CN116790555A - Rna靶向基因编辑工具的开发 - Google Patents
Rna靶向基因编辑工具的开发 Download PDFInfo
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Abstract
紧凑型Cas13蛋白的筛选方法及应用。本公开内容涉及生物技术及医学领域。更具体地，本公开内容涉及新的Cas13家族蛋白、筛选新的Cas13家族蛋白的方法、以及相应的RNA编辑系统及其应用。本公开内容尤其涉及Cas13蛋白及相关的RNA编辑系统。所述新型Cas13蛋白的分子量很低，几乎将具有guide RNA引导的且具有RNase活性的CRISPR‑Cas蛋白推向了极限，并且包含较多的拓展型的HEPN结构域。本公开内容首次提出快速寻找具有超低分子量，依赖guide RNA引导且具有RNase活性的CRISPR‑Cas13蛋白的筛选方法，并获得了多种新的Cas13蛋白及其新的家族，具有广阔的应用前景和巨大的市场价值。
Description
技术领域
本公开内容涉及生物技术及医学领域。更具体地，本公开内容涉及新的Cas13家族蛋白、筛选新的Cas13家族蛋白的方法、以及相应的RNA编辑系统及其应用。本公开内容尤其涉及低分子量Cas13蛋白及相关的RNA编辑系统。
背景技术
CRISPR-Cas系统被称为新一代基因组工程工具的关键组件，在细菌，古细菌等微生物中起着适应性免疫机制的作用，可保护微生物免受病毒和其他外来核酸的侵害。CRISPR-Cas免疫应答主要包括三个阶段：适应阶段、表达和加工阶段和干扰阶段。与其他防御机制类似，CRISPR-Cas系统在与移动遗传元件不断竞争的背景下发展，这导致Cas蛋白序列和CRISPR-Cas基因座结构的极端多样化。
自2011年以来，依据CRISPR-Cas系统的基因组成，基因座结构以及序列相似性聚类等方法1，目前可以将CRISPR-Cas系统分成2大类，其中Class 1类系统具有由多个Cas蛋白质组成的效应器模块，其中一些形成crRNA结合复合物，这些复合物通过额外Cas蛋白来介导pre-crRNA处理和干扰。相比之下，Class 2类系统包含一个单一的具有多功能域结合区的Cas效应蛋白，它能结合crRNA参与干扰所需的所有活动，在某些变体中，还包括参与pre-crRNA成熟过程。目前Class 2类型CRISPR-Cas系统1主要分3个亚型：type II(如Cas9)，type V3(如Cas12a)，和type VI 4(如Cas13d)。其中type II和type V亚型主要靶向DNA1。而type VI效应Cas蛋白则主要靶向RNA。
由于Class 2类CRISPR-Cas系统相较与Class 1类CRISPR-Cas系统具有显著的优势，自其被发现以来，已吸引了大批学者们对它们进行了深入的研究和改造，并开发出多种依赖CRISPR-Cas的基因操作工具，包括CRISPRa，CRISPRi，单碱基编辑技术等。利用这些工具也促进一部分学者们开始从基础研究向临床应用研究发展，特别是目前已有部分基因治疗相关药物上市，这又推进了人类的健康事业的发展。由于很多时候，基因疗法依赖于递送介质，常用的包装工具是逆转录病毒，腺病毒或者腺相关病毒等，但是这类工具的承载容量有限，如腺相关病毒AAV，它是目前最常使用且安全性最高的递送工具，但是它不能容纳超过4.7kb的DNA，不利于大分子量的CRISPR-Cas相关工具的包装。尽管有学者尝试采用共转多个包装不同调控原件的病毒，但是这种处理的结果远不如all-in-one模式的包装体系。
2020年有学者在大型细菌病毒噬菌体中找到了分子量只有Cas9和Cas12a基因组编辑酶的一半的Casφ(也被归为Cas12j亚家族)蛋白，它能在真核生物细胞上发挥切割DNA的功能。近期张锋团队还找到Cas9和Cas12的始祖蛋白IscB(约400个氨基酸)和TnpB家族，它们也是guide RNA依赖的核酸酶，只包含有单一核酸切割结构域(如RuvC)12，暗示自然界存在低分子量的单效应Cas酶能够应用于真核生物的基因编辑。然而当前对于RNA层面的CRISPR-Cas13系统的始祖蛋白还未找到，已知最小的紧凑型Cas13效应蛋白，如Cas13bt，Cas13X等13-14都超过700个氨基酸。因此需要开发新的数据挖掘算法来进一步探索紧凑型的CRISPR-Cas13系统单效应蛋白，如Cas13始祖蛋白等，以便可以更好应用于基因治疗。
既往研究策略主要依据Cas1蛋白的序列保守型来确定临近Cas蛋白10，但是这种方式会遗漏一些不存在Cas1蛋白的单效应蛋白。依据CRISPR-array与Cas蛋白的共存性，促使学者们直接从预测CRISPR array入手，然后寻找临近CRISPR-Cas关联蛋白11，但是受制于当前预测CRISPR array的算法局限性，并没有哪种算法被大家归为金标准。此外，候选蛋白确定问题上，主要依赖DNA和蛋白序列比对，这很容易忽略蛋白空间折叠的影响1。因此，亟需开发的新的寻找自然界分子量更小的Class 2类CRISPR-Cas13系统相关单效应蛋白的计算方法和实验验证方法。
发明内容
针对现有筛选新型CRISPR-Cas蛋白技术的不足和实际需求，本公开内容提供了一种快速寻找包含较多的拓展的HEPN结构域(至少1个)的新型guide RNA引导具有RNase活性的CRISPR-Cas13蛋白的方法并从生物信息分析层面(例如，序列比对、蛋白结构预测等)和实验层面验证了候选蛋白的RNase活性。这些蛋白潜在应用于RNA层面的调控、编辑、检测等方面，具有广阔的学术价值和商业应用价值。
本公开内容所解决的技术问题是如何快速寻找新型的RNA酶切活性结构域(拓展的HEPN结构域)较多的候选CRISPR-Cas13蛋白及其系统；其次是验证候选CRISPR-Cas13蛋白及其系统的活性；并最终获得了多种新型Cas13蛋白。
本公开内容实现了以下技术效果：
(1)开发了快速筛选新型Cas13家族蛋白的分析方法，该方法可以对新更新的原核微生物DNA序列和宏基因组序列进行CRIPSR array系统的分析和相关效应蛋白的筛选；
(2)筛选的低分子量的Cas13家族成员，拓展CRISPR-Cas13的应用范围。由于候选Cas13蛋白低分子量能很好的通过腺相关病毒等递送载体包装从而实现相关疾病诊疗，如神经相关退行性疾病的诊疗，在植物领域则可以开展育种，逆境胁迫等方面的研究，在微生物领域可以进行相关工程菌的改造等；
(3)本方法在筛选过程中，除利用Cas13蛋白的已知HEPN结构域进行筛选外，还将其他种类的蛋白质中具备RNA切割活性的保守型结构域包括在内，从而提供了筛选新的Cas13蛋白的可能，并且由于这些新Cas13蛋白中这些新的功能结构域的鉴定，为进一步改造Cas13蛋白提供了新的思路和可能性。
在本公开内容的一个方面中，提供了Cas13蛋白。
在一个优选的实施方案中，所述Cas13蛋白包含如SEQ ID NO:1-78中任一项所述的氨基酸序列，或具有一个或更多个残基的保守氨基酸取代的SEQ ID NO:1-78中任一项所述的氨基酸序列。
在一个优选的实施方案中，所述Cas13蛋白的RNA切割活性被保留。
在一个优选的实施方案中，所述Cas13蛋白的HEPN结构域或RNA切割结构域经进一步修饰或改造，而使其RNA切割活性降低或消除，成为RNA切割活性降低或消除的dCas13。
在一个优选的实施方案中，所述Cas13蛋白与一个或更多个异源功能性结构域融合。
在一个优选的实施方案中，所述融合在所述Cas13蛋白的N端、C端或者内部。
在一个优选的实施方案中，所述一个或更多个异源功能性结构域具有以下活性：脱氨酶如胞苷脱氨基酶和脱氧腺苷脱氨基酶、甲基化酶、去甲基化酶、转录激活、转录抑制、核酸酶、单链RNA裂解、双链RNA裂解、单链DNA裂解、双链DNA裂解、DNA或RNA连接酶、报告蛋白、检测蛋白、定位信号、或其任意组合。
在本公开内容的另一个方面中，提供了一种核酸分子，其包含编码上述Cas13蛋白的核苷酸序列。
在一个优选的实施方案中，所述核酸分子针对在特定宿主细胞中的表达而进行了密码子优化。
在一个优选的实施方案中，所述宿主细胞是原核或真核生物细胞，优选人细胞。
在一个优选的实施方案中，所述核酸分子包含与编码Cas13的核苷酸序列有效链接的启动子，其为组成型启动子、诱导型启动子、组织特异性启动子、嵌合型启动子或发育特异性启动子。
在本公开内容的另一个方面中，提供了一种表达载体，其包含上述核酸分子，以DNA或RNA或蛋白等形式表达上述氨基酸序列或核苷酸序列。
在一个优选的实施方案中，所述表达载体为腺相关病毒(AAV)、腺病毒、慢病毒、逆转录病毒、单纯孢疹病毒、溶瘤病毒。
在本公开内容的另一个方面中，提供了一种递送系统，其包含(1)上述表达载体，或上述Cas13蛋白；以及(2)递送载体。
在一个优选的实施方案中，所述递送载体是纳米颗粒、脂质体、外泌体、微囊泡或基因枪。
在本公开内容的另一个方面中，提供了一种CRISPR-Cas系统，其包含：(1)上述Cas13蛋白或核酸分子，或者其衍生物或功能片段；(2)用于靶向目标RNA的gRNA序列。
在一个优选的实施方案中，其中所述gRNA序列包含同向重复(DR)序列和靶向靶RNA部分的间隔区域的序列。
在一个优选的实施方案中，其中所述DR序列为表1中所示序列；其中所述间隔区序列为15-60个核苷酸，优选25-50个核苷酸，更优选30个核苷酸。
在一个优选的实施方案中，所述DR序列可以是对应以下任一项的衍生物，其中所述衍生物(i)与表1中所示序列中的任一个相比，具有一个或多个(例如1、2、3、4、5、6、7、8、9或10)个核苷酸的添加、缺失、或取代；(ii)与表1中所示序列中任何一个具有至少20％、30％、40％、50％、60％、70％、80％、90％、95％或97％的序列同一性；(iii)在严格条件下与表1中所示序列任意一个，或与(i)和(ii)中的任意一个杂交；或(iv)是(i)-(iii)中任何一个的互补物，条件是所述衍生物非表1中所示序列中的任何一个，并且所述衍生物编码一个RNA，或本身即是一个RNA，所述RNA与SEQ IDNO：199-397编码的任意RNA基本保持相同的二级结构。
在一个优选的实施方案中，所述CRISPR-Cas系统还包含：(3)靶RNA。
在一个优选的实施方案中，所述CRISPR-Cas系统引起靶RNA序列的降解、切割或序列的改变。
在一个优选的实施方案中，所述靶RNA是mRNA或ncRNA，包括选自lncRNA、miRNA、misc_RNA、Mt_rRNA、Mt_tRNA、rRNA、scaRNA、scRNA、snoRNA、snRNA、sRNA的非编码RNA。
在本公开内容的另一个方面中，提供了一种细胞，其包含上述Cas13蛋白、核酸分子、表达载体、递送系统或CRISPR-Cas系统。
在一个优选的实施方案中，所述细胞为原核细胞或真核细胞，优选人细胞。
在本公开内容的另一个方面中，提供了一种降解或切割目的细胞中靶RNA、修饰目的细胞中靶RNA的序列的方法，其包括使用上述Cas13蛋白、核酸分子、表达载体、递送载体或CRISPR-Cas系统。
在一个优选的实施方案中，所述目的细胞为原核细胞或真核细胞，优选人细胞。
在一个优选的实施方案中，其中所述目的细胞为离体细胞、体外细胞或体内细胞。
附图说明
图1显示出了候选Cas13蛋白DZ4细胞层面验证其RNase活性的流式分析结果。在哺乳动物细胞系检测候选蛋白酶切活性的流式分析实验结果：上图为含有DZ4蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后，流式分析的结果图。可以发现与阴性对照组相比，候选蛋白DZ4的具有RNase活性很强，对应红光被大幅度的敲低。阴性对照为只含有表达mcherry蛋白(红光)和DZ4蛋白(发绿光)的对照组；其中AP459为含有靶向mcherry不同区域的其中一个sgRNA的实验组。
图2展示出了候选Cas13蛋白DZ28细胞层面验证其RNase活性的流式分析结果。在哺乳动物细胞系检测候选蛋白酶切活性的流式分析实验结果：上图为含有DZ28蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后，流式分析的结果图。可以发现与阴性对照组相比，候选蛋白DZ28的具有RNase活性很强，对应红光被大幅度的敲低。阴性对照为只含有表达mcherry蛋白(红光)和DZ28蛋白(发绿光)的对照组；其中AP393为含有靶向mcherry不同区域的其中一个sgRNA的实验组。
图3展示出了候选Cas13蛋白DZ29细胞层面验证其RNase活性的流式分析结果。在哺乳动物细胞系检测候选蛋白酶切活性的流式分析实验结果：上图为含有DZ29蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后，流式分析的结果图。可以发现与阴性对照组相比，候选蛋白DZ29的具有RNase活性很强，对应红光被大幅度的敲低。阴性对照为只含有表达mcherry蛋白(红光)和DZ29蛋白(发绿光)的对照组；其中AP405，AP407为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图4展示出了候选Cas13蛋白DZ30细胞层面验证其RNase活性的流式分析结果。在哺乳动物细胞系检测候选蛋白酶切活性的流式分析实验结果：上图为含有DZ30蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后，流式分析的结果图。可以发现与阴性对照组相比，候选蛋白DZ30的具有RNase活性很强，对应红光被大幅度的敲低。阴性对照为只含有表达mcherry蛋白(红光)和DZ30蛋白(发绿光)的对照组；其中AP411，AP413为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图5展示出了候选Cas13蛋白DZ31细胞层面验证其RNase活性的流式分析结果。在哺乳动物细胞系检测候选蛋白酶切活性的流式分析实验结果：上图为含有DZ31蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后，流式分析的结果图。可以发现与阴性对照组相比，候选蛋白DZ31的具有RNase活性很强，对应红光被大幅度的敲低。阴性对照为只含有表达mcherry蛋白(红光)和DZ31蛋白(发绿光)的对照组；其中AP417，AP419为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图6展示出了候选Cas13蛋白DZ32细胞层面验证其RNase活性的流式分析结果。在哺乳动物细胞系检测候选蛋白酶切活性的流式分析实验结果：上图为含有DZ32蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后，流式分析的结果图。可以发现与阴性对照组相比，候选蛋白DZ32的具有RNase活性很强，对应红光被大幅度的敲低。阴性对照为只含有表达mcherry蛋白(红光)和DZ32蛋白(发绿光)的对照组；其中AP421，AP423为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图7展示出了候选Cas13蛋白DZ33细胞层面验证其RNase活性的流式分析结果。在哺乳动物细胞系检测候选蛋白酶切活性的流式分析实验结果：上图为含有DZ33蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后，流式分析的结果图。可以发现与阴性对照组相比，候选蛋白DZ33的具有RNase活性很强，对应红光被大幅度的敲低。
阴性对照为只含有表达mcherry蛋白(红光)和DZ33蛋白(发绿光)的对照组；其中AP427，AP429为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图8展示出了候选Cas13蛋白DZ35细胞层面验证其RNase活性的流式分析结果。在哺乳动物细胞系检测候选蛋白酶切活性的流式分析实验结果：上图为含有DZ35蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后，流式分析的结果图。可以发现与阴性对照组相比，候选蛋白DZ35的具有RNase活性很强，对应红光被大幅度的敲低。阴性对照为只含有表达mcherry蛋白(红光)和DZ35蛋白(发绿光)的对照组；其中AP441，AP443为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图9展示出了候选Cas13蛋白DZ36细胞层面验证其RNase活性的流式分析结果。在哺乳动物细胞系检测候选蛋白酶切活性的流式分析实验结果：上图为含有DZ36蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后，流式分析的结果图。可以发现与阴性对照组相比，候选蛋白DZ36的具有RNase活性很强，对应红光被大幅度的敲低。阴性对照为只含有表达mcherry蛋白(红光)和DZ36蛋白(发绿光)的对照组；其中AP25，AP27为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图10候选Cas13蛋白DZ37 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ37蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ37能靶向mCherry RNA，具有强RNase活性，对应红光被大幅度的敲低。阴性对照为只含有表达mcherry蛋白(红光)和DZ37蛋白(绿光)的对照组(不带gRNA)；其中AP33，AP35为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图11候选Cas13蛋白DZ38 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ38蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ38能靶向mCherry RNA，具有强RNase活性，对应红光被大幅度的敲低。阴性对照为只含有表达mcherry蛋白(FB132)和DZ38蛋白(绿光)的对照组(不带gRNA)；其中AP38，AP47为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图12候选Cas13蛋白DZ39 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ39蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ39靶向mCherry RNA，具有一定的RNase活性，对应红色荧光略被敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ39蛋白(绿光)的对照组(不带gRNA)；其中AP39，AP43为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图13候选Cas13蛋白DZ40 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ40蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ40能靶向mCherry RNA，具有一定的RNase活性，对应红光被敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ40蛋白(绿光)的对照组(不带gRNA)；其中AP49，AP53为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图14候选Cas13蛋白DZ44 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ44蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ44能靶向mCherry RNA，具有一定的RNase活性，对应红光被敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ44蛋白(绿光)的对照组(不带gRNA)；其中AP59，AP55为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图15候选Cas13蛋白DZ45 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ45蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ45能靶向mCherry RNA，具有强RNase活性，对应红光被大幅度的敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ45蛋白(绿光)的对照组(不带gRNA)；其中AP63，AP65为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图16候选Cas13蛋白DZ46 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ46蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ46能靶向mCherry RNA，具有强RNase活性，对应红光被大幅度的敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ46蛋白(绿光)的对照组(不带gRNA)；其中AP69，AP71为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图17候选Cas13蛋白DZ47 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ47蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ47能靶向mCherry RNA，具有强RNase活性，对应红光被大幅度的敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ47蛋白(绿光)的对照组(不带gRNA)；其中AP91，AP93为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图18候选Cas13蛋白DZ50 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ50蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ50能靶向mCherry RNA，具有一定的RNase活性，对应红光被敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ50蛋白(绿光)的对照组(不带gRNA)；其中AP121，AP125为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图19候选Cas13蛋白DZ51 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ51蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ51能靶向mCherry RNA，具有强RNase活性，对应红光被大幅度的敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ51蛋白(绿光)的对照组(不带gRNA)；其中AP127，AP131为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图20候选Cas13蛋白DZ52 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ52蛋白的质粒(含有对应靶向mCherry的gRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ52能靶向mCherry RNA，具有强RNase活性，对应红光被大幅度的敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ52蛋白(绿光)的对照组(不带gRNA)；其中AP133，AP135为含有靶向mcherry不同区域的其中2个gRNA的实验组。
图21候选Cas13蛋白DZ54 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ54蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ54能靶向mCherry RNA，具有一定的RNase活性，对应红光被敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ54蛋白(绿光)的对照组(不带gRNA)；其中AP153为含有靶向mcherry的gRNA的实验组。
图22候选Cas13蛋白DZ55 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ55蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ55能靶向mCherry RNA，具有一定的RNase活性，对应红光被敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ55蛋白(绿光)的对照组(不带gRNA)；其中AP157为含有靶向mcherry的gRNA的实验组。
图23候选Cas13蛋白DZ57 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ57蛋白的质粒(含有对应靶向mCherry的gRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ57能靶向mCherry RNA，具有一定的RNase活性，对应红光被敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ57蛋白(绿光)的对照组(不带gRNA)；其中AP169，AP171为含有靶向mCherry不同区域的其中2个gRNA的实验组。
图24候选Cas13蛋白DZ62 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ62蛋白的质粒(含有对应靶向mCherry的gRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ62能靶向mCherry RNA，具有强RNase活性，对应红光被大幅度的敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ62蛋白(绿光)的对照组(不带gRNA)；其中AP187，AP191为含有靶向mcherry不同区域的其中2个gRNA的实验组。
图25候选Cas13蛋白DZ63 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ63蛋白的质粒(含有对应靶向mCherry的gRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ63能靶向mCherry RNA，具有强RNase活性，对应红光被大幅度的敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ63蛋白(绿光)的对照组(不带gRNA)；其中AP193，AP197为含有靶向mcherry不同区域的其中2个gRNA的实验组。
图26候选Cas13蛋白DZ65 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ65蛋白的质粒(含有对应靶向mCherry的gRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ65能靶向mCherry RNA，具有强RNase活性，对应红光被大幅度的敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ65蛋白(绿光)的对照组(不带gRNA)；其中AP201，AP203为含有靶向mcherry不同区域的其中2个gRNA的实验组。
图27候选Cas13蛋白DZ68 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ68蛋白的质粒(含有对应靶向mCherry的gRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ68能靶向mCherry RNA，具有强RNase活性，对应红光被大幅度的敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ68蛋白(绿光)的对照组(不带gRNA)；其中AP217，AP219为含有靶向mcherry不同区域的其中2个gRNA的实验组。
图28候选Cas13蛋白DZ86 RNase活性流式分析结果。在哺乳动物细胞系(HEK293T)中检测候选蛋白酶切活性的流式分析实验结果，将含有DZ86蛋白的质粒(含有对应靶向mCherry的gRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后进行流式分析。可以发现与阴性对照组相比，候选蛋白DZ86能靶向mCherry RNA，具有强RNase活性，对应红光被大幅度的敲低。阴性对照为只含有表达mCherry蛋白(FB132)和DZ86蛋白(绿光)的对照组(不带gRNA)；其中AP711，AP713为含有靶向mcherry不同区域的其中2个gRNA的实验组。
图29展示出了候选Cas13蛋白DZ90细胞层面验证其RNase活性的流式分析结果。在哺乳动物细胞系检测候选蛋白酶切活性的流式分析实验结果：上图为含有DZ90蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后，流式分析的结果图。可以发现与阴性对照组相比，候选蛋白DZ90的具有RNase活性很强，对应红光被大幅度的敲低。阴性对照为只含有表达mcherry蛋白(红光)和DZ90蛋白(发绿光)的对照组；其中AP313，AP317为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图30展示出了候选Cas13蛋白DZ91细胞层面验证其RNase活性的流式分析结果。在哺乳动物细胞系检测候选蛋白酶切活性的流式分析实验结果：上图为含有DZ91蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后，流式分析的结果图。可以发现与阴性对照组相比，候选蛋白DZ91的具有RNase活性很强，对应红光被大幅度的敲低。阴性对照为只含有表达mcherry蛋白(红光)和DZ91蛋白(发绿光)的对照组；其中AP319，AP323为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图31展示出了候选Cas13蛋白DZ93细胞层面验证其RNase活性的流式分析结果。在哺乳动物细胞系检测候选蛋白酶切活性的流式分析实验结果：上图为含有DZ93蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后，流式分析的结果图。可以发现与阴性对照组相比，候选蛋白DZ93的具有RNase活性很强，对应红光一定程度上被敲低。阴性对照为只含有表达mcherry蛋白(红光)和DZ93蛋白(发绿光)的对照组；其中AP151为含有靶向mcherry不同区域的其中1个sgRNA的实验组。
图32展示出了候选Cas13蛋白DZ96细胞层面验证其RNase活性的流式分析结果。在哺乳动物细胞系检测候选蛋白酶切活性的流式分析实验结果：上图为含有DZ96蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后，流式分析的结果图。可以发现与阴性对照组相比，候选蛋白DZ96的具有RNase活性很强，对应红光被大幅度的敲低。阴性对照为只含有表达mcherry蛋白(红光)和DZ96蛋白(发绿光)的对照组；其中AP349，AP353为含有靶向mcherry不同区域的其中2个sgRNA的实验组。
图33展示出了候选Cas13蛋白DZ98细胞层面验证其RNase活性的流式分析结果。在哺乳动物细胞系检测候选蛋白酶切活性的流式分析实验结果：上图为含有DZ98蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后，流式分析的结果图。可以发现与阴性对照组相比，候选蛋白DZ98的具有RNase活性很强，对应红光被大幅度的敲低。阴性对照为只含有表达mcherry蛋白(红光)和DZ98蛋白(发绿光)的对照组；其中AP361为含有靶向mcherry不同区域的其中1个sgRNA的实验组。
