataactacc agaagcagtt gtggcgttaa atacttgaac aaattggcta 39960

agatcagtta ctgttttaat ttcattttca ggattgtaaa tgattgattc aggaatagta 40020

acgcttgcgt ctgaagaagt aattccagta gtaccatcac gatgttcttg gtgaagatat 40080

gcgttgaaat tacgtttttc ttcattttcg tcctcgttat tttgagaacg cttatgagga 40140

tctggagctg gattaccctt agctgcttta cggtaaagct taaggtcatc ttcaatactc 40200

cgaacttcct tttcagcagc ttcaatttca gaacgtaatg acttagctcg tgttaagtct 40260

tcatcagttg cgtcttcatt tgagagcagt tggcgcattt cattagtttt ttcgttaatt 40320

aatgctcgct taccttcttt ttgagcaagc aattctttaa ttttttcgcg aaacatttaa 40380

tttcctcctt cgagtgtctt caataattct tgtcgttcta attcacgtaa catcttttta 40440

cgtttctgat cagtagcatc tctattttct tggctctgca tttgcttaac catattgatc 40500

gaacgctgac cgacctgaac ttctgtatcc ggataagcag gtgttgtaac tactgataca 40560

tcgaacaaac gatcaatttg tcgaatggtt cgttcataat caactccatc ctggtcagat 40620

tcttcccaat cttgggcctc atctgtgttg gccaccgtaa aagcaaagct gcattgatta 40680

ataacgcctg ccttgatatt ggcgatcaaa tcacgcgcaa atgatgtatc agttggctta 40740

acagtgaatt tcaagccaat tgcatcagaa gataaagtca tgttcacccc tgatcgtccc 40800

agtacctggt taggatcgtg attgatcgtc gctactacat ttgacatatc agcgtcatcg 40860

aacgctcctg gagcaatttg ctcaataaag cgagtaaagc cacccaaaac ttcggatggc 40920

ttattgtatt tggctgcata accttcaatt actggttcat catcttcatc agttgccgtt 40980

ctcatctgaa ttggcatcat taactgtcga gtttccaagt cgcttgtcat cgattccacc 41040

tcccttcgct gttgattgtt tctgatactc ttctttttta tcaaggaaga cagtattcaa 41100

ggttgattgg aaacggtcta agtctggatt gtcagatttg actaacccca tccgaactcg 41160

tccttcatta ggagtaatga cgttatttgt aactcccttt tgtacatcgt ccattgacat 41220

ccctgtttct ttgcgtgtat caaattcgat gtggcagtta tgacgttgcc gatcagtcag 41280

cattgtcatt tccaaattac ttgcaatcgg cttgaagtaa taaggtaaat cagaagtaat 41340

aaagtcttca tttaattgtt taatcgattg gttaggacta ttaactgcta atttatatgc 41400

tggaatatgc aacgctttgg caatttgagc agttgaatag ttgttgctgt taattagttg 41460

caaaacattg gtatcaatct caattggcga gtaatcgaat gtatcatcag taacgattgg 41520

gctaccggca ttgctattag cttgtgcata ttcaaacgcc tttctagttt taagacgcgc 41580

ttccggactt aacttacctt tagcctttag taatccacct ttaagacctg acttaaagaa 41640

ccgccgtaaa gtcttaattc catcatcctg taatccaatc tcatcagcca aagacaacaa 41700

cggcgaccgg ccatgaatac catcgtaggt aaagaacata aaatgaatta catcttcagc 41760

cggcacaacg attgtttgac caccgccttt ttgattgata ggtgtaaatt cgtatttaat 41820

attcgttaca tcagaatcat caatataggt ttgcgatggt ggaaaatatt gtatttctaa 41880

cggtgctttt gtatgaggat cacgaattat tcttgaaaaa ccatccccag ttaaaattgc 41940

gttaactgtc attaggaaac gccaatgata agcagataac atatcgtttg ggtgcttatt 42000

cagcaaataa tcaacacttt tgatattctt tactgcattt ttaccatcat ccaaaatgac 42060

aatcggaaat cgagctacat tactagcaac atgagagact gcagtcagca catcagagtt 42120

tctaagagca ccgattccac tataagatgg catattgcta aaccccggaa gaatcccctg 42180

gtcaatataa tcttgtgccc attcgcgttt ctcggtatga aataacactc agcttcaccc 42240

cctttcataa gagataaaag ccattacgaa taattcaatt gaaataatta gtaagccaat 42300

cttcgtggaa aataaaaagc ctgtaactgc taaacagata caagctaata tgaacagcaa 42360

aaccggttcg tttagtttcc agaatttcat ggcatcaccc ggagaagaaa tttcaagtct 42420

tcatccgtaa tctcaaattt ctcaatatac ggggcaagct ttctcagtcg ttccaatcgt 42480

cccttgattt cattaagtgc gggagttaca tcaatcttca ttccttcttc accgtcgtat 42540

agatacttag ccatgtctat tcctcctcga ttcctttgat aaattcaatt gcttccttag 42600

ttaaggccaa tggatggaaa tattctccat ctgaacagtc attgatgatc ttccctttta 42660

ataggtgctc cacatcattt ttagtgataa aggtggtcca gttgctatcg ttgccatatt 42720

catctagttg cttaatatca tggctaaatt tatctttttc agaaaccaaa gtcatcatta 42780

aacacatcct catctgttaa ataatcatca atattttcac ggaaacaaat tgcatatgcg 42840

tctagcaatg catcggcagc atcaatctta tttgaataac ggttcttatc aatccggaca 42900

ccattgttat ctgatttaag gatcgcgtta gccattgcgc cagttaaaat ttcattgtca 42960

gaatgtcgaa ctcgcttatc taaaatatca tccctaaatt gcttggttgg cattgaaaga 43020

gttagtgttc cttgacgaac agatatctgt tgccattccg gatgaccttt ttcaatctga 43080

gttaacaagg ttccatattg tgcggggtca taacaaattg cctgaacatc aagattgtgt 43140

tctctgacaa agccatccag ccattcatat acccgttcaa catcaataac tccagattca 43200

agcttcgtaa tttcacactg gcccatccct tgcaaccgga cataatctaa ccggtctgct 43260

ttaatctttg cttccaaacc atatttagta ccgacaaacg cataagaatc cgcataccaa 43320

tacccctctt gaggaattaa ccaactaatt gcataaaggt cagatgattt accaacgtca 43380

atcccaaacc aaactcgttg accatcaatg tcaattggat caatttgagc tgcattccaa 43440

gtatcaatat ccatataact atcctcttcg gcttgtcgcc acatattaaa gttcttaact 43500

agaacagaat tctttgtccc tttctgttta gcttctttcc accgctttgc caaatatcca 43560

tatacctggt cttgcaaagc cgggacactc agaattggat tggatttaat ccaggtactt 43620

ttatcatcaa cttctgatac attatcttgt tcagcaatgt aggcaaaata agtatcatca 43680

gtaatttcac ctttcaaaac cttcgttgca tagggatatt caattgtatg cattggaaca 43740

ttcaaatcaa agccggctgt tgaaataatc atgataagtg agttatgaag taaggcctga 43800

ccggattcta gtaattccat catttcagtg gtcttacttg cagcatactc gtctaggatt 43860

ccaacatgag gttcaaaacc atcaaccgtt cccgtttcct tggaaagaga ccgaacataa 43920

gaataatcat caaggttact aattaaatca cgattaactt ttgtcccacg ttttgtatca 43980

ccatcactgg aacgtagagc attgaggcgt ttcttaatca tattaaaaac gatgttcgct 44040

tgctttttat cgttagcggt acaaaatatt tgccgggaga attcaggtga gttacccatc 44100

aagaactcat acaatgcaat tccggaaata aggatcgact taccattttt tcgtgccatt 44160

gataacattc cttttcggaa gcgccgttca gatggcttgt cttttttcca ccagccatac 44220

atattagcaa taatgaaacg ctgaaaatct gctaaaggat acgcccgcat cgtttttgga 44280

tccggaagta tttccatgaa ttgaataacc ttattagcac gttcattatc ataaaaatat 44340

tcaaagtcat cgttatccgc ctttttaaga tcatttaagt acctttttgc agctaaaatc 44400

acctttttac cagcaactat tttgccggcc acgacctttt cagcataatc tttagcgtaa 44460

ttcatcatga aatcctaaac ttatcacgca aggattgatg ctcttccttg tcagtttgtg 44520

gcatgttcat ctgcagccgc gaattaacat taagcccaag atcagatgca agacccttta 44580

ttgccctggt tgctttatca attgtcttaa ttgcgttatt gattttagca atgttctttc 44640

ttttcttgga ctgttcctta tttaattgaa cggaagcatc tttataaatg ctgtaccagg 44700

tacaatacaa ttccaattcg ctccggtcca agttacgcaa tggcagcttt ccgatagaac 44760

caataattcg ccgatattcc gctttggcca ccttatctaa atgtgccggc ggagtctttt 44820

gcaactcagg taaaccatcg gcagccataa attctgcctt atattttgct tcttgttcaa 44880

ccactctaag gtggcctgtt gactgcgata ataatttttg ctttcttgcc aa 44932

<210> 12

<211> 12924

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 12

ctatacaagc gcccgcaatt gttgaatcat gcttaacact tggtaagggg tatttgacat 60

gtcaatacct ctattttgat accaaaattg cgccaacatc tgaacagcga acgtatattg 120

ttcatactgg gttaagtctg ctgtggggtt tacagcgttt tgtacataag ccttagcgtt 180

ctttagataa tccttgatta attggtcttc ttcttcaccg tcaagatata aggcttgctt 240

aaagttgtct acttcccact gctctacatc aaatttcatt aatttcacct tcttttagaa 300

cacatctagg tctaactttt tagcaacatc tactataaat tgatcacgta acttgtacgc 360

ttttgtagtg ctacaccata tatcgccagt tgctactagg ttctttatcg tcttatgatt 420

agggttaaaa tagcattctt taattatctt ctgggtacag tccccagcgt cttgtaatgc 480

taagtcaata gcttctttct cacgttttaa gcggtgcata tatctatctt ggtcaatggt 540

tattaacata tcatcgtaat aagtccatgg tcgattgata gcacgtccac cacctacatt 600

ttcatctctt ttgcggatag ggtgcattag ttcaaactta cggcttttta gtttcttatt 660

aattgaagga taatctttca gtattactat tactagactt tttacttctt ttcgcattaa 720

tacacctcat aaaaaaagag tgggcgatat tgcccaccct gtgctagtag ctattattta 780

ctttcggtcg ccttagttgt ggttaaagag agattaaaag cagcgtcatt actaattggt 840

tggtagtcat tacggacgat aacagataaa ccttgactgt aactgtcgaa cttgtcccat 900

tgggctgtaa cttgattacg ccggaatact gctaccgctt ctgaaatatc acctaaaatc 960

attggatgtg aaccgtctgt gttatctggt aagaacttat tactaatctt aacaacgggt 1020

tggccaagta aagaaaatcc actaggagcg gttacatctg gttgtaataa gtaacgtcct 1080

tcattgtctt tcaaagtatc taatacttgg aaagctgatt gatttactaa ccatgtagaa 1140

gtactttgta aggctgggtc aaggtcaacg ttgaatactt gcttcaagtc gtcaacatta 1200

gctacggttt tcttagtaaa gttatcgcct tgtaatacct tgataatttc agcgttatca 1260

gtgttatcaa ctaacttttg aagttggttc tttacttcag atacaatatc aacttctgaa 1320

tcgtcaacaa cttcatttga tagtgcaatc ttaccggcac gggtctttac atcaaacttc 1380

acatcttcaa acatattagc gtctacatca gcaatttctg cttgttcttc cttagtagta 1440

agtaccgctg tgttgtaacg ggtagcgatt ggataatgac cggaaccagt ccctacagtc 1500

tttacagtag cgtactttgc taagttataa gcactgttct ttaagtctag gactggagta 1560

acaacttctt ctggaataac ggcttctgcc cccttagttg ttaatccgtc acgggtttca 1620

ccacgtgaac gaatgaagct ttcaaaggcg ttagctggtt gttctgtatt gtttttgggg 1680

tctaagattg tgtgttgcat tttattatgt tcctcacttt cgataaattg gttgtaacta 1740

cgaatgcttg ttttatctac atcaacattt gtctgatcat atgaaggcac tgtaacggtt 1800

gaaacctcaa ataaatcttt aatctgatta acggtgcgtg taatttgtcc gttgtcgtca 1860

cgtgtccaac tgtcagaacc atcatcaaca tcaaaacgga aagacataga attaatgttc 1920

ccattcttaa cattattaag agtgtcactt gcgtaactta cacttgggtc aatagttgcc 1980

ttgaagtgaa gccccttatc gtccacatca agttgtaatg aaccagcctt aacgcttgct 2040

agtggcttag tgtagtcgtg ttgatcaagc ataactacat tggataggtc tacgttatca 2100

agggcgctag ggtcaatcac ttcaacaaaa ccgcctaagt ccttgctggg ttgattaaag 2160

agtaacgcat agccttcaac tgtattaggc tgtgaggctt gcgtttcatc ttggttatct 2220

tgctggtctt ttgttgggtc ttctttaggg cttaccgttt gtaaacctgc gttagttgtt 2280

agtcgtttct ccattggctg taactcctcc ttctttttca ataaaattat ctccattagt 2340

taccggcact aaattaagta aggctcgggc ttcattgcgt gtgtagatac cgttaacata 2400

accatcaact gccatctgct gttgagcgga cgggtcaagg cttaatagct tgctattatc 2460

aaagctaaaa tcttgcccta gcttgtagtt caattcagaa gtaaagcaat cttcatagtg 2520

ctgtaaagta ccttgtaaat actgaacgtt actttgttgt tggttgctat gctcgttttc 2580

tacacctaaa cgctcgggtg gtagtccgaa cgctttcgca atttgacggg tattccagtc 2640

attagaattt actagcttta acacgtctgt attaagtggt aggcttttaa cgtccatacc 2700

atcatcagta acaattgtgt taagtgaatt attgccggta gttgcctcat caaattgttt 2760

tctaatgttc tctttaccat ccgaactaag atcagcttgg tggacttgga caactgttgt 2820

gccatgaatt ccattattaa agaatccagt cattaacctg tttccagccg cctgtatctt 2880

catttcatct tttagcgcat acagtggcga tagtccgact aaaccatcac gggtaaaata 2940

cttaaaatgt aagatgttag aagaggcaat tactcgctta atacgtccat ctgggctgta 3000

tgtgtaagtc agcttttgcg ttacatcatc ttgctggaca gttaatttat tgttaggaat 3060

aaaatttagc gtgtgtgaac ttaaatcaat ttcggcaaac gcattaccat ttaatagcat 3120

ttcaaccgct agggcgtact tgaaacttct gccgtccatt aactcgttag gatgatcatt 3180

aatcatcttg ttatataggc tagaactaga cttaatcgga ttactggcaa gatcaccagc 3240

gatagaacta attgcagcaa atatatcact attacgtaaa gccgaagcac taacgtaact 3300

attagggtca ttactagaaa gacttactag agcgtctaga aaggggtctt gtgcgcttgt 3360

agtcgttgtt ctcttactaa ataggcccat tatctaccct cctttacttg tgattatcag 3420

aatagttaat catgaaagct agaacaatca tttcaatacc aagaaagcca atcccaaaca 3480

accaaccggt taatcctata gcactgatct ccattagaac gcctccggtg atcaacaaaa 3540

ccggtttaat atagctagaa aagattttta gcagtttcat agtattcatt tgtacgttct 3600

tgctcctttc tcttagtgaa gtaatcataa ccagccgccc aagcatccat aagagcagca 3660

aggcaatcta ttttattttg ctttctagtc ttgtctaata acgcttgacc attaaagtta 3720

tatttcaata ttgcatttcc tacgttgtaa gctaacatct gattgttagt gtgaagaatg 3780

ttaccagcaa ttaattcttc cttaaaagtc ccaattggta ccgataaatt cttataggat 3840

tgtgttaatt gaagttgtgg tagtcctgcc cgttcaaaac gattaacaag atcagcccat 3900

ttataagggt caaatactaa gaatttaaca tttagattat acttaccgat taggtccatc 3960

acgaaatcat atacggaatc ataatcaatc aagccacttt ctaatgaact gattgaacat 4020

tctccacgtt tagcagccat ttcataatca attccatcac gttttgattt agtaacaatt 4080

ccaccataac gagatacaaa ggcgtgggaa tcagcgtaaa actttccatc ttctaacgga 4140

actaaccaac taatcgccgt taaatctcgg ctttttgata gatcagcacc aatataaaca 4200

tctctattac taatatctgg agctttttca atcaatccat gttcccaatc ttgcgtacta 4260

atatatgaat tttcattctg ctgtacaaac atattcatat tctttacaag tacgttatta 4320

atattatttt gttttgtggc tacatctaaa tcattttgaa tgtgttccct cattgtctta 4380

gcaatcttag gattactcat caaaggatta gctttaatcc acattgtggg gtcgaatatt 4440

tcttcttttt tatcaagaca atacataacc gcaaagtaac gatcattctt aattttgccg 4500

ttgagtatat cggctgcata gtcccattca tttttgaagg gtgaattaag attaaaacca 4560

gcagtagaaa taatcgctag aataccattg tcttgttgag ccatacctga attcaaggcg 4620

tttagaatag tgttatcttt agcttggtga tattcatcga ctacgcccag tgtcgggtta 4680

taaccgtccg tactgtgggt atcagtagat agtggaattg cgaagctacc tgtctttttg 4740

tcagtcaccc gttgtttatt gattttcact cgttgcctta aatactttga attacgccgt 4800

aagttttcta gttggtcact cataatatca aacgcaaggt gtgcctgctg ggaactgtta 4860

gcagtaaata gtatttgtcg attacgggct ggttgctttt ccataagaag agacacaacg 4920

ccaatcgctg ccagtaagaa gctctttcca tttttacggg cttcacttaa tagaatccga 4980

ttgaagcgca atgaattatc gtctacctgt cgccaaccat agatattaga taagataaac 5040

acttcaaatg gctctagctt caatggttca ccgtttgttt gtggtaagct ctcaacaaat 5100

ttaataacct taccagcata cgtttcagaa taataataag gaaagtcatc ggtcttttgc 5160

tttttcatgt cgtctaagaa gcgctggcaa gcatgaataa ctaacttacc ggcgattaac 5220

tcacctttaa caactgaatt agcatactga atagctctat ccattatgac aacatctctt 5280

caaactcatc agtaggagct tcttcactca ttgccttgtg gacttctagg cgtacccgtg 5340

attgtgggct aagtgccatt tgactatcaa ttgacttcat ttctttaata gccttatcac 5400

gtgccatgta gtaaggtgac tgctgtaatc ctcgttttgt atcaataact accccagtac 5460

gatcaatctc tttagtacat ttgataattg ttgaatacaa agtacagtga gtagcgatta 5520

aactaacatc tagttcgcta ataggggcac tcttctttaa gagtgggaca attctacgcc 5580

attccttttt tgctacatca tcaagccatt taggggcgtg agatgttaag tttgggtatt 5640

ctgccaatgc tttttcagcg tcttcacgtt gtgctttttc tgccttagta atatgacctt 5700

ctggacgggt cttcattggt tgagccaact atttaacctc cttctattta tttggtgaca 5760

ccatgtcact atttgtttaa aaaaataaaa aaaacatcat ttcatattca attatcgtat 5820

tatttaaccc ttattatacc atatttgaat agtttgttaa gttcaatgcc taaaagttat 5880

aataaaaaat ccttgacatt tgaggctcta aattatttaa taaagttgaa aagaaaacca 5940

tacaaaataa aaagaaacgt cttgaaatct tcggatttta aggcgttttt ttgtaaaggg 6000

agaatttttt taaaagaaac ccccaccatt acggctccta ccggtctata ccaatagcgg 6060

gggtattaaa attatttcag ctattcatat ttattttcct ttctcgttga tgatcttttc 6120

ttgtcttttc tcggtgacaa cgatagcaaa gtgattgtag attagttgaa tccaatcttt 6180

tactccaatc ttcacgtata ggtacaatgt ggtcaactaa atcagctttt tttactattc 6240

cctttcgtaa acacataaca caaactgggc ttttctggag cgtttgacgg cttaatttaa 6300

cccacttact cgttttgtag aatctattat agtcatgagg agctttggcg cttctttcca 6360

ttcgcttttt gtatactttt ctattgtatt catgtttctg ttctttggtc atctttttat 6420

agttatctcg ttgccaatgt ttacgcttgt cttcatattc tttttgtaga tattgatgtt 6480

ttttacaaaa atattgacct actggaatca cctctcgaca accagcagca ctacaaaacc 6540

ttactacact catttgataa cctcctatga aaaaagggca tcgcataaca cgacaccctt 6600

agcacttact tattaattaa ttccattgcc ttactatatg cttcaagcct gcttatgtgc 6660

cagcttttaa catcgttaac cgttctctta atgcaagctt caagctccat tgtatcggcg 6720

ctgtaagggt cacactcgct tagtctatac cttgccatca tcacccagtc ttggcgactt 6780

tcctctgcgt ccttgcatac tgccacttgt gcaatacact ttactagctc ttggttgtct 6840

ggtagtccta gatcaatcgc taatggtaat ggactgccat ttactgagac taattttaaa 6900

ctataaagcg tagtatacat atgataatta gaaggatcat caccgcttgc ctcaatcatt 6960

acattaataa agttgttata ctctgggtgc gtgttgactg ttaatttact catccccatt 7020

tcacctctta tttatgacta tcaagcttgc gtaaggttac aatatcatac gccaataact 7080

catcatcaat tgaccaacca gcaactttat agatactgcc attgtattgt gcttctggat 7140

atttcttcat gtccgttaaa tctttcttgt gacgaactac caatataaca tcatcagtaa 7200

ttgaactacc tgcctcacta atagcttgac tagcactgat tgaatagtta ccacaatgta 7260

ctgtcacatc tcctacaaaa cctgtcatga tcataccagt attcgggtta cgtttccccg 7320

taccaccttc atgaccgaat gtgaccttat gctttaatcg tgtaattgcg tatttattac 7380

ttcgcattta attattcctt tctgcctgta cttgcagata caagcaatag tagaacgttt 7440

gcagccgttc tagtgaaatc tactgttact ttatttttaa ctccttaata ggcttttcac 7500

ctatccggtg gtatgtggtc actgcctcaa aataaaagta cggtaaaagc agttttaatt 7560

gttaattttc atctcttcaa ttgaaaatag tttttacttt tatcggttac cccgtttcaa 7620

gtacccacaa tcgcctttat atcaacattt atatggtaac cgataggata taaaggtaac 7680

cgatacattg cagttactta atatagtaac ccgaaactat cggttacctc aaaatgcttt 7740

cggttacctt taaatagctt tatatcaagc tttataagaa cttctgcaaa ggtaaccgat 7800

aaatataaaa ctataactaa tttttaagag caattccttt ataaccttta acatttttac 7860

cgttaatctt aactgtagtg tgtttgtatc ctcgtgctat tagctctttc gcaaacttac 7920

gattactaag acaaggaaca gcagtatttt tacaatactc tttgtaatta tcatatagat 7980

atactgattt ttctttatct ttctcatttg tagtacagca atcattaatc caattaccga 8040

tattgtcatt agcatttacc cagcttcgct tttgctgttc catgtattca gtcactggaa 8100

acttaccaga atcaagtgct tttttataag attttaagca ttcataggca aatgctggca 8160

tttctctttt aaagtcttgc tctttgaatt gctctttaaa atgtttaatc ttatggaacg 8220

ttacaataat tggtcgtctt ttaaatccgt ctgtaaaatc attgaaggct ggtaaatcat 8280

tagcgctaaa aatcaattta gcttcatttt caagtttaaa tggatcttta tatttaaatt 8340

gagctgttgt agtgtcgtca ccagttaatg ttttaataat attagtctgt tccataaatt 8400

tggggcttat atcagcaaaa acattagcag ctttatgata cagctgagaa gtagcaaatt 8460

tagcttcttt ttggtcggat aatgcttcta aagaaacgtt tgaaacgtta ggcttcccta 8520

ttaactccat tagtttattt agaaatgtag atttaccatc cccacccaac ccgtatagaa 8580

tcatataaat ttgaaagtcc ttaaaactac ctgctaaagc aaaaccaata aaggctttta 8640

ctgtttcaat tgctgttagg tcatattcat cttgtagttt tccagtatca tcacggattt 8700