图34展示的阳性对照Cas13d蛋白在细胞层面验证其RNase活性的流式分析结果。在哺乳动物细胞系检测候选蛋白酶切活性的流式分析实验结果：上图为含有Cas13d蛋白的质粒(含有对应靶向mCherry的sgRNA)与含有mCherry蛋白的质粒共转染293T细胞系48h后，流式分析的结果图。可以发现与阴性对照组相比，候选蛋白Cas13d的具有RNase活性很强，对应红光被大幅度的敲低。阴性对照为只含有表达mcherry蛋白(红光)和Cas13d蛋白(发绿光)的对照组；其中px261为含有靶向mcherry不同区域的其中1个sgRNA的实验组。
图35展示的是候选蛋白DZ4敲低内源基因STAT3基因的qPCR结果，可以发现随机设计靶STAT3基因的sgRNA跟DZ4蛋白一起瞬转可以在一定程度上敲低内源基因
图36A和图36B展示的是候选蛋白DZ29敲低内源基因STAT3和EZH2基因的qPCR结果，可以发现随机设计靶STAT3和EZH2基因的sgRNA跟DZ29蛋白一起瞬转可以在一定程度上敲低内源基因。其中不同DR对DZ29敲低内源基因有一定的影响。
图37展示的是候选蛋白DZ32敲低内源基因STAT3和EZH2基因的qPCR结果，可以发现随机设计靶STAT3和EZH2基因的sgRNA跟DZ32蛋白一起瞬转可以在一定程度上敲低内源基因。
图38展示的是候选蛋白DZ47敲低内源基因STAT3和EZH2基因的qPCR结果，可以发现随机设计靶STAT3和EZH2基因的sgRNA跟DZ47蛋白一起瞬转可以在一定程度上敲低内源基因。
图39展示的是候选蛋白DZ51敲低内源基因STAT3和EZH2基因的qPCR结果，可以发现随机设计靶STAT3和EZH2基因的sgRNA跟DZ51蛋白一起瞬转可以在一定程度上敲低内源基因。
图40展示的是候选蛋白DZ54敲低内源基因STAT3和EZH2基因的qPCR结果，可以发现随机设计靶STAT3和EZH2基因的sgRNA跟DZ54蛋白一起瞬转可以在一定程度上敲低内源基因。
图41展示的是候选蛋白DZ68敲低内源基因STAT3和EZH2基因的qPCR结果，可以发现随机设计靶STAT3和EZH2基因的sgRNA跟DZ68蛋白一起瞬转可以在一定程度上敲低内源基因。
图42A和图42B展示的是候选蛋白DZ93敲低内源基因STAT3和EZH2基因的qPCR结果，可以发现随机设计靶STAT3和EZH2基因的sgRNA跟DZ93蛋白一起瞬转可以在一定程度上敲低内源基因。其中不同DR对DZ93敲低内源基因有一定的影响。
图43展示的是候选蛋白DZ98敲低内源基因STAT3基因的qPCR结果，可以发现随机设计靶向STAT3基因的sgRNA跟DZ98蛋白一起瞬转可以在一定程度上敲低内源基因。
图44展示的候选蛋白敲低293T内源基因STAT3的qPCR结果。其中框框部分显示部分蛋白，相对于对照组，潜在具有敲低STAT3的Rnase的效率。
图45A和图45B展示的是候选蛋白DZ806敲低内源基因STAT3和EZH2基因的qPCR结果，可以发现随机设计靶STAT3和EZH2基因的sgRNA跟DZ806蛋白一起瞬转可以在一定程度上敲低内源基因。
图46A和图46B展示的是候选蛋白DZ821敲低内源基因STAT3和EZH2基因的qPCR结果，可以发现随机设计靶STAT3和EZH2基因的sgRNA跟DZ821蛋白一起瞬转可以在一定程度上敲低内源基因。
图47A和图47B展示的是候选蛋白DZ822敲低内源基因STAT3和EZH2基因的qPCR结果，可以发现随机设计靶STAT3和EZH2基因的sgRNA跟DZ822蛋白一起瞬转可以在一定程度上敲低内源基因。
图48A和图48B展示的是候选蛋白DZ825敲低内源基因STAT3和EZH2基因的qPCR结果，可以发现随机设计靶STAT3和EZH2基因的sgRNA跟DZ825蛋白一起瞬转可以在一定程度上敲低内源基因。
图49展示的是筛选DZ825靶向切割RNA的偏好基序分析的实验结果，可以发现它在5’和3’都有很强碱基偏好性。距离target sequence的5’段第1个碱基为C，而3’段则为G；
图50展示的是候选具有guide RNA引导，潜在具有Rnase活性的CRISPR-Cas13与已知Cas13蛋白家族成员的进化关系，可以发现我们候选的蛋白潜在分成2个新的家族。暂且即为Cas13m1和Cas13m2；如DZ30，DZ32；Cas13m2家族的DZ47，DZ29等。
具体实施方式
下面将结合实施例对本发明的实施方案进行详细描述，但是本领域技术人员将会理解，下列实施例仅用于举例说明本发明，而不应视为限定本发明的范围。实施例中未注明具体条件者，按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者，均为可以通过市购获得的常规产品。
如在说明书中所使用的，没有数量词修饰的名词可意指一个/种或更多个/种。如在权利要求书中所使用的，当与词语“包含/包括”结合使用时，没有数量词修饰的名词可意指一个/种或多于一个/种。
权利要求书中术语“或/或者”的使用用于意指“和/或”，除非明确地指出仅指替代方案或替代方案是相互排斥的，尽管本公开内容支持仅指替代方案和“和/或”的限定。如本文中使用的“另一/另一些”可意指至少第二或更多个/种。
在整个本申请中，术语“约”用于表示值包括装置的误差、用于确定该值的方法的固有变化，或者存在于研究对象之间的固有变化。这样的固有变异可以是标注值的±10％的变异。
在整个申请中，除非另有说明，否则核苷酸序列以5’至3’方向列出，并且氨基酸序列以N端至C端方向列出。
通过以下详细描述，本发明的其他目的、特征和优点将变得明显。然而，应理解，尽管表明了本发明的一些优选实施方案，但是详细描述和具体实施例仅以举例说明的方式给出，因为根据该详细描述，在本发明的精神和范围内的多种变化和修改对于本领域技术人员而言将变得明显。
定义
NCBI(https://www.ncbi.nlm.nih.gov/)是指美国国家生物信息中心，是一个面向全世界的公共数据库，本领域技术人员利用该数据库提供的核酸数据库进行下载原核生物的基因组，蛋白质组相关数据库等，也可以利用该数据提供的blast比对软件进行序列比对的分析。
IMG(https://img.jgi.doe.gov/)是指微生物基因组整合数据库，是新一代基因组数据库的代表，不仅能够完整收录现有数据库的内容，还提供了更完善的数据上传、注释和分析服务，将测序数据储存到IMG/M数据库。该数据可以下载纯培养细菌测序基因组、宏基因组、宏基因组组装基因组、单细胞测序基因组的数据。
CRISPR(cluster regularly interspaced short palindromic repeats)是原核生物，主要是指细菌和古细菌体内的一串DNA序列，包括同向重复(direct repeat，DR)区域和非重复间隔区(spacer)区域。而CRIPSR系统除了包含CRISPR array外，还包括相关的Cas蛋白。它们一起构成了细菌低于外来病毒入侵的免疫系统。
Cas13家族是目前已知该能够靶向RNA的CRIPSR酶家族，其成员包括Cas13a、Cas13b、Cas13c、Cas13d、Cas13X和Cas13Y家族。与CRISPR/Cas9切割DNA的活性不同，CRISPR/Cas13能够用于切割细菌细胞中特定的RNA序列。
HEPN结构域是higher eukaryotes and prokaryotes nucleotide domain的简称，是CRISPR-Cas13酶系统中Cas13蛋白发挥切割和抵御外来入侵核酸的重要结构域。
ABE系统是Adenine base editors的简称，即嘌呤碱基转换技术，能够实现A/T到G/C的单碱基改变。最常用的酶是adar酶(adenosine deaminases acting on RNA，一种作用于RNA的腺苷脱氨酶)。主要是通过将腺嘌呤脱氨基成肌苷，在DNA或者RNA中进行读码的时候会被看成G，从而实现A/T到G/C的突变。由于细胞对肌苷的切出修复不敏感，因而这种突变可以维持较高的产物纯度。
CBE系统是Cytidine base editor的简称，即嘧啶碱基转换技术，目前有BE1、BE2和BE3个工具，其中BE3的效率最高，因而在基因治疗，动物模型制作以及功能基因筛选等领域被广泛应用。
真核细胞例如哺乳动物细胞，包括人类细胞(人类原代细胞或已建立的人类细胞系)。所述细胞可以是非人类哺乳动物细胞，例如来自非人类灵长类动物(例如猴子)、奶牛/公牛/家牛、绵羊、山羊、猪、马、狗、猫、啮齿动物(例如兔子、小、大鼠、仓鼠)等。所述细胞来自鱼(例如鲑鱼)、鸟(例如禽鸟，包括小鸡、鸭、鹅)、爬行动物、贝类(例如牡蛎、蛤、龙虾、虾)、昆虫、蠕虫、酵母等。所述细胞可以来自植物，例如单子叶植物或双子叶植物。所述植物可以是粮食作物，例如大麦、木薯、棉花、花生、玉米、小米、油棕果、土豆、豆类、油菜籽或低芥酸菜子、大米、黑麦、高粱、大豆、甘蔗、糖甜菜、向日葵和小麦。所述植物可以是谷物(例如大麦、玉米、小米、大米、黑麦、高粱和小麦)。所述植物可以是块茎(例如木薯和土豆)。在一些实施方案中，所述植物可以是糖料作物(例如甜菜和甘蔗)。所述植物可以是含油作物(例如大豆、花生、油菜籽或低芥酸菜子、向日葵和油棕果)。所述植物可以是纤维作物(例如棉花)。所述植物可以是树木，例如桃树或油桃树、苹果树、梨树、杏树、核桃树、开心果树、柑橘属树(例如橙子、葡萄柚或柠檬树)、草、蔬菜、水果或藻类。所述植物可以是茄属植物；芸苔属(Brassica)植物；莴苣属(Lactuca)植物；菠菜属(Spinacia)植物；辣椒属(Capsicum)植物；棉花、烟草、芦笋、胡萝卜、卷心菜、西兰花、花椰菜、番茄、茄子、胡椒、生菜、菠菜、草莓、蓝莓、覆盆子、黑莓、葡萄、咖啡、可可等。
CRISPR系统
CRISPR(成簇规律间隔短回文重复序列)/Cas13(CRISPR相关蛋白13)介导的RNA编辑正在成为用于疾病诊疗、植物育种等方面的有前景的工具。
CRISPR是包含碱基序列的短重复的DNA基因座。每个重复之后是来自先前暴露于病毒的“间隔区DNA”的短区段。在约40％的测序的真细菌基因组和90％的测序的古细菌中发现CRISPR。CRISPR通常与编码与CRISPR相关的蛋白质的Cas基因相关。CRISPR/Cas系统是原核免疫系统，其赋予对外来遗传元件(例如质粒和噬菌体)的抗性并提供获得性免疫的形式。CRISPR间隔区识别并沉默真核生物体中的这些外源遗传元件(例如RNAi)。
CRISPR重复序列的大小为24至48个碱基对。它们通常显示一些二重对称，这意味着形成二级结构例如发夹，但不是真正的回文结构。重复序列被相似长度的间隔区分开。一些CRISPR间隔区序列与来自质粒和噬菌体的序列准确地匹配，尽管一些间隔区与原核生物的基因组匹配。响应于噬菌体感染，可迅速添加新的间隔区。
指导RNA(gRNA)。作为RNA指导的蛋白，Cas13需要短RNA以指导RNA靶标的识别。
核酸酶
Cas核酸酶。CRISPR相关(Cas)基因通常与CRISPR重复-间隔区阵列相关。截至2013年，已描述了超过四十个不同的Cas蛋白家族。在这些蛋白家族之中，Cas1看来在不同的CRISPR/Cas系统中是普遍存在的。Cas基因和重复序列结构的特定组合已用于限定8种CRISPR亚型(Ecoli、Ypest、Nmeni、Dvulg、Tneap、Hmari、Apern和Mtube)，其中一些与编码重复序列相关神秘蛋白(repeat-associated mysterious protein，RAMP)的另外的基因模块相关。在单个基因组中可存在多于一种CRISPR亚型。CRISPR/Cas亚型的散发性分布(sporadic distribution)表明该系统在微生物进化期间经历水平基因转移。
外源DNA明显地由Cas基因编码的蛋白质加工成小元件(长度为约30个碱基对)，然后以某种方式将其插入到靠近前导序列的CRISPR基因座中。来自CRISPR基因座的RNA是组成型表达的，并且被Cas蛋白加工成由具有侧翼重复序列的单独外源来源序列元件构成的小RNA。RNA指导其他Cas蛋白在RNA或DNA水平上沉默外源遗传元件。证据表明CRISPR亚型之间的功能多样性。Cse(Cas亚型Ecoli)蛋白(在大肠杆菌(E.coli)中称为CasA-E)形成功能性复合体Cascade，其将CRISPR RNA转录物加工成保留Cascade的间隔区-重复序列单元。在另一些原核生物中，Cas6加工CRISPR转录物。有趣的是，大肠杆菌中基于CRISPR的噬菌体灭活需要Cascade和Cas3，但不需要Cas1和Cas2。在激烈火球菌(Pyrococcus furiosus)和另一些原核生物中发现的Cmr(Cas RAMP模块)蛋白与小的CRISPR RNA形成功能性复合体，其识别和切割互补靶RNA。RNA指导的CRISPR酶被分类为V型限制酶。
实施例
实施例1：新型Cas13蛋白从头筛选
我们还进行了从头寻找CRISPR-Cas13其他家族成员。简单来说，该分析系统包括2大块，一部分CRISPR array区域的鉴定，我们首先下载NCBI和IMG截止到2021年7月份的全部细菌，古细菌基因组以及宏基因组的序列，利用CRISPR array鉴定软件(如Pilercr)进行鉴定CRISPR array区域；另一部分是该区域上下游附近Cas相关蛋白的搜寻，即取该区域上下游临近的6个蛋白，共计12个蛋白进行目标结构域分析。最终候选蛋白的氨基酸序列编号、拓展的HEPN结构域和坐标等信息参见表2。
其中本筛选体系的HEPN结构域除了包含过去Cas13家族成员中发现的RxxxxH(R4xH)特征(我们记为早期HEPN结构域)外，还进行了拓展，包括其他具有RNA切割活性的结构域RxxxxxH(R5xH)和RxxxxxxH(R6xH)(我们将RxxxxxH(R5xH)和RxxxxxxH(R6xH)以及RxxxxH(R4xH)总体记为拓展的HEPN结构域)。其中临近R保守氨基酸优选为N、Q、H或D，例如R[NDQH]xxxH、R[NDQH]xxxxH、R[NDQH]xxxxxH等组合；其中R4xH优选R[NDQH]xxxH，而R5xH和R6xH则优选R[NDQH]xxxxH和R[NQDH]xxxxxH；其中x代表任意氨基酸，而中括号内N、D、Q和H为优先考虑保守氨基酸。
实施例2：新型候选Cas13蛋白的功能验证
候选蛋白筛到后将进行功能验证。简而言之，我们首先将候选蛋白的核酸序列，DR序列以及target spacer序列送公司进行合成，然后将其导入表达质粒中构建相应的质粒。然后通过质粒转化在DH-5a大肠杆菌感受态细胞进行质粒扩增培养，然后抽提质粒后进行人源293T(能表达红光)细胞系的转染试验，与此同时我们还设计了对应的阴性和阳性对照组来进一步确认候选蛋白的切割活性。共转染质粒48h后进行流式细胞分析等试验来最终确定候选蛋白的RNA酶切活性。
按照上述研究策略，我们对候选蛋白进行了验证。如图1至34中所示，从DZ4，DZ28，DZ29，DZ30，DZ31，DZ32，DZ33，DZ35，DZ36，DZ37，DZ38，DZ39，DZ40，DZ44，DZ45，DZ46，DZ47，DZ50，DZ51，DZ52，DZ54，DZ55，DZ57，DZ62，DZ63，DZ65，DZ68，DZ86，DZ90，DZ91，DZ93，DZ96，DZ98以及Cas13d的结果表明，我们候选的具有RNase活性的Cas13蛋白在瞬转哺乳动物细胞系能够很好的敲低mCherry(红光)蛋白，而阴性对照组则不能切割，表现为红色荧光高亮；表明我们的候选蛋白可以在哺乳动物等上发挥RNase的作用。
实施例3：新型候选Cas13蛋白的敲低293T内源基因功能验证
为了进一步验证候选蛋白切割内源基因的能力，我们从实施例2中进一步筛选一些切割mCherry的RNase活性比高蛋白如DZ4，DZ29，DZ32，DZ47，DZ51，DZ54，DZ68，DZ93，DZ98等，进行敲低内源基因(STAT3，EZH2)实验验证，我们随机对293T这两个内源基因设计sgRNA。发现还有有一部分蛋白保持比较好的敲低内源基因的作用。如图35到43所示。尽管也有一部分实验组和对照组差异不明显，这可能跟蛋白的PFS有关。因为以往研究报道Cas13蛋白，如Cas13a,Cas13b等在靶向RNA序列的时候有很强的偏好性(PFS)。而此处我们紧紧随机设计靶向目标基因的sgRNA并没有去筛选对应Cas蛋白靶向RNA的PFS偏好性。
从qPCR的结果充分证明我们筛选候选guide RNA引导的具有Rase活性的CRISPR-Cas蛋白的方法的可行性，为了进一步加快筛选潜在切割真核生物内源基因RNA的CRISPR-Cas蛋白，我们将一部分筛选的guide RNA引导的潜在具有RNase活性的CRISPR-Cas蛋白(包括dz806，dz825，dz822，dz821等)直接针对内源基因STAT3和EZH2进行knock down实验。结果如图44到48所示。
可以发现尽管蛋白的PFS未知，还是有一部分候选蛋白显示出一定的knock down内源基因STAT3的作用，包括DZ784，DZ787，DZ788，DZ791，DZ793，DZ794，DZ796，DZ797，DZ798，DZ800，DZ803，DZ805，DZ810，DZ813，DZ814，DZ816，DZ817，DZ821和DZ824。随后对DZ806，DZ810，DZ821，DZ822和DZ825进一步进行EZH2内源基因的knock down实验，结果如图45至48所示。发现多次重复实验都显示它们对内源基因EZH2和STAT3有一定的knock down效果。
实施例4：新型候选Cas13蛋白PFS功能筛选
为了进一步挖掘候选蛋白靶向RNA的偏好性，我们还设计了寻找蛋白PFS的检测实验，简单来说就是首先在抗性基因(如氨苄)前设计添加5’-6N(NNNNNN)-spacer(targetsequence)-抗性基因的library质粒或者spacer(target sequence)-NNNNNN(6N)-3’-抗性基因的library质粒(统一标记为6N library质粒)。同时针对target sequence设计guideRNA的质粒。质粒构建好后，首先针对6N library的质粒转染大肠杆菌。然后将候选蛋白的质粒和对应靶向目标区域的guide RNA一起共转染大肠杆菌。与此同时做1组阴性对照，该对照为候选蛋白相关质粒与非靶向目标区域的guide RNA(即nonTarget)质粒进行共转。然后将活下来的菌全部进行抽提并做deep-seq测序。通过生物信息学方法，分析实验组和对照组差异的5’或者3’偏好序列就可以计算对应蛋白的PFS。
按照该方法，我们测试了DZ825蛋白，如图49所示，它的潜在PFS为5’-C-tareget-G-3’。
实施例5：新型紧凑型候选Cas13蛋白的碱基编辑功能验证
当前用于单碱基编辑的系统主要有两种，一种是ABE系统，另一种是CBE系统。简单来说，通过候选Cas13蛋白的切割结构域(拓展的HEPN结构域)进行突变处理，获得只有结合RNA而没有切割活性的候选dCas13蛋白，然后融合adar酶序列，构建ABE单碱基编辑系统的质粒，然后对特定序列，比如TP53基因的转录本进行定点碱基突变处理的sgRNA设计并构建相应的质粒载体。然后通过共转染人源293T细胞系，48小时后进行流式细胞分选获得共转染的细胞系。然后进行RNA转录本的提取以及建库。然后进deep seq测序。测序结束后通过生物信息方法分析TP53基因转录本的突变情况就可以获得对应的ABE系统的单碱基编辑效能分析。从而通过不断的优化sgRNA来实现构建目标区域的最优单碱基编辑系统。
实施例6：候选Cas13蛋白与已知Cas13蛋白的同源性分析
依据未知蛋白在已知蛋白的覆盖度越高且相似度占比越大则未知蛋白与已知蛋白的同源性越近的原理进行。对所筛选到的候选蛋白后，我们先从NCBI数据库以及专利文献中下载Cas13a，b，c，d，x(e)，y(f)，bt的相关蛋白序列，然后与我们的数据一起合并构建本地blastp的索引文件，然后将候选蛋白序列比对到本地blastp索引库中进行蛋白序列比对分析。对于蛋白之间相似度(identity)小于20％或者没法比对到本地索引库的部分我们统一标注为20％；类似的，对于覆盖度(coverage)小于5％或者没法比对到本地索引库的标记为1％。本发明方法所鉴定出的新Cas13蛋白与已知各家族Cas13蛋白的同源性水平极低。例如，DZ47、DZ51、DZ68、DZ825等与目前已知的各Cas13类别的同源性均在20％以下。
进一步进化树分析显示，我们候选的guide RNA引导的，具有RNase活性的CRISPR-Cas13蛋白具有独立分支，潜在具有两个比较大的紧凑型Cas13新家族，暂且记为Cas13 m1家族和Cas13 m2家族。其中Cas13 m1家族如DZ30，DZ32等；Cas13m2家族的DZ47，DZ29等。
候选Cas13蛋白的DR序列参见下表1。
表1.候选Cas13蛋白的DR序列
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最终候选Cas13蛋白的氨基酸序列编号、拓展的HEPN结构域和坐标等信息参见表2。
表2.候选Cas13蛋白总结表
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序列表
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His Arg Pro Tyr Glu Gly Ser Gln Pro Trp Leu Thr Arg Val Ala Glu
50 55 60
Trp Met His Asn Ser Pro His Arg Pro Tyr Glu Gly Ser Gln His Ala
65 70 75 80
Arg Gly Pro Val Gly Leu Pro Thr Thr Lys Ser Ser Ser Pro Leu Arg
85 90 95
Gly Val Ala Thr Val Thr His Asp Leu Arg Gln Leu Arg His Arg Gly
100 105 110
Pro His Arg Pro Tyr Glu Gly Ser Gln Pro Gly Arg Ser Ala Val Val
115 120 125
Phe Ala Gly Ser Ala Ser Ser Ser Pro Leu Arg Gly Val Ala Thr Gly
130 135 140
Glu Arg Val Pro Ala Gly Cys Pro Glu His Val Leu Ile Ala Pro Thr
145 150 155 160
Arg Gly Arg Asn Arg Ile Asn Ser Ile Ala Arg Ser Gln Phe Cys Gly
165 170 175
Val Leu Ile Ala Pro Thr Arg Gly Arg Asn Arg Val Thr Lys Ile Glu
180 185 190
Gln Glu Ile Lys Gly Ser Pro His Arg Pro Tyr Glu Gly Ser Gln Gln
195 200 205
His Glu Gly Ala Val Val Asp Gly Thr Gln Glu Ser Ser Ser Pro Leu
210 215 220
Arg Gly Val Ala Thr Arg Arg Arg Arg Ser Ser Ser Trp Cys Arg Arg
225 230 235 240
Cys Val Leu Ile Ala Pro Thr Arg Gly Arg Asn Arg Phe Thr Arg Thr
245 250 255
Arg Ser Pro Gly Ser Ser Ala Val Leu Ile Ala Pro Thr Arg Gly Arg
260 265 270
Asn Thr Arg Thr Thr Arg Ser Arg Phe Arg Ser Val Ser Val Leu Ile
275 280 285
Ala Pro Thr Arg Gly Arg Asn Thr Ala Asp Gly Arg Arg Pro Thr Pro
290 295 300
Asp Leu Pro Ser Ser Ser Pro Leu Arg Gly Val Ala Thr Arg Arg Thr
305 310 315 320
Arg Thr Ser Arg Thr Cys Ser Arg Gly Pro His Arg Pro Tyr Glu Gly
325 330 335
Ser Gln Arg Ala Gly Arg Asp Gln Leu Arg His Val Pro Ala Val Leu
340 345 350
Ile Ala Pro Thr Arg Gly Arg Asn Thr Arg Ser Arg Gly Arg Ala Gly
355 360 365
Pro Ser His Gly Val Leu Ile Ala Pro Thr Arg Gly Arg Asn
370 375 380
<210> 6
<211> 359
<212> PRT
<213> 宏基因组(metagenomic)
<400> 6
Met Arg Gly Ser Ala Arg Pro Ser Val Arg Ser Leu Pro Arg Arg Gln
1 5 10 15
Leu Arg Asn Val Ala Gly Asn Ser Ala Asn Cys Ala Cys Ser Ser Leu
20 25 30
Pro Arg Arg Gln Leu Arg Lys Ile Ser Leu Ile Asp Leu Asp Thr Val
35 40 45
Gln Arg Ser Leu Pro Arg Arg Gln Leu Arg Asn Arg Ser Arg His Lys
50 55 60
Ala Arg Trp Ala Ala Gly Ser Leu Pro Arg Arg Gln Leu Arg Asn Arg
65 70 75 80
Gln Ala Arg Arg Arg Pro Thr Asp Ser Arg Ser Leu Pro Arg Arg Gln
85 90 95
Leu Arg Asn Val Val Ser Arg Arg Pro Pro Gln Pro Arg Cys Ser Leu
100 105 110
Pro Arg Arg Gln Leu Arg Lys Pro Thr Thr Gly Val Ala Arg Thr Trp
115 120 125
Val Gly Ser Leu Pro Arg Arg Gln Leu Arg Lys Pro Thr Ser Pro Ser
130 135 140
Arg Met Gln Arg Phe Arg Ser Leu Pro Arg Arg Gln Leu Arg Asn Ile
145 150 155 160
Arg Leu His Ser Val Gln Ile Asn Gln Cys Ser Leu Pro Arg Arg Gln
165 170 175
Leu Arg Lys Ala Ala Pro Leu Gln Thr Val Leu Ser Ala Cys Ser Leu
180 185 190
Pro Arg Arg Gln Leu Arg Asn Pro Arg Pro Leu Glu Gln His Val Gln
195 200 205
Gly Gly Ser Leu Pro Arg Arg Gln Leu Arg Asn Ala Ala Gly Gly Gly
210 215 220
Thr Ser Val Ser Arg Arg Ser Leu Pro Arg Arg Gln Leu Arg Asn Val
225 230 235 240
Gly Ala Asp Gly Asp Asn Thr Gln Ala Gly Ser Leu Pro Arg Arg Gln
245 250 255
Leu Arg Asn Leu Arg His Thr Glu Thr Glu Phe Arg Asp Ser Ser Leu
260 265 270
Pro Arg Arg Gln Leu Arg Asn Lys Gln Arg Val Lys Asn Thr Phe Val
275 280 285
Thr Cys Ser Leu Pro Arg Arg Gln Leu Arg Asn Arg Cys Pro Asp Cys
290 295 300
Arg Ser Gln Asp Leu Cys Ser Leu Pro Arg Arg Gln Leu Arg Asn Leu
305 310 315 320
Trp Gly Asn His Phe Arg Lys Val His Arg Ser Leu Pro Arg Arg Gln
325 330 335
Leu Arg Asn Pro Arg Ala Val Gly Val Ala Pro Cys Phe Gly Ser Leu
340 345 350
Pro Arg Arg Gln Leu Arg Asn
355
<210> 7
<211> 350
<212> PRT
<213> 宏基因组(metagenomic)
<400> 7
Val Leu Ala Asp Lys Pro Ser Arg Phe Leu Val Pro Val Arg Pro Ala
1 5 10 15
Leu Val Asp Ala Glu Val Val Ala Leu Leu Tyr Ala Arg Gly Gln Lys
20 25 30
Arg Ala Glu Glu Glu Thr Cys Ser Gly Leu Ser Ala Ser Gly Pro Thr
35 40 45
Thr Trp Phe Ala Gly Thr Val Asn Pro Lys Lys Ala Gly Pro Ser Pro
50 55 60
Arg Lys Asp His His Ser Lys Ser Tyr Tyr Arg Pro Ser Thr Leu Leu
65 70 75 80
Ala Leu Gly Asp Arg Arg Ser Ser Arg Lys Thr Ala Thr Thr Trp Arg
85 90 95
Gly Tyr Glu Arg His Thr Pro Cys Gly Ser Ser Pro Ser Gly Pro Ala
100 105 110
Gly Cys Asp Pro Gly Ser Pro Arg Gly Arg Pro Pro Pro Gly Gly Gln
115 120 125
Arg Arg Leu Val Leu Asp Ala Val Asp Leu Arg Ser Ser Ala Val Pro
130 135 140
Lys Gly Asp Arg His Val Ser Ser Thr Gly Ser Gly Val Pro Arg Ser
145 150 155 160
Thr Ser Leu Arg Ser Ser Val Val Pro Lys Gly Asp Gly His Pro Ser
165 170 175
Ser Met Leu Arg Pro Ile Gly Lys Pro Ala Phe Ala Leu Lys Gly Asn
180 185 190
Gly His Leu Thr Val Asp Ile Tyr Ala Ile Val Ala Phe Lys Leu Arg
195 200 205
Ser Ser Leu Val Pro Arg Ala Ala Ala Thr Thr Asp His Pro Arg Ile
210 215 220
Ala Thr Ala Ala Arg Gly Cys Asp Pro Arg Ser Pro Arg Arg Ala Thr
225 230 235 240
Ala Thr Phe Ser Pro Ser Arg Gly Val Val Gly Ala Leu Gly Val Ala
245 250 255
Ile Leu Gly Arg Pro Glu Gly Arg Pro Pro Arg Ser Ser Trp Phe Cys
260 265 270
Ser Ser Pro Val Ser Ser Gly Cys Asp Pro Arg Ser Ser Arg Arg Thr
275 280 285
Thr Ala Thr Arg Arg Pro Pro Gly Pro Arg Lys Ala Arg His Arg Gly
290 295 300
Cys Asp Pro Arg Ser Phe Pro Arg Ala Thr Ala Thr Arg Gln Gly Ser
305 310 315 320
Arg Arg His Arg Arg His Gln Gly Val Ala Ile Leu Gly Arg Pro Glu
325 330 335
Gly Arg Pro Pro Arg Arg Arg Gly Arg Ser Ala Ser Ser Thr
340 345 350
<210> 8
<211> 320
<212> PRT
<213> 宏基因组(metagenomic)
<400> 8
Met Arg Thr Gly Arg Pro His Cys Pro Pro Thr Glu Val Asn Pro Cys
1 5 10 15
Ala Asp Arg Thr Gly Ala Asp Arg Arg Arg Gly Arg His Arg Ala Gly
20 25 30
His Leu Arg Leu Arg Gly Glu Asn Pro Ala Ser Gly Thr Trp Ser Leu
35 40 45
Val Ser Gly Gly Leu Pro Pro Pro Ala Arg Arg Glu Arg Gly Leu Arg
50 55 60
Tyr Arg Pro Asp His Pro Asp Arg Thr Thr Ser Ala Cys Ala Glu Arg
65 70 75 80
Thr Val Cys Gly Ser Pro Ala Gly Cys Pro Ala Pro Asp His Leu Arg
85 90 95
Leu Arg Gly Glu Asn Ala Ala Gln Phe Ala Phe Asp Leu Gly Ala Asp
100 105 110
Gly Pro Pro Pro Pro Ala Arg Arg Glu Pro Ser Ala Thr Arg Asp Glu
115 120 125
Phe Leu Asn Asp Arg Thr Thr Phe Ala Cys Ala Glu Arg Thr Arg Arg
130 135 140
Arg Cys Arg Ala Gly Cys Gly Ser Ala Asp His Leu Arg Leu Arg Gly
145 150 155 160
Glu Asn Leu Asp Thr Glu Ser Trp Ala Thr Ser Gly Tyr Gly Pro Pro
165 170 175
Pro Pro Ala Arg Arg Glu Leu Gly Gly Ala Arg Gly Ala Leu Leu Gly
180 185 190
Gly Arg Thr Thr Ser Ala Cys Ala Glu Arg Thr Leu Lys Arg Arg Arg
195 200 205
Asp Leu Ala Gln Val Val Gln Ala Ser Pro Glu Thr Asp Lys Arg Ala
210 215 220
Arg Leu Phe Ser Gly Asp Leu Ala Glu Gly Ala Arg Leu Lys Cys Gly
225 230 235 240
Lys Ala Ile Asp Pro Arg Ile Pro Glu Gly Ser Gly His Cys Ser Gly
245 250 255
Val Asp Glu Val Thr Asp Arg Arg Val Arg Gly Trp Glu Pro Ser Val
260 265 270
Leu Ala Pro Gln Ala Arg Ile Asn Thr Gln His Leu Trp Arg Ala Leu
275 280 285
Arg Arg His Gly Gln Leu Gln Asn Gly Ser Thr Val Ile Ala His Arg
290 295 300
Ile Thr Cys His Arg Gln Asn Val Arg Ser Pro Ser Lys Leu Leu Pro
305 310 315 320
<210> 9
<211> 432
<212> PRT
<213> 宏基因组(metagenomic)
<400> 9
Met Ile Tyr Leu Gln Glu Lys Val Asn Lys Thr Ile Asp Val Arg Asn
1 5 10 15
Tyr Lys Thr Gly Lys Thr Ser Thr Ile Asp Lys Ser Trp Met Met Thr
20 25 30
Thr Phe Tyr Lys Arg Glu Trp Asn Gln Glu Val Gly Lys Gln Leu Thr
35 40 45
Glu Val Lys Leu Pro Asp Asn Leu Ser Gly Ile Pro Phe Thr Leu Arg
50 55 60
Gln Leu Lys Glu Lys Ala Ser Tyr Ser Leu Asp Gln Trp Leu Asn Asn
65 70 75 80
Val Thr Lys Gly Lys Val Ala Gly Asp Gly Lys Arg Pro Ile Asn Leu
85 90 95
Pro Thr Asn Leu Phe Asp Glu Thr Leu Ile Asn Leu Leu Gln Asn Asp
100 105 110
Leu Glu Ala Gln Gln Val Glu Tyr Pro Thr Asp Ala Lys Tyr Asn Glu
115 120 125
Leu Phe Lys Ile Trp Trp Arg Lys Arg Gly Asp Ser Thr Gln Ser Phe
130 135 140
Tyr Asn Ala Glu Arg Glu Tyr Val Ile Phe Asp Glu Lys Val Asn Phe
145 150 155 160
Lys Leu Gln Glu Asn Ala Met Phe Thr Asp Phe Tyr Ser Asp Ser Leu
165 170 175
Lys Lys Ala Phe Arg Ala Lys Gln Asn Thr Arg Arg Ile Glu Gln Arg
180 185 190
Ser Asn Arg Arg Leu Pro Asp Ile Gln Phe Ser Gln Val Glu Lys Val
195 200 205
Phe Lys Arg Ser Ile Ser Asn Thr Glu Lys Gln Ile Arg Leu Leu Lys
210 215 220
Glu Glu Asp Gln Ile Met Leu Leu Met Leu Glu Glu Leu Met Ser Ser
225 230 235 240
Asp Leu Asp Leu Lys Leu Asn Gln Ile Asp Thr Leu Leu Asn Lys Thr
245 250 255
Ile Thr Val Lys Lys Pro Val Thr Gly Asn Leu Ser Phe Gly Asp Lys
260 265 270
Ser Glu Ile Thr Arg Thr Ile Ile Asp Gln Arg Lys Arg Lys Asp His
275 280 285
Ser Met Leu His Lys Tyr Val Tyr Asp Arg Arg Leu Pro Glu Leu Phe
290 295 300
Glu Tyr Phe Glu Glu Asn Glu Ile Pro Leu Gln Asp Leu Lys Asn Glu
305 310 315 320
Leu Glu Ala Tyr Asn Thr Ala Lys Gln Met Val Leu Asp Ala Val Phe
325 330 335
Lys Phe Glu Glu Asp Ile Val Thr Asn Asn Gln Val His Asp Leu Ile
340 345 350
Gly Ser Ala Cys Asp Thr Gly His Ile Gln His Lys Val Tyr Leu Gln
355 360 365
Trp Leu Lys Lys Glu Gly Met Ile Asn Glu Asn Glu Tyr Leu Phe Leu
370 375 380
Asn Arg Val Arg Asn Cys Phe Ser His Asn Leu Phe Pro Gln Lys Arg
385 390 395 400
Thr Met Ser Leu Phe Val Asn Gln Trp Ala Asp Ser Asn Phe Ala Leu
405 410 415
Gln Ile Ala Glu His Tyr Asn Glu Lys Ile Asn Ala Ile Leu Ala Ile
420 425 430
<210> 10
<211> 533
<212> PRT
<213> 宏基因组(metagenomic)
<400> 10
Met Ser Gly Leu Pro Leu Pro Lys Pro Leu Arg Arg Trp Leu Leu Gly
1 5 10 15
Thr Leu Ser Asn Gln Glu Arg Lys Ala Gln Asp Trp Leu Gln Ser Lys
20 25 30
Leu Glu Glu Asn Leu Arg Leu Leu Leu Glu Ala Lys Arg Gln Leu Arg
35 40 45
Lys Leu Ala Glu Pro Pro Asn Arg Pro Phe Leu Arg Ala Gly Asp Leu
50 55 60
Ala Thr Trp Ile Ala Arg Asp Ile Ala Tyr Trp Gln His Pro Asp Gly
65 70 75 80
Arg Gln Leu Asn Ser Glu Glu Tyr Gly Glu Leu Gln Ala Ala Leu Ala
85 90 95
Phe Phe Gly Arg Glu Gly Ala Glu Gln Val Tyr Glu Leu Phe Glu Lys
100 105 110
Leu Glu Leu Val Gly Gln Pro Asn Arg Asp His Pro Phe Leu Phe Arg
115 120 125
Leu Lys Asn Arg Met Cys Pro Ala Arg Ser Arg Lys Arg Gln Pro Lys
130 135 140
Pro Ile Arg Gly Ile Leu Glu Phe Tyr Ile Ala Tyr Leu Arg Ala Arg
145 150 155 160
Glu Lys Phe Leu Thr Arg Leu Gln Gly Arg Leu Lys Ser Gly Ser Asn
165 170 175
Pro Thr Pro Glu Met Glu His Trp Ile Pro Pro Leu Arg Ser Arg Ser
180 185 190
Leu Pro Pro Arg Met Asn Glu Arg Ala Val Val Leu Pro Arg Gly Leu
195 200 205
Phe Arg Arg Arg Phe Val Glu Leu Leu Pro Pro Pro Asp Gly Ser Pro
210 215 220
Ala Asp Thr Pro Leu Glu Gln Leu Thr Leu Met Lys Leu Leu Glu Pro
225 230 235 240
Tyr Leu Asp Thr Pro Gln Ala Phe Tyr Arg Tyr Glu Arg Arg Tyr Ala
245 250 255
Arg Asn Glu His Gln Tyr Thr Thr Asp Gly Cys Asn Leu Ser Glu Leu
260 265 270
Ile Pro Gln Ile Glu Asn Glu Ala Thr Gln Gly Ser Glu Glu Glu Asp
275 280 285
Pro Asp Phe Arg Arg Lys Asp Leu Leu Cys Leu Ala Arg Thr Ile Asn
290 295 300
Asp Thr Glu Arg Arg Leu Arg Arg Tyr Gln Leu Gln Asp Arg Leu Leu
305 310 315 320
Trp Met Ile Val Arg Gln Glu Leu Glu Arg Glu Lys Asn Met Asn Leu
325 330 335
Gln Gly Trp Thr Leu Glu Asn Val Gly Phe Asp Gln Pro Asp Ser Pro
340 345 350
Leu Lys Gln Glu Ile Thr Phe Gln Leu Glu Tyr Pro Leu Glu Gly Arg
355 360 365
Asn Glu Pro Ile Arg Ile Glu Ala Lys Leu Arg Pro Arg His Tyr Gly
370 375 380
Lys Met Arg Arg Thr Leu His Asp Arg Arg Leu Pro Gly Leu Leu Gln
385 390 395 400
Trp Leu Asn Thr Glu Ser Val Asp Trp Glu Arg Leu Gln Gln Glu Leu
405 410 415
Thr Lys Thr Ile Thr Arg Gln Asp Glu Val Leu Gln Leu Ile Phe Glu
420 425 430
Leu Glu Lys Ala Val His Ser Arg Phe Glu Lys Glu Cys Thr Ala Lys
435 440 445
Lys Asn Arg Asp Tyr Leu Lys His Arg Asn Tyr Leu Glu Val Leu Arg
450 455 460
Glu His Gln Ile Leu Arg Ser Asp Glu Glu Phe Glu His Leu Leu Asn
465 470 475 480
Leu Arg Asn Arg Phe Ala His Asn Glu Val Pro Ser Pro Ala Glu Leu
485 490 495
Pro Leu Trp Lys Asp Glu Arg Pro Gly Asn Gly Asn Leu Met Glu Trp
500 505 510
Leu Ile Glu Lys Ser Leu Gln Leu Tyr Arg Asp Leu Leu Glu Arg Val
515 520 525
Gln Gly Lys Gly Asp
530
<210> 11
<211> 688
<212> PRT
<213> 宏基因组(metagenomic)
<400> 11
Met Gly Phe Ile Ser Val His Asp Leu Arg Lys Leu Leu Leu Met Glu
1 5 10 15
Leu Leu Cys Glu Gly Ser Phe Ser Arg Met Gln Ser Gly Phe Leu Arg
20 25 30
Lys Ala Asn Arg Ile Leu Asp Glu Thr Ala Glu Gly Lys Leu Gln Phe
35 40 45
Ser Ala Leu Phe Pro Glu Met Arg His Arg Phe Ile Pro Pro Gln Asn
50 55 60
Pro Lys Ser Lys Asp Arg Arg Glu Lys Ala Glu Thr Thr Leu Glu Lys
65 70 75 80
Tyr Lys Gln Glu Ile Lys Gly Arg Lys Asp Lys Leu Asn Ser Gln Leu
85 90 95
Leu Ser Ala Phe Asp Met Asn Gln Arg Gln Leu Pro Ser Arg Leu Leu
100 105 110
Asp Glu Trp Met Asn Ile Arg Pro Ala Ser His Ser Val Lys Leu Arg
115 120 125
Thr Tyr Val Lys Gln Leu Asn Glu Asp Cys Arg Leu Arg Leu Arg Lys
130 135 140
Phe Arg Lys Asp Gly Asp Gly Lys Ala Arg Ala Ile Pro Leu Val Gly
145 150 155 160
Glu Met Ala Thr Phe Leu Ser Gln Asp Ile Val Arg Met Ile Ile Ser
165 170 175
Glu Glu Thr Lys Lys Leu Ile Thr Ser Ala Tyr Tyr Asn Glu Met Gln
180 185 190
Arg Ser Leu Ala Gln Tyr Ala Gly Glu Glu Asn Arg Arg Gln Phe Arg
195 200 205
Ala Ile Val Ala Glu Leu His Leu Leu Asp Pro Ser Ser Gly His Pro
210 215 220
Phe Leu Ser Ala Thr Met Glu Thr Ala His Arg Tyr Thr Glu Asp Phe
225 230 235 240
Tyr Lys Cys Tyr Leu Glu Lys Lys Arg Glu Trp Leu Ala Lys Thr Phe
245 250 255
Tyr Arg Pro Glu Gln Asp Glu Asn Thr Lys Arg Arg Ile Ser Val Phe
260 265 270
Phe Val Pro Asp Gly Glu Ala Arg Lys Leu Leu Pro Leu Leu Ile Arg
275 280 285
Arg Arg Met Lys Glu Gln Asn Asp Leu Gln Asp Trp Ile Arg Asn Lys
290 295 300
Gln Ala His Pro Ile Asp Leu Pro Ser His Leu Phe Asp Ser Lys Ile
305 310 315 320
Met Glu Leu Leu Lys Val Lys Asp Gly Lys Lys Lys Trp Asn Glu Ala
325 330 335
Phe Lys Asp Trp Trp Ser Thr Lys Tyr Pro Asp Gly Met Gln Pro Phe
340 345 350
Tyr Gly Leu Arg Arg Glu Leu Asn Ile His Gly Lys Ser Val Ser Tyr
355 360 365
Ile Pro Ser Asp Gly Lys Lys Phe Ala Asp Cys Tyr Thr His Leu Met
370 375 380
Glu Lys Thr Val Gln Asp Lys Lys Arg Glu Leu Arg Thr Ala Gly Lys
385 390 395 400
Pro Val Pro Pro Asp Leu Ala Ala Asp Ile Lys Arg Ser Phe His Arg
405 410 415
Ala Val Asn Glu Arg Glu Phe Met Leu Arg Leu Val Gln Glu Asp Asp
420 425 430
Arg Leu Met Leu Met Ala Ile Asn Lys Met Met Thr Asp Arg Glu Glu
435 440 445
Asp Ile Leu Pro Gly Leu Lys Asn Ile Asp Ser Ile Leu Asp Glu Glu
450 455 460
Asn Gln Phe Ser Leu Ala Val His Ala Lys Val Leu Glu Lys Glu Gly
465 470 475 480
Glu Gly Gly Asp Asn Ser Leu Ser Leu Val Pro Ala Thr Ile Glu Ile
485 490 495
Lys Ser Lys Arg Lys Asp Trp Ser Lys Tyr Ile Arg Tyr Arg Tyr Asp
500 505 510
Arg Arg Val Pro Gly Leu Met Ser His Phe Pro Glu His Lys Ala Thr
515 520 525
Leu Asp Glu Val Lys Thr Leu Leu Gly Glu Tyr Asp Arg Cys Arg Ile
530 535 540
Lys Ile Phe Asp Trp Ala Phe Ala Leu Glu Gly Ala Ile Met Ser Asp
545 550 555 560
Arg Asp Leu Lys Pro Tyr Leu His Glu Ser Ser Ser Arg Glu Gly Lys
565 570 575
Ser Gly Glu His Ser Thr Leu Val Lys Met Leu Val Glu Lys Lys Gly
580 585 590
Cys Leu Thr Pro Asp Glu Ser Gln Tyr Leu Ile Leu Ile Arg Asn Lys
595 600 605
Ala Ala His Asn Gln Phe Pro Cys Ala Ala Glu Met Pro Leu Ile Tyr
610 615 620
Arg Asp Val Ser Ala Lys Val Gly Ser Ile Glu Gly Ser Ser Ala Lys
625 630 635 640
Asp Leu Pro Glu Gly Ser Ser Leu Val Asp Ser Leu Trp Lys Lys Tyr
645 650 655
Glu Met Ile Ile Arg Lys Ile Leu Pro Ile Leu Asp His Glu Asn Arg
660 665 670
Phe Phe Gly Lys Leu Leu Asn Asn Met Ser Gln Pro Ile Asn Asp Leu
675 680 685
<210> 12
<211> 420
<212> PRT
<213> 宏基因组(metagenomic)
<400> 12
Met Asn Thr Val Pro Ala Ser Glu Asn Lys Gly Gln Ser Arg Thr Val
1 5 10 15
Glu Asp Asp Pro Gln Tyr Phe Gly Leu Tyr Leu Asn Leu Ala Arg Glu
20 25 30
Asn Leu Ile Glu Val Glu Ser His Val Arg Ile Lys Phe Gly Lys Lys
35 40 45
Lys Leu Asn Glu Glu Ser Leu Lys Gln Ser Leu Leu Cys Asp His Leu
50 55 60
Leu Ser Val Asp Arg Trp Thr Lys Val Tyr Gly His Ser Arg Arg Tyr
65 70 75 80
Leu Pro Phe Leu His Tyr Phe Asp Pro Asp Ser Gln Ile Glu Lys Asp
85 90 95
His Asp Ser Lys Thr Gly Val Asp Pro Asp Ser Ala Gln Arg Leu Ile
100 105 110
Arg Glu Leu Tyr Ser Leu Leu Asp Phe Leu Arg Asn Asp Phe Ser His
115 120 125
Asn Arg Leu Asp Gly Thr Thr Phe Glu His Leu Glu Val Ser Pro Asp
130 135 140
Ile Ser Ser Phe Ile Thr Gly Thr Tyr Ser Leu Ala Cys Gly Arg Ala
145 150 155 160
Gln Ser Arg Phe Ala Asp Phe Phe Lys Pro Asp Asp Phe Val Leu Ala
165 170 175
Lys Asn Arg Lys Glu Gln Leu Ile Ser Val Ala Asp Gly Lys Glu Cys
180 185 190
Leu Thr Val Ser Gly Leu Ala Phe Phe Ile Cys Leu Phe Leu Asp Arg
195 200 205
Glu Gln Ala Ser Gly Met Leu Ser Arg Ile Arg Gly Phe Lys Arg Thr
210 215 220
Asp Glu Asn Trp Ala Arg Ala Val His Glu Thr Phe Cys Asp Leu Cys
225 230 235 240
Ile Arg His Pro His Asp Arg Leu Glu Ser Ser Asn Thr Lys Glu Ala
245 250 255
Leu Leu Leu Asp Met Leu Asn Glu Leu Asn Arg Cys Pro Arg Ile Leu
260 265 270
Tyr Asp Met Leu Pro Glu Glu Glu Arg Ala Gln Phe Leu Pro Ala Leu
275 280 285
Asp Glu Asn Ser Met Asn Asn Leu Ser Glu Asn Ser Leu Asn Glu Glu
290 295 300
Ser Arg Leu Leu Trp Asp Gly Ser Ser Asp Trp Ala Glu Ala Leu Thr
305 310 315 320
Lys Arg Ile Arg His Gln Asp Arg Phe Pro Tyr Leu Met Leu Arg Phe
325 330 335
Ile Glu Glu Met Asp Leu Leu Lys Gly Ile Arg Phe Arg Val Asp Leu
340 345 350
Gly Glu Ile Glu Leu Asp Ser Tyr Ser Lys Lys Val Gly Arg Asn Gly
355 360 365
Glu Tyr Asp Arg Thr Ile Thr Asp His Ala Leu Ala Phe Gly Lys Leu
370 375 380
Ser Asp Phe Gln Asn Glu Glu Glu Val Ser Arg Met Ile Ser Gly Glu
385 390 395 400
Ala Ser Tyr Pro Val Arg Phe Ser Leu Phe Ala Pro Arg Tyr Ala Ile
405 410 415
Tyr Asp Lys Arg
420
<210> 13
<211> 558
<212> PRT
<213> 宏基因组(metagenomic)
<400> 13
Met Ser Pro Ile Ile Trp Val Ile Ile Cys Ile Ala Thr Leu Val Ile
1 5 10 15
Gly Phe Val Leu Gly Val Ile Tyr Arg Lys Lys Val Ala Glu Ala Gln
20 25 30
Ile Ala Gly Ala Glu Glu Glu Ala Lys Arg Ile Ile Asn Asp Ala Leu
35 40 45
Lys Ser Ala Glu Ser Lys Lys Arg Glu Ala Leu Val Glu Ala Lys Glu
50 55 60
Glu Ile Gln Lys Ser Arg Thr Glu Phe Glu His Glu Glu Lys Ser Arg
65 70 75 80
Arg Ala Glu Leu Gln Lys Gln Glu Arg Arg Leu Gln Gln Lys Glu Glu
85 90 95
Thr Leu Asp Arg Lys Thr Asp Ala Leu Asp Arg Lys Thr Glu Ser Leu
100 105 110
Asn Ala Lys Leu Ala Asn Val Glu Ala Gln Gln Asn Glu Ile Arg Thr
115 120 125
Ile Arg Lys Thr Gln Gln Glu Met Leu Glu Lys Ile Ser Gly Tyr Thr
130 135 140
Ile Glu Glu Ala Lys Ala Tyr Leu Ile Glu Thr Leu Glu Ala Glu Val
145 150 155 160
Thr His Glu Lys Ala Ala Lys Leu Lys Glu Ile Glu Ala Gln Tyr Lys
165 170 175
Asp Glu Ala Asp Arg Met Ala Arg Glu Ile Val Ala Thr Ala Ile Gln
180 185 190
Arg Cys Ala Ala Asp His Ala Ser Glu Val Thr Val Ser Val Val Pro
195 200 205
Leu Pro Asn Asp Glu Met Lys Gly Arg Ile Ile Gly Arg Glu Gly Arg
210 215 220
Asn Ile Arg Ala Ile Glu Thr Leu Thr Gly Cys Asp Leu Ile Ile Asp
225 230 235 240
Asp Thr Pro Glu Ala Ile Thr Val Ser Ser Phe Asp Pro Val Arg Arg
245 250 255
Glu Val Ala Arg Leu Ala Leu Glu Lys Leu Ile Ala Asp Gly Arg Ile
260 265 270
His Pro Ala Arg Ile Glu Glu Met Val Ala Lys Ala Gln Lys Glu Val
275 280 285
Asn Ala Thr Ile Lys Ala Glu Gly Glu Arg Ala Val Phe Glu Thr Asn
290 295 300
Ile His Gly Ile His Pro Glu Leu Val Arg Leu Leu Gly Arg Met Lys
305 310 315 320
Tyr Arg Thr Ser Tyr Gly Gln Asn Val Leu Gln His Ser Ile Glu Val
325 330 335
Ser His Ile Ser Gly Leu Ile Ala Ser Glu Leu Gly Ala Asn Val Ala
340 345 350
Ile Ala Lys Arg Ala Gly Leu Leu His Asp Leu Gly Lys Ser Ile Asp
355 360 365
His Glu Val Glu Gly Ser His Val Thr Ile Gly Val Asn Met Ala Arg
370 375 380
Lys Tyr Lys Glu Asn Glu Ala Val Thr Arg Glu His Arg Lys Leu Arg
385 390 395 400
Gln Ala Ser Gly Glu Ala Arg Gly Gly His Glu Val Leu Pro Arg His
405 410 415
Arg Gln Leu Leu Cys His Pro Gly Arg Thr Arg Asp Pro Arg Asp Gly
420 425 430
Gln Ala Arg Gly Gly Gln Arg Gly Ser Asp Asp Ala Ala Arg Ala Arg
435 440 445
His Arg Gln Glu Asp Arg Arg Arg Ala His Leu Ser Arg Pro Asp Gln
450 455 460
Gly Pro His His Pro Arg Asp Gln Gly Arg Arg Leu Arg Glu Ile Lys
465 470 475 480
Pro Asn Val Cys Pro Arg Lys Gly Asn Ala Ser Ala Gly Val Phe Leu
485 490 495
Ser Lys Gly Asn Val His Glu Ser His His Gly Asp Arg Ala Glu Gly
500 505 510
Arg Glu Ala Arg His Leu Tyr Arg Ala Gly Leu Ser Arg Arg Ala Leu
515 520 525
Glu Ala Asp Asp Leu Ser Gly Gln Gln Gly Ala Gly Ala Val Arg Pro
530 535 540