ctttagttga gggtactaat gattgtttta accatttatt ccacgttgta gcttttcctt 8760

ttgtttctaa ttcatatggt ctattttgta aaagatagtt ctctggtgaa ttatcttcta 8820

aagtgtccgt catcaagttg taagtgtgtt taccaaacac tgctttataa ggtgctggtt 8880

gatcaaatgg attgtcttct acaatttctt gtgaatctaa ggcaattaat ttagaaaccg 8940

atgctaataa gttcgttctc catattccat gaggtacaat ttcatttttg attgattttt 9000

caatgactga atcagcgtaa tttttccatt gtctactttt aggatcgaac caatagccgt 9060

gttctataaa ttggttggat gtaaagtgat actttttttc taaatattta gccaacttaa 9120

tttcatcaac cagtttattt cctttatcac ttacagttaa ccattctgga aaactattgt 9180

tatttactaa gtcatcaata ttagccatat ttttcctctc tttacatggt aatttagcgt 9240

ttgtatttaa gactgataat ctctacttcg ccttttgttt tgatatattc aaagcttttc 9300

ccgtctaaac ttgtttcttc gtatatattg tcccaatcga ctattgattg atcttcgttt 9360

tgactagcat tattccaata cttaaagagt ggtgtacggt gcagtattcg gttaacttgt 9420

ttaacgtcat cccctgtgta gtagagaaga taaaaacaca ttgcaccaat agcaggttta 9480

acgctatgat atttatttag gtagtaggaa attatttcct cattattttt ataaagagca 9540

ataaattttc tactactttt actactaaca gctcgttcta taaccttgta atcatcgtta 9600

tagtattcaa gtgctttctg aaactgtttc tttgcgtact cattaagggg atgtgacgtt 9660

gaattaataa ttttttccat cttgtcacta tctaataaag gattaacgga tagtttttct 9720

tcaaaactta acactattaa tcacgtccca cctattgcaa ttgatcatga atttgatcta 9780

acttatccaa caaaatacta gatagtgcta acaatgcttt cttacgtgat actactgttt 9840

gagcaaccgt agacacttgt tcatcactca ttgctgcata attgtctact cgtacaattg 9900

gagcaaaaac atcatttaac tgatcataca gtgcttgggc tattccggtt gtttcttcaa 9960

cattggaatc tgcttcccac aactgatttt caaacgctag ttcttggttc ttatcttttt 10020

ccattgttta aaccctccac tcatgttaga atgaaagaga attaaatatt agtaattcgt 10080

ccaacatccc ccgtaaagat tgttggcttt tttattagct aaatttaatt ctctaagttt 10140

tatttagtcg ttgttagtgt ctgaattggc gttctcacta tcaacggctt ttttattacc 10200

atccataaag tcgttttcaa aatcagttaa agttttatat tgaattgcta aatctttaat 10260

ttctttagcg accttaataa tttcatctgc tgaggaaatc ggatcatctg caattatttc 10320

tccattttct ttagtgatag taccttcaaa acaatcttca gtaattgaat ttatataact 10380

gccagcttta acaatcctgt cttctatatc aataattctt ttacttaata tttcttattc 10440

aaatttttta gtgtataaac tcatttctta ttactcctta aaataagtta ctaattgcaa 10500

caacaatacc cagcaatggt aaaaacgttg ctagataagc ccaaaggcgt tggtctggtg 10560

aagtccacca gtaagctaaa atcttaccat caattttttg taacatatgt ctcacctcct 10620

cctatttatt tctctacaat cacttgccag ccagctgtgc gattattact tagaattttc 10680

ttttacccaa cggtcaacat cttcacgttt atatacatat tttcctccaa cctttgatga 10740

tggaaaattt cgggtcttaa tccaacgatc aatagttgtt cgagaaacac ttagataatc 10800

cgcagtttgg cccttattca taaactctgg ttcggtttta acagtcttag catcttgtaa 10860

cttattgttc tctagctttt tagccacagc ttcagctaac tgatcaatga actgatctaa 10920

aatagctgta ctcattatta tcacctcttt atttcatatc gtaattacaa tatataacat 10980

aatgaaatat tatgcaacaa caaaataaaa tatattgtgt tttgtattgt tattttcggt 11040

tatgtataat atattaacta tacaaggagg aattaatgtg aaagaaattt catctgacca 11100

gaaaataggt tctaacattg cactaattcg taaagctaaa aagatgactc aaaaagactt 11160

agcaagaaaa attggaatat cacaaccagc cttggggaat tatgaacgtg gccagcggat 11220

tatacctgct tcgatttttc acagtttacc tcaaatatta gatgttcctc tagattttct 11280

agtctatgga attcccaaca atattacctt tcatgattta gcaacgctcg attccgtatc 11340

tctcaaaaag aaagctgctc atattgaaaa tgatagaagt attattgaac gtgggttagg 11400

gcgtgacgac ataaaaagac atttattagc tgctgttttg cttgataatt atggttatct 11460

tgtgtaccca gatgaatcta gcaatgacaa agaacatgaa caacttaaaa aatttgtttg 11520

gaatacatta gatggattac ttaccggcgt accaattacc aatgaaataa attttaaaca 11580

aattaatgaa caatttgaat tacttgcaaa tacgtattta agcctaatta ttgacgaaga 11640

ggcaaatcgc tatcctgctt tccgcaagga tttaccaccc ttcaaagaaa aacttacaca 11700

gtttttaaat gataacggat attttaaaat tttaaaaaaa taattttaaa cctttctcta 11760

caatctcttg ccagccagct gtgcgattgt aggaggaaag taaaaatgtc agtatacaag 11820

gataaaagaa atggtacgtg gtatgttaaa taccgcatta atggtaaatc aaccagtaag 11880

cgaggtttca aattaaaatc ggacgctgta gcgtgggaac aagaaaaagc tgtggagctt 11940

aaaaaatatg gtcaaatgaa taaccataac atgaccgttg aagagctatc aaaaaaatgg 12000

ttgccagtat atgaacagac tgggatagaa aattctacta ttcataaaac aaaacaaatc 12060

attaaaaatc atattctacc ccgcattgga aactataaag ttgctgatct tagcattgag 12120

ttattaactg aagttgctgc taaatggaaa gaagaattag taaagagtga cccttttaac 12180

tatactaagc gaatgttaga ctacgcagta caattaagag cgatcccaac taatccaatg 12240

aattctgtag taaagcctcg taaaaaacat gacaaaacat ttactaacaa taatttcttt 12300

aatgaaaagc aacttcacaa ttttatcaag tgtataaaaa aagactacga agaaaaaaat 12360

cctcgtgctt tcatggttct atggttagcc ctttttactg gcatgagaaa acaagaactt 12420

caagccttga cgtggcaaga tgttaaattt acaaaaagcg gtggaataat ccatattaac 12480

aaggctatta agaacgcaaa acatccctat cttggtggcc ctaaaaccgc taactcttat 12540

cgctggattt caatagataa aaaaactgct tcctatctaa aaagatggaa acagcaacag 12600

atagatattt taactaaact aggttttaat ccacaccaaa aagagcaatt gattttctct 12660

acctacacta aaaataaaat tgttttaggt gcagaattgg acaagccgtt aaataaagta 12720

attgttagaa ataatctaaa gaaagttaca ttccatggat tacgtcatac ccatgctaca 12780

catttggctt ctattggtac ccagccaaaa ttaatagcag atagattagg cgatacaatg 12840

gaaactgttc ttgaagttta tattaatgca gatactgaac cagaacaagg aatcgccgac 12900

aagttcgccg atagtttagt ataa 12924

<210> 13

<211> 36497

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 13

ttaaaattga ctaagtgcct ggacgatttt gtcatcagtt ttacttttat attcatcaat 60

caagtaggcg tatgtgttca ttgtggttgt gatatcatta tgtccaagtc tcttgctaat 120

tgcataaata tctattccgt tacttaacag taaagcaacg tgactatgac gtaaactatg 180

gaaatgaaaa ttcttctttg ataattttgt atcactcata atttgtcgaa gcgttttatt 240

taatgctgta ctagtaggga tagtattaaa ttgatccata aaaacaaggt tacttgatga 300

attattgcgc aattgtttaa gtagtaatag tagtttttca ttaactttta ttttacgtgt 360

agatgattca ttcttggtgg gtttaaagcg atgtgtatta aaatgttcgt ctgtttcgtc 420

tcttcctttt acctctctcc atgctttatc aatattaatg gtatgatgaa tgaaatcaat 480

atcattccag gtaagagctt gaatttcttc ttttctcatt ccagtatata tcgcagtgac 540

tatcatataa cggcttgtat atcttctatt agtaattcca ttgattgtag tagttaataa 600

tttccttatt tcagttacat tagggtattc taccttcatt gttttgctag tattggcagc 660

taaagtaaca cctttagtaa aatctttaag cagataatca tcaaggatag cagattgtac 720

gcaagaccgt attattgaat taagcttctt tacacttgct atagcatgag ttgctccata 780

cttgttaata aattcttgat atttagaacg gttaatttct ttaatagaag cttctttgaa 840

ataatcatta attagattac cgattataat ataacgattc agagttacac ttgatatttt 900

aggttgctta taagtattaa cccattcttc gtaataatca ctaaaagcaa tttttttctc 960

aatgtgtatt ccgcgattaa gattagtttc catttcagct gcccactttt tagctagtgc 1020

ttttgttgca aaaccagctt tggactttga aaaacgtttt ccttcagtat ctttccactg 1080

aattcttaca gaccattttc cgcttctttt atatatttga gccatacttg tccctccatt 1140

attaaattgt gtataatgaa agggttgata gaatttgttt ggacgcaatt aatatcaacc 1200

cttggtccac tgacagttgc cgctgtcggt gggctttttt agtttaaata gctttttacg 1260

tcatcaattt agttggacga gaaatttagc ttttgtttag cttctttaat tgattaataa 1320

tttcttgatt ttgcttaatt gttatccaat tttgctgttt gattattctt aattcatgaa 1380

aaagggcttt tgtatcatta cttgtaaaag taggataggc acctgttaat ttttcggcaa 1440

ctgctatatc ttttacagat tcgagaattt cgtcatctaa actttctaaa cgattttctt 1500

taatatattt atctttgtct ttagcaaatc catctttacc attgaaaata ctcattttta 1560

gctttctcct atctttttaa tttttacatt gtaaagagcg ttatttaaat cattacagtt 1620

cctaagatag gaaaaagcgt tttttaaaag tttattcatc ctttgactcc ttatattcct 1680

aaaatttgtt tctttttagc atcaaattct tctttgttaa tgattccatc atcaagtagt 1740

tgttttaatt ctcttaattg cgaaaggtca ttactagaat tggattgtgc tgatgtctgc 1800

ttaaaatctg cagctgcctg tttaatagtc tcagccatct ttttagcagg gtagggttgc 1860

atattttcta tttgagttgt aatagctccg ttggttactg caatagatcc caacattaaa 1920

cctttagaat atgaaacgcc attaatcata tcaaggggaa tgtcagttga tttactacca 1980

taaatcaggc cgtggtcaag aaaaattacg cgcttattag tacaaaccgc aagaatggaa 2040

ttagtttcaa cgaaagcatt agcagcataa agtatttttt ctccaccatc gacatctatt 2100

atgtctggca aggctttgat ttccttttta gtaccgaata gattttctac attggcatcc 2160

tttaactgtt gcttaataac atccaatttt tcttgttcgc ctttgttgat ttcgtcaatt 2220

gctgcttgta aagtatcagc ttttttacca gcgctatcag caattgaatc aagtttctca 2280

gttactttag atagctttga tttatacaaa tcaggatgtt ctttgtaagg tacggtagaa 2340

tgaatcccta ataaactttc aatttcacta gcggtatggg aactaatgaa attgctaagt 2400

ttaattggaa cacggtaagt atttgctgga cgtttgggat caagtctgtt ggcatctttg 2460

ccacaaatta aatatccatc tttggtaatg caagaagcct ttaacattcc cagggcatca 2520

ttacatatga agcatctctt tttactcatt tggatcacct tattccattg tcactgaata 2580

tttaactaat ttaccaacaa ttcgacctgg atgctcttca tcaagaatta ttggatcata 2640

gtttttatta tctggcatta gcataactaa attgccttga tgcttaatac gttttaaagt 2700

agcttcgtta tcatcgtcaa caagaactgc agctatctca ccatcttcaa cttcaggttg 2760

ctctctaata atggctaatg cgccattagg aatggtaggc tccatgctat cacctttaca 2820

acgaagacca aaaagtgttc cactaggaac tggtttttca aagatttcat cagtataacc 2880

ttcgatattt tcttcagcag taatcggatc gccacaagca atttcaccaa tgattggaat 2940

tgaaacaact tctaatcctt ctttaggata aataatatta gaaggtttct tttctctggt 3000

agggaaaaag tcatcaacat ttacattaaa aatttcagct aatttaaata aaacatcttg 3060

atttgccttt ctagatccta actcatatct agtcacagtt gctcttgtcg tatcaagttt 3120

gtctgcaagt tgttctactg aaaggccacg ttgttcacga aaatttctga ttttttgacc 3180

aatgaattta cttagttcca ttttatgtac ctctcttatg taactaatta tataacttgc 3240

gttaccgaaa tggaatattt tttatctttt tcgaaaaaaa tagttgcatt tgttccagaa 3300

tggttttaat atataattgt tccaaattgg aacggaaagg agagataata tgcagtcaag 3360

attatatgat ttacgaaaac atgttaaagg aatgactcaa caacaaatgg ctgattatct 3420

aaatattagt gtcaaagctt atcgtgataa ggaaaatggt aaaaatcaat ttactcaaga 3480

tgaaatgttt gctattagta aattatttga tcttaatatt gacactattt ttttgcctcg 3540

taagttccat attggaacaa agggctaaga gaggtgacac aaaatgcaag catcgctaga 3600

tgaacaagat taccaagtaa ttactaatga ggttctcagg cgtattaagg aatgttacaa 3660

cttagtgcct aaacaagatg tccaaactga caaatgggtc ggtatcaagg aattcacgag 3720

taagttgccc gttataaagg acaaggaatg ggtcagaatg tttctcctta cacttccagt 3780

ctttaagaac tgggttatca atttgaatgc aggtcaaggt catcgaacta aggtaaatgt 3840

gaccaaggca ttgccttgga ttatgtctca tcaagctgat attgattgga accagtcatt 3900

gccacgttag gaggtgaaca gatggcacaa ccgatatttg taaaaggaat ttttggaaga 3960

cgaattatag tttctcaatt tgaacaaacg tctgatttta ttttgcaaaa gaaaaacgct 4020

gacctaataa aagaagccag cggattttca aaaaacaaga aattgtttac tcgttatcaa 4080

ggtaaagact ttcaaattat ggagcaatta aagtggcaac ccagtgttct ttttgatgat 4140

gccagtaaga accgtagtag ccacagattt aaggacacca atggaaactg actttagagt 4200

gttagtagct ttctttgttt gtttccaaac ctcatcatct cgaatggaat caagtagttc 4260

atgcccagac caagtgatgt catagacagt taattggcga accattttat taataaatgg 4320

tttggcacta atgaaaccgc ctgagtagag ctgataaagg gtataaacaa tatcattttt 4380

atcaaaggtc tcattagcaa taacatttgc aatttgagaa tctgtaaaac cttggtaaag 4440

catttgcaat tgctgttttt ctaatgataa aagaacatac ctaacacagt cttgcttcaa 4500

tttcacaatt ttcaccacct ttattttgaa ttaatcaaag tatagcagaa agggagagca 4560

gttaaatggc gttggtaatt ggattaccaa tattaattat tgcaatgtgt ttgttatatg 4620

cattggtgta cagcttattg tatgagcgga ataaaccatt gctattgaaa gaaaagtatc 4680

ggaagaaaca ttgaaaggag gtgatatgaa ctttggtaaa tttatttctt ttgttgtttt 4740

ttatctttgt tataatgccg cttttgtatc aatttttgaa ttacatcaga aggcaaattg 4800

aaaatttctt ttggaaaaaa agccataaaa ataagaacga atgaaatgaa agtagctgta 4860

aattcaaatt gttctttggt ctgagttatt aatagcccag taatagtaga aagaataatt 4920

gctatttttg aaagatcatc caagtatttt tcgcttattt gaaatcctaa agtttctgta 4980

agccagccaa ggataaacaa tggaaaagaa aagaagccag ttagaaaaag aatagtaatt 5040

aagcaatcaa atattctgag tttttggaat tcaaaaatta cagcatctgg attagcagca 5100

tatgtaatat gtagagctaa ataagtaaaa cagatgaggc agaagataca aaccataaag 5160

tcggttagtt tcttactata gtaattaaat ttggttaaaa acttaaaaat tttacgcata 5220

atgattttcc ttttgtactg agttaatcaa agtatagcaa atgaaaggat ggtagtttaa 5280

tgagtaaatt aattgcgcta atcgttggtg cttggatcat gtattgttca atgattggct 5340

cttatgatgg agcgatggcg atcctagcag tttatcttct gctagtggtt cttgatccac 5400

taaataaaaa aggtacgact gccgcaaaca gtcgtaccga taaataaata atctacgagg 5460

taattatata tgaatgagtt taatttatca aaattaaatg ccagggttgg cgataattgt 5520

gtatttgtat cgaatttggc agttcgctat caaagcgctg ctactcctga agagcgaatg 5580

gcaatggcca tcaaaatgga gaatgctgct acgatgttac gcatttcagc tgagcgtttg 5640

gccactgaaa ctaagaatgt ttacggaggt aagaacaatg actaacgaag aaaagattaa 5700

ggcaattaaa caaattcttg gaccagaata cgaggaggta gccattttcg ctgctaaaaa 5760

atcagaaatt cgtgacgaac gtactaattc attagttgat gcggatcccg gaacggtagc 5820

agccatgatt atgaattggt tacatactaa tccgatagca gcttcaattg tcaaagcaac 5880

tattgatact ttggaaactg atccaactgc tgatttattt gcacagattt ttctaggagg 5940

taagaactaa tgaatttatt tgaattaaac gacaactaca aaacacttgc tagtcgggat 6000

gacttagatc caactattct aaaggataca ttggaatcaa ttaaggatga tcggaagacc 6060

aaattggaca atcttgcttc atgggcagat catttgaagt cagaaattga ttttatgacc 6120

gacaaaaaga agtcatggga agaagaaatt acttaccgaa aaaataaact tacttggatc 6180

aagaaatata ttactgaagt tcttgatgat gccggtatta agaaaataac tactgaaaat 6240

cacttactta gtgcccggaa ctttaaggcc tcaaccatta ttgatagtga taagaagctt 6300

ccggataagt tcaaaattac tgagacgact actaagccag ataagcaggc catttaccaa 6360

gcactcaaag ctggagaaga agtaccagga gcacatttaa aagctaaccg taacacggtg 6420

attaaataat gtttgaactc cgtgattatc agcaagaaac gattgataac atcatgaatt 6480

ctataagtgc tggtcaccgt tctatcatgg ttcaacagcc gccacgaacg gggaagacag 6540

ttattatggc cgagattgct agacgagcaa cggcaaaggg taaccgtatc ttgttcgtgg 6600

ttcatcggca agaaattgtc cagcaggtta tcaagacttt caaagctaat gatgtaaata 6660

tggatttagc taaaatcggc atggttcaaa cgattacccg acacgttaat aatttggacc 6720

caccggcgat aatttttgtt gatgaggccc atcatgttct ggctaaatca tatcgaagga 6780

ttcttgatgc ttttccgaaa gcttataagt tactgtttac cgctactcct tatcggttag 6840

gtggacaggg ttttactgat gtggctgatg atttaattat tggcaaatca gtcccctggt 6900

taattgacca tcacttttta gcaccagttg attattacgc tccttcttac attgatactg 6960

ccaagttaaa agtaaaacga actggtgaat atgacactga ttcaatcaaa aaagccatga 7020

agcctaaaat ctacgggaat gcggttaagc actatttgaa acttgctacg ggaatgcaag 7080

ccattgccta tacctataac gttgatagtg caattaagtt agccaatgca tttaatggct 7140

atgggataac tgcaagcgcc gtttccggaa aaacgcccaa agaagaacgg aataaaatca 7200

ttgaggacta tcgccaagga aaaattcaaa ttgtaaccaa tgcagaatta tttacagagg 7260

gcctagattt accaaatgtt gattgtgtca ttatgttacg gccaactcaa tcattatcgt 7320

tgtacttaca atttgcaatg cgctcaatga atccacggga aggtaagact gcaataatta 7380

tcgatcacgt gggaaatgtt gaacgattcg ggctgccgac tgatgagcgg caatggacat 7440

tagaaggtag cggaaaaaat aaacaacaac caggaacaac gcttaaacct gtatcagtat 7500

gtccgatatg ttttgcttcg ttttatcgta caagtgatat ttgcccttat tgcggggcgg 7560

cattaggaga agaaaaagaa attgaagtcg ttgatgatgt tcaactaaaa aaagttacta 7620

agtcacggct agcgattatt aagaaaattc aatcgtcagc aattatgaat aatgttgctg 7680

gcaagcgtcc aaacgaattg aagaatctga aagaaataca agcctatgcc aaattaaaag 7740

gttacaaacc aggctgggct taccactacg ctaaacagcg tggatttatt aagaagtgag 7800

gttgatatta tgagtatttt gccaccaaat aagccacaga aggcacggcg agttccaaga 7860

aattacttta tctacggaga tacaatgtcc ggaaagtcat atctagctga acgttttcca 7920

agtccgctat ttcttaacac cgatggtaat agtgagatga acaccgcacc aagtattcaa 7980

ttaaagaatg tccgaaagag cgatggaagc ttgaaagagt cagtgattga tcaactagac 8040

aagattattc ttgctcttgg tactgaaaat catggttaca aaacagtagt tattgatgtg 8100

attgatgatg tagtaacact aattgaacag gccatctgtt atgacaatgg agtagaaacg 8160

ctgggggatg ttccttacgg caagggatat gcacaattta ataccgtctt tcaagcattt 8220

gtcactgagc taaaagcctt accactgaat acggtttaca ttagccggtt aatgatgcta 8280

actgatgaat cttctggcca caccgaagac cgaccatcac taaaacagaa atattacaac 8340

gtggttaacg gaaattgtga tttagtgatt gaaactaagc gctatggtga ccgttatatc 8400

cggatggtta aagatcgacg aattcattat gtcaaagatg atattactga tccggcaatc 8460

ttacgggtac ttgaacatgt aaatggcgtc tttgataagc caaagcagac tactacaaaa 8520

gagcagaatg aaattgttaa caaaattaaa aagcaaaatg taaaggaagg ttaatgaatt 8580

atgagtttac gagatgcaat gaataaagct actgaaggtt ttgatccaaa gaatgattca 8640

gttaataaat ttaagggact ggaaagtggt aaatataccg ttgtagttgc aaaagtagaa 8700

aaccatgaaa ctccttggaa tgctgaacag cttaactttg agttagaagt tgtcgatgga 8760

gaatcagccg gccaaaaaga attcttacaa attggattag atgaattaac ttctaagggt 8820

aatcccaatc caatgctaga aactaattta cgattggttt ctaagttagc agcaattcta 8880

ggtgttgaaa ttcccgatga agtttgggat gacgatactt taatctacga gaacttggct 8940

aaagcatttg caccggcagt aggaaagacc atgattatgg atttgaaggt tcgaccaaac 9000

aagaagaacc cccaatatcc ataccgtaat tatgactttg atgaagcgga acagccggaa 9060

acgccagaag ttacagatga tgaaatgccc ttttaagtaa atatttgagt cagtgaactt 9120

ataacaccgt atggctggga ggccattaag gaggaaaaat gaaaaatcta gttaattacg 9180

ccttagccta tcaagcaaag ggtttaagcg tcctcccaat tgctggcaag cgtccactaa 9240

ttaagtttgc tgatcgtgat ccacttaccg ccgaagaaat aaagaccatc tggatagaac 9300

atccatatgc tcaaattgcg ttgcggactg ataagttctt cgttgttgat atagaccgca 9360

accatgctga taacattgat ggttttgaat caattaagca attaccagcg gaatattttc 9420

cggaaacttt aacccaaacc accaagcatg gtggccgaca gttattttat ttgaaacgtt 9480

cagatatgcg agttaatcag ttaattggtt atttaccagg tgttgatgta aaggcccacc 9540

aaaataatta tgttgtggtt gcgccatcag aaggttacca atggctgaat aagaagccga 9600

ttgttacggc tcctaagtcg ttagtagtga atattaacca aatgcgagcc agtaaccggc 9660

gaagttctcc agatgattta gtttttaaac cacgtgaacg caactcaact actgacttac 9720

ttgagaccat cgcaaatgga ttgggagata aaggaataag aaataaaaac ttagctggaa 9780

tgattggtgc tttattgttt cggggagtag aaccaaagtc tgcttatcaa ttagcgatga 9840

tttgtaatga gaatacgccc gatccactac cagaagaaga agtgaaccgg acatttcaat 9900

caatgctaag acgtgatttg agaaacgggg gtgaaatacg tggcggataa tataattcgc 9960

aaaccaattg aatttgaatt aaatactcaa ggcaatccta aaactaatag tttgaagaat 10020

attggtttaa tccttgatgg cgatccacta ctgcatggca ccttcaaata taacgagttt 10080

gcttattcaa ttgatgttgt taaggacatt ccacagctat ttattgaaaa ggggcaactt 10140

gatgatagct attcagcaat tatgctccgt tacattgagg atgagtatgg ggtgatgttt 10200

caagaaaaat tgttaaatat ggcaatcact gttgaagcaa aaagccaccc atataatccg 10260

gttaaagagt atatggaaaa gtgctataag aattgggacc acaaagaacg aatcaaagac 10320

ttcctaccag tctatttagg agtacccagt ggtgaagtaa caacgctgca gacaaaatta 10380

ttcttagtcg gagcggtgat gaaagtctat aagccggaaa gtaaatttga ttgggtgttt 10440

gatttagttg gtggccaagg cgttggtaaa actactttac ttaaaaagtt agcccatggt 10500

tggtatacag accaatttac ggacttcaaa gacaaggata attttgccaa tatgctgcgg 10560

gcattgattg ttaatgacga tgaaatgacg gccacaaata attctgactt tgaaaatttg 10620

aagaaattta tctcagctga agaattagag ttccggccac catatggacg acatacaatc 10680

cgccggccaa agaattttgt tatggcccgg actactaacg aatcaaccta tttgaaagat 10740

aaaaccggtg agcggcgttt cttacctaac atggctgata agtcccaagc aatggctaat 10800

ccggtaactg atcttgatga tacgatggtc aatcacattt ggggtgaagc tgttggcctc 10860

tacaaagaag gcttttcttt catattgacg aaggagcagc agaagctcat tgaggataat 10920

cgaaagatat ttatgtacat tgatgaaact gaaaatcaaa ttgaacgggt tctcagtact 10980

tgggacgatg actggattga aagctcagaa attgctcatc aattaggtga agataatctg 11040

gttaagaatc gttcattagc caaaaagatt aagtatgtaa tggataaccg gcatgattgg 11100

aaagcaggac aacggcgaat taaaggaatt agtcatcgtg gttatagaaa agtgcataca 11160

gataatacac tatgaatata gttaagtgta tgcagaaaaa acctattaca tcaacgttta 11220

ttaaggtttg catacactac tacactattt taataataaa aaataaatat atataaatac 11280

tatatatgcg ttataaaaag ttgaaagtta gtgtatgcgt gtatgcagta actaaatccg 11340

ttgagagagt aagaattaag ctgtatacac tactgtatgc atagtgtatg caaggaggaa 11400

aatatgaatg aattaaaaaa catcagcaca tcagaattac ttgatgaact gattgaacgt 11460

aatgcattat ttcgtgtaga ttgtggtctt tatcggaatt gggagttgaa gggaaaatat 11520

caatttagcg atattaaatt gcctagcgca tatcctattt atgtaggaaa ttcaattatt 11580

gaccgcatga ttaagtggga gtgtgagcat tgacgagtga acataaaatt caaaacgata 11640

tccgggttgc actatcaaaa catcagtgta cagtgttccg ggtaaatgtc ggttcggtaa 11700

aaacaccaga tggaagattt ttttcagctg gtgtacctag tggtcacccg gacttatatg 11760

gatttcgttg gtcggatcat caagtatttt atattgaagt gaaaaacgaa aaaggtaagc 11820

ctagagcgga tcaaattaga tttcatgaaa tgctaactaa acgagaaatt attcatggaa 11880

tcgctaggtc tgctggagat gcagtaaaaa ttgttgagga aggattgatt ggttatggat 11940

atgaaaataa aaaagaacca ccacaacaag taaaattagt tgaagtaatt caggttataa 12000

cctctcgtgg agctggaaca aaggaggatc cgataagaaa gattattcag tattggagca 12060

aagaaggcac attattagca gaaagttttg gaaactaatt tttctttgag tcacgtttct 12120

ctatgtgctt tagttgtttt tcaggaagac tattataaaa cttatcaatt tgatttggct 12180

gtgttattaa attgtctgta attatattta acagatttaa aagacttttc gccatttctt 12240

ggttatcagt taagtctatc tcaccaggat gaacagcatt atttccaaca attcttacaa 12300

tatcaaacgc ttgttgtaat ttagccggaa gtcccttaga tactagatca ccaatacgat 12360

tatctaatga atgtttattg gaaaattctt ttgttagttt atctattgcc agacgagaca 12420

aagcagcaga tgctcttggt gaatctctaa gtacttttgc cgcttcaata taaatttgtt 12480

taatatcatc gggcatatct ggattaggag ctgttaattt taaaggaaaa ctaggaaaga 12540

taagtatatc ttcattggat tgccgattat gaaaccaaat agaatctaaa taacatgaag 12600

tgcattttgc aactattaaa ttatattcgt attgttttga ttttattttt tctgcatatt 12660

tccaatcaaa ttgagcatat acatgacaat aaggacaaat aataggagct ttccctgtaa 12720

aatcattact aaaatattta gtcattaaat tcacctcaaa taagattata gcgtaaagga 12780

gtagatgtaa tgctacacaa atatagaaag ttaccaattg tggaagctga acagtttgat 12840

gggtcagatg aaatgattga aaggtattca gtgcatgtat ttaatcctaa tttagctaaa 12900

aacatcttct ttataggtat gaacgttcta gctattggtg attggattgt taaggatgaa 12960

tatgggaatt atcaagtggt agctgatgat atattccgta aaagttatga gaggtgcgac 13020

taatgcacat ttatgaagta atcgttgtag ctgtatttgg cacagatatt agccactttg 13080

ttgttgctaa gaatgccgat aatgctaaga aaattattct tgattattac agcactcgtg 13140

atgatggtat caggccaact gtgacaatgt atgacctaac aacaaaatta atcaatctta 13200

ataactacat tgatgaggtg atgcttggat gagattaagt gacaaaatta ttatgacttc 13260

ctttttactg ttattaattg tttcagtcgt cttatcaata gttacgggaa gtaaagtctg 13320

gatatggatt tttctttttc tgatgacact tgagatgctt tacaaaattt ggcgttagga 13380

gcgtgactaa tgaaagcaag atgtggcgat tggagtactg atgtttatcc agttgaaccg 13440

tttgtttatg aagtagcagc tgatggtaag aaagaatttt atcaatcagc atgggatgct 13500

tttagttctg aattgcatat gaaaaatgtg acggcgaaga ttatcaaaat accagttgtt 13560

ccaatgagta atgatgaaat taaagcagag agtgttgcat ttgaattagg cgaaaaaagg 13620

cattatccac atgtggaaga attcactaat agtaaaccga ggaggaaagt tacatttctt 13680

caggttgcgg aagcttttgc aattataacg tggattattt taataattct tataataagt 13740

acggcgttta tgtaggaggt aagttactaa tgacatttga agaggcgtta aaacatgaag 13800

aaaataatgt gccagtattc tatagcgatc gaaaatatta tgtaatcggg cacaatgagt 13860

taaccgaaca atttacaatt cgtgagctaa gtggtaatcc gttatttacg gtgccggttg 13920

atgtacaagc ggaggaattg tcatgagtat taaaattaat gcgcaaacag ttgtatttaa 13980

gggacaaagt tttattccta gcaaaagtaa ctgtgagtat tgtcaatttc catttaagaa 14040

gttaatggtt actaaagcat ctccagatcc atcagttaag acagaagtac caatcaaagt 14100

tgacggtgag gtatttaatt attgtcctca ttgtggacgg aatttacaag ggtgaggcaa 14160

tgttactttt aattgtatta atgatgattg tctttattgc tgggttccta ttgggtaaga 14220

aaaatccatg acgaaaaaag gacccaccgt aaaagatgtg tcctcactaa aaatatttaa 14280

ccataattat tatatcagat agcgagggta cgtcatgcaa acaagtttga atttagatat 14340

tgattgtctg aaaactgcaa gaaaggttac tgactttctt gataagaagc tggatcgcta 14400

tctggcttta tcggggaagc aacgttttga tttgaagtca ccagggatgg acggaatgcc 14460

taaagctccc agccatggta acggtagtga aaatcgaatg ctgaatatct ggttggcaga 14520

agaggttgtt gattgtgtgg gttgcgctat gcggaatatg acaaaggaat cacaacggat 14580

tttactaaat cggtattcgg atcagatgtt aacgtataac attgctaggg agctaagtat 14640

tagttcagca acgtatagtc gaaagcagga aaaagcgctg tgtgagtttg cggaccggtt 14700

tgaatttcag ttagttaagc acggtattca tactgaaatt gatgatctac acgtttatct 14760

tgacgaggaa gattgataaa ttgatgatag attattgagc gagcaattca tgatagaaat 14820

gtgataatag tattgtcgaa tgattcgata ttcatataat aatcttccca aatgaagtct 14880

agctattatg gctaggcttt tgtattatgt ttaattgggt gatttttata tgattgaata 14940

cttaaaaaca tttttggagg ctttgagtat gaagccaaaa actaaatttg ttggagttat 15000

ttttggaata gtacttttat gcttaaaacc atttttaatt caatataata tgaaatggtt 15060

ttacaataag ttttcttgga ttattatttt aattacttta ttttttgtag cctcattaat 15120

aattgaagta atagttgaag tatacgaatg gggtagaaac aagtataaca aacataaagt 15180

tgaaagagat tacgaaaaat atatcttagg cttgtctgat aaaaagttgg caattgtaaa 15240

gaaactttac gctaatgaac atcaccaagg atatttaaga caaaacgata ctaatgttat 15300

tgaattggtt aatatgtacg taattatgca acttaataat gaaattatag taagagaaag 15360

ccaagttgaa gatataaacg atcctgaatt tctttttgta ttacagccac cggctttaca 15420

catcatagaa aagaattcag aaaaatttaa ataaatttaa ttagtttagc ttaacggctg 15480

actttttact ttttaggagg tgagtagcat tactcaaaaa ttaacacaga agcaacaacg 15540

atttgtcgat gagtacatta tttcgggtaa tgctactcag gcggcaatta aagctggata 15600

ttctaagaag acagctaagc agtctggtgc tgaaaaccta gcaaaacctt acttaaaagc 15660

tgcaatcgaa aaacgcaatg ctgaaattca atccgagaaa acagctgata tgacagaggt 15720

gatggaatat cttacttcag ttatgcgtgg tgagcaaaca gaatcggttg ctactgctaa 15780

gggtatttat gaagacgttg aagtgtcggc aaaagatcgt attaaagctg ctgaattaat 15840

tggaaagcgt cacggcgcct ggactgataa aaaggttatt tctggtgatg ttcagattga 15900

tgtgggaatg ggggattatg atgatgaaga gtgaagaaca atggagaaaa atcaaagatc 15960

atcctcatta cttggttagc aataaaggta atgtttacag tgagtataaa ggcggcttgc 16020

ttaaacagat gaaagatgct tatggatatt ctcaggttaa tttaaaccgc cgctccaaaa 16080

aggtgcatcg tttagtagcg gaagctttta tcccaaaccc agacaaattg cctgaagtta 16140

atcataaaga cgaagataaa aataataacc aggtggataa cttggaatgg tgtactagca 16200

agtacaacat gaattatggt gacgtggaga aaaggtcaat tctttcacaa caaagccata 16260

gtacttggaa aatttatcaa tatgatttaa acggtaattt ggtaaaagta tggaattcag 16320

cgagagaagc cgacagacat ggattcaacc gtagaagtgt gtatcgctgt tgtgatgggg 16380

aaataaaatc tttcaaagga tacatatggt caagacaaaa gaaggtgata ccatgccaaa 16440

catcaaacta aattttccta aaccatacaa cgttttcaat aaacaaattt ttgataactt 16500

gtttgattac agtcatttcg ttgaggtttg gtactgactt atggcggtgc atcttctggt 16560

aaatcgcatg gtgtggtaca gaaagttgta cttaaatcac tccaacactg gaaacatccc 16620

cgcaaagtgc tatggctgcg gaaagttgat cgaacaattc aagaatctat cttcgctgac 16680

gtaattgact gtttatcaaa ttggcaattg ttatcgttat gtagagtaaa taaatcaaac 16740

cgtactgttc atttaccgaa cggtgcggtt ttcctgttta agggtatgga tgacccagaa 16800

aagattaagt caatcaaagg gttatctgat gtggtaatgg aagaagcttc cgaatttaca 16860

caggacgact tcacgcagct taccctacgt ctccgtgaac ctaagcataa gaaacgacaa 16920

ttgttttgta tgtttaatcc agttagcaaa ttgaactgga cttataagca atggtttgat 16980

ccgaaagtga aagttaatcc ggaacgagta tcaattcacc aatcaactta caaggataat 17040

cactttttgg acgctgataa cattgcaacg attgagaact taaaacaaac caacccggcc 17100

tactataaaa tctatacgct gggcgagttt gctacattgg ataagctggt ctttccagag 17160

tttgaaaaac gtcggttaag tattcgaacc ttatcacagc ttccctcgta cttcggcttg 17220

gactttgggt acactaacga tgaaacagcc tttatgcacg ttaaagtgga tgagagtacc 17280

cgtaaaattt acgtgatgga agagtacgct aagcacggta tgttgaacga tgatattgcc 17340

cgaataatta aacaaatggg ttatagcaag gaagttatta ctgctgatgc tgctgagcct 17400

aaatcgattg ctgaaattaa acgcgatggt atctcgcgga ttcgtccagc taaaaagggg 17460

aaggacagca ttatacaggg actttcattt atgcagcaat atcacttagt cgttgatgac 17520

cggtgtgtga aaacgattga ggaattggaa aattatacat acaagaagga caaacaaact 17580

ggtgaataca ccaatgagcc tgtcgatagt tacaaccacg aaatcgatgc tatcaggtat 17640

gctttatctg aaatcaacgg aatggctagt ccaaaggcaa ctgtaatgaa aaatatttat 17700

atttaggtgg tgattgaatg gaaacagtaa acggtaaagg acaaatttta gatggccata 17760

tttttatcta tccagctgat gaagaagaac ttgatccgca tgatttactg tcgttcatga 17820

gaagaaatat tcagtatgct aaggattaca agcataatat gcaaatgtat ctaggtaatc 17880

acgatatctt agatcaacag cggcggatgt atgggccaga taatcggcta gtagcaaatt 17940

taccgcatta tattgttgat acttataatg gattctttac tggaatccca cctaagatta 18000

ctttagatga taagaatgag aacgaagcat tacagcaatg gaatgacact aattcgttcc 18060

aggacaaatt gagtgaaatt agtaagcaaa cggatatcta cggacgttcg tttgctttta 18120

tttatcaaga tgagaacgca gacacttgta ttgcttatgc ttctcctaca gatgccttca 18180

tggtttacga tgatacggtt gctagaaaac cttttgcttt tgttcgttac tggaaagata 18240

ctgaaagcgg attatggacc ggaatggttt attacgctaa taaaattaaa acctttaaag 18300

gtagtgttgt tgaagattca gatcaaaata atatgtataa tttagtgcca gcagttgaat 18360

tttatggaaa tgaagagcgg caaggtgttt ttgataatgt gaaaacctta atcgacgaat 18420

tagacagagt gttatcacag aaagctaacc aagtggaata ttttgataat gcttacctta 18480

aaattcttgg tcttgattta gatgaggatg gtgatggtag accggatgct aatttaattg 18540

gtaatcaaat gatttattcg cctaatgctg atgctgctaa tgccgatgtc gaattcattt 18600

caaaaccaga tggtgataat atgcaagaac atattattga ccggcttgtt tcaatgattt 18660

accaggtaag tatggttgct aaccttaatg atgaagcgtt tgctggtaat agttctgggg 18720

tggctttgca atataagtta cttccaatgc gaaatatggc agctaataaa gagcgtaaat 18780

ttactcaggc actccggaag ttatatcgaa tagtgtttag tgctgatcaa gtagtcaaag 18840

ataaggaagc ctggcaagac ttgctctttg atttcaaaca aaacttaccg attgatgttt 18900

ctgaagaagc tgatacttta caaaaactat caggggttgt gtcaaaagaa actgcattcc 18960

gaaatagtcg tttaattgat gatcctaaaa aagaagttga gcgtatgcaa aaagagaagc 19020

aggaagaaat aaaccaagcg cttcaacatt ctgcttctgc tacagatcaa atgctaatgg 19080

atgatcaaaa agaaaatgat aaagagatag ttggtttccg gaagaacggt gaatccgatg 19140

acgaagaaga ataattattg ggctgatcgt attgctcggg aacgtaaatg gcaagaagag 19200

caattaagta aagatgctca atttaatcag cgccttcaac agtattatga tcaagcaatt 19260

gtccagatta ataaagacat tgaagatcag ataaattctt tagctgtccg gaataaagtt 19320

tcttatgctg aagctcaaaa agaagtgtcc actaccgata ttgctgatta tgaaacagaa 19380

gctaagaagg tagttcagga agctaatcgt ttaagagcac aagggaagca tgttacttac 19440

aatgatttct ctgatgaagt taatgaacga ttgaggaatt ataatacggc gatgcgatat 19500

aaccgattga atttattgaa atctaaaatt ggtttatcga tggttgaagc cggaatgaat 19560

attgatgctg atatgcaagc taaaattggt aaagattata ctgatgagct aaaacgtcag 19620

tctggtattc tagatcattc taccgaaaat agttcgtttt ggacttctaa agatgttgcg 19680

aaacaagtaa tgaagcaaat taatggagca acttttagtc aacgaatttg ggctaatcaa 19740

gatactttga aagctcaact tgatacggtt atcaccaacg gaattttaac tggtaagaat 19800

ccgcgagttg tagcaagaca attaagagat aaagtaaaag tcactgttaa aaatcacagt 19860

tatgttactg aacgtattgc caggacagaa tcagcacggg ttcagtattc tgctcagatt 19920

gaattaatca aaaagaatgg ttatcaattt gtccactgga ttgcggagcc aagagcctgt 19980

gatgagtgtc gaaagattgc gacgcaagat aatggctttg gtgatggtat ttatcgaatt 20040

aataaagttc ctaaaatacc agacgatact catcctaatt gtcgctgttc aattagtgag 20100

acatgggtcg atggtcaacg caatatagca ttatctgatg atgaacaggc ggcattgaat 20160

aattatatta gttcagattc atataagatt aatgatgatt tgaggcgtaa taagatttct 20220

aaaaataaga aacaatttat tgaaaactta gatgccgcat tggctaaaat gccaatttat 20280

catagtagca agccactcca gcgtgattat ttctttgata aacaagaagc attggatgat 20340

tttattagta attttgaaat tggtggagtc ttcactgatt catcatacat ttcaacttct 20400

aaaatttatt atggacaggg caaagagaca attcatgtta ttattaaatc aagtaagaca 20460

ggaagagata tctctgagtt taattttaat gagcaggaag tattattccc cagaaatagt 20520

aagtttagga ttgatgatgc atacgttgat gataacggga agatgacaat ggtttggagt 20580

gaattagatg agtaacaagc cttttactga taaacgttgg cgagataatt ctttggaagg 20640

cgttaaattt gataattcaa aagtaacgtc agaacaaaag aagaagacag aagaattcca 20700

tgagttattc aaaaagacgt ttgctaaaca attaaaagaa aaacactcac ataaaaagta 20760

ggtgatccaa tgggaaataa tgatttcttt acggtaacgt acaaaatact aagttatctt 20820

aagtattgtt atgaaaatgg aattaatcct gatcctaata ttcttaatgc tgatacattt 20880

aatattagta aagttcaatt tggaagaact ctacaaatgt taagcgagca tggttatatt 20940

tcaggagtga gatttacaca agccaaaatc gaaggtactg ttgttggtgg actccacaat 21000

acgtcaataa cggttgaagg tctgcaatat ttagctgaaa actcaatgat gaaaaaagca 21060

tatcgaattt ttaaagaagt cagagattgg cttccgggtt tctaagcatt cacaaattag 21120

tgagtgcttt ttgttttgga ctttttactt gttgcagtcg ttaaagaaca acccggatat 21180

tacagtccac cggactataa acgaggtgta tttatgtttg aaaaattacc aatgcgttta 21240

caattctttg ctgaagatcc aacgccagat ccagataatg atggtgcacc tgaaggaact 21300

gatgatggag ataacggtaa aagtgaaaag acatttactc aagcagaatt aaacgatatt 21360

gtcaaagccc gagtcaatcg agccttgaag aataagcaag aggaaattga ccaggctaag 21420

agtgaagcta ctaaacttgc caagatgaat aaggatcaaa agcaagaata taagcttcaa 21480

caaactgaaa aacgtgccca agatgctgaa gcagaattgg cccgttataa aatgcgtgat 21540

acagcgaagc aacaattaat tgatggcggt tatgacaatc caactgatga agatatcgat 21600

ttaattgtta ctgataaagc agaaacaact aaagaacgtg gtgaagcatt tcttaaagct 21660

tataaccgaa ttaaagaaaa tgttcgtcaa gaactattaa agggaaagtc accacgaatt 21720

aatggtgctc ctgctactgc aatgactaaa gaacaaattg caaagatcaa ggatcccgtc 21780

aaacgggtcc aagccattcg ggataactta tcccaatatg aaaaataaaa ggaggaatat 21840

aaaatggctg aaactaattt aacgacaagt acagacctag ttgcacaatc tatcgacttt 21900

gtagaacaat tctctggagg aatccaaact ttattgaatg cattgggagt tattcgtatg 21960

cagccaatga ctactggttc acagattaag atttacaagt cagaagtaac taaggtcgat 22020

ggtaatgttg ctgaagggga agttattccg ttaagtaagg ttactcgcaa gctagctaat 22080

actttgacat taggatttaa aaagtatcgt aaggtaacta ccattgaagc tattcagtca 22140

gctggtggtg ctacacctgc tatcgtggat actgataata agctacttcg agaaattcaa 22200

aaggatgtta agaaggactt atttaattac attaccaagt ctgatgcaaa caagactacc 22260

gcctctggtg acgattttca aaaggcaatg gcggcagctt taggacaact ttcagttaag 22320

tgggaagact acgacacaca aactgtcgcc tttgctaatc cgcttgatct atatgcatgg 22380

ttaggtaacc aaactcttac tgttcaatct gcctttggtt tgcaatacat tcagaatttc 22440

cttggctttg acactattat tctaagtgct gaagtaccac agggaacgat tgctacaaca 22500

gttgcagata atatcaatta cttctacgct ccaatttcat ccgttggtca gttatttaac 22560

atgacttctg atgaaactgg tttaattggt gtaactcatg atgcagttaa taacaatttg 22620

tcatacgaaa ctgttgtaac aatggctaat gtattgacta cagaacgttt ggacggcatt 22680

gtattgtcaa caatcagtgg tgctaagtct gctggtaagt aggtgattga atggaccaga 22740

atacggtttt gcaaaactta aaagtaatgc ttgaaattaa aaatgatgac cgtgatgctt 22800

tgctgaaact aatcattgat aatacagacc aagcattgcg atttaagcta gaactaactg 22860

aagacgaaaa tttacctgga gaactaggtt atattgaatt agaagtttca gttcgacgat 22920

tcaatcgact gcaaaacgaa gggatgagtc aatatagtca agaaggggaa agtattactt 22980

ttaattcttc agattttgat gatttccttg atgatattga tttgtggaaa cgacgacacc 23040

agaaagatgt taaatcttta ggtgccgttt cttttattaa tccttatgcg gggatgagta 23100

aaaatgcgaa aaacacagat aattaagttt tattttcaag atgaaaatgg ctataatcca 23160

tatgcagaag aagacacgat tactagtcct aagctagtag cacaacggta tgccaatgtt 23220

actgatgtag ggacaaatcg cctagttgaa ttatttagca ggctagatca gaacgccaag 23280