Leu Val Ser His Ala Val Arg His Gly Gln Arg Leu Pro Val
545 550 555
<210> 14
<211> 465
<212> PRT
<213> 宏基因组(metagenomic)
<400> 14
Met Thr Asp Leu Ser Arg Ile Gly Ala Val Leu Ile His Pro Arg Gly
1 5 10 15
Asp His Met Gly Asn Leu Leu Trp Pro Gly Val Leu Pro Ala Asp Gln
20 25 30
Ala Ala Ala Leu Ala Ala Ser Ile Ala Ala Val Arg Ala Gln Gly Tyr
35 40 45
Trp Ala Ser Pro Phe Pro Glu Gly Asp Gly Val Thr Phe Lys Arg Tyr
50 55 60
Gly Glu Ser Ser His Asp Ala Ala Ala Cys Leu Ala Asp Phe Glu Ala
65 70 75 80
Ala Phe Ala Phe Leu Glu Ile Glu Glu Leu Asp Ala Ala Glu Gly Arg
85 90 95
Ala Ser Ala Leu Ala Arg Leu Ala Ala Gly Arg Thr Ile Met Cys Thr
100 105 110
Tyr Leu Val Pro Val Glu Ala Leu Arg Leu Glu Arg Pro Phe Leu Leu
115 120 125
Gly Asp Thr Arg Phe His Ala Pro Val Asp Gly Glu Glu Val Arg Leu
130 135 140
Ala Asn His Ala Trp Gln Ser Leu Cys Asp Val Pro Gly Ala Asp Val
145 150 155 160
Asp Pro Ser Trp Val Pro Ser Ala Arg Ala Ser Gly Thr Thr Glu Leu
165 170 175
Leu Gly Tyr Pro Leu Ile Glu Arg Ser Val Glu Val Pro Leu Ser Val
180 185 190
Phe Tyr Glu Ala Gln Ala Ser Phe Asp Arg Gln Ser Pro Leu Leu Arg
195 200 205
Leu Val Met Glu Asp Ala Asp His Ala Leu Asp Pro Ile Arg Tyr Asp
210 215 220
Leu Cys His Tyr Arg Arg Leu Glu Tyr Leu Pro Ala Lys Pro Gly Trp
225 230 235 240
Ile Gly Glu Thr Ala Ile Ala Tyr Val Met Pro Glu Gly Arg Val Val
245 250 255
Lys Pro Arg Leu Leu Gln Gly Lys Pro Tyr Val Leu Arg Val Ser Asn
260 265 270
Asn Trp Leu Gly Leu Glu Val Asp Pro Ser Gly Phe Ser Trp Ala Glu
275 280 285
Pro Leu Ala Ser Met Val Val Asp Asp Lys Ser Gly Asp Glu Val Ala
290 295 300
Leu Ala Leu Lys Ala Ala Leu Arg Ala Phe Asn Arg Ala Phe Tyr Leu
305 310 315 320
Val Glu Leu Glu Ala Ser Phe Leu His Leu Val Tyr Ala Ile Asp Ala
325 330 335
Leu Cys Glu Pro Asp Lys Leu Arg Gly Asp Arg His Arg Leu Trp Ile
340 345 350
Ser Ala Phe Gly Ala Ser Gly Asp Pro Gly Arg Phe Ala Thr Leu Leu
355 360 365
Gly Asp Phe Asp Arg Arg Tyr Gly Val Arg Asn Arg Ile Val His Glu
370 375 380
Gly Ala Thr Phe Ala Gly Leu Gly Leu Lys Gly Glu Glu Gln Cys Gln
385 390 395 400
Phe Met Leu Gly Leu Leu Gly Ala Cys Ile Gln Ile Phe Val Arg Glu
405 410 415
Gly Phe Thr Thr Arg Arg Glu Ala Ala Asp Tyr Ala Phe Asp Phe Leu
420 425 430
Thr Ser Ser Pro Val Thr Thr His Val Thr Ala Met Ala Ser Glu His
435 440 445
Phe Lys Leu Pro Leu Ala Thr Asp Lys Gly Phe Arg Glu His Met Gln
450 455 460
Pro
465
<210> 15
<211> 485
<212> PRT
<213> 宏基因组(metagenomic)
<400> 15
Met Leu Glu Thr Leu Ala Ala Glu Tyr Ile Ala Ala Ser Phe Ser Val
1 5 10 15
Leu Glu Ala Glu Asn Ala Ile Pro Thr Ser Lys Met His Arg His Leu
20 25 30
Gln Ala His Leu Asp Phe Asp Gly Ser Ala Ile Glu Ala Met Pro Glu
35 40 45
Phe Ala Ala Leu Ala Gln Ala Val Ile Lys Thr Gly Pro Glu Gly Leu
50 55 60
Pro Ala Ser Asp Thr Asn Pro Gly Pro Glu His Thr Leu Arg Leu Leu
65 70 75 80
Leu Phe Ser Leu Leu Glu Leu Cys Val His Arg Cys Ala Asp Val Asp
85 90 95
Asp Tyr Asp Pro Ala Ser Thr Glu Val Ser Ala Val Ile Thr Glu Phe
100 105 110
Cys Asp Val Val Gly Gly Ser Ser Tyr Asn Leu Val Val Ala Arg Val
115 120 125
Val Ser His Leu Val Pro Ala Thr Gly Thr Glu Val Asp Ile Ala Gly
130 135 140
Ile Thr Leu Val Ala Glu Thr Pro Thr Gln Asp Leu Arg Arg Cys Ile
145 150 155 160
Val Ala Ala Ile Pro Asp Ala Asp His Ala Trp Lys Arg Arg Gly Pro
165 170 175
Met Leu Asp Arg Pro Pro His Ser Leu Leu Val Val Arg Asp Pro Val
180 185 190
Glu Gly Val Gly Tyr His Gly Ser Trp Asn Leu Glu Gly Val Leu Asp
195 200 205
Arg Leu Gln Leu Ala Val Cys Leu Leu Thr Gly Ala Asn Val Gln Ser
210 215 220
Val Tyr Gln Val Ser Gly Val Ser Ser Arg Ile Ser Ala Arg Pro Ala
225 230 235 240
Arg Val Glu Phe Leu Thr Arg Asp Glu Glu Val Pro Leu Val Arg Arg
245 250 255
Val Ala Arg Leu Asp Glu Thr His Val Pro Val Leu Ala Glu Leu Ser
260 265 270
Ala Leu Ile Asp Asn Val Arg Ser Glu Asn Gly Gln Glu Ile Trp Leu
275 280 285
Ser Ser Phe Gly Glu Ala Leu Thr Lys Phe Ala Arg Leu Asp Asp Thr
290 295 300
Ser Ser His Phe Asp Gln Ile Val Gln Leu Ser Thr Ala Leu Glu Gly
305 310 315 320
Val Ile Leu Gly Ala Asn Glu Gly Glu Gly Leu Thr Leu Arg Leu Cys
325 330 335
Thr Arg Ala Thr Ala Leu Leu Ala Tyr Asp Asp Glu Pro Ala Ala Glu
340 345 350
Leu Phe Glu Asp Leu Lys Arg Leu Tyr Ser Phe Arg Ser Thr Ile Val
355 360 365
His Gly Gly Glu Met Thr Thr Lys Gln Leu Arg Lys Asp Leu Arg Ala
370 375 380
Met Arg Cys Val Pro Asp Glu Ser Ser Gly Pro Gly Leu Leu Pro Arg
385 390 395 400
Val Gly Phe Ala Val Asp Arg Leu Arg Asp Ile Val Arg Arg Ala Ile
405 410 415
Leu Ala Arg Leu Cys Leu Ser Ser Gly Asp Thr Pro Leu Trp Ser Ile
420 425 430
Phe Ser Asn Glu Ala Pro Lys Leu Asp Ile Val Leu Cys Asp Asp Ser
435 440 445
Asn Arg Ala Gln Trp Arg Ser His Trp Arg Asp Val Leu Ala Ser Ile
450 455 460
Gly Ala Glu Thr Ala Gly His Arg Ala Ala Ala Pro Val His Ser Leu
465 470 475 480
Ser Asp Asp Asp Met
485
<210> 16
<211> 164
<212> PRT
<213> 宏基因组(metagenomic)
<400> 16
Met Pro Ile Lys Glu Asn Asn Leu Ile Phe Pro Tyr Pro Pro Ser Ile
1 5 10 15
Asn Asn Pro Asn Glu Glu Ser Ile Asn Tyr Lys Lys Phe Tyr Thr Glu
20 25 30
Met Gly Lys Ser Val Gln Lys Tyr Asn Leu Thr Asn Thr Phe Asn Ala
35 40 45
Ala Val Arg Met Lys Val Ile Leu His Ala Lys Asp Asn Thr Lys Arg
50 55 60
Asp Val Asp Asn Pro Val Lys Ala Leu Leu Asp Ala Phe Thr Tyr Ala
65 70 75 80
Gly Val Trp Val Asp Asp Asn Gln Val Arg Asp Leu Arg Ile Ile Tyr
85 90 95
Gly Lys Ser Asp Ser Lys Gly Glu Gly Tyr Ile Glu Val Ser Met Asn
100 105 110
Glu Ala Thr Asp Leu Ser Asn Leu Ile Cys Asp Glu Val Lys Gln Lys
115 120 125
Ile Lys Ile Lys Asn Gln Lys Ile Lys Asn Lys Pro Thr Pro Ser Pro
130 135 140
Ala Ala Gln Gln Asn Gln Lys Lys Arg Lys Thr Arg Leu Val Ile Asn
145 150 155 160
Asn Asn His Gln
<210> 17
<211> 177
<212> PRT
<213> 宏基因组(metagenomic)
<400> 17
Met Lys Met Phe Tyr Ile Glu His Lys Thr Pro Tyr Leu Tyr Val Asp
1 5 10 15
Cys Glu Thr Thr Gly Leu Lys Pro Gly Thr Asp Glu Val Leu Ser Ile
20 25 30
Ser Val Ile Asp Gln Thr Gly Lys Ile Ala Phe Asn Ser Leu Ile Arg
35 40 45
Pro Glu Arg Arg Lys Lys Trp Val Glu Ala Gln Asn Ile Asn His Ile
50 55 60
Ser Pro Glu Met Val Arg Asp Ala Pro Thr Tyr Arg Asp Val Val Pro
65 70 75 80
Gln Leu Gln Arg Ile Phe Arg Gly Ala His Ile Val Thr Tyr Asn Leu
85 90 95
Glu Phe Asp Tyr Lys Phe Leu His Glu Ala Met Cys Thr Ala Ser Ser
100 105 110
Phe His Cys Cys Met Lys Ala Tyr Ala Lys Phe Gln Lys Glu Ile Asp
115 120 125
Pro Tyr Arg Gly Thr Tyr Arg Trp His Lys Leu Thr Ala Ala Tyr Asp
130 135 140
Asn Leu Phe His Asn Thr Phe Asn Ala His Asp Ser Leu Ala Asp Val
145 150 155 160
Gln Ala Thr Leu Arg Val Trp Asn Arg Ile Met His Asp Tyr Lys Val
165 170 175
Arg
<210> 18
<211> 199
<212> PRT
<213> 宏基因组(metagenomic)
<400> 18
Met Lys Ile Ala Tyr Leu Asp Ile Glu Thr Asn Tyr Thr Gly Lys Phe
1 5 10 15
Thr Asp Gln Arg Leu Phe Ser Asp His Glu Asn His Leu Ile Thr Val
20 25 30
Leu Gly Met Arg Leu Phe Gln Glu Asp Lys Asp Ser Phe Leu Gln Leu
35 40 45
Val Arg Lys Asp Val Ser Lys Gly Ala Leu Leu Ser Ala Leu Arg Gly
50 55 60
Thr Asp Leu Ile Val Thr Tyr Asn Gly Arg Ser Ile Pro Asp Lys Met
65 70 75 80
Lys Gly Tyr Thr Gly Phe Asp Phe Pro Val Ile Ala Ala Gln Leu Gly
85 90 95
Val Val Leu Asp Glu Glu Phe Lys His Val Asp Leu Cys Pro Val Cys
100 105 110
Trp Gln Arg Gly Leu Trp Gly Gly Gln Lys Thr Ile Glu Arg Ser Leu
115 120 125
Gly Leu Lys Arg Lys Leu Pro Gly Arg Asp Gly Ala Trp Ala Asp Val
130 135 140
Thr Trp Lys Lys Tyr Glu Thr Thr Gly Tyr Tyr Asn Phe Leu Glu Glu
145 150 155 160
Leu Leu Glu Tyr Asn Arg Glu Asp Val Phe Met Leu Arg Arg Ile Gln
165 170 175
Glu Ala Leu Arg Cys Thr Glu Asn Tyr Gln Arg Pro Arg Arg Ser Cys
180 185 190
Ser His Ala Ala Pro Leu Leu
195
<210> 19
<211> 252
<212> PRT
<213> 宏基因组(metagenomic)
<400> 19
Met Thr Val Arg Ile Met Gln Ser Leu Arg Leu Val Val Ala Cys Val
1 5 10 15
Leu Ala Phe Leu Leu Thr Asn Gln Gly Thr Ala Ile Ala Asp Asn Cys
20 25 30
Pro Pro Lys Gly Cys Ala Gln Lys Tyr Thr Leu Ala Leu Ser Trp Gln
35 40 45
Pro Gly Phe Cys Glu Thr His Gly Tyr Lys Ala Glu Cys Gln Glu Gln
50 55 60
Thr Ala Thr Ser Tyr Asp Ala Asn Asn Phe Thr Leu His Gly Leu Trp
65 70 75 80
Pro Asp Lys Leu Glu Tyr Cys Asn Val Ser Ala Ser Tyr Ile Glu Asn
85 90 95
Asp Lys Thr Gly Asn Trp Lys Asp Leu Pro Ile Val Glu Val Asp Thr
100 105 110
Glu Thr Thr Asp Glu Leu Asp Glu Val Met Pro Gly Phe Gly Glu Ser
115 120 125
Ser Leu Glu Arg His Glu Trp Ile Lys His Gly Thr Cys Asp Gly Arg
130 135 140
Asn Ser Asp Asn Tyr Tyr Asp Leu Ser Val Asp Leu Leu Lys Glu Phe
145 150 155 160
Asn Asp Ser Gln Val Arg Asn Leu Phe Ile Glu His Ile Gly Glu Thr
165 170 175
Val Ser Leu Asp Gln Val Lys Thr Ala Phe Glu Gly Thr Phe Gly Ser
180 185 190
Gly Ser Ser Ser Ser Leu Asn Leu Lys Cys Asp Ser Thr Asn Asn Leu
195 200 205
Ala Thr Glu Ile Arg Leu Lys Ile Lys Arg Pro Gln Ser Gly Gln Lys
210 215 220
Leu Ala Asp Leu Leu Leu Pro Ser Gly Gly Gln Ser Cys Asn Gln Val
225 230 235 240
Val Val Asp Gly Phe Gly Val Ser Asn Leu Thr Gln
245 250
<210> 20
<211> 302
<212> PRT
<213> 宏基因组(metagenomic)
<400> 20
Met His Ala Val Ile His Thr Arg Thr Val Ser Gly Leu Ala Asp Asp
1 5 10 15
Ala His Thr Val Ala Ala Leu Ala Trp Ser Ala Ala Asp Gly Glu His
20 25 30
Arg Ser Gln Leu Phe Ile Arg Pro Gly Ala Ser Cys Glu Ala Arg Tyr
35 40 45
Ala Ala Gly Leu Asp Ala Phe Glu Met Ala Leu Ala Leu Cys Glu Thr
50 55 60
Asn Asp Asp Thr Val Met Leu His Ile Asn Asp His Gly Leu Arg Ala
65 70 75 80
Asn Ile Asp Met Val Leu Asp Ala Phe Pro His Ile His Leu Ala His
85 90 95
Gly Thr Arg Gly Asn Leu Leu Ala Leu Leu Glu Thr Ala Met His Arg
100 105 110
Ile Asp Glu His Thr His Val Ala Ala Ala Val His His Glu Arg Leu
115 120 125
Glu Arg Ala Arg Ile Glu Arg Ala Gln Ala Leu Ala Glu Leu Pro Glu
130 135 140
Leu Val Ile Ala Thr Asp Ala Ser Lys Ala Thr His Arg Arg Gly Ala
145 150 155 160
Gly Ile Ala Tyr Ala Thr Ala Asp Gly Arg Tyr Arg Gln Glu Phe Leu
165 170 175
Ser Gly Val Arg His Ile Ser Val Gly Glu Leu Leu Ala Ile Glu Ala
180 185 190
Ala Val Ala Asp Phe Pro Gly Arg Arg Leu His Ile Leu Thr Asp Ser
195 200 205
Arg Asp Ala Met Gln Ala Ile Gly Val Glu Trp Glu Pro Arg Trp Arg
210 215 220
Val Thr Gly Glu Thr Ala Thr Leu Gly Ala Arg Ile Arg Ala Leu Met
225 230 235 240
Ser Gly Arg Asp Val Arg Ile Ser Trp Val Lys Gly His Ser Gly His
245 250 255
Glu Leu Asn Glu Ile Ala Asp Arg Leu Ala Val Ala Ala Arg Arg His
260 265 270
His Glu Ser Lys Val Pro Ala Ala Ser Arg Arg Ser Val Ala Asp Asn
275 280 285
Ile Val Ala Pro Leu Arg Ala Ala His Val Gln Ala Ala Ala
290 295 300
<210> 21
<211> 267
<212> PRT
<213> 宏基因组(metagenomic)
<400> 21
Met Asp Ile Thr Met Leu Gly Thr Gly Asn Ala Leu Ala Thr Glu Cys
1 5 10 15
Tyr Asn Thr Cys Phe Val Ile Asp Asp Gly Gly Arg Leu Phe Met Val
20 25 30
Asp Gly Gly Gly Gly Asn Gly Val Leu Arg Gln Leu Arg Ala Ala Gly
35 40 45
Phe Asp Trp Met Gly Val Arg Glu Ile Phe Val Thr His Lys His Ile
50 55 60
Asp His Leu Leu Gly Ile Val Trp Met Met Arg Val Ile Cys Gln Ser
65 70 75 80
Met Asp Gln Gly Val Tyr Glu Gly Glu Ala His Ile Tyr Ala His Asp
85 90 95
Glu Val Ile Ser Leu Leu Arg Trp Leu Ala Asp Glu Leu Leu Val Pro
100 105 110
Lys Gln Thr Arg Phe Ile Gly Asp Arg Leu Gln Leu Ile Pro Val Val
115 120 125
Asp Gly Glu Gln Arg Ile Ile Asn Gly Arg Arg Val Thr Phe Phe Asp
130 135 140
Ile His Ser Thr Lys Ala Lys Gln Phe Gly Phe Cys Met Glu Leu Asp
145 150 155 160
Gly Gly Glu Lys Leu Thr Cys Cys Gly Asp Glu Pro Cys Ser Pro Tyr
165 170 175
Gly Glu Gln Tyr Ala Asn Asn Ser Ala Trp Leu Leu His Glu Ala Phe
180 185 190
Cys Leu His Ala Gln Ala Asp Val Phe Arg Pro Tyr Glu Lys His His
195 200 205
Ser Thr Val Lys Asp Ala Cys Glu Leu Ala Glu Arg Leu His Val Arg
210 215 220
Asn Leu Leu Leu Tyr His Thr Glu Asp Ser Asn Leu Ala Glu Arg Lys
225 230 235 240
Ala Arg Tyr Ala Ala Glu Gly Glu Arg Tyr Phe Ser Gly Lys Leu Trp
245 250 255
Val Pro Asp Asp Leu Glu Thr Ile Thr Ile Gln
260 265
<210> 22
<211> 464
<212> PRT
<213> 宏基因组(metagenomic)
<400> 22
Met Asp Tyr Pro Gln Arg Lys Ala Lys Pro Gln Gln His Phe Lys Gly
1 5 10 15
Lys Ser Ser Lys Pro Asn Arg Phe Ser Pro Gly Thr Arg Thr Gly Arg
20 25 30
Arg Pro Ser Ser Glu Tyr Asp Phe Phe Thr Gly Gly Gly Thr Lys Ile
35 40 45
Thr Asn Thr Asp Lys Glu Ser Thr Lys Thr Ala Asp Phe Thr Ile Phe
50 55 60
Tyr Asn Ile Ala Phe Glu Asn Val Glu Lys Val Lys Thr Gln Leu Gln
65 70 75 80
Asn Lys Thr Gln Ser Gln Gln Asn Ala Phe Trp Ala Glu Tyr Phe Trp
85 90 95
Lys Ala His Phe Leu Ser Lys Asn Thr Asn Gly Tyr Glu Leu Thr Gln
100 105 110
Gln Asp Asp Arg Ile Ile Thr Gln Leu Ile Lys Lys Val Glu Glu Ile
115 120 125
Arg Asn Tyr His Ser His Ile Trp His Asp Asn Ser Val Leu Val Phe
130 135 140
Ser Asp Glu Leu Lys Arg Phe Val Glu Gln Lys Tyr Asn Glu Ala Leu
145 150 155 160
Val Gln Leu Ser Val Asp Phe Pro Gly Ala Val Ser Asp Tyr Gln Phe
165 170 175
Leu Lys Gln Lys Asn Tyr Glu Lys Glu Ser Lys Leu Phe Asn Pro Ile
180 185 190
Gly Phe Gly Gly Asp Ala Lys Asn Phe Ile Thr Ile Glu Gly Arg Ile
195 200 205
Phe Phe Leu Ser Phe Phe Leu Thr Thr Gly Gln Met Asn Gln Phe Leu
210 215 220
Gln Gln Arg Thr Gly Tyr Lys Arg Ala Asp Met Pro Gln Phe Lys Ile
225 230 235 240
Lys Arg Leu Leu Tyr Thr Phe Tyr Cys Asn Arg Asp Gly Ala Ala Ile
245 250 255
Thr Asp Phe Asn His Glu Asp Arg Phe Ile Asp Thr Leu Ala Pro Glu
260 265 270
Val Arg Gln Asn Val Phe Lys Ala Arg Thr Ala Phe Lys Leu Ile Ser
275 280 285
Tyr Leu Met Asp Tyr Pro Asp Tyr Trp Gly Ser Asn Asp Ala Met Pro
290 295 300
Leu Phe Asp Asn Asn Asn Glu Leu Ile Lys Asn Val Glu Gln Leu Lys
305 310 315 320
Asp Tyr Ile Glu Ser Lys Asn Ile Leu Pro Glu Leu Lys Phe Thr Leu
325 330 335
Ile Asp Arg Lys Ile Lys Ala Ser Leu Glu Leu Glu Glu Asp Ala Glu
340 345 350
Thr Leu Lys Lys Glu Gln Glu Asp Lys His Ser Thr Gly Thr Ile Ala
355 360 365
Phe Thr Tyr Asp Glu Leu Ser Gly Phe Ser Phe His Ile Asn Phe Glu
370 375 380
Ala Leu His Arg Leu Val Leu Leu Gln Thr Leu His His Asn Met Val
385 390 395 400
Asp Leu Pro Ala Pro Leu Ala Ile Leu Ala Ile Glu Leu Lys Lys Gln
405 410 415
Ala Asn Asn Arg Thr Thr Leu Tyr Asp Ile Leu Ile Lys Pro Ile Ala
420 425 430
Glu Arg Thr Asp Asp Glu Gln Val Tyr Leu Leu Thr Lys Glu Asn Gln
435 440 445
Tyr Leu Arg Gly Gly Arg Lys Val Thr Glu Leu Gly Ile Ile Phe Phe
450 455 460
<210> 23
<211> 509
<212> PRT
<213> 宏基因组(metagenomic)
<400> 23
Met Pro Lys Trp Thr Tyr Ser Ile Thr Ile Val Asn Asn Met Asp Arg
1 5 10 15
Pro Leu Glu Leu Ile Ser Ser Ser Ile Ala Trp Gly Lys Lys Ala Lys
20 25 30
Ala Phe Pro Thr Glu Ile Ala Ala Gly Glu His Gly Glu Phe Ser Val
35 40 45
Tyr Ala Lys Ala Gly Thr Ser Thr Gly Ile Glu Phe Tyr Phe Thr Met
50 55 60
Gly Asp Lys Val Glu Lys Lys Gly Asp Pro His Tyr Gly Asn Phe Ser
65 70 75 80
Phe Ser Val Asp Met Pro Tyr Trp Lys His Ala Asn Lys Ser Ser Phe
85 90 95
Asp Cys Thr Gly Ile Leu Thr Gln Ser Gly Phe Lys Lys Ile Pro Asp
100 105 110
Gly Ala His Asp Phe Ala Ile Thr Ala Thr Val Ser Thr Ser Leu Cys
115 120 125
Ser Ile Phe Leu Pro Gly Glu Lys Thr Trp Lys Gly Asn Gly Tyr Gln
130 135 140
Met Glu Tyr Cys Asn Leu Tyr Asp Trp Glu Glu Ile Gln Lys Leu Lys
145 150 155 160
Val Val Glu Pro Asp Glu Lys Arg Ile Asp Asp Phe Ile Pro Val Glu
165 170 175
Asn Ile Phe Ser Lys Arg Lys Thr Asp Ser Arg Thr Glu Gln Val Ser
180 185 190
Val Pro Lys Lys Leu Trp Lys Glu Ile Ile Asp Arg Lys Tyr Pro Asp
195 200 205
Asp Tyr Ser Lys Arg Asn Phe Val Lys Asp Tyr Phe Thr Val Ser Ala
210 215 220
Tyr Glu Ile Arg Lys Asn Val Thr Val Ser Ile Ala Ala Asn Gln Ser
225 230 235 240
Tyr Glu Lys Thr Leu Glu Val Ser Asn Arg Ser Thr Val Arg Arg Glu
245 250 255
Thr Arg Glu Glu Phe Gln Ile Glu Asn Thr Ile Ser Gly Glu Gly Thr
260 265 270
Gly Glu Lys Phe Thr Leu Ser Glu Thr Leu Arg Met Gln Tyr Gln Ile
275 280 285
Ser Arg Leu Asp Glu Tyr Cys Glu Glu Asn Met Glu Ser Val Arg Glu
290 295 300
Val Phe Asn Tyr Asp Ala Ile Glu Gln Asp Arg Asp Val Val Leu Trp
305 310 315 320
Asp Leu Ala Glu Val Leu Met Leu Tyr Arg Val Asn Ile Lys Gly Lys
325 330 335
Val Glu Leu Val Gly Ile Gly Asp Tyr Tyr Val Ala Ser Thr Gln Lys
340 345 350
Thr Tyr Leu Arg Asn Gly Ile Gln Gly Glu Glu Ser Pro Tyr Asp Thr
355 360 365
Arg Glu Glu Glu Leu Pro Cys Ala Asp Ser Glu Ser Asn Tyr Arg Leu
370 375 380
Tyr Asp Asp Ser Asp Glu Glu Leu Leu Gly Glu Glu Phe Glu Ser Trp
385 390 395 400
Asp Asp Gly Asp Gly Val Pro Val Lys Ser Leu Gly Val Val Gln Gly
405 410 415
Asn Ile Arg Ile Asn Asn Val Asn Tyr Arg Trp Met Glu Val Arg Arg
420 425 430
Gly Thr Asn Arg Gly Met Leu Phe Asn Pro Gly Glu His Arg Tyr Ser
435 440 445
Phe Arg Pro Asn Pro His Asp Asp Pro Trp Tyr Asn Arg His Gln Thr
450 455 460
Asp Trp Tyr Glu Ala Met Ala Arg Glu Phe Gln Arg Val Gly Asn Ala
465 470 475 480
Asn His Trp Thr Gln Asp Asn Trp Ser Asp Gly Gln Asp Ile Arg Asn
485 490 495
Leu Arg Val Asn Asn Glu Thr Tyr Thr Ala Arg Val Tyr
500 505
<210> 24
<211> 413
<212> PRT
<213> 宏基因组(metagenomic)
<400> 24
Leu Ser Ser Leu Ile Ser Lys Phe Gln Gln Ile Glu Ala Phe Asn Gly
1 5 10 15
Asp Ile Arg Asn Leu Lys Leu Arg Asn Pro Glu Leu Asn Leu Asp Ile
20 25 30
Met Asp Asp Gly Lys Ile Gly Tyr Asn Ser Lys Lys Tyr Ser Ile Phe
35 40 45
Glu Asn Ser Gly Gln Ile Ala Asn Asp Leu Asp Lys Leu Arg Gly Val
50 55 60
Ile Lys Met Asp Ile Asn Asp Leu Asp Ala Tyr Glu Pro Met Ile Lys
65 70 75 80
Asp Ala Leu Arg Val Ile Gly Val Asp Gly Ser Asp Leu Asp Ser Ile
85 90 95
Tyr Lys Asp Tyr Phe Asn Lys Lys Asp Lys Lys Asn Ser Val Ala Gly
100 105 110
Phe Phe Arg Asn Asn Ile Ile Asn Ser Arg Arg Phe Arg Tyr Ile Ile
115 120 125
Lys Tyr Ile Asn Pro Ser Asp Ala Tyr Arg Ile Ile Gln Asn Glu His
130 135 140
Val Arg Asn Tyr Val Leu Gly Arg Met Asn Asp Ala Ile Ile Asp Arg
145 150 155 160
Tyr Ala His Ser Val Gly Ile Glu Asp Lys Val His Asp Lys Arg Lys
165 170 175
Val Leu Ser Asp Ile Leu Ser Lys Val Lys Phe Asp Asn Phe Thr Lys
180 185 190
Leu Thr Tyr Ile Asn Pro Lys Asp Lys Asn Lys Gly Glu Lys Ala Lys
195 200 205
Glu Arg Glu Lys Pro Lys Ala Ile Leu Gly Leu Tyr Leu Thr Ile Val
210 215 220
Tyr Leu Val Val Lys Ser Leu Val Arg Ile Asn Ser Gln Tyr Val Met
225 230 235 240
Ala Val Tyr His Leu Glu Arg Asp Ser Arg Leu Cys Pro Gly Val Ser
245 250 255
Ser Asn Asn Pro Leu Ser Met Thr Ser Met Thr Lys His Tyr Cys Asp
260 265 270
Arg Arg Asn Asn His Leu Leu Lys Glu Lys His Ile Val Lys Leu Glu
275 280 285
Arg Tyr Gln Asn Thr Pro Gln Lys Ile Cys Thr Ala Tyr Arg Asn Ala
290 295 300
Ile Ala His Leu Ser Ala Val Arg Lys Gly Val Lys Tyr Ile Gly Asp
305 310 315 320
Ile Gln Lys Ala Asp Ser Tyr Phe Gly Ile Tyr His Tyr Cys Met Gln
325 330 335
Lys Phe Leu Tyr Ser Ala Asp Ser Ala Thr Arg His Pro Phe Ala Glu
340 345 350
Phe Val Arg Ser Ile Phe Gly Asp Glu Lys Glu Leu Asp Lys Leu Arg
355 360 365
Asn Gly Ser Tyr Ser Gln Ala Ile Leu Arg Ala Leu Asn Tyr Pro Phe
370 375 380
Gly Tyr Asn Pro Ala Arg Tyr Lys Asn Leu Ser Tyr Glu Lys Ile Phe
385 390 395 400
Met Arg Ala Trp Gln Asp Glu Asp Thr Asn Lys Lys Thr
405 410
<210> 25
<211> 581
<212> PRT
<213> 宏基因组(metagenomic)
<400> 25
Met Asn Tyr Arg Leu Gly Leu Asp Leu Gly Thr Asn Ser Ile Gly Trp
1 5 10 15
Ala Val Val Arg Leu Asp Glu Asn Gly Asp Ala Cys Gly Ile Leu Asp
20 25 30
Met Gly Val Arg Ile Phe Pro Asp Gly Arg Glu Asn Ser Gln Ser Leu
35 40 45
Asp Pro Leu Gly Gln Ser Leu Ala Val Asn Arg Arg Ile Pro Arg Gly
50 55 60
Gln Arg Arg Arg Arg Asp Arg Tyr Leu Lys Arg Arg Ser Asp Leu Leu
65 70 75 80
Asp Lys Leu Thr Ala Cys Glu Leu Met Pro Gln Asp Leu Glu Thr Arg
85 90 95
Ala Ala Ile Ala Ala Leu Asp Pro Tyr Pro Ile Arg Lys Arg Ala Leu
100 105 110
Glu Glu Pro Leu Ser Pro Tyr Asp Leu Gly Arg Ala Leu Phe His Leu
115 120 125
Asn Gln Arg Arg Gly Phe Lys Ser Asn Arg Lys Ala Gln Pro Lys Asp
130 135 140
Asp Glu Lys Gln Ser Arg Glu Asn Leu Ser Ala Leu Asp Ile Ala Leu
145 150 155 160
Lys Glu Ser Gly Gln Thr Leu Gly Glu Phe Leu Ala Pro Arg His Glu
165 170 175
Gln Ser Glu Pro Val Arg Phe Arg Pro Gly Val Gly Leu Tyr Pro Asp
180 185 190
Arg Ala Met Tyr Glu Ala Glu Phe Glu Ala Ile Arg Lys Ala Gln Glu
195 200 205
Leu His His His Leu His Pro Gly Gln Trp Asp Ser Leu Arg Asp Ile
210 215 220
Ile Phe Phe Gln Arg Pro Leu Lys Pro Val Asp Pro Gly Trp Cys Leu
225 230 235 240
Leu Glu Asp Gly Glu Arg Arg Ala Ala Arg Ala Met Pro Ser Ala Gln
245 250 255
Glu Phe Arg Met Thr Gln Glu Val Asn Asn Leu Arg Ile Arg Val Asp
260 265 270
Ile Glu Pro Glu Arg Ser Leu Asn Asp Ala Glu Arg Glu Arg Val Leu
275 280 285
Gln Arg Leu Arg Glu Gly Lys Asp Ile Asp Leu Gln Lys Pro Thr Lys
290 295 300
Ser Leu Gln Leu Glu Gly Ser Phe Asn Leu Ala Arg Gly Gly Arg Lys
305 310 315 320
Lys Leu Gln Gly Asp Gln Thr Thr Ala Arg Leu Ala Lys Pro Glu Leu
325 330 335
Phe Gly Lys Arg Trp Phe Asp Leu Pro Leu Glu Glu Arg Asp Ala Val
340 345 350
Ala Ala Arg Leu Leu Asp Thr Glu Asp Pro Lys Thr Ile Arg Gln Val
355 360 365
Ala Val Asn Glu Trp Arg Leu Asn Lys Glu Gln Ala Ala Arg Val Ala
370 375 380
Asp Val Thr Leu Pro Asp Gly His Thr His Leu Ser Leu Lys Ala Ile
385 390 395 400
Arg Asn Ile Leu Pro His Leu Glu Asn Gly Met Arg Tyr Asp Glu Ala
405 410 415
Val Gln Ala Arg Tyr Pro His His Ser Asp Phe Arg Asn Ala Glu Ala
420 425 430
His Asp Arg Leu Pro Tyr Tyr Gly Val Val Leu Glu Arg Asp Val Val
435 440 445
Gly Ala Asp Pro Asn Ala Pro Asp Gly Asp Glu Pro Arg Arg Tyr Gly
450 455 460
Arg Phe Pro Asn Pro Thr Val His Leu Gly Leu Asn Gln Leu Arg Arg
465 470 475 480
Val Val Asn Arg Leu Ile Asp Val Tyr Gly Lys Pro Ala Glu Thr Val
485 490 495
Val Glu Leu Ala Arg Asp Leu Lys Ser Asn Lys Glu Gln Arg Glu Arg
500 505 510
Tyr Val Arg Gln Gln Arg Ser Asn Glu His Glu Asn Asp Arg Arg Arg
515 520 525
Glu Glu Leu Arg Glu Cys Gly Val Glu Pro Asn Gly Tyr Val Met Glu
530 535 540
Lys Leu Arg Leu Cys Asn Tyr Lys Pro His Arg Ala Leu Ile Val Ile
545 550 555 560
Ser Leu Ser Asp Gly Arg Ser Gly Arg Asn Arg Asn Pro Ser Cys Cys
565 570 575
Ser Ser Arg Ser Pro
580
<210> 26
<211> 523
<212> PRT
<213> 宏基因组(metagenomic)
<400> 26
Met Ser Ser Arg Gly Gly Gly Gln Lys Ser Ala Val Cys Ala Arg Ala
1 5 10 15
Leu Gln Arg Pro Asn Phe Phe Thr Tyr Thr Pro Asn Arg Arg Cys Ala
20 25 30
Arg His Ala Val Gln Pro Ile Gly Ala Leu Thr Ala Ala Arg Tyr Gly
35 40 45
Cys Lys Met Arg Pro Tyr Arg Lys Gly Thr Met Ser Glu Gln Glu Ala
50 55 60
Ala Gln Glu Gly Thr Lys Leu Leu Ala Lys Thr Leu Thr Phe Gly Leu
65 70 75 80
Gly Asn Pro Met Gly Phe Lys Ser Lys Gly Ser Val Leu Val Glu Leu
85 90 95
Thr Glu Asp Gln Arg Lys Ala Ile Tyr Asn Gly Leu Arg Asp Ala Ser
100 105 110
Thr Val Val Ala Arg Ile Ile Asn Leu Leu Asn Ser Arg Glu Tyr Ile
115 120 125
Arg Gln Ile Met Lys Val Pro Glu Glu Leu Val Ala Gln Phe Lys Pro
130 135 140
Asn Tyr Ser Leu Val Lys Gly Pro Leu Lys Arg Leu Gly Ile Glu Glu
145 150 155 160
Ala Glu Gln Val Ala Gly Ser Val Leu Ser Gln Thr Phe Ala Leu Gly
165 170 175
Val Lys Pro Asp Phe Gln Gly Glu His Gly Lys Gly Leu Leu Leu Lys
180 185 190
Gly Glu Arg Gln Ile Pro Leu His Arg Thr Asp Gly Thr His Pro Ile
195 200 205
Pro Gln Arg Ala Thr Glu Thr Arg Leu Phe Gln Val Glu Lys Asn Phe
210 215 220
Tyr Val Ala Met Gln Val Phe Ala Glu Thr Trp Ala Lys Lys Gln Glu
225 230 235 240
Leu Pro Ser Gly Trp Leu Ala Phe Pro Ile Lys Val Lys Pro Arg Asp
245 250 255
Lys Thr Met Ala Gly Gln Leu Leu Lys Thr Ile Gly Gly Glu Trp Lys
260 265 270
Leu Lys Asn Ser Arg Leu Met Arg Asn Pro Arg Thr Gly Gly Asn Arg
275 280 285
Trp Leu Gly Gln Ile Val Val Ala Phe Ala Pro Glu Pro Phe Lys Lys
290 295 300
Met Thr Arg Ser Val Val Met Gly Ile Asp Leu Gly Val Asn Val Pro
305 310 315 320
Ala Cys Leu His Ile Ser Glu Asn Gly Lys Pro Leu Pro Trp Ala Met
325 330 335
Met Val Gly Arg Gly Arg Asp Met Leu Asn Thr Arg Asn Leu Ile Arg
340 345 350
Ser Glu Ile Val His Ile Ile Lys Ala Leu Lys Ser Lys Asp Ser Pro
355 360 365
Leu Asp Gly Lys Ala Arg Ala Ile Tyr Arg Asp Lys Leu Arg Asp Leu
370 375 380
Arg Lys Arg Glu Arg Arg Val Met Lys Met Ala Ser Gln Thr Val Ala
385 390 395 400
Ala Arg Ile Ala Asp Thr Ala Lys Arg His Gly Ala Gly Thr Trp Gln
405 410 415
Met Glu Asp Leu Ser Pro Asp Ile Lys Thr Asp Gln Pro Trp Leu Ala
420 425 430
Arg Asn Trp Ala Pro Gly Met Leu Leu Asp Ala Val Arg Trp Gln Ala
435 440 445
Arg Gln Cys Gly Ala Glu Leu Val Met Val Asn Pro Ala Tyr Thr Ser
450 455 460
Gln Arg Cys Ala Arg Cys Gly His Ile Asp Pro Gln Asn Arg Pro Lys
465 470 475 480
Gln Thr Asp Phe Lys Cys Met Ala Cys Gly His Glu Asp Asn Ala Asp
485 490 495
Lys Asn Ala Ala Arg Asn Leu Ser Val Val Gly Ile Glu Lys Leu Ile
500 505 510
Ala Asp Phe Lys Ala Pro Asn Gly Ala Val Gln
515 520
<210> 27
<211> 191
<212> PRT
<213> 宏基因组(metagenomic)
<400> 27
Met Met Leu Tyr Asn Trp Ile Glu Gly Ser Ile Thr Glu Ala Ile Asp
1 5 10 15
Glu Ile Phe Glu Ala Ile Thr Gln Gln Lys Val Arg Phe Asp Glu Leu
20 25 30
Ile Pro Ala Tyr Gln Arg Leu Trp Leu Ser Tyr Gln His Gly Leu Val
35 40 45
Lys Ile Ala Ala Glu Ser Ala Lys Ala Lys Asp Pro Thr Lys Asn Lys
50 55 60
Ile Gly Lys Ser Leu Pro Asn Val Leu Gln Gln Val Gln Tyr Phe Lys
65 70 75 80
Ile Leu Thr Phe Thr Asp Lys Glu Gly Ile Gln Tyr Lys Asn Tyr Arg
85 90 95
Gly Tyr Leu Lys Val Val Asp Thr Ala Asp Leu Ser Gly Asn Leu Asp
100 105 110
Ala Arg Lys Met Arg Glu Leu Ala Ser Lys Tyr Ala Phe Ser Val Pro
115 120 125
Glu Arg Cys Asp Asp Leu Leu Lys Ile Lys Asn Ile Arg Asn Gln Leu
130 135 140
Ala His Gly Glu Ile Ile Phe Ser Glu Ala Gly Thr Glu Pro Ile Ser
145 150 155 160
Arg Leu Met Glu Ile Lys Gly Asn Val Leu Glu Tyr Leu Glu Ala Ile
165 170 175
Leu Leu Asn Ile Asn Glu Phe Ile Glu Lys Asp Gly Phe Lys Ile
180 185 190
<210> 28
<211> 789
<212> PRT
<213> 宏基因组(metagenomic)
<400> 28
Met Glu Asn Ile Lys Leu Glu Lys Gln Lys Ala Ala Phe Tyr Phe Asn
1 5 10 15
Gln Ala Glu Leu Asn Leu Lys Ala Ile Glu Gly Asn Ile Phe Asp Lys
20 25 30
Gly Arg Arg Lys Thr Leu Phe Asp Asn Pro Lys Ile Leu Ser Lys Val
35 40 45
Glu Asn Phe Ile Phe Asn Phe Lys Asp Val Thr Lys Asn Ala Lys Gly
50 55 60
Glu Ile Asp Cys Leu Leu Ser Lys Leu Met Glu Leu Arg Asn Phe Tyr
65 70 75 80
Ser His Tyr Val His Lys Pro Asp Val Lys Glu Leu Ser Lys Gly Glu
85 90 95
Lys Pro Leu Leu Glu Arg Tyr Tyr Gln Ile Ala Ile Glu Ala Thr Gly
100 105 110
Ser Glu Asn Val Lys Leu Glu Ile Ile Glu Asn Asp Lys Trp Leu Thr
115 120 125
Asp Ala Gly Val Leu Leu Phe Leu Cys Met Phe Leu Lys Lys Ser Gln
130 135 140
Ala Asn Lys Leu Ile Ser Gly Ile Ser Gly Phe Lys Arg Asn Asp Thr
145 150 155 160
Phe Gly Gln Pro Arg Arg Asn Leu Phe Asn Tyr Phe Ser Val Arg Glu
165 170 175
Arg Tyr Lys Val Val Pro Asp Met Gln Lys His Phe Leu Leu Phe Val
180 185 190
Leu Val Asn His Leu Ser Glu Gln Asp Asp Tyr Ile Glu Lys Ala Gln
195 200 205
Gln Pro Tyr Asn Ile Gly Glu Gly Leu Phe Phe His Arg Ile Ala Ser
210 215 220
Thr Phe Leu Asn Val Ser Gly Ile Leu Arg Asn Met Glu Phe Tyr Thr
225 230 235 240
Tyr Gln Ser Lys Arg Leu Lys Glu Gln Arg Gly Glu Leu Lys Arg Glu
245 250 255
Lys Asp Ile Phe Thr Trp Glu Glu Pro Phe Gln Gly Asn Ser Tyr Phe
260 265 270
Glu Ile Asn Gly His Lys Gly Val Ile Gly Glu Asp Glu Leu Lys Glu
275 280 285
Leu Cys Tyr Ala Leu Leu Ser Tyr Asn Lys Ser Lys Tyr Ala Val Glu
290 295 300
Gln Ile Glu Lys Phe Leu Lys Gly Phe Gly Glu Val Lys Ser Glu Gln
305 310 315 320
Glu Ile Arg Asp Ser Asp Ile Leu Asn Glu Ser Tyr Phe Pro Thr Asn
325 330 335
Tyr Phe Ala Glu Ser Asn Ile Gly Ser Ile Lys Glu Lys Ile Leu Asn
340 345 350
Arg Leu Gly Lys Thr Asp Asp Ser Tyr Lys Lys Thr Gly Thr Lys Ile
355 360 365
Lys Pro Tyr Asp Met Met Lys Glu Val Met Glu Phe Ile Asn Asn Ser
370 375 380
Leu Pro Ala Asp Glu Lys Leu Lys Arg Lys Asp Tyr Arg Arg Tyr Leu
385 390 395 400
Lys Met Val Arg Ile Trp Asp Ser Glu Lys Asp Asn Ile Lys Arg Glu
405 410 415
Phe Glu Ser Lys Glu Trp Ser Lys Tyr Phe Ser Ser Asn Phe Trp Met
420 425 430
Ala Lys Asn Leu Glu Arg Val Tyr Gly Leu Ala Arg Glu Lys Asn Ala
435 440 445
Glu Leu Phe Asn Lys Leu Lys Ala Val Val Glu Lys Met Asp Glu Arg
450 455 460
Glu Phe Glu Lys Tyr Arg Gln Ile Asn Ser Ala Glu Asp Leu Ala Ser
465 470 475 480
Leu Arg Arg Leu Ala Asn Asp Tyr Gly Val Lys Trp Glu Glu Lys Asp
485 490 495
Trp Gln Glu Tyr Ser Gly Gln Ile Lys Lys Gln Ile Ser Asp Arg Gln
500 505 510
Lys Leu Thr Ile Met Lys Gln Arg Ile Thr Ala Glu Leu Lys Lys Lys
515 520 525
His Gly Ile Glu Asn Leu Asn Leu Arg Ile Thr Ile Asp Ser Asn Lys
530 535 540
Ser Arg Lys Ala Val Leu Asn Arg Ile Ala Val Pro Arg Gly Phe Val
545 550 555 560
Lys Glu His Ile Leu Gly Trp Gln Gly Ser Glu Lys Val Ser Lys Lys
565 570 575
Thr Arg Glu Ala Lys Cys Lys Ile Leu Leu Ser Lys Glu Tyr Glu Glu
580 585 590
Leu Ser Lys Gln Phe Phe Gln Thr Arg Asn Tyr Asp Lys Met Thr Gln
595 600 605
Val Asn Ser Leu Tyr Glu Lys Asn Lys Leu Ile Ala Phe Met Ala Val
610 615 620
Tyr Leu Met Gly Gln Leu Asn Ile Arg Phe Asp Lys Pro Thr Arg Leu
625 630 635 640
Asn Glu Leu Glu Lys Ala Glu Val Asp Phe Lys Ile Ser Asp Lys Val
645 650 655
Thr Ala Lys Ile Pro Phe Ser Gln Tyr Pro Ser Leu Val Tyr Ala Met
660 665 670
Ser Ser Lys Tyr Ala Asp Ser Val Gly Ser Tyr Lys Phe Glu Asn Asp
675 680 685
Glu Lys Asn Lys Pro Phe Leu Gly Lys Ile Asp Ile Ile Glu Lys Gln
690 695 700
Arg Met Glu Phe Ile Lys Glu Val Leu Gly Phe Glu Glu Tyr Leu Phe
705 710 715 720
Glu Lys Lys Ile Ile Asp Lys Ser Lys Phe Ala Asp Thr Ala Thr His
725 730 735
Ile Ser Phe Arg Glu Ile Cys Asp Glu Leu Ile Gln Lys Gly Trp Asp
740 745 750
Glu Asn Lys Leu Thr Asn Leu Lys Asp Ala Arg Asn Ala Ala Leu His
755 760 765
Gly Glu Ile Pro Ala Glu Thr Ser Phe Arg Glu Ala Lys Pro Leu Ile
770 775 780
Asn Gly Leu Lys Lys
785
<210> 29
<211> 643
<212> PRT
<213> 宏基因组(metagenomic)
<400> 29
Met Lys Asn Arg Lys Glu Leu Leu Ile Asn His Phe Ser Asp Ile Lys
1 5 10 15
Ser Ala Cys Thr Ser Leu Ala Gln Val Cys Gln Ile Tyr Met Thr Glu
20 25 30
Tyr Asn Gln Gln Asn Asn Gln Ile Lys Lys Val Arg Ser Ser Asn Asp
35 40 45
Ser Ile Phe Asp Gln Pro Ile Tyr Gln His Tyr Lys Val Leu Leu Lys