gtaattcgat tggagtctcc agtaaatgac tcttggtcat atctgactat tgatgattgt 23340

cctatcaagt atcgtcttga aacctgtcgg aaaccattaa aaggcacaac gctgattgta 23400

ggtgaagcca gtggctaatt cttttaaggt tgatgttaaa ggtactaaag aactagcgaa 23460

tttcttaaag aagaataaag atttgactcc agttaaacgg atagtcgcaa aacatggagc 23520

aggtctcaaa aagcaaacgc agcaaaatat gaacaatttg tataagggcc actacgaatg 23580

gaagaagggg gctgggctaa caatggttag ccctaccggg aatactagac ggtccgtaac 23640

aaatacaatt tctaataatg gtttaacagc aacggttgct ccacaaactg aatatttccc 23700

atatcttgaa tatggaactc gctttatggc agcacgaccg acattacatc cagcgtttgc 23760

aattgagtct atgaaatttg ctaatgattt gaataagtta tttaagtagg tgagaaaatg 23820

tcaccaagta ttgaaattta tgatgcagtt tttgctcagg tccaaaagca ttatcagacc 23880

tacgatcacc caccgcagtt aaatgaacca gttacctatc cattcgtggt tgttgatgat 23940

agccagtcga ttttgacgaa ctataaaaca gccacaggaa tgcgggtaac tttaatagtc 24000

catgtgtggg ggaagtctaa ccaacgtaag actgttacta agatggttga tgaaattagt 24060

cgtctgggga tgcaagcagt tcggacgaaa cattatgctt ggcaaggacg acctaatgag 24120

caagaacaac aattattaac tgatacgagt gttccgaata ctgtgttaaa gcacggttat 24180

ttaacactcg tttttgattt gaaataaagg aggataaata tggcaacata tccagtattg 24240

gaagggaaga atgcagttct ttttgaacga ctattagaaa atgcaaggaa agagccggca 24300

caattgatcc cgtatcaaac atcactaagt tatgatccta aacgggatac tgattcaaca 24360

actacgaaga tggggaatgt tcctactgct tctaatattg aaacagattt agaagtagag 24420

ttcctaaatg caatttccaa agctgcagat gatgtttatg attctttgta ctttaataag 24480

aagattgaag tatggaaggt tcatattgat cgagtccggt cagatggcaa agtttatgcc 24540

gaatatatgc aaggaattgt gtcagaagac tctaatgata acgacccaga cgatcattca 24600

actcgggatg tgacctttac gattgatggc gtggccaaac gtgggtgggt cactctaccg 24660

ccagaaatca aggaagaaat tgactatgta ttccgtggct tggcacagct taaaggcgat 24720

gacgacaacg gtgaaggtga agcttttgct gatggcgatc gtggtgctgg tgcaaatgaa 24780

gcagtaacaa ctgaatagga ggaagattat gaagttaaaa attaatggtc aagaccaatc 24840

atttgtattc ggagttaagt ttttgcgaaa acttgatgct tatcggggcg ctgaacaaga 24900

aatccaagga gttaaggtta agctaggaat ggggctagcc atgatgcttc cccaattaat 24960

gactaaggat gcggccgctt tggcagacgt gttgtactgt gcggctaagt ctagcattaa 25020

gttagataca attgatgatt atattgataa ttgcaaggac ttggattcat tatttaatcg 25080

ggtaatgaat gaaattaagg caagtaatgc cgctaagccg attgcaaaaa atctaaaagc 25140

ctagatggtc ctgagcttag ttcagaacaa agctatcacg aaattctttt gaattcgttg 25200

gcttatctag gctttcataa tatttcagaa attgaagaaa tgggattggc tgaatatcag 25260

ctccggatgg aagcctataa cctccaacgg gttagccagg aacgagactt agcattgcaa 25320

gctttcctta atcagtcggt acaagcgacg aaagggagcg aaaagcaccc aattccgaag 25380

tataagaagt ttagccaatt ttttgattat gataaatttg ttgatgatgt tcgtgggcac 25440

tatgagcctg actatcagcc aacaagcaag gccagccttg aaaagaaacg aaatgatcta 25500

atcgtcaagc ggtggcgtga attccggaag atgaaacaaa aacagagagg aggtaatggc 25560

tagtgtcaca atcaatgaga gttgaagcgg tattatcagc atacgatgag agttttagcg 25620

caaccttaga taaggcgctt aaatcgatta ataatttagg ccgtgaaacc cagtcaacct 25680

ctcaaactgt tagtgcaggt ggttctagta tttccagtac ctttaaatcg atggctggag 25740

caatgggtgt agttgcgatt gctggtaaag catgggacgt tgttaaagat tcaatgagtg 25800

gcgccattaa ccggtttgat acattaaaca agtatccggt agtaatgaag gctttgaatt 25860

attcaactaa ggatgttgca aagtcaaccg ctatcttatc taagggaatt gatggattac 25920

ctacttcttt gcaagacgtt acaagtgttg cccaacaatt agcgccatta actggtagtg 25980

caactaaggc ttctaagtcg gcgattgcct tgaataatgc cttccttgcc tccggtgcta 26040

gtgttgccga tacctctcgt ggacttcaac aatacacaca aatgctttca actggtaaag 26100

tcgatttaat gtcttatcga acattgatgg aaaccatgcc aattgcatta cgtaaagtcg 26160

ccaattcatt tggttttact ggtaagtctg ctgaacaaga cctttataaa gctttgcagt 26220

caggacaaat tacggtagat cagttgaatg atcgttttat caagctgaat ggtggagtta 26280

atggttttgc tcaattagca aagaaaaata gtgaaggtat cggtacatct tttgcaaact 26340

taaaaaatgc cgttgtcaaa aatctggcaa atatgttatc ggcaattgac aatggtttta 26400

agcaagcggg ctttggaagt attgcacaag tcctagacaa catgaagggt agtattaatt 26460

ctgcttttca agttattgga ccagttgtta ctaatgctac tgttgtagtt cttaattttg 26520

caaaggttat aggcggagcg cttaaatctg ctttcagtaa tgatattttt aaaacagcag 26580

ttgtgggaat attaggcttt gtgggtgcag ttatggcagc ccataaggtt atttcaatat 26640

ttacaacatt aagatctgca atagttggtt taagtgtgat tacaaaagct ggtaatttgg 26700

caatggcgtt tagtgaagca atgtcaacac ttgctaaaac ttctaagatt gctggtggag 26760

cgatgaaagc attcagtgcg gtggcctcat taggtccctg gggaattatt gctgttgcaa 26820

ttgcagctgt ggttgcagcc ttaacttatt tctttaccca aacgaaaacc ggtagggctt 26880

tatggcaaag ttttactacg tggttatctg gagtatggca gagtttggtt ggagtggcta 26940

ctactgtttg gaatgcaatt ggtaatgcta ttaatgcagt agttaatttt attaaacctt 27000

attggcaagg attattaaca ttctttaccg gaatctggac atcaattgtg gcgggtgttg 27060

ctccaatttg gcaagggtta gttaatgtct ttaatagcat tatcagtgca attttagccg 27120

tttggcaggc tttagctcca attattgttc cgattgtagc tggtgtagtt gctatcattg 27180

gggcaaccct aattacgatt gttaccgtct ttcaaactgt gtgtaatatg cttgtaccca 27240

ttgttcaagt tgtatggcaa ttaatttcaa cagttgtatc tactgctatt acgatgctag 27300

gtacaataat ccaaacaggc ttggcaatta tcgttgctat ttggaatgtg gtctggaata 27360

cattcagtat tgttgtaagt acggtatgga acgttattac tactattata tctaccgtgc 27420

tgaacgttat tgcgggaata attcaagcta tcactgctgc aatacaaggc gattggtcag 27480

gagcttggaa tgcaattcag aatgttgtat caactgtttg gaatgcaatt gccagtatta 27540

cttctagtgt attgaatgga ataaaaggaa tctttgatgg tgtaatgaat ggtttaaaga 27600

gtattacttc tagtagttgg aatggtatta aatcgctatt cagtgaaggt gttaatttca 27660

ttaaatcagt tgttcatata gatttaggtg ctgctggtag agctatcatg aattcacttt 27720

ggaatggaat gaaatccatt tggaatagta ttaagaattg ggttagcggt attgctgatt 27780

ggattaaaga acataaagga ccaatcagtt atgaccgtaa gttacttatt ccagctgggc 27840

aagcaattat gaatggtctt aataacggat tgattaatgg attcagtgaa gttcaatcaa 27900

acgttagtga tatggctaat caaattcagc aagctattac taatccaggc tttgatattg 27960

gagcaagtat tggtaacttg ggttcaatta attcaaatta tactggtagc ctggcaattc 28020

aagatagtca gttacaaatg cagaataatg ctttgcttcg tcaattactt aataaagaca 28080

cgacaatggt tcttgacgat ggcactcttg ttggctatac agcggatcaa tacgattatc 28140

gcttgggtca aaatacagca ttgaaggata ggtggagccg atgaaattct taaataatga 28200

ctattctttt cgtggattag gacccacaaa ggatgatcca gaatacttag aaaatgcaga 28260

atatatcgac tttgccggtt ttaattcttc tgattatgat tggtggttga ttgatcgaac 28320

agcaactacg ccagaagaac aagaaattac agaaagcgtc ccttacatgc aaggagaata 28380

tgatttctca atgtatgatc aggaacgttt ttttaagacc cgtgagttga cttataagtt 28440

tgtatatttt ggtgaagttt atcaggatcg taaagcttac gaagaggagc ttaaacggca 28500

attactgcca catggtttca ctaaactaat tgattctcat gatcctgttt actactggtc 28560

agctaagtgt actagtgttg aggttgagga tgaccaagaa aagggaatgc tcacagcaac 28620

tattactttc aaggcttatc cttttgctta tactaatcat aacgagggca ccgattattg 28680

ggatgatgtc gcatttgatc attggatttg gcaaccagtt aaattcaatg ttaacggtga 28740

tcaggatgtt aatgttaaga atatcggctc acgaccagtc gaatgctcat ttcaattgac 28800

agggtccata actttgaaga acgattcaat tggtgaagta ggtttaactc aagacaattt 28860

taaaacaacc acgattgtat tagagatggg tgacaataaa atgcatctat ccggaaacgg 28920

gacaattgaa tttcaattta agcgtgagga gatgatttag tgtaccgaat tattggttat 28980

aatgaaccaa cagataaagc aggatttatt gtactggatc cccgagttaa tcgtcatatt 29040

agttcgggaa aactcacgct taaagaatct aatattgatg atttgactat tacggttaat 29100

caagcaagtc cattatggga caacgtaagg ccttatcata ctcatgttaa cgtttatgat 29160

gataatgaac ttatttttcg tggacgagct atcaaaccta aaaagtcgat ggaagaaagc 29220

ggacaattca ttcgtgaata tgtttttgaa gatattgaag catatctcat ggatagcacc 29280

caaagatttt atgaaggtgt tggtcaaacg cccaaagaat ttttacaaac tttaatcgat 29340

gttcataatt cacaggttcc tgactataaa aagtttcaag tccggaatgt aaatgtcact 29400

aataataagg atgaccaata tcgacaaatt gattatccca aaactagcga tgctattaat 29460

gataaattag ttaaatctct tggtggttat attgtgacta cttacaacgc taacggaata 29520

aactacattg actacttaac ggatattggg gttgatcata aagatgatac tcctattcag 29580

ttagctaaaa atatgaagtc tgcaagtatg caaattgatc ctactaaggt gattacaaga 29640

ctgattccac tgggaaagac actagaacca tcaaaagttg atgtaagtga tgatgatgga 29700

gagggcggtt ctggatcatt agatagccct gaagaatttt gtaaatcaga aattaatgct 29760

acttggggta gtgatattaa taatatgaaa caagattttg ccgctcgttc ttcgagagtt 29820

cgggcttggg gagtggacgt taatcgttta tatgatgtgg tgaaaaatgc tggagtaagt 29880

cctgaatggt tctttgctta tgaacttcaa gaacaaggaa cttactatgg atggcttaac 29940

catacttatc gacacggtga tgcgtatagt gatgcgcaat ctgtttgtga gtggattaaa 30000

aattgttcaa atagtaattc cattaatcca gcatggagcg caccggaagg atcaatggcg 30060

ccgaatcaag cattagcgga taaatggaat caagagtttg gaaaaggtac tattggccgc 30120

gtttatttac aagggactgc cgctgctgtt tgggatttag ctggtcaaac gcctaatcca 30180

gctattggaa agccaattag tggatgcatt tcttgtatta aacgttgggg tggtcattct 30240

aatgcagctg gtggtacatg gggatggcct tttcctgatg ttggggaagg tcatttttct 30300

caagttcaga gtttcggaaa tgatggcgga tatcgtcaaa atagttatca cgatggtgtg 30360

gattttggat caatagatca tcctggtaga gaagtgcatt gtattcatgg tggaacggta 30420

actatcaaat cagctatggg tggcttaggt aattttgtgg ttattcatac gccggaagga 30480

ttcaatatcg tttatcaaga agcttttagt tctccctcta atattattgt tagtgttggg 30540

caaaaagtaa aaactggtga tgtaattgga tatcgtgata cagaccatgt tcatattggc 30600

gtaactaagc aagattttta tcaagcagtt cgaaattctt tttctcctgc aggtggttgg 30660

ctagatccag taaaactaat taaagaaggt ggcgatgggt ctaaaccaca agaaggaaag 30720

aaagatcaaa ctgttgataa tagtaatgct gcacgtccta aattaaccat tactactgtc 30780

aataacggta gagactatat tgatattcct gatttacaaa aagaattcgg tattattgag 30840

ggaactgttg aatttgataa tgtagatgat ccgaatgttt taatgcaaca agctcaaaca 30900

tggataaagg ctcaaagaat acctcaaagt tgggaagtta cagctttaga attacatatg 30960

acaaacttca aatcttttaa ggttgctgat aggtacatgt ttattaatcc aaatgttgca 31020

aaaccccaat tattacgaat tactcaaaaa gaaattgatt tactaaagcc ccatgcgtct 31080

tcattaacga ttggtgataa gacgatgggg cttactgatt atcagttaga aaatcaagtc 31140

aattttcaac aatttaagga aattcgagtg atggttaatc aggttgtcca aacccaagag 31200

caatctgcta ataacaataa taaggttatg caaaattttg ctagtagtgc tgatcttgca 31260

caaatgagac aggatctaag aaatcttcaa gatgataacg atcgtgctcg caaaggaatg 31320

gtttccttag aagaattcaa taaactaaag gaacaagtag aaaaactaac aacaggaggc 31380

gatgataatg gcaagtgaaa cctatgatta tgagtcattt gataatacgg atcatactat 31440

gaaacaaatc gctgacgcta ttcgtcacaa gggttatgga aaagatgtgc gtgaggcaat 31500

tgcacagggc ttcgaaaact tagataaaca tttaagtagt attgaagaag aactgaaaca 31560

acaagaaaag aaaaagtcat cgtctatgga tgatattttt aattcttttg gtaagaagga 31620

gtgatgataa atggcaaaag agattagtaa tctgattacc tttaacacct acaaatttga 31680

acgaggcggc cttttggttg atatgtttaa ccaatttaat gctcgtgtag gagatcaagg 31740

aacggaatta gccatccagt gggaaactag taagactgaa actaaaatta atttaaaaga 31800

acgaggatta catttctttg ggacaggttc agttggacag taccttgaaa aattagaaga 31860

tggaactggc tttaagatgt ctgcagatgc atctaccgtt gaatgggaag ataaagatga 31920

agctggtagt ttagacgatg gaattacggt tgttaagttg ccaaaacaat ttttccctca 31980

aaaaggtatt ttctttggtt actttggcct aaaagataga caaggtaata tctttactag 32040

tgttaatgtt tggttccgtg ttcttggcgg tgttccaacg atgggggctg ctattcctta 32100

ctttgttact gaatttgatg aagtattaga gcgatgcaat ggtaagatta ttgacgcttt 32160

agcagaatta cgtgaaaagt accaggcaga agttaagaag aatgaggata tgtctgcgga 32220

aacaagagca gcgttgagta aacttgctga tgctgttggt gcaattcaag cgcagattga 32280

tgcaggtaat gtaattacac gtaaagaata taataatctt gctaatcaga ttgataatcg 32340

ccttagtaaa atgacacaga acattgagag tttttcatca ctcgatgact taaaagctca 32400

atatcctaat ggaaaagatg gactctttgt tactaatgac aataatcata agtatcaata 32460

caagaatgga tcatgggtag atgaaggtat atggacagtt actacctttg atccagaaac 32520

gcgacgacgc cttacatacc ttgatacatc taattctatt ttgcaaaaat cactcagtga 32580

attgactaaa gaagtcgttg atataaattg gtttttaggc gaaattgatg caaatactgg 32640

taaaattaca cctcacgaaa gttttaatcg tgcatactca tcactcaaag taattggtaa 32700

atcacatgct tttgaatttt tgcttaatac tgattatttg caatatatta gtatttttga 32760

attcaattca aatggggaga ttattaagca cgatagtgtt ggcgataaag cgatatatac 32820

atttgaaaag gatacgactg cggtaaggtt tcagattact agtactgctg ctactatgaa 32880

gcaaaatgat ttacgcgata ctattcaaaa tagtgggcta aaaattatcg atcgaggaca 32940

ccgttcagta attaatgata ttgatcattt atatatccgt gaacatttaa tgccgattga 33000

agcaattgaa ggctatacaa ttaacactaa tgttgattat ggtgaaactg ttgatgtatc 33060

acatccaata gtaacgtctg cgtttcaata tattaatcaa gtatgtaagc cgggcgatat 33120

tttcgtaatt aataatctat ctggaggctt taacgcaatg gcttgggcct ttattgatag 33180

cgagaaccgg ttaattcaaa aatctgaagt aaacttatct caatctcagg ttactttata 33240

tgcgccaagt aatgctgcaa aattaattat caataatatg gatagtaact gtacagcata 33300

tagatatact ccggataaag aacatttagc taaaattaat attgctttaa ctaccaatat 33360

tcatcgaaat atcaagttag cttatcaaac aggagaaaca gtctcattaa ttcctgaaaa 33420

agtaagtaat tacaaatata ttatattaga ttgtaatttt aatgatgctt tcagaataaa 33480

gggttatggt ggattgaatc cacgattata tggatttatt tctgagcaaa acacattaat 33540

gaatgttgcg cctgcaaatg ttaatgatca attaactgat gtctttatca aaacgccaaa 33600

aggcgccaaa aaattagtag ttaatttcaa tcttgatcaa caagtaaatc ccaaacaaat 33660

tgctgaactt tataagcttc cagacattaa tgttattact gagcaaaaaa taaaaaagaa 33720

tttattgaat caacaaacag tcgatacatt aatttctagt tatatgaatg gtatattact 33780

tggtaaagat ttagccaatc atcttgcgga ttttgctaaa agcggcgata aaatggttca 33840

tgtatcaacg ttttataaag ttaacgatac tctttttatg tcttattatg ctaatacaag 33900

atcggcttac gaagatccaa cccagcatac agcacgatta gtttatgcac cgtttgagaa 33960

tctaaatcaa caaacgtaca tcgatgtagc cgacattgga caagaatata atggtcaaaa 34020

gattgaagca atatacgatt ctttgcttct taaaacaagt gatgattcat atatgatcta 34080

tgcctttact gctaaggtag gtggtaagtt ctatatgctt taccgtcgtt ttgatcctaa 34140

gactaagctg ttgagcgata ttcatacaat gaattttaaa gttggagtaa tgacgtcgac 34200

atttgataca gtaagcgttc atgatttgtt agctaaggca ggaattgact atgattatga 34260

agatcgtgat atttcctttg tccaaaaatt aagtccacgg attgaagatg gcgtagtgca 34320

atactatgca ggtatcggaa tcttacattt ttgctttgta gtaaaatcaa gcgacttaat 34380

caactggact tttgtcagta ctccagactt tatgtacaag ccagaatttg aaccttctgt 34440

atatgttaag ggagacaatg tttattattt ttgtcgtcag cgagggacag aaggtaatgc 34500

agttttagct aagtataata tccctaatgg tcaatggtcg aatccgattc ttgtaccaga 34560

tactcaatca cgctatgact ttttcgagaa taatagtcag ttgtaccttg ttcattcacc 34620

gctagaccgt aatcatatta gtttgatgca gattgatcag aatgttctgg aaaagagcta 34680

tgaagtagca acagcagtag tacaagattg cttctaccca tttactcaaa atattgatgg 34740

tcagatgtat atgagcttta cacaaagtag gcaacatatt tggctaaata aatttaatcc 34800

ccacagtctg ctagatagtg atgtagcaac tattttcagt aatttaattg agtagcaaaa 34860

catagtcgcc ttataaatac acaatacata gaaagttgca ttgcttaaat ggaaacttaa 34920

tgggcggctt atagactttt attatttaaa agagtatact ggaattattc tttttattgg 34980

gaggaaatat aaaagtgaca actaaagtga gagataaatc gcttgatatt atccggggga 35040

gcatccttct agttgtcttg ggtcatatat ctgggatacc ttttgagtta aaaaagtaca 35100

tatattcttt tcatatacca ctgttctttt ttgtttcggg atatttgttc aactttgcta 35160

aatacaggta tttttcttat aaagagttta taaaatataa agctaaaaaa tatatcctac 35220

cttatttcag aatggggttg atatgtttgc ttctatttgg catagtttat ccactatttg 35280

ctgaagggtt tagtaaacag tatatgcttc aatctacaaa atatgtttta ggtttactat 35340

actcgcgtgg aggtcctaat tatatggcct ggagttcgcc actttggttt ctaacagcct 35400

tatttattgc agagattatt ttttttgtgg ttctaaaatt taattttaaa tatccattaa 35460

tagtgtttgg aatcttagct atattgagtt atatttactc gattacaatt aaaattccgt 35520

taccgtggaa tattgatgtt gcaatgtttg cagtgttatt catgtaccta ggctttatta 35580

cacacaaata taatttaact aagcacatta acttaccggt tttcttactt ttgattgtta 35640

tttttgtatt atctgtagct tacaataatg agattgacat gaatttaaga aactatggca 35700

atggattttt aacaattatt agtggaacaa taggtactgt tatatgcttg caaattgcgc 35760

gattgttaaa agagaataag atattagagt tttatggtaa aaacacatta tttatcatgg 35820

gctacacgta tgctgtgttt aattgtattt tggctttaag cagccatttt agtactgtaa 35880

agaatgttgt agcttcgttt ttgattcaaa ttataatttt aactttatta atagtactga 35940

agaatttatt caaacaaata aaaaagccca tttatgcata tactaggaaa attaacaatt 36000

aaaaaaacta atacgtccta ctcagggcgt ttttatttta ccttcaaagg agatgatcaa 36060

cattgcccta tcatttattt atgctgcacc aaatgcagac attagttgat gataaactaa 36120

tgtgggcttt tacaatcgtg atgattgtag atttaattac gggaatgatt aaaccgtatt 36180

atgcaaagaa aacaattaag aaaactaata gttcagttgg aatccctggg ataattaaac 36240

acacagtaat ttatttagtg gtagtaattg cttatccata tctttatacg attggagcaa 36300

gcacgatggc taccactttt ttaattgctt ggatttatca atatttaatt tcaattgtag 36360

aaaattggac agagatgggg tggtggttgc ctaaaccaat catggatttc tttgaagcca 36420

aattagctaa ggatcaagaa gattatgacc catctaagta caattttctt ggtaaatata 36480

aaggaggtaa aaagtaa 36497

<210> 14

<211> 300

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 14

ttaaagtatt aaaatagatg taaaatttat ttttttcaaa agaaatttta attgtacact 60

gttggtattg aacggggtta aacaaaggta aattagcatt tctgcggatt aagataaata 120

gaaaaatgtt aaagaacacc ttaaaaagat taatttttta taattggacc gtatcaattt 180

gtaaaaaggt tgactttttg aaaaaaaagt ttatcattaa cattgtaaat ttaatgattt 240

acgttatgtt gttatagagc acaggacgta ttgatttata tagaaggagt gtttattaga 300

<210> 15

<211> 300

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 15