50 55 60
Lys Ala Ile Ala Asn Ala Phe Ala Ala Tyr Leu Lys Asn Asn Lys Asp
65 70 75 80
Leu Phe Gly Phe Ile Gly Lys Pro Phe Lys Ala Asn Glu Ile Arg Glu
85 90 95
Ile Asp Lys Glu Gln Phe Leu Pro Asp Trp Thr Ser Arg Lys Tyr Glu
100 105 110
Ser Leu Cys Ile Glu Val Ser Gly Ser Gln Glu Leu Gln Lys Trp Tyr
115 120 125
Ile Val Gly Lys Phe Leu Asn Ala Arg Ser Leu Asn Leu Met Ala Gly
130 135 140
Ser Met Arg Ser Tyr Ile Gln Tyr Val Thr Asp Ile Lys Arg Arg Ala
145 150 155 160
Ala Ser Ile Gly Asn Glu Leu His Val Ser Val Gln Asp Val Glu Lys
165 170 175
Val Glu Lys Trp Val Gln Val Ile Glu Val Cys Ser Leu Leu Ala Phe
180 185 190
Arg Thr Ser Asn Gln Phe Glu Asp Tyr Phe Asn Asp Lys Asp Asp Tyr
195 200 205
Ala Arg Tyr Leu Lys Ser Tyr Val Asp Phe Ser Asn Ala Asp Met Pro
210 215 220
Ser Glu Tyr Ser Ala Leu Val Asp Phe Ser Asn Glu Glu Gln Ser Asp
225 230 235 240
Leu Tyr Val Asp Pro Lys Asn Pro Lys Val Asn Arg Asn Ile Val His
245 250 255
Ser Lys Leu Phe Ala Ala Asp His Ile Leu Arg Asp Ile Val Glu Pro
260 265 270
Val Ser Lys Asp Asn Ile Glu Glu Phe Tyr Asp Gln Lys Ala Glu Ile
275 280 285
Val Tyr Cys Lys Ile Lys Gly Lys Asp Ile Thr Ala Glu Glu Gln Lys
290 295 300
Ala Val Leu Lys Tyr Gln Lys Leu Lys Asn Arg Val Glu Leu Arg Asp
305 310 315 320
Ile Val Glu Tyr Gly Glu Ile Ile Asn Glu Leu Leu Gly Gln Leu Ile
325 330 335
Asn Trp Ser Phe Met Arg Glu Arg Asp Leu Leu Tyr Phe Gln Leu Gly
340 345 350
Phe His Tyr Asn Cys Leu Arg Asn Asn Ser Glu Lys Lys Glu Ala Tyr
355 360 365
Gln Lys Ile Glu Val Asn Gly Asp Ser Ile Lys Asp Ala Ile Leu Tyr
370 375 380
Gln Ile Ile Gly Met Tyr Val Asn Gly Val Ala Val Tyr Ala Pro Asp
385 390 395 400
Glu Lys Gly Asp Ile Leu Lys Glu Gln Cys Thr Lys Gly Gly Ala Gly
405 410 415
Gly Lys Ile Ser Ala Phe Asn Arg Tyr Ser Glu Tyr Leu Gly Leu Asp
420 425 430
Lys Leu Thr Leu Tyr Asn Ala Gly Leu Glu Ile Phe Glu Val Val Ala
435 440 445
Glu His Asp Asp Ile Ile Asn Leu Arg Asn Gly Ile Asp His Phe Lys
450 455 460
Tyr Tyr Leu Gly Asp Tyr Arg Ser Met Leu Ser Ile Tyr Ser Glu Val
465 470 475 480
Phe Asp Arg Phe Phe Thr Tyr Asp Ile Lys Tyr Gln Lys Asn Val Leu
485 490 495
Asn Leu Leu Gln Asn Ile Leu Leu Arg His Asn Val Ile Val Glu Pro
500 505 510
Val Leu Glu Ser Gly Phe Lys Thr Ile Gly Lys Gln Thr Thr Ser Gly
515 520 525
Ala Lys Leu Ser Ile Arg Ser Ile Lys Ser Asp Thr Phe Gln Tyr Lys
530 535 540
Val Lys Gly Gly Ser Leu Ile Thr Asp Ala Lys Asp Glu Arg Tyr Leu
545 550 555 560
Lys Thr Ile Arg Lys Ile Leu Tyr Tyr Val Glu Asn Glu Glu Asp Asn
565 570 575
Leu Lys Lys Ser Val Val Val Thr Asn Ala Asp Lys Tyr Glu Lys Asn
580 585 590
Lys Glu Ser Asp Asp Gln Asn Lys Gln Lys Glu Lys Lys Asn Lys Asp
595 600 605
Asn Lys Gly Lys Lys Asn Glu Glu Thr Lys Ser Asp Ala Gly Lys Asn
610 615 620
Asn Asn Glu Arg Leu Ser Tyr Asn Pro Phe Ala Asn Leu Asn Phe Lys
625 630 635 640
Leu Pro Asn
<210> 30
<211> 410
<212> PRT
<213> 宏基因组(metagenomic)
<400> 30
Leu Phe Asp Gln Ile Thr Asp Trp Arg Asp Ile Gly Ala His Ala Glu
1 5 10 15
Asp Ile His Lys Lys Lys Leu Thr Ala Lys Phe Val Glu Asp Leu Arg
20 25 30
Lys Ile Tyr Asp Ala Ser Trp His Lys Thr Lys Glu Gly Leu Ser Leu
35 40 45
Ser Val Ser Gln Met Asp Gln Leu Val Arg Leu Lys Gly Arg Gln Thr
50 55 60
Lys Asn His Asp Lys Leu Gln Val Gln Lys Asp Arg Pro Asn Phe Lys
65 70 75 80
Tyr Gly Leu Ile Asn Asp Ser Asn His Phe Ser Glu Gln Gly Leu Ala
85 90 95
Phe Leu Ile Cys Leu Phe Leu Gln Glu Ser Glu Ala Asn Leu Phe Leu
100 105 110
Lys His Val Glu His Val Trp Ser Asn Gln Asp Phe Asn Ser Asn Ser
115 120 125
Thr Asn Leu Asn Ser Ser Gln Ala Ile Lys Leu Phe Thr Ala Tyr Arg
130 135 140
Leu Arg Leu Pro Phe Pro Lys Leu Gln Ser Ser His Asp Ser Gln Ser
145 150 155 160
Leu Phe Leu Asp Met Ile Asn Ser Leu Lys Lys Cys Pro Ser Ile Leu
165 170 175
Phe Asn His Ile Ser Asn Leu Asp Lys Lys Lys Phe Lys Lys Asp Ile
180 185 190
Lys Asn Ser Glu Val Asn Phe Gly Ser Glu Glu Leu Ile Thr Asn Glu
195 200 205
Asn Ile Glu Thr Val His Glu Val Arg Lys Arg Asp Arg Phe Val Ile
210 215 220
Gln Ala Leu Glu Tyr Phe Asp Leu Thr Asn Thr Phe Asn Tyr Met Arg
225 230 235 240
Phe His Ile Asp Leu Gly Lys Phe Ile Tyr Lys Thr Tyr Thr Ser Pro
245 250 255
Ile Asp Gly Thr Asp Arg Ile Arg Arg Leu Glu Lys Asn Leu His Ala
260 265 270
Phe Asp Lys Ile Gln Glu Val Ile Gly Glu Lys Arg Pro Glu Ile Trp
275 280 285
Lys Asn Leu Ile Lys Glu Pro Asn Thr Ile Pro Glu Asp Ser Leu Asp
290 295 300
Cys Tyr Ile Val Asp Thr Glu Pro His Tyr His Ile Ser Asp His Lys
305 310 315 320
Ile Gly Phe Lys Val Phe Gln Asn Val Leu Pro Ser Phe His Asp Ile
325 330 335
Val Lys Ile Gly Glu Asn Asn Phe Lys Phe Ser Glu Asn Pro Lys Pro
340 345 350
Asp Gly Phe Ile Ser Ile Tyr Glu Leu Pro Ala Met Val Phe Tyr Glu
355 360 365
His Ile Thr Gly Asn His Gly Lys Ser Glu Glu Ile Ile Arg Glu His
370 375 380
Ile Lys Gln Arg Arg Lys Ile Ile Thr Glu Val Leu Asn Asp Ser Asn
385 390 395 400
Leu Pro Leu Tyr Lys Thr Ser Ile His Leu
405 410
<210> 31
<211> 423
<212> PRT
<213> 宏基因组(metagenomic)
<400> 31
Val Glu Glu Val Phe Asn Leu Leu Arg Arg Lys Ser Asn Pro Pro Gln
1 5 10 15
Ser Lys Ser Gln Leu Asp Cys Asp Ile His Glu Tyr Val Glu Lys His
20 25 30
Arg Lys Asp Lys Ser Lys Glu Trp Ala Glu Asp Pro Thr Ala Leu Met
35 40 45
Arg Lys Gln Ala Lys Gln Val Glu Gln Thr Glu His Ala Ile Arg Arg
50 55 60
Cys Gln Ile Glu Asp Ile Val Met Leu Tyr Ala Ala Arg Asp Met Leu
65 70 75 80
Tyr Ala Ala Arg Asp Ile Leu Ser Ala Lys Asn Asn Arg Thr Pro Asn
85 90 95
Gly Thr Asp Thr Pro Gln Pro Gln Lys Phe Lys Leu Lys His Val Gln
100 105 110
Lys Asp Asp Gly Leu Leu Glu Arg Thr Ile Asp Phe Asp Trp Val Val
115 120 125
Asp Ile Asp Gly Gln Gln Lys Thr Ile Arg Gln Gln Asn Met Lys Met
130 135 140
Lys Asp Tyr Gly Lys Phe Tyr Lys Phe Ala Ser Asp Gly Glu Arg Leu
145 150 155 160
Lys Ser Leu Leu Ala His Leu Gly Gly Asn Glu Phe Gln Arg Ala Asp
165 170 175
Ile Glu Ala Glu Tyr Ala Asn Tyr Asp Val Ser Arg Ser Gln Val Phe
180 185 190
Arg Tyr Val Tyr Met Leu Glu Ser Lys Ala Tyr Gln Leu Leu Ala Lys
195 200 205
Arg Lys Asp Asn Pro Ile Asp Leu Leu Asn Asp Gln Lys Pro Ile Pro
210 215 220
Asp Ala Phe Trp Phe Val Ser Lys Glu Gly Ile Arg Asn Glu Phe Arg
225 230 235 240
Val Phe Lys Glu Asn Leu Phe Asn Glu Tyr Lys Ala Asp Val His Asp
245 250 255
Tyr Lys Ala Asn Asp Pro Asp Ala Glu Ala Leu Val Asn Ala Ile Cys
260 265 270
Asn Phe Thr Gly Thr Ser Ile Glu Glu Trp Asp Ala Asn Gln Asp Thr
275 280 285
Leu Met Lys Gln Val Lys Ser Leu Leu Thr Asp Glu Asn Pro Asn Glu
290 295 300
Lys Asp Lys Ala Leu Ile Leu Gln Val Lys Asp Tyr Cys Met Lys Lys
305 310 315 320
Asp Ile Ala Arg Lys Ala Ile Arg Asn Asn Phe Gly Glu Leu Ile Glu
325 330 335
Ile Leu Leu Arg Gly Asp Glu Pro Ile Phe Thr Asp Asp Asp Lys Tyr
340 345 350
Ile Ile Gln His Ile Arg Asn Ala Phe Gly His Asn His Tyr Leu Lys
355 360 365
Lys Glu Asp Glu Tyr Asn Thr Val Phe Arg Gly Lys Glu Ala Lys Leu
370 375 380
Lys Leu Pro Glu Val Ala Lys Thr Ile Lys Asp Trp Met Gly Glu Lys
385 390 395 400
Thr Thr Lys Ala Leu Ser Leu Thr Pro Asp Thr Gln Arg Ala Leu Pro
405 410 415
Pro Lys Ser Gln Ala Ala Glu
420
<210> 32
<211> 619
<212> PRT
<213> 宏基因组(metagenomic)
<400> 32
Met Arg Ile Ile Lys Pro Tyr Gly Arg Ser His Val Asp Ala Ala Asp
1 5 10 15
Leu Thr Glu Arg Lys Arg Val Leu Arg Leu Asn Asp Gly Ala Lys Thr
20 25 30
Ser His Asp Ile Lys Ser Phe Ala Leu Gly His Asp Glu Leu Ile Ile
35 40 45
Ala Gln Trp Ile Ser Ser Ile Asp Lys Ile Ala Thr Lys Pro Ser Gly
50 55 60
Asn Lys Gly Ala Thr Pro Ile Gln Arg Lys Phe Arg Asp Gln Leu Gly
65 70 75 80
Lys Thr Ser Trp Ala Leu Leu Val Glu Lys Asp Leu Leu Pro Gly Leu
85 90 95
Lys Asp Pro Asp Lys Arg Ala Gln Leu Glu Lys Leu Trp Glu Phe Lys
100 105 110
Val Ala Pro Tyr Gly Lys Ala Glu Phe Arg Asp Lys Leu Lys Ala Pro
115 120 125
Asn Pro Lys Gly Arg Trp Phe Lys Arg Phe Ala Gly Asp Val Asp Val
130 135 140
Thr Thr Val Asp Ala Asp Glu Val Ala Arg Arg Ile His Glu His Leu
145 150 155 160
Tyr Met Ala Glu Tyr Arg Leu Glu Gly Gly Asn Lys Arg Val Gly Arg
165 170 175
Ile Glu Ala Arg Ala Lys Ser Ile Glu Arg Asn Val Leu Arg Pro Ala
180 185 190
Lys Thr Ser Ser Val Ala Asp Asn Gly Trp Ser Asp Ala Asp Trp Asn
195 200 205
Thr Tyr Ile Gly Ala Gly Asp Val Ala Lys Gln Ile Trp Leu Ala Ala
210 215 220
Arg Ala Arg Gln Glu Gly Arg Asp Gly Ala Gly Thr Arg Arg Val Thr
225 230 235 240
Ala Ser Val Ala Ala Ile Ala Leu Ala Glu Gln Tyr Ala Lys Leu Phe
245 250 255
Ala Gly Ile Asp Gly Lys Pro Leu Gly Ile Lys Gln Ala Lys Ser Ala
260 265 270
Ala Pro Gly Leu Phe Asn Leu His Met Ala Ile Lys Asp Ala Tyr Ser
275 280 285
Arg Ile Leu Lys His Arg Lys Lys Asp Gln Arg Gln Ser Gly Lys Pro
290 295 300
Gln Arg Lys Val Ser Asn Ile Leu Pro Pro Asp Met Cys Arg Leu Ile
305 310 315 320
Ile Leu Val Lys Asn Lys Thr Lys Asn Gln Arg Leu Asn Ala Leu Val
325 330 335
Arg Glu Gly Lys Val Ile His Tyr Glu Ala Ser Gln Ala Gly Gly Lys
340 345 350
Asp Ser Pro Ser Val Leu Thr Asp Asn Trp Pro Arg Asp Thr Ser Asn
355 360 365
Ser Phe Tyr Trp Thr Ser Asp Gly Gln Ala Glu Ile Lys Arg Asn Glu
370 375 380
Ala Phe Val Arg Ile Trp Arg His Val Leu Ala Leu Ala Ala Arg Thr
385 390 395 400
Leu Lys Asp Trp Ala Asp Pro Asn Asp Lys Ile Lys Ile Asp Ile Leu
405 410 415
Asp Arg Gly Ser Ile Glu Ser Ala Ile Gly Asp Ser Phe Val Ile Ile
420 425 430
Gln His Asn Gln Lys Leu Ala Leu Leu Phe Gly Lys Arg Ala Ser Val
435 440 445
Phe Asn Asp Ala Gly Asp Glu Asp Val Gln Arg Glu Val Leu Glu Leu
450 455 460
Ala Leu Lys Gly Ile Ala Ser Leu Arg His Ser Ala Phe His Phe Lys
465 470 475 480
Gly Leu Asp Ser Phe Arg Glu Ala Leu Thr Lys Ser Asp Trp Thr Ser
485 490 495
Ser Gly Ala Ala Arg Gly Ala Leu Glu Ala Leu Trp Gln Thr Asp Val
500 505 510
Lys Ala Arg Ala Leu Gln Arg Lys Lys Thr Leu Ser Gly Ala His Val
515 520 525
Glu Tyr Phe Leu Asp Glu Ser Gln Ala Arg Lys Leu Met Ala Ala Leu
530 535 540
Glu Gln Ala Ala Pro Ala Ser Ile Pro Leu Pro Arg Phe Ala Arg Val
545 550 555 560
Leu Leu Arg Thr Ser His Met Glu Thr Lys Gln His Asp Gly Leu His
565 570 575
Leu Pro Met Pro Ala Asn Arg Ala Asp Leu Glu Asp Pro Ala Arg Leu
580 585 590
Cys Gln Tyr Thr Val Leu Lys Leu Leu Tyr Glu Gln Pro Phe Arg Lys
595 600 605
Trp Leu Glu Thr Arg Ser Thr Gln Thr Leu Asn
610 615
<210> 33
<211> 550
<212> PRT
<213> 宏基因组(metagenomic)
<400> 33
Val Arg Glu Arg Asp Gly Glu Arg Arg Arg Val Ser Ile Arg Ala Arg
1 5 10 15
Leu Gly Ser Arg Ala Ile Pro Ser Leu Gln Gly Asp Phe Gly Ala Ile
20 25 30
Leu Glu Val Ser Ile Arg Ala Arg Leu Gly Ser Arg Ala Ile His Ile
35 40 45
Gly Gln His Ala Asp Cys Glu Arg Glu His Val Ser Ile Arg Ala Arg
50 55 60
Leu Gly Ser Arg Ala Ile Leu Asp Val Asp Glu Glu Ser Gly Ile Gln
65 70 75 80
Leu Glu Val Ser Ile Arg Ala Arg Leu Gly Ser Arg Ala Ile His Tyr
85 90 95
Leu Ala Lys Leu Gly Glu Ser Gly Lys Val Phe Gln Ser Ala Pro Gly
100 105 110
Trp Glu Ala Gly Arg Phe Asn Cys Ser Ala Met Thr Pro Ala Gln Leu
115 120 125
Leu Glu Phe Gln Ser Ala Pro Gly Trp Glu Ala Gly Arg Phe Gln Val
130 135 140
Ala Ala Ala Leu Ser Ile Ser Gln Ser Val Phe Gln Ser Ala Pro Gly
145 150 155 160
Trp Glu Ala Gly Arg Phe Pro Val Thr Ala Ala Gly Val Ile Thr Leu
165 170 175
Arg Leu Phe Gln Ser Ala Pro Gly Trp Glu Ala Gly Arg Phe Asp Tyr
180 185 190
Leu Ala Lys Leu Gly Glu Ala Gly Lys Val Phe Gln Ser Ala Pro Gly
195 200 205
Trp Glu Ala Gly Arg Phe Gly Arg Ala Gly His Gly Asn Arg Arg Cys
210 215 220
Pro Met Phe Gln Ser Ala Pro Gly Trp Glu Ala Gly Arg Phe Gly Ala
225 230 235 240
Ser Cys Thr Met Ser Pro Ser Arg Pro Met Phe Gln Ser Ala Pro Gly
245 250 255
Trp Glu Ala Gly Arg Phe Leu Ser Arg Leu Ala Pro Ala Arg Thr Ser
260 265 270
Thr Trp Phe Gln Ser Ala Pro Gly Trp Glu Ala Gly Arg Phe Met Ser
275 280 285
Arg Pro Pro Thr Asn Arg Pro Lys Met Met Phe Gln Ser Ala Pro Gly
290 295 300
Trp Glu Ala Gly Arg Phe Gly His Ala Leu Gly Tyr Arg Asp Gly Arg
305 310 315 320
Glu Glu Phe Gln Ser Ala Pro Gly Trp Glu Ala Gly Arg Phe Ala Leu
325 330 335
Thr Gly Lys Ala Asn Thr His Ala Asn Leu Phe Gln Ser Ala Pro Gly
340 345 350
Trp Glu Ala Gly Arg Phe Arg Ala Ala Trp Gly Gly Ala Ala Thr Asp
355 360 365
Thr Val Val Ser Ile Arg Ala Arg Leu Gly Ser Arg Ala Ile His Arg
370 375 380
Asn Ala Lys Tyr Gly Ala Thr Ala Met Leu Phe Gln Ser Ala Pro Gly
385 390 395 400
Trp Glu Ala Gly Arg Phe Arg Lys Ile Pro Pro Pro Ala Val Gly Arg
405 410 415
Ser Cys Phe Asn Pro Arg Pro Ala Gly Lys Pro Gly Asp Ser Ala His
420 425 430
Pro Arg Pro Ser Arg Glu Gly Ala Gly Val Ser Ile Arg Ala Arg Leu
435 440 445
Gly Ser Arg Ala Ile Leu His Ala Ala Glu Ala Glu Gln Glu Pro Ala
450 455 460
Glu Cys Phe Asn Pro Arg Pro Ala Gly Lys Pro Gly Asp Ser Arg Ala
465 470 475 480
Ile Ala Glu Gly His Asp Ile Ile His Val Ser Ile Arg Ala Arg Leu
485 490 495
Gly Ser Arg Ala Ile Leu Val Ser Thr Arg Asp Trp Lys Arg Ser Pro
500 505 510
Lys Phe Gln Ser Ala Pro Gly Trp Glu Ala Gly Arg Phe Gly Arg Pro
515 520 525
Arg Val Arg Ala Gly Cys Gly Cys Ser Phe Asn Pro Arg Pro Ala Gly
530 535 540
Lys Pro Gly Asp Ser Arg
545 550
<210> 34
<211> 129
<212> PRT
<213> 宏基因组(metagenomic)
<400> 34
Met Ala Phe Asp Leu Pro Thr Ile Asp Pro Ser Leu Asp Ile Glu Lys
1 5 10 15
Ile Thr Thr Glu Glu Leu Arg Arg Lys Gly Ile Val Leu Ile Gln Lys
20 25 30
Ala Phe Trp Thr Arg Gly Tyr Tyr Arg Lys Val Ser Glu Arg Glu Cys
35 40 45
Arg Leu Tyr Phe Phe Leu Cys Ala Phe Met Arg Ile Glu Asp Phe Thr
50 55 60
Val His Leu Glu Trp Gly Ala Met Glu Asp Tyr Leu Glu Ser Asn Arg
65 70 75 80
Arg Gly Val Arg Val Ala Leu Arg Arg Leu Ile Arg Lys Arg Leu Ile
85 90 95
Ser Ile Ile Ser His Arg Leu Ala Asp Asp Arg Asp Pro Gln Lys Asp
100 105 110
Ala Leu Thr Ile Arg Ile Trp Arg Leu Arg Tyr Gly Lys Lys Glu Lys
115 120 125
Lys
<210> 35
<211> 123
<212> PRT
<213> 宏基因组(metagenomic)
<400> 35
Met Ala Ile Ile Leu Gln Ala Pro Leu Val Ser Cys Trp His Gln Arg
1 5 10 15
Gly Ala Phe Gly Asn Pro Ile Phe Gly Arg Asp Phe His Pro Ala His
20 25 30
Leu Gly Thr Ser Arg His Lys Leu Ser Met Gln Glu Cys Ser Val Arg
35 40 45
Ser Ser Leu Arg Ser Cys Ile Leu Arg Ile Glu Trp Glu Leu Trp Gly
50 55 60
Lys Asn Ser His Arg Asn Gly Ser Pro Val Ile Tyr Gly Val Phe Ile
65 70 75 80
Arg Phe Trp Pro Thr Leu Tyr Ile Cys Ala Phe Pro Ala Lys Asn Thr
85 90 95
Val Tyr Lys Leu Tyr Ile Tyr Gly Ser Cys Gln Pro Tyr Ile Tyr Thr
100 105 110
Glu Tyr Ile Arg Tyr Cys Trp Gln Gly Ser His
115 120
<210> 36
<211> 158
<212> PRT
<213> 宏基因组(metagenomic)
<400> 36
Met Ala Arg Leu Arg Lys Leu Lys Arg Lys Phe Ser Thr Ser Phe Arg
1 5 10 15
Ala Val Leu Phe Arg Met Val Pro Lys Leu Gly Lys Met Ala Glu Gly
20 25 30
Leu Thr Gln Arg Phe Arg Ala Val Leu Phe Arg Met Val Pro Lys Leu
35 40 45
Leu Ser Phe Gln Leu Ile Glu Cys Ile Cys Phe Arg Ala Val Leu Phe
50 55 60
Arg Met Val Pro Lys His Leu Glu Ala Val Lys Lys Ala Gln Tyr Gly
65 70 75 80
Phe Arg Ala Val Leu Phe Arg Met Val Pro Lys His Gly Thr Phe Asn
85 90 95