atgaatggac agatgtttta atcgctagaa tagaaggaaa gaaagtcgca acaaatacgg 60

tttctagtac gtggcaggaa cgactaggta agcagattga cgaattaata gaaaaacatt 120

agtcaaatac atttacaaat gaacagatag ttgatattat atttaagaat tcttcttcag 180

agcctaagat taaagctttc aattggcgaa aagaagttgt acaatatgta taaaggtatg 240

tcagtcaccg aatcagatga tctggcatta tacttgtaaa ttatcaggag gttttcatta 300

<210> 16

<211> 300

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 16

atctcacgtg cgatccatta cactaagggc gcgtcaacaa atattatact atcttaaata 60

agaatgaatt gcaagcatta tttgaaaatt ttaattaaaa taacgcttac atcagaaaaa 120

tgttgtgatt gaatagacaa tttttttgaa gatggtatca taagtatcgt aggagttgta 180

ttattgctta gaccttacca ctgcgtcact tacaatggtt gagagttgcg atgctgatgt 240

aatgtgataa actaagcaag tacactaatt atgttttttc ctaaaggagg aatttgcagt 300

<210> 17

<211> 300

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 17

ttgtttaaga tatctttcaa agctgcggaa tttttcccag cttttttagt tagttttgtt 60

ttcataagct ataattttaa ccgattccaa atttctttta aaagtttttt tgatctagac 120

cattaattga taaacgctta ccaaagacta atcaacaagc catttagcgg tagtggtcca 180

ttttaacttt ctaagacatc ttctcagaaa acgtttcctt tgatagtgca gattgtgctt 240

taagagtata taattgtcac ggtataagaa ttttctgaaa tttcagaagg agtgaacatt 300

<210> 18

<211> 449

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 18

ctcctctatt attattcctg atcaatttta aattaatctc cctagatagg tatattttag 60

cacaggtcac caacgttcca aagtttaatc tatgtttaaa ctttaatttt caaaaaaatg 120

ctatactatg ttcacgatac tttaaggaaa ggtgattaca atagtgagtc tcttaattgc 180

tattcttatc tgctggttgc tatggaagat tgggggttta acggttaagt tcattggtct 240

aatccttctt attctattaa tcgggacatt aattcatgtt ttactttggc cagcgatcct 300

tttagcagtt attatcttag gagcaggttt attcactaac taatttatct ataaaatctt 360

atagtaattt ttctgcggaa tgttataatc attactgtga gagaaatctc aaataatgta 420

tacataagat gaaagggaga ctgtttatt 449

<210> 19

<211> 250

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 19

acaaatacgg tttctagtac gtggcaggaa cgactaggta agcagattga cgaattaata 60

gaaaaacatt agtcaaatac atttacaaat gaacagatag ttgatattat atttaagaat 120

tcttcttcag agcctaagat taaagctttc aattggcgaa aagaagttgt acaatatgta 180

taaaggtatg tcagtcaccg aatcagatga tctggcatta tacttgtaaa ttatcaggag 240

gttttcatta 250

<210> 20

<211> 3017

<212> PRT

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 20

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

1 5 10 15

Arg Lys Gln Arg Trp Gly Ile Arg Lys Leu Ser Val Gly Val Ala Ser

20 25 30

Val Leu Leu Gly Thr Thr Phe Met Leu Tyr Gly Asn His Ala Val Leu

35 40 45

Ala Asp Thr Val Thr Ser Pro Ser Asp Asp Val Thr Arg Ser Thr Thr

50 55 60

Thr Gln Gly Gly Asn Lys Asp Lys Val Thr Glu Gly Thr Thr Glu Gly

65 70 75 80

Thr Thr Ser Thr Pro Gln Thr Ser Gly Asp Ser Thr Asp Lys Gln Ala

85 90 95

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

100 105 110

Ala Thr Asn His Gln Asp Thr Pro Gln Gly Gln Asp Thr Thr Gln Asn

115 120 125

Thr Thr Asn Val Asp Lys Lys Asp Thr Glu Val Thr Pro Ala Asn Asp

130 135 140

Ala Thr Thr Pro Thr Thr Gln Lys Ile Thr Ala Lys Phe Thr Thr Ala

145 150 155 160

Lys Phe Thr Thr Ala Lys Phe Thr Ala Ala Lys Phe Lys Val Leu Ala

165 170 175

Ala Arg Pro Val Met Lys Val Ala Gly Thr Ala Ser Leu Pro Ile Ser

180 185 190

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

195 200 205

Asn Lys Pro Thr Asp Asn Trp Val Tyr Asn Asn Leu Lys Trp Tyr Gln

210 215 220

Asp Thr Ser Thr Glu Lys Ile Lys Glu Ile Leu Gln Asn His Thr Ala

225 230 235 240

Asn Asp Glu Ser Gly Arg Tyr Tyr Phe Ala Gly Val Ala Asn Tyr Asn

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

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

305 310 315 320

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

325 330 335

Met Val Val Gly Phe Ala Tyr Gly His Asn Thr Gly Ile Lys Gly Asp

340 345 350

Pro Ala Ser Met Gly Asn Gly Phe Val Met Thr Pro Ile Pro Thr Lys

355 360 365

Met Thr Thr Thr Ile His Tyr Ile Asp Gln Ala Thr Gly Asp Glu Ile

370 375 380

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

385 390 395 400

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

405 410 415

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

420 425 430

Phe Arg Leu Asp Lys His Val Val Ile Lys Gln Thr Val Ile Asp Ala

435 440 445

Gln Gly Leu Val Arg Val Thr Ala Tyr Tyr Asp Gly Glu Val Leu Asn

450 455 460

Asn Thr Thr Arg Tyr Leu Gly Asn Lys Leu Asn Val Asn Asp Arg Met

465 470 475 480

Ser Phe Ile Ser His Gly Lys Trp Tyr Thr Tyr Ile Asn Gln Ile Thr

485 490 495

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

500 505 510

Asp Lys Ser Glu Val Arg Val His Tyr Ile Asp Val Thr Gly Ser Lys

515 520 525

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

530 535 540

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

545 550 555 560

Thr Asp Tyr Asn Leu Ala Thr Asn Gln Ala Asn Thr Val Asn Gly Thr

565 570 575

Tyr Thr Ile Asp His His Asp Glu Tyr Val Tyr Val Val Lys Lys Thr

580 585 590

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

595 600 605

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

610 615 620

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

625 630 635 640

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

645 650 655

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

660 665 670

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

675 680 685

Leu Thr Ala Asp Glu Lys Lys Thr Ile Glu Asp Lys Ile Val Glu Ala

690 695 700

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

705 710 715 720

Gly Lys Ala Thr Ile Thr Tyr Pro Asp Lys Ser Val Asp Thr Ile Glu

725 730 735

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

740 745 750

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

755 760 765

Asp Glu Lys Lys Thr Ile Glu Asp Lys Ile Val Glu Ala Asn Lys Asp

770 775 780

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

785 790 795 800

Thr Ile Thr Tyr Pro Asp Lys Ser Val Asp Thr Ile Glu Gly Asn Gln

805 810 815

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

820 825 830

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

835 840 845

Lys Thr Ile Glu Asp Lys Ile Val Glu Ala Asn Lys Asp Lys Phe Pro

850 855 860

Glu Gly Thr Gly Val Thr Val Ala Asn Asp Gly Lys Ala Thr Ile Thr

865 870 875 880

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

885 890 895

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

900 905 910

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

915 920 925

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

930 935 940

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

945 950 955 960

Lys Ser Val Asp Thr Ile Glu Gly Asn Gln Leu Val Glu Glu Lys Thr

965 970 975

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

980 985 990

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

995 1000 1005

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

1010 1015 1020

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

1025 1030 1035

Ser Val Asp Thr Ile Glu Gly Asn Gln Leu Val Glu Glu Lys Thr

1040 1045 1050

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

1055 1060 1065

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

1070 1075 1080

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

1085 1090 1095

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

1100 1105 1110

Asp Lys Ser Val Asp Thr Ile Glu Gly Asn Gln Leu Val Glu Glu

1115 1120 1125

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

1130 1135 1140

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

1145 1150 1155

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

1160 1165 1170

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

1175 1180 1185

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

1190 1195 1200

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

1205 1210 1215

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

1220 1225 1230

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

1235 1240 1245

Pro Glu Gly Thr Gly Val Thr Val Ala Asn Asp Gly Lys Ala Thr

1250 1255 1260

Ile Thr Tyr Pro Asp Lys Ser Val Asp Thr Ile Glu Gly Asn Gln

1265 1270 1275

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

1280 1285 1290

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

1295 1300 1305

Glu Lys Lys Thr Ile Glu Asp Lys Ile Val Glu Ala Asn Lys Asp

1310 1315 1320

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

1325 1330 1335

Ala Thr Ile Thr Tyr Pro Asp Lys Ser Val Asp Thr Ile Glu Gly

1340 1345 1350

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

1355 1360 1365

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

1370 1375 1380

Ala Asp Glu Lys Lys Thr Ile Glu Asp Lys Ile Val Glu Ala Asn

1385 1390 1395

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

1400 1405 1410

Gly Lys Ala Thr Ile Thr Tyr Pro Asp Lys Ser Val Asp Thr Ile

1415 1420 1425

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

1430 1435 1440

Asp Pro Thr Val Pro Ala Lys Thr Lys Val Asp Asn Pro Thr Ser

1445 1450 1455

Leu Thr Ala Asp Glu Lys Lys Thr Ile Glu Asp Lys Ile Val Glu

1460 1465 1470

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

1475 1480 1485

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

1490 1495 1500

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

1505 1510 1515

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

1520 1525 1530

Thr Ser Leu Thr Ala Asp Glu Lys Lys Thr Ile Glu Asp Lys Ile

1535 1540 1545

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

1550 1555 1560

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

1565 1570 1575

Val Asp Thr Ile Glu Gly Asn Gln Leu Val Glu Glu Lys Thr Ser

1580 1585 1590

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

1595 1600 1605

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

1610 1615 1620

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

1625 1630 1635

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

1640 1645 1650

Lys Ser Val Asp Thr Ile Glu Gly Asn Gln Leu Val Glu Glu Lys

1655 1660 1665

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

1670 1675 1680

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

1685 1690 1695

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

1700 1705 1710

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

1715 1720 1725

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

1730 1735 1740

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

1745 1750 1755

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

1760 1765 1770

Thr Ile Glu Asp Lys Ile Val Glu Ala Asn Lys Asp Lys Phe Pro

1775 1780 1785

Glu Gly Thr Gly Val Thr Val Ala Asn Asp Gly Lys Ala Thr Ile

1790 1795 1800

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

1805 1810 1815

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

1820 1825 1830

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

1835 1840 1845

Lys Lys Thr Ile Glu Asp Lys Ile Val Glu Ala Asn Lys Asp Lys

1850 1855 1860

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

1865 1870 1875

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

1880 1885 1890

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

1895 1900 1905

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

1910 1915 1920

Asp Glu Lys Lys Thr Ile Glu Asp Lys Ile Val Glu Ala Asn Lys

1925 1930 1935

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

1940 1945 1950

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

1955 1960 1965

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

1970 1975 1980

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

1985 1990 1995

Thr Ala Asp Glu Lys Lys Thr Ile Glu Asp Lys Ile Val Glu Ala

2000 2005 2010

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

2015 2020 2025

Asp Gly Lys Ala Thr Ile Thr Tyr Pro Asp Lys Ser Val Asp Thr

2030 2035 2040

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

2045 2050 2055

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

2060 2065 2070

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

2075 2080 2085

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

2090 2095 2100

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

2105 2110 2115

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

2120 2125 2130

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

2135 2140 2145

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

2150 2155 2160

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

2165 2170 2175

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

2180 2185 2190

Ser Val Asp Thr Ile Glu Gly Asn Gln Leu Val Glu Glu Lys Thr

2195 2200 2205

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

2210 2215 2220

Asp Asp Pro Thr Lys Leu Thr Asn Asp Glu Lys Lys Glu Val Glu

2225 2230 2235

Asp Asn Ile Arg Asp His Asn Thr Gly Leu Pro Glu Gly Thr Lys

2240 2245 2250

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

2255 2260 2265

Lys Ser Val Asp Thr Ile Glu Gly Asn Gln Leu Val Glu Glu Lys

2270 2275 2280

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

2285 2290 2295

Val Asp Asp Pro Thr Lys Leu Thr Asn Asp Glu Lys Lys Glu Val

2300 2305 2310

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

2315 2320 2325

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

2330 2335 2340

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

2345 2350 2355

Lys Ser Asp Ala Ala Lys Asn Glu Pro Lys Val Pro Gly Asp Lys

2360 2365 2370

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

2375 2380 2385

Glu Val Val Lys Ala Val Glu Asp Ala Asn Lys Asp Glu Asn Gly

2390 2395 2400

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

2405 2410 2415

Gly Asp Val Thr Val Thr Tyr Pro Asp Gly Ser Lys Asp Thr Ile

2420 2425 2430

Pro Gly Asp Lys Val Val Glu Gly Lys Gly Thr Glu Gly Gln Thr

2435 2440 2445

Asp Ala Asp Lys Asn Glu Pro Lys Val Pro Gly Asp Lys Val Lys

2450 2455 2460

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

2465 2470 2475

Val Lys Ala Val Glu Asp Ala Asn Lys Asp Glu Asn Gly Lys Ser

2480 2485 2490

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

2495 2500 2505

Val Thr Val Thr Tyr Pro Asp Gly Ser Lys Asp Thr Ile Pro Gly

2510 2515 2520

Asp Lys Val Val Glu Gly Lys Gly Thr Glu Gly Gln Thr Asp Ala

2525 2530 2535

Asp Lys Asn Glu Pro Lys Val Pro Gly Asp Lys Val Lys Val Asp

2540 2545 2550

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

2555 2560 2565

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

2570 2575 2580

Pro Glu Gly Ser Lys Val Thr Val Gly Asp Asn Gly Asp Val Thr

2585 2590 2595

Val Thr Tyr Pro Asp Gly Ser Lys Asp Thr Ile Pro Gly Asp Lys

2600 2605 2610

Val Val Glu Gly Arg Gly Thr Glu Gly Gln Thr Asp Ala Asp Lys

2615 2620 2625

Asn Glu Pro Lys Val Pro Gly Asp Lys Val Lys Val Asp Asp Pro

2630 2635 2640

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

2645 2650 2655

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

2660 2665 2670

Gly Ser Lys Val Thr Val Gly Asp Asn Asp Asp Val Thr Val Thr

2675 2680 2685

Tyr Pro Asp Gly Ser Lys Asp Thr Ile Pro Gly Asp Lys Val Val

2690 2695 2700

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

2705 2710 2715

Pro Lys Val Pro Gly Asp Lys Val Lys Val Asp Asp Pro Asn Lys

2720 2725 2730

Leu Met Glu Asp Glu Lys Ser Asp Val Glu Gln Ala Ile Lys Asp

2735 2740 2745

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

2750 2755 2760

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

2765 2770 2775

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

2780 2785 2790

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

2795 2800 2805

Lys Val Lys Val Asp Asp Pro Thr Lys Leu Thr Asp Asn Glu Lys

2810 2815 2820

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

2825 2830 2835

Asp Gly Thr Lys Val Thr Val Gly Asp Asp Gly Thr Thr Thr Ile

2840 2845 2850

Thr Tyr Pro Asp Gly Ser Thr Asn Thr Ile Ser Gly His Asp Leu

2855 2860 2865

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

2870 2875 2880

Gln Ala Val Asn Val Val Asp Pro Asn His Leu Thr Gln Ala Glu

2885 2890 2895

Gln Asp Gln Val Lys Glu Ala Ile Gln Thr Thr Asn Pro Thr Ala

2900 2905 2910

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

2915 2920 2925

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

2930 2935 2940

Lys His Val Thr Glu Ala Thr Thr Gly Ser Ala Ile Lys Pro Gly

2945 2950 2955

Val Gly Thr Asn Gly Gly Gln Thr Lys Gly Ala Thr Ser Thr Asn

2960 2965 2970

Gln Thr Ala Thr Lys Gln Gln Ala Gln Gln His Leu Pro Gln Thr

2975 2980 2985

Gly Asp Gln Pro Ala Thr Trp Ala Met Leu Ser Gly Leu Gly Val

2990 2995 3000

Ala Phe Leu Gly Leu Leu Gly Leu Lys Lys Lys Arg Glu Asp

3005 3010 3015

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<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 21