Arg Leu Ser Ala Ser Cys Phe Arg Ala Val Leu Phe Arg Met Val Pro
100 105 110
Lys His Gly Gln Lys Ser Lys Lys Thr Lys Ser Cys Phe Arg Ala Val
115 120 125
Leu Phe Arg Met Val Pro Lys Gln Leu Asn Lys Arg Ile Val Phe Leu
130 135 140
Ala Cys Phe Arg Ala Val Leu Phe Arg Met Val Pro Lys Pro
145 150 155
<210> 37
<211> 187
<212> PRT
<213> 宏基因组(metagenomic)
<400> 37
Met Ala Arg Leu Arg Lys Leu Lys Arg Lys Phe Ser Thr Ser Phe Arg
1 5 10 15
Ala Val Leu Phe Arg Met Val Pro Lys Leu Gly Lys Met Ala Glu Gly
20 25 30
Leu Thr Gln Arg Phe Arg Ala Val Leu Phe Arg Met Val Pro Lys Leu
35 40 45
Leu Ser Phe Gln Leu Ile Glu Cys Ile Cys Phe Arg Ala Val Leu Phe
50 55 60
Arg Met Val Pro Lys His Leu Glu Ala Val Lys Lys Ala Gln Tyr Gly
65 70 75 80
Phe Arg Ala Val Leu Phe Arg Met Val Pro Lys His Gly Thr Phe Asn
85 90 95
Arg Leu Ser Ala Ser Cys Phe Arg Ala Val Leu Phe Arg Met Val Pro
100 105 110
Lys Arg Asn Pro Ser Phe Pro Ser Val Pro Ser Cys Phe Arg Ala Val
115 120 125
Leu Phe Arg Met Val Pro Lys Leu Arg Ile Leu Asp Thr Leu Leu Glu
130 135 140
Cys Cys Phe Arg Ala Val Leu Phe Arg Met Val Pro Lys Gln Asp Leu
145 150 155 160
Lys Ser Ile Arg Ile Asp Phe Ser Phe Arg Ala Val Leu Phe Arg Met
165 170 175
Val Pro Lys Pro Ser Leu Glu Thr Glu Gln Asn
180 185
<210> 38
<211> 289
<212> PRT
<213> 宏基因组(metagenomic)
<400> 38
Met Ala Tyr Phe Leu Phe Ile Asp Glu Ser Gly Gln Asp His Arg Asp
1 5 10 15
Ser Pro Tyr Glu Val Leu Ala Gly Val Ala Val Lys Asp Thr Ala Leu
20 25 30
Trp Ser Leu Ile Leu Ala Ile Gln Glu Ala Glu Ile Arg His Phe Gly
35 40 45
Met Arg Tyr Ser Gln Gly Asn Arg Glu Leu Lys Gly Lys Lys Ile Leu
50 55 60
Lys Arg Lys Val Phe Thr His Ala Ala Leu Leu Pro Glu Ile Gln Glu
65 70 75 80
Ser Asp Arg Met Pro Leu Ala Lys Ser Cys Leu Glu Asn Gly Ala Gly
85 90 95
Ala Gly Lys Arg Glu Met Val Ala Leu Ala Gln Ala Lys Leu Ala Tyr
100 105 110
Val Gln Glu Val Phe Glu Ile Cys Ser Lys His Glu Cys Arg Leu Phe
115 120 125
Ala Ser Ile Val Ala Asp Arg Glu His Cys Asp His His Ala Asp Tyr
130 135 140
Leu Arg Lys Asp Tyr Ser Tyr Leu Phe Glu Arg Phe Tyr Tyr Phe Leu
145 150 155 160
Glu Asp Val Asp Pro Ala Ala Ala Gly Val Val Val Phe Asp Glu Leu
165 170 175
Glu Lys Ser Lys Ser His Ile Leu Leu Gly Gln Met Asp Ser Tyr Phe
180 185 190
Lys Leu Thr Ala Lys Gly Arg Lys Arg Ala Lys Gln Ile Ile Pro Glu
195 200 205
Pro Phe Phe Val His Ser Asp Leu Thr Thr Gly Ile Gln Val Ala Asp
210 215 220
Leu Ala Ala Tyr Ile Ile Ser Trp Gly Phe Arg Thr Ala Arg Leu Leu
225 230 235 240
Glu Pro Lys Arg Glu Glu Leu Lys Ser Leu Val His Ile Ile Ser Gln
245 250 255
Leu Arg His His Ser Glu Arg His Val Glu Gly Gln Thr Asn Phe Lys
260 265 270
Val Trp Ser Phe Gly Ile Ile Gln Asp Leu Arg Gly Gly Ser Ala Arg
275 280 285
Cys
<210> 39
<211> 205
<212> PRT
<213> 宏基因组(metagenomic)
<400> 39
Met Glu Gly Asp Ile Tyr Arg Met Leu Arg Ile Gln Asn Lys Ile Ile
1 5 10 15
Ser Ile His Ser Leu Arg Met Glu Gly Asp Ile Tyr Arg Met Leu Arg
20 25 30
Ile Gln Asn Lys Ile Ile Ser Ile His Ser Leu Arg Met Glu Gly Asp
35 40 45
Pro Asp Thr Thr Thr Glu Val Gln Thr Thr Glu Tyr Phe Asn Pro Leu
50 55 60
Pro Pro His Gly Gly Arg Gln Thr Asn His Tyr Ser Glu Thr Leu Thr
65 70 75 80
Ser Pro Phe Gln Ser Thr Pro Ser Ala Trp Arg Glu Thr Ser Ser Asp
85 90 95
Trp Met Ser Arg Arg Arg Pro Asp Ile Ser Ile His Ser Leu Arg Met
100 105 110
Glu Gly Asp Val Val Ser Val Ala Val Ser Gly Lys Pro Leu Tyr Phe
115 120 125
Asn Pro Leu Pro Pro His Gly Gly Arg Gln Lys Thr Ser Arg His Pro
130 135 140
Ala Arg Glu Ser His Phe Asn Pro Leu Pro Pro His Gly Gly Arg Arg
145 150 155 160
Gln Val Leu Ala Ala Val Cys Leu Phe Gln Asp Ile Ser Ile His Ser
165 170 175
Leu Arg Met Glu Gly Asp Ile Val Met Arg Val Gly Phe Gly Tyr Ser
180 185 190
Ala Phe Ile Ser Ile His Ser Leu Arg Met Glu Gly Asp
195 200 205
<210> 40
<211> 269
<212> PRT
<213> 宏基因组(metagenomic)
<400> 40
Met Glu His Tyr Phe Pro Val Lys Ser Ser Lys Arg Arg Asn Ser Thr
1 5 10 15
Ala Glu Ile Lys Tyr Tyr Leu Gln Thr Asp Thr Ile Lys Cys Cys Leu
20 25 30
His His Gln Ile Ile Asn Ala Val Arg Gly Leu Ala Leu Arg Glu Gly
35 40 45
Lys Phe Asn Leu His Gly Phe Asp Asp Lys Leu Ile Pro Asn Glu Arg
50 55 60
Asn Val Ser Ser Ser Ile Leu Asn Glu Leu Lys Thr Ser Glu Gly Phe
65 70 75 80
Val Leu Asn Met Leu Gly Gly Cys Ala Phe Ala Ala Asn Cys Leu Arg
85 90 95
Asn Ile Val Asp Ala Thr Gln Arg Ser Asp Leu Leu Gly Phe Arg Cys
100 105 110
Phe Glu Val Ser Leu Lys Lys Gly Lys Ser Asn Ser Asp Leu Phe Ala
115 120 125
Leu Phe Phe Gly Phe Gly Arg Glu Asp Met Asp Asp Asp Ser Glu Trp
130 135 140
Glu Lys His Leu Tyr Ala Ala Arg Tyr Ser Val Ser Glu Ile Arg Asn
145 150 155 160
Arg Val Ala His Tyr His Lys Ser Ala Ile Glu Asn Ile Tyr Asn Ile
165 170 175
Thr Asp Phe Lys Tyr Arg Glu Asn Ser Met Cys Ser Tyr Thr Asp Thr
180 185 190
Lys Phe Thr Thr Ala Leu Gln Asn Glu Ile Tyr Asn Thr Pro Lys Ala
195 200 205
Leu Ser Leu Gln Leu Met Thr Gly Lys Val Leu Glu Tyr Tyr Pro Lys
210 215 220
Glu Lys Leu Val Ser Phe Phe Gln Lys Tyr Lys Phe Ser Leu Tyr Arg
225 230 235 240
Ser Val Val Pro Phe Ala Pro Gly Phe Lys Asn Ile Met Arg Thr Gly
245 250 255
Val Asn Tyr Gln Asn Ala Thr Gln Asn Ser Leu Phe Leu
260 265
<210> 41
<211> 126
<212> PRT
<213> 宏基因组(metagenomic)
<400> 41
Met Gly Asn Asp Tyr Asn Asn Tyr Gln Asn Lys Arg Gln Gly Ile Lys
1 5 10 15
Pro Lys Gln Pro Asn Lys Val Arg Arg Ile Gln Ser Arg Asp Phe Arg
20 25 30
Ile Ser Leu Thr Gly Lys His Val Phe Gly Ala Tyr Phe Asn Met Ala
35 40 45
Arg Thr Asn Phe Val Lys Thr Ile Asn Tyr Val Leu Pro Ile Ala Gly
50 55 60
Val Arg Gly Asn Tyr Ser Glu Asn Gln Ile Asn Lys Met Leu Gln Ala
65 70 75 80
Leu Phe Leu Ile His Ser Asn Arg Ser Ala Glu Leu Thr Gln Asp Ser
85 90 95
Asn Asn Gly Lys Gly Ser Ser Arg Ser Leu Pro Asn Asn Arg His Gly
100 105 110
Tyr Asn Ala Cys Ser Ser Ser Ile Ser Pro Tyr Leu Val Gln
115 120 125
<210> 42
<211> 197
<212> PRT
<213> 宏基因组(metagenomic)
<400> 42
Met Gly Arg Leu Ser Gly Arg Arg Gln Pro Arg Gln Arg Asp Ala Asp
1 5 10 15
Pro Ala Leu Ala Arg Leu Asn Pro Ser Arg Pro Met Ala Gly Tyr Cys
20 25 30
Trp Arg Pro Ala His Ser Trp Gln Cys Arg Arg Glu Ile Asn Ala Thr
35 40 45
Glu Leu Ala Arg Pro Thr Ala Leu Pro Ala Arg Gln Arg Ala Ala Cys
50 55 60
Ala Ala Ala Asn Glu Thr Ile Arg Gly Gln Val Phe Asn Asn Glu Phe
65 70 75 80
Leu Ser Cys Leu Arg Gly Ser Glu Leu His Ala Thr Ala Phe Pro Gln
85 90 95
Gly Gly Gly Phe Leu Ser Cys Leu Arg Gly Ser Glu His Gln Gly Gln
100 105 110
Gln Arg Glu Gly Glu Ala Val Phe Leu Ser Cys Leu Arg Gly Ser Glu
115 120 125
Arg Gly Ile Pro Arg Leu Glu Arg Cys Arg Lys Phe Leu Ser Cys Leu
130 135 140
Arg Gly Ser Glu Pro Asp Asp Gly Gly Ile Ser Pro Ile Pro Glu Phe
145 150 155 160
Leu Ser Cys Leu Arg Gly Ser Glu Arg His Gln Arg Ser Arg Trp Ala
165 170 175
Cys Arg Met Phe Leu Ser Cys Leu Arg Gly Ser Glu His Glu Ser Tyr
180 185 190
Gly Cys Arg Gly Ser
195
<210> 43
<211> 144
<212> PRT
<213> 宏基因组(metagenomic)
<400> 43
Met Gly Tyr Asp Arg Lys Leu Lys Asn Ala Val Ala Gln Ser Val Ile
1 5 10 15
Glu Leu Val Ala Arg Glu Gly Leu His Leu Asp Trp Gly Met Gln Val
20 25 30
Leu Asp Gly Ala His Val Leu Val Asp Ala Arg Val Arg Gly Lys Gln
35 40 45
Ala Ala His Leu Gly Asn Leu Gly Leu Phe Glu Lys Gly Gln Gly Glu
50 55 60
Arg Glu Lys Lys Pro Ala His Arg Leu Thr Glu Thr Leu Ser Gly Pro
65 70 75 80
His Leu Ala Glu Met Ala Ala Thr Leu Phe Ala Gly Arg Val Ala His
85 90 95
Ala Thr Asp Ala Ala Ala Leu Pro Arg Ala Ala Ile Asp Trp Gln Arg
100 105 110
Ser Ala Ala Pro Glu Thr Asn Thr Lys Gln Ala His Asn His Pro Gln
115 120 125
Arg Pro Pro Lys Pro Arg Ile Gln Arg Pro Pro Leu Gln Ala Ala Lys
130 135 140
<210> 44
<211> 181
<212> PRT
<213> 宏基因组(metagenomic)
<400> 44
Met Ile Pro Gly Ile Ala Tyr Cys Arg Asp Phe Asn Pro Arg Ser Pro
1 5 10 15
Asn Gly Glu Arg Gln Pro Ala Ile Leu Ser Phe His Gly Ser Phe Gln
20 25 30
Ile Ser Ile His Ala Pro Arg Met Gly Ser Asp Leu Pro Val Ser Val
35 40 45
Ile Val Ser Pro Ala Gly Ile Ser Ile His Ala Pro Arg Met Gly Ser
50 55 60
Asp Glu Ser Cys Arg Ala Arg Val Phe Trp His Gly Asp Phe Asn Pro
65 70 75 80
Arg Ser Pro Asn Gly Glu Arg Leu Gly Ser Cys Pro Gly Leu Pro Ile
85 90 95
Ser Arg Phe Gln Ser Thr Leu Pro Glu Trp Gly Ala Thr Arg His Pro
100 105 110
Asp His His Pro Arg Gly Gly Ala Phe Gln Ser Thr Leu Pro Glu Trp
115 120 125
Gly Ala Thr Ala Lys Leu Trp Gln Trp Gly Gly Ala Asp Glu Tyr Phe
130 135 140
Asn Pro Arg Ser Pro Asn Gly Glu Arg Arg Arg Arg Arg Pro Gln Ile
145 150 155 160
Val Ala Asp Gln Ala Ile Ser Ile His Ala Pro Arg Met Gly Ser Asp
165 170 175
Phe Ser Ser Arg Leu
180
<210> 45
<211> 257
<212> PRT
<213> 宏基因组(metagenomic)
<400> 45
Met Lys Ile Pro Gln Ile Ile Glu Asp Asn Lys His Leu Phe Gly Thr
1 5 10 15
Tyr Ser Thr Met Ala Leu Ala Asn Ile Arg Asn Ile Leu Tyr His Ile
20 25 30
Ala Thr Leu Ala Cys Ile Glu Asn Asp Phe Asn Ala Asp Ser Asp Asp
35 40 45
Phe Trp His His Pro Cys Met Glu Ile Ile Asn Pro Gln Asn Leu Cys
50 55 60
Asn Asp Val Thr Lys Ala Asp Phe Val Thr Glu Lys Leu Lys Ser His
65 70 75 80
Phe Pro Phe Val Val Ile Met Ala Glu Ala Lys Arg Gln Lys Asp Ile
85 90 95
Ala Trp Ala Lys Asn Gln Ala Lys Lys Ala Phe Glu Asn Arg Asp Phe
100 105 110
Gln Lys Gln Gln Glu Phe Asn Asn Lys Gln Lys Ser Phe Leu Ser Ile
115 120 125
Thr Asn Ala Asp Ile Tyr Arg Met Leu Asn Asn Leu Leu Arg Val Leu
130 135 140
Thr Ser Tyr Arg His Tyr Thr Ala His Tyr Leu Ile Asn Tyr Ile Tyr
145 150 155 160
Phe Asn Glu Glu Ser Asn Leu Leu Lys Tyr Asn Glu Gln Pro Leu Ser
165 170 175
Tyr Asn Leu Asn Asp Tyr Phe Thr Ile Ala Leu Arg Asp Thr Ala Gln
180 185 190
Lys Tyr Ser Tyr Ser Pro Glu Ala Leu Ser Phe Ile Gln Ser Ser Arg
195 200 205
Tyr Lys Ile Glu Asn Arg Arg Lys Ile Leu Asp Thr Asp Phe Phe Leu
210 215 220
Ser Ile Gln His Arg Asn Gly Asp Ser Asn Pro Lys Asn Leu His Ile
225 230 235 240
Ser Gly Val Gly Val Ala Leu Leu Ile Cys Leu Phe Leu Glu Lys Asn
245 250 255
Met
<210> 46
<211> 289
<212> PRT
<213> 宏基因组(metagenomic)
<400> 46
Met Lys Lys Ile Asp Asp Leu Thr Gly Glu Ile Tyr Ala Lys Ile Ala
1 5 10 15
Thr Arg Ile Arg Glu Arg Lys Ser Gln Arg His Lys Lys Arg Asn Glu
20 25 30
Ile Thr Asp Asp Asn Ser Val Gln Leu Leu Ser Asn Ile Met Asn Asn
35 40 45
Lys Arg Leu Ser Ser Arg Asn Pro Tyr Leu Leu Asn Ser Lys Met Thr
50 55 60
Tyr Asp Ile Val Thr Asn Leu Asp Phe Lys Ser Ser Tyr Glu Leu Ile
65 70 75 80
Trp Gly Asn Gly Lys Asp Leu Asp Glu Met Leu Arg Ile Val Phe Glu
85 90 95
Tyr Ser Leu Glu Tyr Leu Gln Asn Lys Ser Asn Asp Tyr Gly Lys Ile
100 105 110
Ile Glu Asp Cys Leu Leu Asn Phe Tyr Pro Tyr Ala Arg Leu Ser Ala
115 120 125
Glu Tyr Asp His Ala Ile Glu Pro Phe Lys Pro Glu Ile Pro Asp Ile
130 135 140
Gly Leu Ala Tyr Asp Phe Ala Lys Lys His Leu Tyr Phe Glu Ile Ser
145 150 155 160
Asp Asp Phe Lys Ser Lys His Arg Lys Tyr Phe Glu Thr Leu Glu Thr
165 170 175
Lys Lys Leu Pro Asp Lys Ile Ile Thr Phe Val Glu Lys Asp Val Phe
180 185 190
Glu Ile Leu Lys Glu Tyr Leu Lys Lys His Ala Glu Gly Ile Thr Thr
195 200 205
Tyr Glu Leu Ile Ser Lys Ile Ile Gly Tyr Glu Thr Glu Asp Met Tyr
210 215 220
Glu Asp Met Ile His Gly Pro Glu Trp Ser Ala His Gln Pro Leu Thr
225 230 235 240
Tyr Thr Gly Glu Thr Tyr Lys Lys Val Arg Gln Glu Thr Ile Asp Ala
245 250 255
Gly Arg Ser Tyr Ile Asp Ala Ile Ile His Glu Gln Glu Glu Thr Asp
260 265 270
Tyr Phe Tyr Gln Ile Tyr Pro Tyr Pro Leu Thr Asn Gly Asp Phe Asp
275 280 285
Tyr
<210> 47
<211> 292
<212> PRT
<213> 宏基因组(metagenomic)
<400> 47
Met Lys Val Trp Tyr Met Pro Ser Trp Asn Gly Asp Leu Arg Leu Glu
1 5 10 15
Pro Leu Ser Ser Asp Pro Lys Arg Thr Ala Leu Ser Ile Ile Asp Pro
20 25 30
Thr Asp Ala Glu Lys Ile Ala Leu Gly Ala Ile Gly Ala Thr Leu Leu
35 40 45
Asp Lys Gly Trp Leu Lys Glu Pro Ile Leu Asp Leu Leu Ser Gly Glu
50 55 60
Ser Tyr Arg Glu Ser Gln Lys Val Glu Val Val Leu Asp Gly Pro Leu
65 70 75 80
Glu Glu Val Gly Pro Val Val Ile Ala Ala Leu Lys Pro Gly Pro Ala
85 90 95
Val Leu Thr Ala Val Ile Leu Arg Gly Gly Lys Val Glu Ile Val Glu
100 105 110
His Arg Asp Pro Gly Lys Ala Gly Ile Lys Glu Thr Ser Lys Glu Leu
115 120 125
Ala Ala Val Ala Lys Gln Pro Ala Glu Ala Ala Ala Thr Val Lys Arg
130 135 140
Pro Thr Pro Cys Cys Pro Asp Cys Tyr Val Asp Ala Ile Lys Pro Ala
145 150 155 160
Thr Gln Thr Leu Leu Ala Phe Leu Ser Pro Glu Gln His Glu Thr Trp
165 170 175
Ser Arg Glu Arg Tyr Ile Val Cys Arg Gly Gly Leu Ser Gly His Arg
180 185 190
Tyr Ile Leu Ala His Arg Ser Ser Pro Met Ala Ala Arg Asn Thr Arg
195 200 205
Ile Cys Phe Asp Ala Asp Asp Arg Ser Ile Leu His Phe His Asp Gln
210 215 220
Thr Val Pro Pro Glu Glu Glu Val Leu Ser Ala Met Leu Ile Leu Gln
225 230 235 240
His Arg Glu Asp Trp Leu Arg Asn Glu Ala Thr Cys Leu Gly Gly Gly
245 250 255
Leu Arg Arg Val Phe Lys Asn Pro Phe Gly Asp Ile Met Asp Gly Val
260 265 270
Ala Asp Ser Asn Leu Thr Arg Thr Ile Gly Leu Leu Ala Lys Ser Leu
275 280 285
Thr Pro Gly Pro
290
<210> 48
<211> 200
<212> PRT
<213> 宏基因组(metagenomic)
<400> 48
Met Leu Phe Gln Leu Phe Glu Asp Asp Lys Met Ile Glu Lys Ile Arg
1 5 10 15
Arg Lys Leu Pro Tyr Met Phe Phe Ile Ala Glu Ile Glu Met Ser Lys
20 25 30
Ser Gly Lys Val Gly Met Glu Val Gly Thr Leu Arg Glu Gln Ile Leu
35 40 45
Ile Ala Leu Leu Ile Tyr Lys Phe Gly Tyr Glu Asn Val Lys Leu Asn
50 55 60
Pro Ile Ala Asn Glu Asp Phe Asp Leu Glu Leu Tyr Gly Phe Ser Gly
65 70 75 80
Ser Gly Ile Lys Leu Val Trp Thr Val Asp Trp Glu Lys Val Glu Asn
85 90 95
Phe Tyr Arg Asn Tyr Glu Pro Arg Ser Glu Ile Ile Leu Val Glu Val
100 105 110
Ser Trp Tyr Lys Asn Gly Gly Leu Tyr Tyr Ile Pro Leu Glu Val Gln
115 120 125
Arg Glu Ile Phe Glu Glu Ile Gly Arg Asp Arg Tyr Leu Tyr Arg His
130 135 140
Lys Lys Gly Thr Asn Pro Arg Gly Val Glu Ile Ser Asn Glu Gly Ile
145 150 155 160
Lys Met Leu Ile Glu His Pro Gln Ala Lys Lys Ile Asp Ile Phe Trp
165 170 175
Glu Lys Pro Glu Ile Gly Ala Asp Ile Ile Tyr Lys Pro Tyr Gln Arg
180 185 190
Trp Val Glu Leu Trp Ala Glu Glu
195 200
<210> 49
<211> 252
<212> PRT
<213> 宏基因组(metagenomic)
<400> 49
Met Leu Pro Arg Ile Val Phe Phe Tyr Asp Met Gly Leu Thr Ala Phe
1 5 10 15
Gln Asp Lys Asp Arg Tyr Ile Asp Asp Val Glu Ala Trp Asn Ala Arg
20 25 30
Tyr Phe Phe Met Lys Leu Ile Tyr Asn Asn Leu Phe Leu Pro Pro Arg
35 40 45
Phe Thr Asn Asp Lys Asn Asp Leu Phe Arg Lys Thr Val Glu Gln Val
50 55 60
Leu Glu Val Asn Arg Ser His Met Glu Lys Val Gly Lys Asp Asp Arg
65 70 75 80
Gln Ala Phe Gly Glu Val Arg Gln Met Arg Glu Asn Glu Thr Ala Asn
85 90 95
Asp Tyr Met Ala Tyr Leu Gln Ser Ser Leu Arg Met Glu Lys Asn Lys
100 105 110
Lys Glu Leu Glu Gly Asn Gly Ala Lys Arg Ile Asn Phe Glu Lys Phe
115 120 125
Ile Leu Gln Met Phe Val Lys Gly Phe Asp Ser Phe Leu Lys Asp Gly
130 135 140
Ala Phe Glu Phe Val His Ser Pro Glu Asn Cys Leu Thr Gly Ser Thr
145 150 155 160
Glu Lys Glu Arg Ala Lys Glu Arg Asn Ser Leu Cys Glu Glu Ile Gly
165 170 175
Asn Tyr Val Arg Leu Val Ser Thr Val Ile Asp Pro Gly Lys Gly Ser
180 185 190