Met Glu Ile Lys Lys His Phe Lys Leu Tyr Lys Asp Gly Lys Lys Trp

1 5 10 15

Cys Cys Ala Ala Ile Ala Thr Thr Val Leu Gly Ile Gly Leu Ala Ile

20 25 30

Gly Ser Pro Ser Val Leu Ala Asp Ala Asp Thr Ile Thr Ser Thr Ser

35 40 45

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

50 55 60

Ser Asn Ser Glu Ser Thr Phe Thr Asp Thr Asn Lys Asn Ser Thr Asn

65 70 75 80

Glu Lys Glu Ile Asn Glu Asn Lys Asn Ile Asp Ser Ser Gln Gln Ile

85 90 95

Asn Gln Glu Gln Thr Lys Ser Asn Asn Ser Glu Glu Gln Thr Thr Pro

100 105 110

Val Asn Val Lys Ala Glu Asn Thr Asp Ile Lys Asp Ser Ile Pro Glu

115 120 125

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

130 135 140

Tyr Gly Glu Asn Gly Asp Leu Tyr Arg Asn Gln Phe Tyr Asn Asn Trp

145 150 155 160

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

165 170 175

Phe Tyr Asn Asn Trp Gly Arg Thr Tyr Tyr Phe Gly Ser Asp Gly Ala

180 185 190

Arg Trp Asp Asn Arg Phe Tyr Asn Asn Trp Gly Arg Thr Tyr Tyr Phe

195 200 205

Gln Asn Asp Gly Ser Arg Leu Asp Asn Ser Phe Tyr Asn Asn Trp Gly

210 215 220

Arg Thr Tyr Tyr Phe Gly Val Asp Gly Ala Arg Trp Asp Asn Arg Tyr

225 230 235 240

Met Val Lys Trp Gly Arg Ala Tyr Tyr Phe Gly Asn Asp Gly Ala Leu

245 250 255

Leu Gln Asn Gln Leu Lys Ser Ile Asn Gly Ile Asn Tyr Trp Ile Asn

260 265 270

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

275 280 285

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

290 295 300

Phe Tyr His Asn Trp Gly His Thr Tyr Tyr Phe Gly Ser Asp Gly Ala

305 310 315 320

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

325 330 335

Asp Asn Gln Gly Ile Met Tyr Gln Asp Gln Tyr Tyr Lys Asn Trp Gly

340 345 350

His Thr Tyr Tyr Phe Gly Ser Asp Gly Ala Arg Tyr Thr Asp Gln Phe

355 360 365

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

370 375 380

Tyr Gln Asp Gln Tyr Tyr Lys Asn Trp Gly His Thr Tyr Tyr Phe Gly

385 390 395 400

Ser Asp Gly Ala Arg Tyr Thr Asp Gln Phe Leu Asn Arg Asp Gly Lys

405 410 415

Val Tyr Tyr Phe Asp Asn Gln Gly Ile Met Tyr Gln Asp Gln Tyr Tyr

420 425 430

Lys Asn Trp Gly His Thr Tyr Tyr Phe Gly Ser Asp Gly Ala Arg Tyr

435 440 445

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

450 455 460

Gln Gly Ile Met Val Thr Asn Gln Val Arg Val Ile Asp Gly Lys Gly

465 470 475 480

Tyr Glu Phe Asn Asp Asn Gly Glu Ala Thr Glu Thr Ser Asp Met Gly

485 490 495

Gln Thr Arg Asp Thr Val Ala Lys Glu Val Ala Gln Ala Leu Thr Asn

500 505 510

Gln Gly Ile Lys Gly Val Lys Tyr Asp Trp Arg Asn Thr Asn Asn Asp

515 520 525

Tyr Gln Glu Leu Ala Leu His Asp Ile Ala Gln Glu Val Ala Gln Gly

530 535 540

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

545 550 555 560

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

565 570 575

Asn Asp Asp Pro Gln Lys Ile Thr Asn Thr Phe Met Asn Ser Tyr Asp

580 585 590

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

595 600 605

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

610 615 620

Gln Pro Gln Ala Thr Ser Thr Ile Ser Ala Ser Ile Ser Asp Ile Phe

625 630 635 640

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

645 650 655

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

660 665 670

Ala Glu Leu Leu Lys Gly Asp Lys Gly Thr Ile Ile Ser Gln Glu Val

675 680 685

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

690 695 700

Gly Thr Lys Asn Tyr Tyr Asn Gly Asn Asp Ala Tyr His Tyr Glu Phe

705 710 715 720

Trp Leu Glu Gly Gln Ser Ala Asp Asp Lys Leu Asn Asn Phe Leu Ala

725 730 735

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

740 745 750

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

755 760 765

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

770 775 780

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

785 790 795 800

Lys Ile Pro Gly Met Thr Asp Asp Met Val Arg Gly Val Asp Val Ser

805 810 815

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

820 825 830

Gly Gln Glu Ser Asn Leu Phe Lys Ile Leu Lys Asp Ser Gly Val Asn

835 840 845

Trp Val Arg Leu Arg Val Trp Asn Asp Pro Tyr Asn Ala Gln Gly Gln

850 855 860

Pro Tyr Ala Gly Gly Asp Asn Asn Glu Glu Asn Leu Ile Lys Met Ala

865 870 875 880

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

885 890 895

Ser Asp Phe Trp Thr Asp Pro Ala Gln Gln Ile Leu Pro Lys Ala Trp

900 905 910

Arg Asn Leu Ser His Gly Glu Met Ser Gln Glu Val Tyr Leu Tyr Thr

915 920 925

Ser Lys Ile Leu Asn Asp Leu Gln Lys Ala Gly Ala Ser Val Lys Met

930 935 940

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

945 950 955 960

Gly Arg Asn Gly Gly Gly Asn Trp Ala Ser Leu Trp Glu Thr Ser Asp

965 970 975

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

980 985 990

Arg Ile Asp Pro Thr Ile Lys Val Ala Ile Gln Leu Glu Thr Pro Glu

995 1000 1005

Ile Asn Lys Tyr Arg Gly Ile Met Asn Val Leu Lys Lys Asn Asn

1010 1015 1020

Val Asp Tyr Asp Tyr Leu Gly Thr Ser Tyr Tyr Pro Phe Trp Ser

1025 1030 1035

Thr Thr Gln Gly Asn Gly Trp Tyr Asp Asn Val Asp Leu Gly Tyr

1040 1045 1050

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

1055 1060 1065

Trp Asn Glu Phe Gly Lys Arg Thr Val Ile Leu Glu Ser Gly Trp

1070 1075 1080

Leu Asn Asn Thr Asn Asp Ala Asp Gly Thr His Asn Ser Val Gly

1085 1090 1095

Glu Asn Asn Glu Thr Thr Asn Ile Asp Arg Tyr Ser Ala Asp Pro

1100 1105 1110

Gln Gly Gln Val Asp Glu Ile Glu Asp Met Tyr Asn Ala Ile Ile

1115 1120 1125

Ala Gln Lys Gly Leu Gly Ala Phe Tyr Trp Glu Pro Ala Trp Ile

1130 1135 1140

Pro Val Lys Ala Gly Trp Asn Asn Trp Gln Tyr Asn Lys Leu Met

1145 1150 1155

Ser Asn Ile Tyr Gly Ser Gly Trp Ala Ser Gln Tyr Ala Lys Gly

1160 1165 1170

Tyr Ala Pro Asp Ser Val Leu Tyr Tyr Asp Gly Lys Glu Ala Trp

1175 1180 1185

Gly Gly Ser Ser Trp Asp Asn Ile Ser Leu Phe Asp Asp His Gly

1190 1195 1200

His Pro Leu Gln Ser Leu Asn Val Tyr Asn Gly Met Leu Asn Gly

1205 1210 1215

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

1220 1225 1230

Val Lys Ile Trp Asn Glu Thr Asp Val Ile Pro Asn Asp Gly Leu

1235 1240 1245

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

1250 1255 1260

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

1265 1270 1275

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

1280 1285 1290

Thr Ala Ala Asn Gly Ala Arg Tyr His Tyr Ile Tyr Trp Leu Glu

1295 1300 1305

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

1310 1315 1320

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

1325 1330 1335

Val Val Asp Ser Glu Pro Gly Thr Gln Val Ala Thr Ser Pro Leu

1340 1345 1350

Gln Ile Lys Ile Ser Gln Val Trp Asn Thr Val Asn Asn Glu Glu

1355 1360 1365

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

1370 1375 1380

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

1385 1390 1395

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

1400 1405 1410

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

1415 1420 1425

Ser Lys Thr Tyr Thr Thr Ala Asp Gly Asn Gln Tyr Tyr Tyr Asp

1430 1435 1440

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

1445 1450 1455

Pro Ile Ile Val Asn Tyr Thr Ala Ser Leu Lys Trp Leu Gly

1460 1465 1470

<210> 22

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<212> PRT

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 22

Met Glu Lys Thr Met Lys Lys Lys Ala Leu Val Ala Thr Thr Ala Val

1 5 10 15

Ala Gly Ile Thr Leu Val Gly Glu Val Thr Thr Val His Ala Ala Asp

20 25 30

Asn Val Gln Gln Pro Val Asn Glu Gln Asn Val Asn Gln Ser Ser Gln

35 40 45

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

50 55 60

Asn Thr Ala Thr Glu Ala Asn Ser Asn Ala Gln Asp Asn Leu Ala Ser

65 70 75 80

Ala Asn Asn Asn Leu Ser Asn Ala Lys Lys Ala Val Ser Asp Gln Ala

85 90 95

Ala Lys Val Ala Asp Ala Thr Lys Ala Gln Ser Asp Ala Ser Thr Lys

100 105 110

Val Asp Asn Asp Asn Lys Val Val Ala Asp Ala Gln Gln Lys Ala Asp

115 120 125

Gln Ala Thr Pro Ala Asn Ile Glu Asn Ala Lys Gln Ala Ile Glu Gly

130 135 140

Gln Asn Lys Val Ile Asp Gln Asp Asn Glu Asn Ile Lys Tyr Ser Asn

145 150 155 160

Thr Asp Gln Asp Lys Ala Gln Asn Thr Leu Asn Asn Ala Gln Ser Asn

165 170 175

Glu Asp Lys Ala Asn Ala Thr Leu Ser Asn Lys Lys Ser Ser Gln Ala

180 185 190

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

195 200 205

His Leu Val Glu Ala Gln Asn Ala Phe Asn Gln Ala Gln Ser Asn Val

210 215 220

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

225 230 235 240

Ala Gln Lys Lys Val Thr Thr Asn Gln Asn Asp Leu Thr Ala Lys Asn

245 250 255

Lys Ala Leu Asp Asn Ile Asn Asn Gln Val Asp Thr Asp Gln Asn Asn

260 265 270

Val Asn Ser Asn Gln Ala Thr Ala Asp Ser Ala Ser Ser Ala Thr Gln

275 280 285

Val Ala Gln Asn Ala Val Asp Gln Thr Lys Gln Ser Leu Asp Lys Val

290 295 300

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

305 310 315 320

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

325 330 335

Ala Asp Gln Ser Gln Leu Asp Ser Leu Ala Lys Lys Met Tyr Asp Thr

340 345 350

Leu His Gln Gly Gln Gly Thr Asn Gly Ile Asn His Phe Asn Ser Ser

355 360 365

Lys Tyr Asp Gln Asn Gln Leu Val Asp Val Asp His Leu Thr Thr Asp

370 375 380

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

385 390 395 400

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

405 410 415

Gln Gly Val Ser Glu Met Ala Gln Gln Ile Ala Lys Gly Tyr Val Ser

420 425 430

Asp Asn Trp His Ile Ser Gln Gly His Asp Val Lys Arg Val Thr Ala

435 440 445

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

450 455 460

Ala Ser Glu Gly Tyr Val His Ala Trp Pro Trp Glu Lys Asp Ser Tyr

465 470 475 480

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

485 490 495

Leu Phe Ala Asp Asp Asn Ser Gly Asn Gly His Met Thr Asp Leu Leu

500 505 510

Gly Leu His Val Asn Arg Lys Glu Asp His Gln Tyr Phe Gly Leu Ser

515 520 525

Thr Asn Met Cys Pro Gly Ser Tyr Met Gly Gln Leu His Phe Ile Ile

530 535 540

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

545 550 555 560

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

565 570 575

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

580 585 590

Asp Leu Ala Thr Lys Gln Asp Ala Leu Asn Lys Ala Asn Gln Asn Leu

595 600 605

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

610 615 620

Ala Gln Gln Asn Arg Asp Ser Ala Gln Lys Ala Leu Asn Asp Ala Thr

625 630 635 640

Ala Lys Val Ser Asn Leu Gln Ala Thr Val Asn Ser Leu Ser Gln Asp

645 650 655

Leu Asn Ser Ala Lys Ala Thr Leu Asp Gln Ala Lys Lys Thr Leu Glu

660 665 670

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

675 680 685

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

690 695 700

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

705 710 715 720

Asp Gln Lys Lys Gln Ala Val Thr Asp Ala Gln Asn Lys Leu Ala Asn

725 730 735

Asp Gln Glu Tyr Leu Ala Thr Leu Lys Gln Asn Leu Ala Asp Leu Gln

740 745 750

Asn Ala Pro Gln Asn Leu Gln Lys Ala Lys Asp Gln Leu Ala Lys Asp

755 760 765

Gln Ile Ala Leu Asp Asn Ala Asn Lys Asp Leu Gln Asn Gln Lys Asp

770 775 780

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

785 790 795 800

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

805 810 815

Ala Gln Ala Lys Leu Ser Asp Ala Glu Ala Thr Trp Lys Glu Leu His

820 825 830

Asn Asp Ala His Arg Tyr Gly Asn Val Val Lys Val Thr Pro Ile Thr

835 840 845

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

850 855 860

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

865 870 875 880

Asp Ser Ser Asn Asn Asn Ile Pro Gln Gly Thr Lys Ala Ser Trp Ala

885 890 895

Asn Arg Ser Lys Ala Leu Thr Asp Ser Gln Asn Ala Gly Ser Tyr Ser

900 905 910

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

915 920 925

Val Asp Leu Thr Val Thr Ala Lys Lys Ile Thr Glu Asp Gln Lys Ala

930 935 940

Thr Glu Gly Gly Tyr His Ile Val Asn Gly Ser Val Val Asp Lys Gln

945 950 955 960

Asn Asn Leu Val Ser Gly Trp Thr Val Lys Asn Gly Gln Met Val Asp

965 970 975

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

980 985 990

Thr Ile Glu Lys Asn Asn Ser Lys Ser Gly Asn Thr Lys Thr Asn Met

995 1000 1005

Ile Gln Thr Ser Leu Thr Ile Ala Asn Asn Lys Ala Thr Thr Asn

1010 1015 1020

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

1025 1030 1035

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

1040 1045 1050

Met Phe Gly Tyr Lys Lys Arg Gln His Asn

1055 1060

<210> 23

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<212> PRT

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 23

Met Lys Ser Thr Thr Lys Lys Ile Leu Ala Ser Ser Leu Gly Val Ala

1 5 10 15

Gly Ala Met Ala Met Gly Thr Val Thr Ala Lys Ala Asp Thr Thr Val

20 25 30

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

35 40 45

Val Ser Ala Asp Asp Ile Ala Thr Ala Asn His Leu Gln Asn Lys Glu

50 55 60

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

65 70 75 80

Glu Thr Pro Ala Asn Asn Ala Glu Lys Lys Asp Gln Ala Ser Lys Asn

85 90 95

Ser Gln Ser Leu Gln Asp Ser Val Asn Lys Ala Met Ser Tyr Leu Gly

100 105 110

Thr Pro Tyr Val Trp Gly Gly Asn Lys Pro Gly Gly Phe Asp Cys Ser

115 120 125

Gly Leu Val Gln Tyr Cys Tyr Gly Ile Pro Gln Arg Thr Thr Tyr Glu

130 135 140

Gln Gln Ala Leu Gly Pro His Ile His Asp Asn Val Leu Asn Ala Pro

145 150 155 160

Tyr Gly Ala Leu Val Phe Tyr Gly Ser Asp Asp Ala Pro Tyr His Val

165 170 175

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

180 185 190

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

195 200 205

Val Val Met His

210

<210> 24

<211> 1029

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 24

atggaaagcc gtaataacaa taacctgaag ggcatcgatg tgagcaactg gaagggcaac 60

atcaattttc aaagcgtcaa aaatgacggt gttgaagttg tttacattaa ggcaaccgaa 120

ggcaactact tcaaagacaa atatgctaag caaaactacg agcgcgctaa agaacagggt 180

ctgcgtgtgg gcttctacca ctttttccgc gcaaacaaag gtgccaaaga tcaggcgaac 240

ttcttcgtga attacctgaa cgaaatcggt gcggtcaatt atgactgtaa actggcactg 300

gacatcgaga ctaccgaagg cgtcggtgcg cgtgacctga cctctatgtg catcgagttc 360

ctggaagagg tgaagcgtat tacgggtaag gaagttgtcg tgtacaccta taccagcttc 420

gcgaacaata atctggattc ccgtctgtct agctatccgg tgtggattgc gcactatggc 480

gtcaacaccc cgggtgcgaa caatatctgg agcgagtggg tgggtttcca gtacagcgag 540

aatggctccg tcgccggtgt cagcggtggc tgcgatatga acgaatttac caatggtatc 600

tttattgact cgaacaattt cacgttggac aatgcaacga ccaaaaatgt tagcattaag 660

ctgaacattc gcgccaaggg tacgaccaac agcaaagtta ttggtagcat tccggcgaac 720

gaaaagttta agatcaaatg ggttgatgaa gattacctgg gttggtatta cgttgagtat 780

aacggtatcg tgggttacgt taacgccgat tacgtcgaga aactgcaaat ggcgaccacg 840

cataatgtta gcacctttct gaatgtacgc gaggagggtt ccttgaatag ccgtattgtg 900

gacaagatca acactggcga catctttcgt attgactggg ttgatagcga tttcattggt 960

tggtatcgtg tgacgacgaa aaacggcaag gtcggctttg ttaatgcaga gtttgtgaaa 1020

aagttgtaa 1029

<210> 25

<211> 738

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 25

atgacaacag taaatgaagc attaaataat gtaagagctc aggttgggtc cggtgcgtct 60

gttggcaacg gcgaatgcta cgctttggct agttggtacg agcgcatgat tagtccggat 120

gcaactgtcg gacttggcgc tggtgtgggc tgggtcagcg gtgcaatcgg cgatacaatc 180

tctgccaaaa acatcggctc atcatacaac tggcaagcta acggctggac agtttccaca 240

tctggtccat ttaaagcagg tcagattgtg acgcttgggg caacaccagg aaacccttac 300

ggacatgtgg taatcgtcga agcagtggac ggcgatagat tgactatttt ggagcaaaac 360

tacggcggga aacgttatcc cgtccgtaat tattacagcg ctgcaagcta tcgtcaacag 420

gtcgtgcatt acatcacacc gcctggcacg gtcgcacagt cagcacccaa ccttgcaggc 480

tctcgttcct atcgcgagac gggcactatg actgtcacgg tcgatgctct caatgttcgc 540

agggcgccaa atacttcagg cgagattgta gcagtataca agcgtggtga atcatttgac 600

tatgatactg tcatcatcga tgtcaatggc tatgtctggg tgtcttacat aggcggcagc 660

ggcaaacgta actacgttgc gacgggcgct accaaagacg gtaagcgttt cggcaatgct 720

tggggtacat ttaaataa 738

<210> 26

<211> 543

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 26

atgtcgaaga tttttggttt agatgcgggt cattgtacga gcggcgcaga tacgggtgcg 60

cagggcaatg gttacaaaga acaagacttg acccgtcaag ttgttaccta tctgagcgaa 120

tacttggaga aagagggcca cactaccaag tactgccatt gcaatagcgc gagcacggtt 180

aacgaatccc tgcgctatcg tgtgaacaaa gccaactcca tcggtgtcga ctacttcgtg 240

agcatccacc tgaacgccgg tggcggcgtt ggtaccgaaa cgtacatctg cgcgcgtggc 300

ggcgaggccg agcgcgtggc gaaacgcgtc aattctaaac tggtgcagta cggttatcgt 360

gaccgtggtg tcaaggttgg taatctgtat gtgattaaga acaccaatgc accggctatc 420

ctggttgaga tctgtttcat tgacagcagc agcgatgtgg caaagtttaa cgcgaaggca 480

atcgcgaaag cgattgctga gggtctgctg gataaaacca ttggtgaagt cgagaataag 540

taa 543

<210> 27

<211> 1107

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 27

atgcgtaatc aattcatcga tgtttcaagt tatcaaccag atactgttgc ctttttccaa 60

gctgctaaag ctcagggtgc attaggggtc gttgttaagt taacggaagg gtccgaagat 120

ggttcggctt atgttaatcc acgtgcggcc gctcaaattc gtaatgcctt agcggttggc 180

ttgcgcgttt cctgttacca ctttgctcgt tatacatcag tgactgatgc acaaaatgaa 240

gctcgattct tcgttaaaat cgctaagcaa tttggtatgt atgacgatac tttgatgatt 300

gatgatgcgg aagttcattc aactgcagat tatcaatcag tatccttagc ctttcttcaa 360

gaagtagaag ctcttggtta caagaatact gggatttact ccatgaagtc cttcttcact 420

ggcggtattc ttaattcaca tggctttgat tcccggaaga tttggattgc tggctatggt 480

gtgactgaac tggggattga taatgcaagt gcttggcaat attctgatca tagcatcatg 540

ggaattgata ctagttatga ctttgacggt gcctttacga ctggtttagt atcaggcaat 600

gttccgcaag ctgttattcc agcaccacag ccggttcaac atattggtca cccagctact 660

ggaacctaca ttgttcagcc gggcgataca ttgagtggaa ttgcagaaaa atacgggact 720

acttatcaga acctagcagc aatcaatggt attggtaatc caaaccagat caatgtcggc 780

caagtcctca aagtcaccgg aaaagtatca aacgaaaata cttactttgt tcaatcaggc 840

gatacgttat ccggaattgc caccaaattc ggcaccactg tctcagacct cgtaagccgt 900

aatcacatta ctaacccgaa tgtgatctac gttgggcaaa aactctactt agccggcaac 960

ggacaatcca atgcttatac tgtccaagca ggggacacac taagcggaat tgcggctaag 1020

tttggcaaga cctggcaagc attagctcaa aagaatggca tcgcaaatcc taatatgatt 1080

ttcattggtc aaacaattca gatttaa 1107

<210> 28

<211> 2439

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 28

gtgtaccgaa ttattggtta taatgaacca acagataaag caggatttat tgtactggat 60

ccccgagtta atcgtcatat tagttcggga aaactcacgc ttaaagaatc taatattgat 120

gatttgacta ttacggttaa tcaagcaagt ccattatggg acaacgtaag gccttatcat 180

actcatgtta acgtttatga tgataatgaa cttatttttc gtggacgagc tatcaaacct 240

aaaaagtcga tggaagaaag cggacaattc attcgtgaat atgtttttga agatattgaa 300

gcatatctca tggatagcac ccaaagattt tatgaaggtg ttggtcaaac gcccaaagaa 360

tttttacaaa ctttaatcga tgttcataat tcacaggttc ctgactataa aaagtttcaa 420

gtccggaatg taaatgtcac taataataag gatgaccaat atcgacaaat tgattatccc 480

aaaactagcg atgctattaa tgataaatta gttaaatctc ttggtggtta tattgtgact 540

acttacaacg ctaacggaat aaactacatt gactacttaa cggatattgg ggttgatcat 600

aaagatgata ctcctattca gttagctaaa aatatgaagt ctgcaagtat gcaaattgat 660

cctactaagg tgattacaag actgattcca ctgggaaaga cactagaacc atcaaaagtt 720

gatgtaagtg atgatgatgg agagggcggt tctggatcat tagatagccc tgaagaattt 780

tgtaaatcag aaattaatgc tacttggggt agtgatatta ataatatgaa acaagatttt 840

gccgctcgtt cttcgagagt tcgggcttgg ggagtggacg ttaatcgttt atatgatgtg 900

gtgaaaaatg ctggagtaag tcctgaatgg ttctttgctt atgaacttca agaacaagga 960

acttactatg gatggcttaa ccatacttat cgacacggtg atgcgtatag tgatgcgcaa 1020

tctgtttgtg agtggattaa aaattgttca aatagtaatt ccattaatcc agcatggagc 1080

gcaccggaag gatcaatggc gccgaatcaa gcattagcgg ataaatggaa tcaagagttt 1140

ggaaaaggta ctattggccg cgtttattta caagggactg ccgctgctgt ttgggattta 1200

gctggtcaaa cgcctaatcc agctattgga aagccaatta gtggatgcat ttcttgtatt 1260

aaacgttggg gtggtcattc taatgcagct ggtggtacat ggggatggcc ttttcctgat 1320

gttggggaag gtcatttttc tcaagttcag agtttcggaa atgatggcgg atatcgtcaa 1380

aatagttatc acgatggtgt ggattttgga tcaatagatc atcctggtag agaagtgcat 1440

tgtattcatg gtggaacggt aactatcaaa tcagctatgg gtggcttagg taattttgtg 1500

gttattcata cgccggaagg attcaatatc gtttatcaag aagcttttag ttctccctct 1560

aatattattg ttagtgttgg gcaaaaagta aaaactggtg atgtaattgg atatcgtgat 1620

acagaccatg ttcatattgg cgtaactaag caagattttt atcaagcagt tcgaaattct 1680

ttttctcctg caggtggttg gctagatcca gtaaaactaa ttaaagaagg tggcgatggg 1740

tctaaaccac aagaaggaaa gaaagatcaa actgttgata atagtaatgc tgcacgtcct 1800

aaattaacca ttactactgt caataacggt agagactata ttgatattcc tgatttacaa 1860

aaagaattcg gtattattga gggaactgtt gaatttgata atgtagatga tccgaatgtt 1920

ttaatgcaac aagctcaaac atggataaag gctcaaagaa tacctcaaag ttgggaagtt 1980

acagctttag aattacatat gacaaacttc aaatctttta aggttgctga taggtacatg 2040

tttattaatc caaatgttgc aaaaccccaa ttattacgaa ttactcaaaa agaaattgat 2100

ttactaaagc cccatgcgtc ttcattaacg attggtgata agacgatggg gcttactgat 2160

tatcagttag aaaatcaagt caattttcaa caatttaagg aaattcgagt gatggttaat 2220

caggttgtcc aaacccaaga gcaatctgct aataacaata ataaggttat gcaaaatttt 2280

gctagtagtg ctgatcttgc acaaatgaga caggatctaa gaaatcttca agatgataac 2340

gatcgtgctc gcaaaggaat ggtttcctta gaagaattca ataaactaaa ggaacaagta 2400

gaaaaactaa caacaggagg cgatgataat ggcaagtga 2439

<210> 29

<211> 154

<212> PRT

<213> 人工序列

<220>

<223> 合成结构

<400> 29

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Arg Thr Gly Ser Leu Tyr

20 25 30

Ser Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val

35 40 45

Ala Ala Ile Thr Trp Arg Pro Ser Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Val Tyr

65 70 75 80

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

85 90 95

Ala Ala Arg Pro Arg Gly Gly Leu Ser Pro Thr Pro Gln Ala Tyr Asp

100 105 110

Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser Ala Ala Ala Ser

115 120 125

Gly Ser Leu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Asn Gly Ala

130 135 140

Ala His His His His His His Gly Ala Ala

145 150

<210> 30

<211> 141

<212> PRT

<213> 人工序列

<220>

<223> 合成结构

<400> 30

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Ala Thr Ile Asn

20 25 30

Asp Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Gln Arg Asp Trp Val

35 40 45

Ala Thr Ile Val Ser Asp Gly Ser Thr Ala Tyr Ala Asp Ser Val Lys

50 55 60

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

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ser

85 90 95

Ala Arg Arg His Tyr Gly Gln Gly Thr Gln Val Thr Val Ser Ser Ala

100 105 110

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

115 120 125

Asn Gly Ala Ala His His His His His His Gly Ala Ala

130 135 140

<210> 31

<211> 462

<212> DNA

<213> 人工序列

<220>

<223> 合成结构

<400> 31

gaggtgcagc tcgtggaaag tggcggaggt cttgttcagg ctgggggatc gctccgtctg 60

agctgtgcgg ggtctggcag aacaggtagt ctctattcca tgggttggtt tcggcaggcc 120

ccgggtaagg agcgggagtt cgttgcagcg attacgtgga ggcccagctc tacctactac 180

gcggacagcg taaagggacg attcaccatt agtagagacg acgcaaagaa tactgtatat 240

ttgcagatga attcgttgaa gcctgaggac accgctgtct atttttgcgc ggcgcgaccg 300

aggggcggtc tctccccgac acctcaagca tatgattact ggggacaagg gacccaagtc 360

actgtatcca gtgcggccgc gagcggcagc cttgaacaaa agctgataag cgaggaggat 420

ctcaatggtg ctgcacatca tcatcaccat cacggggcag cg 462

<210> 32

<211> 423

<212> DNA

<213> 人工序列

<220>

<223> 合成结构

<400> 32

gaagttcagc ttgtagagtc cggtgggggt cttgtacagc ccggcgggag cttgcgactc 60

tcatgcgctg cttccggaag cattgcgaca ataaatgata tgggttggtt tagacaagcc 120

cccgggaagc agcgtgactg ggtcgcgact attgtgagtg acggcagcac ggcttatgcg 180

gactcagtga aagggagatt tacgatttcg cgagataacg cgaaaaacac tgtatacctg 240

cagatgaatt cactcaagcc ggaagataca gctgtgtatt attgttctgc ccgacggcac 300

tacggacagg ggacccaggt cacagtctcg agcgctgccg ccagtgggtc actcgagcag 360

aagctgatat cagaggagga ccttaacggt gcggcgcacc atcaccacca tcatggtgcg 420

gcg 423

<210> 33

<211> 148

<212> PRT

<213> 人工序列

<220>

<223> 合成结构

<400> 33

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Phe Ser Thr Asn

20 25 30

Val Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Phe Val

35 40 45

Ala Gly Ile Thr Ile Gly Gly Thr Ala Arg Tyr Pro Asp Ser Val Lys

50 55 60

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

65 70 75 80

Gln Met Asn Asn Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn

85 90 95

Ala Val Leu Pro Ser Asp Gln Arg Arg Trp Ser Trp Gly Gln Gly Thr

100 105 110

Gln Val Thr Val Ser Ser Ala Ala Ala Ser Gly Ser Leu Glu Gln Lys

115 120 125

Leu Ile Ser Glu Glu Asp Leu Asn Gly Ala Ala His His His His His

130 135 140

His Gly Ala Ala

145

<210> 34

<211> 143

<212> PRT

<213> 人工序列

<220>

<223> 合成结构

<400> 34

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Thr Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Thr Ile Asp Met Thr Tyr

20 25 30

Gly Leu Ile Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val

35 40 45

Ala Ser Ile Arg Arg Asp Gly Arg Thr Asn Tyr Ala Asp Ser Val Lys

50 55 60

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

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Asp Asp Thr Ala Arg Tyr Tyr Cys Asn

85 90 95

Ser Pro Tyr His Ala Leu Trp Gly Gln Gly Thr Gln Val Thr Val Ser

100 105 110

Ser Ala Ala Ala Ser Gly Ser Leu Glu Gln Lys Leu Ile Ser Glu Glu

115 120 125

Asp Leu Asn Gly Ala Ala His His His His His His Gly Ala Ala

130 135 140

<210> 35

<211> 444

<212> DNA

<213> 人工序列

<220>

<223> 合成结构

<400> 35

gaggtacaac tggttgagag tgggggtggt ttggtgcaag ccggaggttc cttacgtttg 60

tcttgcgcgg ctagtgggag catcttttca acaaacgtaa tggggtggta ccgccaagcc 120

ccaggtaagc agcgggaatt tgtggccggg ataacgatcg gaggaactgc gaggtatcct 180

gatagtgtga aagggcgttt cacaattagt cgagataata cacagaatac tgtctatctc 240

caaatgaata atctcaagcc cgaagacaca gcagtttatt attgtaatgc cgttctcccc 300

tctgatcagc gtcgatggag ctggggacaa ggcacccagg ttacggttag cagcgcggca 360

gcgtctggtt cgctcgagca aaagctcata tctgaggagg acctgaacgg ggcagcccac 420

catcaccacc atcacggagc agct 444

<210> 36

<211> 429

<212> DNA

<213> 人工序列

<220>

<223> 合成结构

<400> 36

gaggtacagc tggtggagtc cggcggtggt ttggtgcaaa ccgggggtag tctgcggctt 60

agttgcacgg cgtctgggac aatagacatg acttatggtc tcatatggta caggcaagcg 120

cctgggaaag agagggaact cgttgcgagt atcagaaggg acggccgcac aaattacgct 180

gattcagtga aagggcgctt cactatctcg atcgataatg cgaaaaacag tattcacctt 240

caaatgaact cccttaagcc cgatgatacc gccaggtatt attgcaacag cccatatcac 300

gcactttggg gtcagggtac gcaggtaaca gtgtctagtg cggcagcctc tggtagtttg 360

gagcaaaagt tgataagtga ggaggactta aatggggcgg cacatcacca ccaccatcat 420

ggggcggct 429

<210> 37

<211> 1001

<212> DNA

<213> 人工序列

<220>

<223> 合成结构

<400> 37

aaattaaaag gctggatttt ttcggccttt ttttagtgca aataattatt ttttacgtat 60

ttatattata gggctaatca ctaaactaat aattagtggt tgaagcgctg aaaattttct 120

gctattttat taatagtttg ataataaaat aatgatattt aatataaaga gggataaacg 180

aaataatgaa atcaacaaca aagaaaattc ttgcatcgtc gttaggggta gctggcgcaa 240

tggcaatggg cacggtaact gcaaaggctg atacgaccgt tacggtcaat gctggcgata 300

gtttgaatgg gattgctcaa aagtataatg ttagtgcgga tgatattgca accgctaatc 360

acttgcaaaa taaagagttg atttttgtgg gacaaaagtt gacaattcca accaaagata 420

aaaatgaaac agaagttcaa ttagttgaaa gtggtggtgg tttagttcaa ccaggtggta 480

gtttacgttt atcatgtgct gcaagtggtt caattgcaac tattaatgat atgggttggt 540

ttcgtcaagc accaggaaag caacgtgatt gggttgctac tattgttagt gatggttcaa 600

ctgcttatgc tgatagtgtt aaaggtcgtt ttactatttc acgtgataat gctaagaata 660

ctgtttacct tcaaatgaat agtcttaagc cagaagatac tgcagtttac tattgttcag 720

ctcgtcgtca ttatggtcaa ggtactcaag ttactgttag ttcagctgca gctagtggtt 780

cattagaaca aaaattaatt tcagaagaag atttaaatgg tgcagctcat catcatcatc 840

atcatggtgc agcttacttc cctggaaatt actatgttgt gatgcattaa tattcccttt 900

cacctcacct ttaataattt aaattagtaa ttatcttgcg catcacaaaa gagtgctata 960

tactatttca gattagaaag ttttatgagg gagacaaatt g 1001

<210> 38

<211> 1393

<212> DNA

<213> 人工序列

<220>

<223> 合成结构

<400> 38

gaagtacaaa gttactttaa ctataatgaa aaacaagaca atataaagaa aacaacatat 60

aaggttcagt tcataactga ttagatttat aataaatatt gtaaatcgga caaaaataaa 120

ttaattttca attaattcaa aaaaaccata tttttttcgt tttggcatat ttggatttgc 180

tacactaaag atgatcaaga aaggggaaaa gataatcttc aatcttgtgt acttagtttg 240

ttaattaatt tataaattta gggaggaaac ctatcatgga aaagactatg aaaaagaaag 300

ctttagttgc aactactgct gtagccggta ttactttagt aggagaggtt actactgttc 360

atgccgctga caatgtacaa caagaagttc aattagttga aagtggtggt ggtttagttc 420

aaccaggtgg tagtttacgt ttatcatgtg ctgcaagtgg ttcaattgca actattaatg 480

atatgggttg gtttcgtcaa gcaccaggaa agcaacgtga ttgggttgct actattgtta 540

gtgatggttc aactgcttat gctgatagtg ttaaaggtcg ttttactatt tcacgtgata 600

atgctaagaa tactgtttac cttcaaatga atagtcttaa gccagaagat actgcagttt 660

actattgttc agctcgtcgt cattatggtc aaggtactca agttactgtt agttcagctg 720

cagctagtgg ttcattagaa caaaaattaa tttcagaaga agatttaaat ggtgcagctc 780

atcatcatca tcatcatggt gcagctaaga agattacaga agatcaaaaa gcaacagaag 840

gcggttatca tattgttaat ggatctgttg tagataagca gaataacttg gttagtggtt 900

ggactgttaa gaatggtcaa atggttgatc ctgaaggtaa cgttatcaaa acaacaatgt 960

ctacagccca aggtgttact attgaaaaaa ataatagcaa gtccgggaat acaaagacaa 1020

acatgattca aacttcttta actattgcta acaacaaggc aacaacaaac aaagacaacc 1080

agttaccaca aactggcaat tacaacaaca atacaaaggt attaggatta gctggtattg 1140

cacttgcatc tgctttaact atgtttggat acaagaagcg ccaacataac taattttctt 1200

acttgatggg tttctaaata aaaaatggac tacttcagct caaggtagcc catttttatt 1260

attatagtga agcagttatt cttacgtata accagactaa aatataataa aaatctatta 1320

tatattaatc aacatctcgg tttaatcgtt aaaactcctc tgagagctaa ttgttaatat 1380

tgagttgtat agt 1393

<210> 39

<211> 1419

<212> DNA

<213> 人工序列

<220>

<223> 合成结构

<400> 39

gcaatgcaca agatgctgaa acaaaggcac aacaaaatgc agatcaagct tcaccagcta 60

atattcaaaa ggcacaagat gctattgcta atcaagaaac tcaaattagt aaagacaccg 120

atgctattaa tgacgctaac aaagccgtta gcgatgcaca aagcacagtt gatgcagcgc 180

aaaaaaagtt aatgatgcaa ctactgctcg tgacaatcaa caaaagaatg ttgatactgc 240

tagtgatgca gttaagaatg ctcaagctat tcttgacaac agtgatcagg ctaaaaagga 300

agcccaagat gctttgaaca aggctaacca aaaagttgct gatgatacta ctgccgttaa 360

caacaaacaa actgatgtta acaatgcagc agaagctaag aagaatgcag atgaggcatt 420

gaagaacgcc aatgatgcgc aaacttctgc acaaaagaat aaagatgcta agcaagcaat 480

tgctgatgag gcaagtgtag aagttcaatt agttgaaagt ggtggtggtt tagttcaacc 540

aggtggtagt ttacgtttat catgtgctgc aagtggttca attgcaacta ttaatgatat 600

gggttggttt cgtcaagcac caggaaagca acgtgattgg gttgctacta ttgttagtga 660

tggttcaact gcttatgctg atagtgttaa aggtcgtttt actatttcac gtgataatgc 720

taagaatact gtttaccttc aaatgaatag tcttaagcca gaagatactg cagtttacta 780

ttgttcagct cgtcgtcatt atggtcaagg tactcaagtt actgttagtt cagctgcagc 840

tagtggttca ttagaacaaa aattaatttc agaagaagat ttaaatggtg cagctcatca 900

tcatcatcat catggtgcag ctgctgaaaa ggaagctaaa gataatggct accatatcga 960

aaataaccaa gttgttgacg ctaaaggtaa tagtgtcaat ggctggacag ttaagggcaa 1020

ccaaattgtt agtccaacta atgctactgt agatcccgct gtttctgtaa ccaccaatgt 1080

caatgttgat agtaaaggtc aagtacaacc acaaactagt gttactgcta atagtgttaa 1140

gactgtagct gcaactgaat cagcaaatcc agtagcaact actactgtgc aaacccgcga 1200

acaatacaag caacaattga agagcaataa tcaattacca caaactggta ataatgatag 1260

tgctgttctt tcacttgctg gagtagcact tgcagcaatg ttgagtttgt tcggtattaa 1320

gaaacgtgaa tactaattta gaaaatgtaa gtattattat gtaaaaaggt tcaaccaaat 1380

tggctgaacc tttttgtcta aaatttaagg agaagtttt 1419

<210> 40

<211> 739

<212> PRT

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 40

Met Asn Lys Ala Asn Gln Lys Val Ala Asp Asp Thr Thr Ala Val Asn

1 5 10 15

Asn Lys Gln Thr Asp Val Asn Asn Ala Ala Glu Ala Lys Lys Asn Ala

20 25 30

Asp Glu Ala Leu Lys Asn Ala Asn Asp Ala Gln Thr Ser Ala Gln Lys

35 40 45

Asn Lys Asp Ala Lys Gln Ala Ile Ala Asp Glu Ala Ser Val Ala Leu

50 55 60

Ala Asp Ala Asn Thr Ala Val Lys Asp Ala Gln Ala Lys Val Asp Ala

65 70 75 80

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

85 90 95

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

100 105 110

Tyr Asn Gln Asp Glu Ala Asn Asn Leu Ala Gln Asp Leu Leu Lys Tyr

115 120 125

Arg Asp Gln Ala Met Ser Gln Asn Lys Phe Lys Asp Asn Leu Ser Asp

130 135 140

Asp Arg Val Val Asp Ile Asp Asn Leu Asn Ser Thr Asp Arg Ala Glu

145 150 155 160

Leu Ser Gln Phe Val Ala Ser Leu Ile Asn Gln Val Arg Thr Gln Met

165 170 175

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

180 185 190

Gln Val Ser Gln Asn Tyr Asn Lys Asp Asn Trp Asn Ser Ala Asp Asn

195 200 205

Gly Lys His Asp Gln Ser Ala Leu Asn Asn Ala Thr Asp Gln Leu Asn

210 215 220

Ile Ser Trp Asn Gly Glu Asn Met Gly Leu Asp Gln Ser Ile Phe Thr

225 230 235 240

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

245 250 255

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

260 265 270

Ser Met Met Phe Asp Asp Ala Asp Ser Ala Trp Gly His Ala Thr Asn

275 280 285

Phe Ala Gly Ile Asp Asn Phe Ala Ala Glu Lys Gln Ala Val Gly Phe

290 295 300

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

305 310 315 320

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

325 330 335

Thr Gln Lys Leu Ala Asp Ala Lys Asp Asp Leu Ser Ile Lys Gln Thr

340 345 350

Asp Gln Ala Ser Lys Gln Lys Ala Asn Asp Asp Ala Gln Asn Ala Leu

355 360 365

Ser Ser Ala Asn Gln Val Leu Val Ala Ala Gln Asn Asp Val Lys Asp

370 375 380

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

385 390 395 400

Asn Asp Leu Ala Thr Leu Gln Asn Gln Leu Ser Ala Asp Gln Ala Asn

405 410 415

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

420 425 430

Thr Lys Gln Ala Asn Leu Gln Lys Ala Thr Asp Ser Leu Lys Ala Glu

435 440 445

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

450 455 460

Ala Leu Ser Asp Ala Asn Asn Asn Leu Ala Gln Lys Lys Gln Val Val

465 470 475 480

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

485 490 495

Leu Gln Asn Asn Leu Ser Asp Leu Gln Asn Ala Pro Lys Leu Leu Ala

500 505 510

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

515 520 525

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

530 535 540

Thr Ala Ala Gly Thr Thr Thr Asn Val Ser Lys Ala Gln Gln Ala Leu

545 550 555 560

Ala Glu Ala Lys Asn Asn Glu Asp Ala Ala Lys Glu Val Leu Asp Gln

565 570 575

Ala Gln Gln Ala Leu Thr Glu Leu Arg Gln Lys Glu Ala Leu Ala Lys

580 585 590

Gln Val Ala Glu Glu Gln Ala Lys Leu Ala Ala Glu Lys Glu Ala Lys

595 600 605

Asp Asn Gly Tyr His Ile Glu Asn Asn Gln Val Val Asp Ala Lys Gly

610 615 620

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

625 630 635 640

Thr Asn Ala Thr Val Asp Pro Ala Val Ser Val Thr Thr Asn Val Asn

645 650 655

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

660 665 670

Ser Val Lys Thr Val Ala Ala Thr Glu Ser Ala Asn Pro Val Ala Thr

675 680 685

Thr Thr Val Gln Thr Arg Glu Gln Tyr Lys Gln Gln Leu Lys Ser Asn

690 695 700

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

705 710 715 720

Ala Gly Val Ala Leu Ala Ala Met Leu Ser Leu Phe Gly Ile Lys Lys

725 730 735

Arg Glu Tyr

<210> 41

<211> 185

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 41

aaattaaaag gctggatttt ttcggccttt ttttagtgca aataattatt ttttacgtat 60

ttatattata gggctaatca ctaaactaat aattagtggt tgaagcgctg aaaattttct 120

gctattttat taatagtttg ataataaaat aatgatattt aatataaaga gggataaacg 180

aaata 185

<210> 42

<211> 275

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 42

gaagtacaaa gttactttaa ctataatgaa aaacaagaca atataaagaa aacaacatat 60

aaggttcagt tcataactga ttagatttat aataaatatt gtaaatcgga caaaaataaa 120

ttaattttca attaattcaa aaaaaccata tttttttcgt tttggcatat ttggatttgc 180

tacactaaag atgatcaaga aaggggaaaa gataatcttc aatcttgtgt acttagtttg 240

ttaattaatt tataaattta gggaggaaac ctatc 275

<210> 43

<211> 313

<212> DNA

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 43

gcaatgcaca agatgctgaa acaaaggcac aacaaaatgc agatcaagct tcaccagcta 60

atattcaaaa ggcacaagat gctattgcta atcaagaaac tcaaattagt aaagacaccg 120

atgctattaa tgacgctaac aaagccgtta gcgatgcaca aagcacagtt gatgcagcgc 180

aaaaaaagtt aatgatgcaa ctactgctcg tgacaatcaa caaaagaatg ttgatactgc 240

tagtgatgca gttaagaatg ctcaagctat tcttgacaac agtgatcagg ctaaaaagga 300

agcccaagat gct 313

<210> 44

<211> 46

<212> PRT

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 44

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

1 5 10 15

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

20 25 30

Ser Lys Gly Tyr Trp Val Gly Ile Tyr Glu Ser Val Asp Lys

35 40 45

<210> 45

<211> 64

<212> PRT

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 45

Leu Ala Ser Thr Leu Gly Ile Ser Thr Ala Ala Ala Lys Lys Ala Ile

1 5 10 15

Asp Ile Ile Asp Ala Ala Ser Thr Ile Ala Ser Ile Ile Ser Leu Ile

20 25 30

Gly Ile Val Thr Gly Ala Gly Ala Ile Ser Tyr Ala Ile Val Ala Thr

35 40 45

Ala Lys Thr Met Ile Lys Lys Tyr Gly Lys Lys Tyr Ala Ala Ala Trp

50 55 60

<210> 46

<211> 38

<212> PRT

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 46

Glu Phe Ser Gly Gly Gly Gly Ala Glu Gln Arg Gly Ile Ser Gln Gly

1 5 10 15

Asn Asp Gly Lys Leu Cys Thr Leu Thr Trp Glu Cys Gly Leu Cys Pro

20 25 30

Thr His Thr Cys Trp Cys

35

<210> 47

<211> 35

<212> PRT

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 47

Ser Asp Ala Thr Pro Met Thr Val Thr Pro Thr Thr Ile Thr Ile Pro

1 5 10 15

Ile Ser Leu Ala Gly Cys Pro Thr Thr Lys Cys Ala Ser Ile Val Ser

20 25 30

Pro Cys Asn

35

<210> 48

<211> 36

<212> PRT

<213> 人工序列

<220>

<223> 基于罗伊氏乳杆菌的序列

<400> 48

Ser Glu Ala Thr Pro Met Thr Val Thr Pro Thr Thr Ile Thr Ile Pro

1 5 10 15

Ile Ser Leu Ala Gly Cys Pro Thr Thr Lys Cys Ala Ser Ile Val Ser

20 25 30

Pro Cys Asn Asp

35

<210> 49

<211> 154

<212> PRT

<213> 人工序列

<220>

<223> 合成结构

<400> 49

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly His Thr Gly Ser Leu Tyr

20 25 30

Ser Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val

35 40 45

Ala Ala Ile Thr Trp Arg Pro Ser Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Val Tyr

65 70 75 80

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

85 90 95

Ala Ala Arg Pro Arg Gly Gly Leu Ser Pro Thr Pro Gln Ala Tyr Asp

100 105 110

Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser Ala Ala Ala Ser

115 120 125

Gly Ser Leu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Asn Gly Ala

130 135 140

Ala His His His His His His Gly Ala Ala

145 150

<210> 50

<211> 143

<212> PRT

<213> 人工序列

<220>

<223> 合成结构

<400> 50

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Thr Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Thr Ile Asp Met Thr Tyr

20 25 30

Gly Leu Ile Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val

35 40 45

Ala Ser Ile Arg Arg Asp Gly His Thr Asn Tyr Ala Asp Ser Val Lys

50 55 60

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

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Asp Asp Thr Ala Arg Tyr Tyr Cys Asn

85 90 95

Ser Pro Tyr His Ala Leu Trp Gly Gln Gly Thr Gln Val Thr Val Ser

100 105 110

Ser Ala Ala Ala Ser Gly Ser Leu Glu Gln Lys Leu Ile Ser Glu Glu

115 120 125

Asp Leu Asn Gly Ala Ala His His His His His His Gly Ala Ala

130 135 140

Claims (47)

1.一种活体递送平台，包括含有表达盒的遗传修饰微生物；其中所述表达盒包括以下一种或多种：用于转录表达的启动子、编码用于分泌的信号序列的核苷酸序列、编码细胞壁锚的核苷酸序列、至少一个编码所需生物分子的异源编码区、编码用于检测的表达肽标签的核苷酸序列、以及用于终止翻译和转录的终止子。

2.根据权利要求1所述的活体递送平台，其中所述遗传修饰微生物是选自芽孢杆菌属、乳杆菌属、乳球菌属和肠球菌属的细菌。

3.根据权利要求1所述的活体递送平台，其中所述遗传修饰微生物是罗伊氏乳杆菌菌株。

4.根据权利要求1所述的表达盒，其中用于转录表达的启动子选自SEQ ID NO：14、SEQID NO：15、SEQ ID NO：16、SEQ ID NO：17、SEQ ID NO：18、SEQ ID NO：19、SEQ ID NO：41、SEQID NO：42和SEQ ID NO：43。

5.根据权利要求1所述的表达盒，其中所述编码用于分泌的信号序列的核苷酸序列编码至少20个氨基酸、至少25个氨基酸、至少30个氨基酸、至少35个氨基酸、至少40个氨基酸、至少44个氨基酸、至少50个氨基酸、至少55个氨基酸、至少60个氨基酸氨基酸、或至少65个氨基酸。

6.根据权利要求1所述的表达盒，其中所述编码用于分泌的信号序列的核苷酸序列编码20～65个氨基酸、25～65个氨基酸或25～40个氨基酸。

7.根据权利要求5或6所述的编码用于分泌的信号序列的核苷酸序列，其中所述用于分泌的信号序列包含选自以下组的氨基酸序列：SEQ ID NO：20、SEQ ID NO：21、SEQ ID NO：22、SEQ ID NO：23和SEQ ID NO：40。

8.根据权利要求1所述的表达盒，其中所述编码细胞壁锚的核苷酸序列编码100～250个氨基酸、125～175个氨基酸或125～150个氨基酸。

9.根据权利要求8所述的编码细胞壁锚的核苷酸序列，其中所述细胞壁锚包含选自以下组的氨基酸序列：SEQ ID NO：20、SEQ ID NO：21、SEQ ID NO：22、SEQ ID NO：23和SEQ IDNO：40。

10.根据权利要求1所述的表达盒，其中所述编码所需生物分子的异源编码区编码有抗感染活性的生物分子、益生菌因子、免疫调节因子或促进生长的生物分子。

11.根据权利要求10所述的生物分子，其中所述抗感染活性对致病菌或寄生虫具有活性。

12.根据权利要求11所述的抗感染活性，其中所述寄生虫是艾美球虫种。

13.根据权利要求11所述的抗感染活性，其中所述致病菌选自沙门氏菌、梭菌、弯曲杆菌、葡萄球菌、链球菌和大肠杆菌。

14.根据权利要求10所述的生物分子，其中所述抗感染活性选自杀菌肽、酶、溶素、噬菌体和抗体。

15.根据权利要求14所述的生物分子，其中所述杀菌肽是包含选自SEQ ID NO：2和SEQID NO：4序列的美杀菌素样分子。

16.根据权利要求14所述的生物分子，其中所述杀菌肽是包含选自由SEQ ID NO：44、SEQ ID NO：45、SEQ ID NO：46、SEQ ID NO：47和SEQ ID NO：48组成的组的序列的杆菌素。

17.根据权利要求14所述的生物分子，其中所述酶包含选自SEQ ID NO：5和SEQ ID NO：6的序列。

18.根据权利要求14所述的生物分子，其中所述溶素包含选自SEQ ID NO：24、SEQ IDNO：25、SEQ ID NO：26、SEQ ID NO：27和SEQ ID NO：28组成的组的序列。

19.根据权利要求14所述的生物分子，其中所述噬菌体包含选自下组的序列：NO：11、SEQ ID NO：12和SEQ ID NO：13。

20.根据权利要求14所述的生物分子，其中所述抗体是单链抗体。

21.根据权利要求20所述的抗体，其中所述单链抗体特异性地识别来自产气荚膜梭菌的细菌蛋白质。

22.根据权利要求21所述的抗体，其中所述细菌蛋白是产气荚膜梭菌α毒素或产气荚膜梭菌NetB毒素。

23.根据权利要求20所述的单链抗体，其中所述单链抗体包含选自由SEQ ID NO：29、SEQ ID NO：49、SEQ ID NO：30、SEQ ID NO：33、SEQ ID NO：50和SEQ ID NO：34组成的组的序列。

24.根据权利要求10所述的生物分子，其中所述益生因子是包含选自由SEQ ID NO：9和SEQ ID NO：10组成的组的序列的凝集素受体。

25.一种表达盒，包含用于转录表达的启动子和至少一个编码所需生物分子的异源编码区；其中所述所需生物分子选自杀菌肽、酶、溶素、噬菌体和抗体。

26.根据权利要求25所述的表达盒，其中所述启动子包含选自SEQ ID NO：14、SEQ IDNO：15、SEQ ID NO：16、SEQ ID NO：17、SEQ ID NO：18、SEQ ID NO：19、SEQ ID NO：41、SEQ IDNO：42和SEQ ID NO：43组成的组的序列。

27.根据权利要求25所述的表达盒，其中所述所需生物分子包含选自由SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：5、SEQ ID NO：6、SEQ ID NO：24、SEQ ID NO：25、SEQ ID NO：26、SEQ ID NO：27、SEQ ID NO：28、SEQ ID NO：29、SEQ ID NO：49、SEQ ID NO：30、SEQ ID NO：33、SEQ ID NO：34、SEQ ID NO：50、SEQ ID NO：44、SEQ ID NO：45、SEQ ID NO：46、SEQ IDNO：47和SEQ ID NO：48组成的组的序列。

28.根据权利要求25所述的表达盒，其中所述表达盒位于质粒上。

29.根据权利要求28所述的表达盒，其中所述质粒包含SEQ ID NO：8。

30.根据权利要求25所述的表达盒，其中所述表达盒位于细菌染色体上。

31.根据权利要求30所述的表达盒，其中所述表达盒插入到转座酶基因座中。

32.根据权利要求30所述的表达盒，其中所述表达盒插入到尿嘧啶磷酸核糖(UPP)转移酶基因座中。

33.根据权利要求30所述的表达盒，其中所述表达盒插入到pyrE基因座中。

34.一种减少动物被致病菌定殖的方法，所述方法包括用权利要求1～24任一项所述的活体递送平台处理动物。

35.根据权利要求34所述的方法，其中所述动物是鸟、人类或非人类哺乳动物。

36.根据权利要求34所述的方法，其中，所述致病菌选自由沙门氏菌、梭菌、弯曲菌、葡萄球菌、链球菌和大肠杆菌组成的组。

37.根据权利要求34～36任一项所述的方法，其中所述治疗是经口、肠胃外、经鼻或经粘膜给药。

38.根据权利要求34～36任一项所述的方法，其中所述动物是鸟类并且其中在卵内给药治疗。

39.权利要求1～24任一项所述的活体递送平台在治疗中的应用。

40.权利要求1～24任一项所述的活体递送平台在减少动物被致病菌定殖中的应用。

41.权利要求1～24任一项所述的活体递送平台在制备减少动物被致病菌定殖的药物中的应用。

42.权利要求25～33任一项所述的表达盒在制备动物被致病菌定殖的药物中的应用。

43.一种抗体，包含选自以下组的氨基酸序列：SEQ ID NO：29、SEQ ID NO：49、SEQ IDNO：30、SEQ ID NO：33、SEQ ID NO：34和SEQ ID NO：50。

44.根据权利要求43所述的抗体，其中所述抗体结合产气荚膜梭菌产生的毒素。

45.权利要求43的抗体在治疗中的应用。

46.权利要求43所述的抗体在减少动物被致病菌定殖中的应用。

47.权利要求43所述的抗体在制备减少动物被致病菌定殖的药物中的应用。

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