His Val Ala Phe Tyr Val Phe Cys Lys Met Leu Asn Ser Val Tyr Leu
195 200 205
Ser Asn Ile Arg Asn Glu Met Ile Lys Tyr His Lys Val Val Arg Thr
210 215 220
Asp Glu Leu Ala His Leu Ile Glu Ile Ile Glu Leu Cys Leu Leu Ser
225 230 235 240
Ala Asp Ala Val Ser Gln Lys Asn Ser Glu Cys Arg
245 250
<210> 50
<211> 203
<212> PRT
<213> 宏基因组(metagenomic)
<400> 50
Met Leu Pro Val Phe Gly Trp Met Ile Gly Ala Ala Val Val Gly Ala
1 5 10 15
Gly Lys Phe Leu Leu Glu Ile Leu Glu Asp Glu Ala Ser Ser Ala Arg
20 25 30
Ser Glu Trp Arg Glu Arg Ser Glu Glu Leu Ala Gln Lys Thr Asp Glu
35 40 45
Tyr Asn Arg Thr Leu Glu Arg Gln Leu Thr Thr Ser Ala Gln Ser Ile
50 55 60
Glu Phe Lys Arg Leu Thr Asn Leu Tyr Arg Glu Ser Arg Arg Lys Ala
65 70 75 80
Asp Leu Val His Arg Ser Leu Gln Glu Gly Arg Thr Leu Leu Ser Arg
85 90 95
Leu Asp Cys Ala Val Arg Gln Ser Ala Met Asp Lys Thr Ala Leu Ile
100 105 110
Gln Ala Thr Gln Gln Arg Asn His Lys Ala Arg Gln Ala Ala Arg Ser
115 120 125
Gln Leu Glu Ala Leu Asn Phe Leu Arg Ser Gly Leu Phe Ala Glu Arg
130 135 140
Thr Arg Ile Arg Glu Ser Gln Glu Asn Leu Leu Gly Arg Val Arg Glu
145 150 155 160
Leu Asn Gln Asn Thr His Gln Leu Lys Leu Gln Met Arg Asp Asn Cys
165 170 175
Gly His Ala Gly Arg Cys Trp Phe Asp Lys Ile Glu Gly His Arg Leu
180 185 190
Ala Pro Pro Gly Glu Ile Asn Lys Arg Lys Gly
195 200
<210> 51
<211> 247
<212> PRT
<213> 宏基因组(metagenomic)
<400> 51
Met Asn His Ser Glu Val Ile Val Ala Arg Ala Asp Lys Leu Gly Asp
1 5 10 15
Leu Gly Glu Gly Gln Thr Ile Gly Glu Phe Phe Phe Glu Leu Ser Ala
20 25 30
Ala Thr Ala Ser Glu Met Arg Ile Gln Arg Ala Tyr Glu Pro Asp Arg
35 40 45
Glu Lys Ala Asn Glu Gln Ala Gly Tyr Ile Glu Lys Leu Lys Trp Asp
50 55 60
Val Val Ala Gln Ala Phe Glu Thr Tyr Leu Lys Glu Lys Gly Phe Asp
65 70 75 80
Phe Val Leu Gln Leu Gly Ser Gly Thr Pro Leu Pro Glu His Ala Gln
85 90 95
Phe Ser Phe Asn Asp Leu Pro Ala Ala Glu Met Asn Leu Pro Ala Glu
100 105 110
Asp Trp Gln Ile Ser Leu Tyr Phe Leu Val His Leu Val Pro Val Ala
115 120 125
Asp Ile Gly Lys Leu Leu His Gln Ile Arg Lys Trp Asn Ile Leu Ala
130 135 140
Gly Lys Gly Gly Leu Thr Glu Ser Glu Arg Ala Asp Asp Thr Ala Ala
145 150 155 160
Lys Leu Asn Thr Val Phe Gly Leu Tyr Leu Asp Met His Asp Ala Lys
165 170 175
Phe Glu Gly Gly Ala Ala Leu Thr Gly Cys Asp Arg Phe Ser Ser Ile
180 185 190
Phe Glu Ser Glu Arg Ala Phe Asn Thr Val Phe Pro Ala His Gly Ser
195 200 205
Gly Asp Asp Arg Thr Pro Lys Arg Gly Leu Arg Glu Ile Met Arg Phe
210 215 220
Gly His Phe Ser Ser Leu Arg Asp Ile Phe Glu Gln Asn His Val Thr
225 230 235 240
Glu Asn Asp Ile Asn Ala Leu
245
<210> 52
<211> 221
<212> PRT
<213> 宏基因组(metagenomic)
<400> 52
Met Pro Leu Lys Arg Thr Ser Glu Phe Gln Ser Thr Arg Leu Ser Arg
1 5 10 15
Ala Ser Thr Cys Arg Leu Leu Pro Tyr Arg Asn Leu Pro Asp Ile Ser
20 25 30
Ile His Lys Ala Leu Thr Ser Leu Asp Arg Ile Leu Asp Asp Arg Lys
35 40 45
Lys Ile Leu Gly Asn Phe Asn Pro Gln Gly Ser His Glu Pro Arg Pro
50 55 60
Met Ser Gly Arg Asn Ile Pro Ala Ile Asn Asn Phe Asn Pro Gln Gly
65 70 75 80
Ser His Glu Pro Arg Leu Phe Asn Ala Leu Pro Arg Leu Ile Lys Trp
85 90 95
His Phe Asn Pro Gln Gly Ser His Glu Pro Arg Gln Gln Lys Tyr Thr
100 105 110
Ile Ile Leu Ala Ser Met Pro Asn Leu Tyr Ile Leu Ile Ile His Leu
115 120 125
Gln Gln Ile Ile Gln Gln Val Lys Lys Phe Ser Arg Ile Leu Ile Leu
130 135 140
Phe Ser Gly Arg Leu Gln Cys Glu Ser Pro Trp Asn Phe Met Cys Asn
145 150 155 160
Tyr Tyr Ser His Gln Ser Phe Leu Thr Lys Asn Gln Pro Ser Phe Lys
165 170 175
Leu Pro Pro Leu Ser Ile Cys Ala Lys Phe Ser Arg Thr Ile Leu Asn
180 185 190
Asn His Leu Lys Trp Arg Ser Gln Leu Thr Val Tyr Pro Ala Val Tyr
195 200 205
Tyr Val Leu Ser Arg Ser Lys Asn Ser Asn Val Phe Thr
210 215 220
<210> 53
<211> 116
<212> PRT
<213> 宏基因组(metagenomic)
<400> 53
Met Arg Asp Ser Gly Ser Ala Arg Ile Glu Phe Gln Ser Thr Pro Ser
1 5 10 15
Gly Gly Lys Ala Thr Leu Phe Ser Ser Val Thr Thr Pro Tyr Arg Asn
20 25 30
Ser Phe Asn Pro Arg Leu Pro Gly Gly Arg Arg Arg Glu His Pro Leu
35 40 45
Asp Arg Asp Ala Arg Ala Val Val Ser Ile His Ala Phe Arg Gly Glu
50 55 60
Gly Asp Leu Leu His Asp Ala Thr Lys Asn Leu Ser Glu Val Ser Ile
65 70 75 80
His Ala Phe Arg Gly Glu Gly Asp Glu Glu Ile Gly Trp Phe Glu Asp
85 90 95
Val Ile Asp Gly Phe Asn Pro Arg Leu Pro Gly Gly Arg Arg Pro Gln
100 105 110
Arg Ala Lys Ser
115
<210> 54
<211> 273
<212> PRT
<213> 宏基因组(metagenomic)
<400> 54
Met Arg Gly Lys Pro Cys Ser Leu Ile Pro Thr Met Ser Ala Ala Arg
1 5 10 15
Ile Ile Pro Ala His Ala Gly Gln Thr Arg Arg Arg Glu Lys Pro Ser
20 25 30
Cys Thr Met Pro Asp His Pro Arg Thr Cys Gly Ala Asn Ala Val Ala
35 40 45
Arg Ala Cys Ala Val Leu Pro Val Gly Ser Ser Pro His Met Arg Gly
50 55 60
Lys Leu Pro Gln Arg Arg Phe Phe Ala Gly Arg Leu Arg Ile Ile Pro
65 70 75 80
Ala His Ala Gly Gln Thr Phe Cys Val Ala Thr Cys Ile Ile Lys Ile
85 90 95
Ser Asp His Pro Arg Thr Cys Gly Ala Asn Thr Pro Pro Ser Arg Ser
100 105 110
Met Pro Cys Ala Phe Gly Ser Ser Pro His Met Arg Gly Lys Pro Pro
115 120 125
Ala Cys Phe Arg Gln Val Trp Leu Leu Arg Ile Ile Pro Ala His Ala
130 135 140
Gly Gln Thr Arg Val Pro Tyr Ala Thr Gly Pro Leu Arg Gly Asp His
145 150 155 160
Pro Arg Thr Cys Gly Ala Asn Ala Val Ser Cys Cys Gly Tyr Gln Gly
165 170 175
Met Gly Gly Ser Ser Pro His Met Arg Gly Lys His Val Leu Ser Tyr
180 185 190
Asp Asp His Ser Leu Leu Arg Ile Ile Pro Ala His Ala Gly Gln Thr
195 200 205
Arg Arg Arg Ala Gly Val Pro Arg Asp Arg Thr Asp His Pro Arg Thr
210 215 220
Cys Gly Ala Asn Asp Ala Ala Ala Val Arg Glu Phe Asp Ser Leu Gly
225 230 235 240
Ser Ser Pro His Met Arg Gly Lys Leu Phe Ala Ala Leu Tyr Leu Arg
245 250 255
Cys Leu Arg Arg Ile Ile Pro Ala His Ala Gly Gln Thr Cys Arg Arg
260 265 270
Arg
<210> 55
<211> 295
<212> PRT
<213> 宏基因组(metagenomic)
<400> 55
Met Arg Ser Thr Gly Lys His Pro His Lys Asn Asn Met Asn Asp Lys
1 5 10 15
Pro Asn Phe Pro Gly Trp Gly Val Asn Arg Val Lys Val Asp Leu Glu
20 25 30
Ser Ser Glu Glu Thr Leu Cys Phe Arg Ala Gly Val Thr Leu Asp Gly
35 40 45
Lys Leu Val Gly His Ala Phe Asn Glu Gly Gly Gly Gly Pro Thr His
50 55 60
Val Thr Leu Thr Lys Ala Ala Leu Lys Asn Ile Ala Lys Pro Thr Gly
65 70 75 80
Ile Ala Asp His Val Asp Arg Leu Val Asp Asn Val Val Ala Leu Lys
85 90 95
Glu Leu Ala Lys Thr His Lys Arg Trp Cys Lys Glu Ile Gly Val Gly
100 105 110
Cys Val Tyr Ala Leu Gln His Asp Glu Leu Lys Ser Trp Glu Ala Gly
115 120 125
Met Asp His Gln Glu Ser Thr Met Pro Pro Phe Arg Ala Tyr Ser Ser
130 135 140
His Glu Ala Ala Ala Lys Ala Leu Ala Gly Val Glu His Phe Ser Tyr
145 150 155 160
Ala Lys Gly Ser Ser Lys Ala Leu Trp Glu Ile Met Val Gln Cys His
165 170 175
Asp Lys Trp Thr Ala Leu Glu Asp Ala Arg Thr Asp Lys Met Arg Ala
180 185 190
Glu Tyr Glu Ala Lys Leu Gln Ala Gln Leu Ala Lys Ala Gln Glu Gly
195 200 205
Lys Thr Ser Gly Pro Glu Glu Asn Glu Asp Tyr Phe Arg Lys Lys Pro
210 215 220
Ser Ala Glu Glu Ala His Arg Val Glu Ile Gly Lys Ile Leu Gln Arg
225 230 235 240
His Leu Ala Ala Asn Leu Pro Leu Pro Thr Thr Val Arg Leu Glu Ala
245 250 255
Leu Lys Leu Leu Ser Glu Ala Cys Val Ile Ile Asp Asp Glu Asp Leu
260 265 270
Ser Pro Ala Ala Leu Leu Ala Tyr Tyr Val Gln Tyr Val Val Pro Glu
275 280 285
Leu Leu Leu Met Pro Asn Asp
290 295
<210> 56
<211> 184
<212> PRT
<213> 宏基因组(metagenomic)
<400> 56
Met Arg Val Ala Lys Gly Ser Arg Phe Asn Ser Ala Thr Ala Val Met
1 5 10 15
Pro Trp Lys Arg Leu Ser Phe Arg Cys Gly Cys Ser Ala Gly Ser Gly
20 25 30
Phe Asn Ser Ala Thr Ala Val Met Pro Trp Lys Arg Cys Ser His His
35 40 45
Asp Arg Arg Arg Ser Ser Ser Gln Leu Gln Phe Gly His Gly Cys Asp
50 55 60
Ala Val Glu Thr Thr Ala Trp Gly Phe Gly Ser Ala Arg Pro Thr Ser
65 70 75 80
Phe Asn Ser Ala Thr Ala Val Met Pro Trp Lys Arg Cys Arg Val Gly
85 90 95
Arg Gly Pro Arg Val Gln Arg Arg Phe Asn Ser Ala Thr Ala Val Met
100 105 110
Pro Trp Lys Arg Arg Arg Ile Thr Pro His Thr His Pro Arg Ile Gly
115 120 125
Leu Gln Phe Gly His Gly Cys Asp Ala Val Glu Thr Gly Arg Thr Gly
130 135 140
Thr Glu Ile Gln Arg Gln Gln Glu Val Leu Gln Phe Gly His Gly Cys
145 150 155 160
Asp Ala Val Glu Thr Leu Thr Cys Ser Ser Ile Ser Ser Leu Asn Arg
165 170 175
Ser Ala Ser Ile Arg Pro Arg Leu
180
<210> 57
<211> 248
<212> PRT
<213> 宏基因组(metagenomic)
<400> 57
Met Ser Arg Pro Thr Ile Pro Ile Asp Leu Tyr Arg Asp Ala Leu Glu
1 5 10 15
Val Phe His Trp Ala Thr Lys Arg His Phe Gln Leu Trp Phe Thr Gly
20 25 30
Thr Ser Gln Asn Arg His Arg Arg Thr Glu Ser Val Leu Cys Arg Leu
35 40 45
Ala Lys Asn Gly Glu Ala Arg Ala Val Arg Tyr Gly Ser Gly Leu Ile
50 55 60
Tyr Ser Leu Arg Arg Lys Lys Ser Glu Glu Phe Arg Glu Leu Glu Lys
65 70 75 80
Val Ala His Gly Leu Ala Cys Thr Glu Gly Leu Val Arg Ala Tyr Arg
85 90 95
Ser Ser Leu Asp Gly Glu Val Ile Ala Glu Lys Phe Phe Ala Gly Pro
100 105 110
Gly Ser Val Pro Glu Trp Gly Ile Arg Tyr Pro Asp Gly Lys Met Leu
115 120 125
Leu Phe Glu Phe Gly Thr Lys His Asn Phe Tyr Phe Ser Gly Met Met
130 135 140
Arg Gly Lys Leu Ser Ala Tyr Glu Ala Asn Leu Glu Lys Ile Glu Glu
145 150 155 160
Lys Phe Gln Ala Lys Ala Ile Val Leu Phe Val Leu Asp Val Pro Arg
165 170 175
Pro Ala Val Glu Arg Tyr Val Gly Ser Leu Lys Arg Asp Ala Gly Ser
180 185 190
Val Ala Asp Gly Val Pro Ser Ala Ser Ser Gly Gly Asp Arg Phe Pro
195 200 205
Ser Asp Pro Phe Phe Phe Thr Asp Tyr Lys Thr Phe Leu Glu Val Pro
210 215 220
Ile Gly Gly Gln Leu Lys Ala Pro Ile Tyr Ile Trp Ser Asp Gly Glu
225 230 235 240
Ser Tyr Pro Leu Arg Asn Asn Val
245
<210> 58
<211> 178
<212> PRT
<213> 宏基因组(metagenomic)
<400> 58
Met Ser Val Thr Val Tyr Asn Ser Leu Arg Asn Gln Pro Gly Ser Ala
1 5 10 15
Lys Arg Leu Gly Ser Gln Leu Pro Glu Leu Thr Ala Ala Arg Leu Ile
20 25 30
Val Glu Ser Gln Leu Val Asn Ala Gln Gln Leu Gln Glu Ser Val Gln
35 40 45
Ser Ile Glu Ala Asn Ala Gly Trp Ala Met Trp Arg Asp Ala Leu Val
50 55 60
Leu Cys Thr Thr Leu Asp Ser Ala Arg Gln Asp Val Ile Glu Gly Gln
65 70 75 80
Trp Cys Asn Glu Gln Val Ser Leu Ser Ala Arg Leu Val Ala Pro Gln
85 90 95
Gln Tyr Arg Leu Val Arg Met His Arg Glu Thr Asp Ala Gly Ala Asp
100 105 110
Gln Val Tyr Arg Asp Gln Arg Val Leu Leu Arg Asp Gly Leu Asp Glu
115 120 125
Gln His Ala Ile Tyr Arg Leu Trp Tyr Gln Lys Gln Asn Thr Pro Leu
130 135 140
Leu Ser Glu Thr Ala Glu Pro Thr Glu Gln Trp Ala Trp Arg Pro Val
145 150 155 160
Ala Gln Gln Phe Met Gly Trp His Thr Ser Gln Ala Leu Gln Lys Glu
165 170 175
Ile Ser
<210> 59
<211> 266
<212> PRT
<213> 宏基因组(metagenomic)
<400> 59
Met Thr Glu Gln Asn Glu Arg Pro Tyr Asn Gly Thr Tyr Tyr Thr Leu
1 5 10 15
Glu Asp Lys His Phe Trp Ala Ala Phe Leu Asn Leu Ala Arg His Asn
20 25 30
Ala Tyr Ile Thr Leu Ala His Ile Asp Arg Gln Leu Ala Tyr Ser Lys
35 40 45
Ala Asp Ile Thr Asn Asp Glu Asp Ile Leu Phe Phe Lys Gly Gln Trp
50 55 60
Lys Asn Leu Asp Asn Asp Leu Glu Arg Lys Ala Arg Leu Arg Ser Leu
65 70 75 80
Ile Leu Lys His Phe Ser Phe Leu Glu Gly Ala Ala Tyr Gly Lys Lys
85 90 95
Leu Phe Glu Ser Gln Ser Ser Gly Asn Lys Ser Ser Lys Lys Lys Glu
100 105 110
Leu Thr Lys Lys Glu Lys Glu Glu Leu Gln Ala Asn Ala Leu Ser Leu
115 120 125
Asp Asn Leu Lys Ser Ile Leu Phe Asp Phe Leu Gln Lys Leu Lys Asp
130 135 140
Phe Arg Asn Tyr Tyr Ser His Tyr Arg His Pro Glu Ser Ser Glu Leu
145 150 155 160
Pro Met Phe Asp Gly Asn Met Leu Gln Arg Leu Tyr Asn Val Phe Asp
165 170 175
Val Ser Val Gln Arg Val Lys Glu Ile Thr Ser Thr Thr Thr Arg Ser
180 185 190
Ile Pro Thr Ala Thr Ser Thr Thr Leu Ser Gly Lys Ala Arg Lys Ile
195 200 205
Gly Ala Ala Ile Thr Thr Ile Pro Ser Ser Ser Thr Ile Leu Ser Ile
210 215 220
Glu Arg Gly Arg Ser Arg Lys Leu Asp Cys Cys Phe Ser Ser Pro Ser
225 230 235 240
Phe Leu Lys Lys Gly Met Pro Ser Gly Cys Lys Arg Arg Ser Glu Ala
245 250 255
Ser Arg Ala Ala Pro Lys Pro Thr Ser Arg
260 265
<210> 60
<211> 150
<212> PRT
<213> 宏基因组(metagenomic)
<400> 60
Met Val Ala Thr Gln Arg Lys Val Ser Thr His Ala Pro Ala Lys Ala
1 5 10 15
Arg Pro Ser Tyr Glu Phe Ala Gly His Ala Leu Thr Gly Phe Asn Ser
20 25 30
Arg Ala Arg Glu Gly Ala Thr His Ser Asp Ala Gln Asp His Val Leu
35 40 45
Arg Gly Phe Asn Ser Arg Ala Arg Glu Gly Ala Thr Gln Met Ile Pro
50 55 60
Val Ile Tyr Asp Val Asp Ser Phe Asn Ser Arg Ala Arg Glu Gly Ala
65 70 75 80
Thr Cys Arg Arg Asn Arg Arg Arg His Ser Leu Cys Phe Asn Ser Arg
85 90 95
Ala Arg Glu Gly Ala Thr Ser Tyr Ala Pro His Gln Pro Lys Phe Leu
100 105 110
Gln Phe Gln Leu Thr Arg Pro Arg Arg Arg Asp Tyr Leu Lys Phe Ala
115 120 125
Val Met Ser Ala Ser Ser Val Ser Thr His Ala Pro Ala Lys Ala Arg
130 135 140
Gln Ala Ser Ser Lys Tyr
145 150
<210> 61
<211> 191
<212> PRT
<213> 宏基因组(metagenomic)
<400> 61
Met Val Phe Thr Tyr Ile Leu Gly Val Ile Gly Ile Ile Val Thr Leu
1 5 10 15
Lys Val Ile Gln Met Thr Phe Ile Gly Leu Gln Met Leu Lys Thr Ala
20 25 30
Val Cys Asp Pro Ser Thr Asn Asp Asp Gly Leu Ala Val Glu Ser Phe
35 40 45
Thr Asp Leu Leu Glu Arg Ala Arg Ser Asn Met Thr Val Tyr Asp Asp
50 55 60
Gly Asn Asp Met Lys Gly Ser Ile Tyr Met Gln Gln Glu Val Ile Asp
65 70 75 80
Ala Val Lys Glu Lys Leu Glu Asn Asn Lys Arg Phe Ser Leu Arg Cys
85 90 95
Tyr Phe Asn Cys Asn Glu Pro Thr Leu Phe Arg Lys Glu Phe Asp Ser
100 105 110
Tyr Ser Lys Val Asp Ile Lys Thr Arg Pro Ser Gly Asp Thr Arg Pro
115 120 125
Asp Asp Thr His Tyr Lys Ile Ile Asp Asn Gly Arg Met Ala Tyr Leu
130 135 140
Ser Arg His Glu His Gly Ser Lys Lys Arg Glu Phe Gln Val Val Asp
145 150 155 160
Cys Thr Arg Val Asn Arg Phe Ala Leu Glu Gly Val Thr Asp Glu Leu
165 170 175
Leu Gly Glu Tyr Lys Val Asp Ile Lys Gln Lys Phe Pro Ser Ala
180 185 190
<210> 62
<211> 75
<212> PRT
<213> 宏基因组(metagenomic)
<400> 62
Met Gly Asp His Lys Ile Ile Ser His Glu Arg Asp Lys Pro Ile Tyr
1 5 10 15
Arg Arg Leu Asp Ser His Pro Phe Arg Leu Gln Asn Ile Ile Gly Tyr
20 25 30
Pro Arg Gln Arg Phe Asp Leu Leu Trp Asn Arg Arg Leu Leu Pro His
35 40 45
Lys Asp Met Glu Pro Ile Gly Asp Phe Ala Val Asp Asp Phe Gln Arg
50 55 60
Cys Asn Leu Tyr Asn Cys Val Phe Leu Asn Val
65 70 75
<210> 63
<211> 72
<212> PRT
<213> 宏基因组(metagenomic)
<400> 63
Met Pro Leu Arg Ala Ala Ala Gly Gly Val Asp Phe Ser Ala Ala Asp
1 5 10 15
Ala Pro Gly Ala Glu Arg Ala Ala Val Leu Arg Pro His Gly Arg Pro
20 25 30
Glu Thr Gly Ala Glu Glu Ala Pro Thr Asp Gln Ala Val Pro Gly Leu
35 40 45
Arg Ser Arg Leu Phe Arg Ala His Arg Gly Ala Val Lys Arg Glu Cys
50 55 60
Leu Ala Asp Val Thr Val Val Ser
65 70
<210> 64
<211> 87
<212> PRT
<213> 宏基因组(metagenomic)
<400> 64
Met Val Ala Tyr Ile Asp Glu Phe Lys Asp Arg Phe Arg Val Gly Pro
1 5 10 15
Ile Arg Arg Val Leu Ala Ala Ser Leu Asp Cys Gly Phe Ile Thr Pro
20 25 30
Arg Gly Tyr Arg Met Phe Lys Ser Arg Pro Val Ser Arg Met Ala Ala
35 40 45
Arg His Glu Ala Leu Ala Arg Asp Ile Leu Glu Ile His Ala Asp Ser
50 55 60
Phe Met Ala Val Tyr Gly Tyr Arg Lys Thr Arg Ala Arg Ser Leu Ala
65 70 75 80
Arg Gly Arg Asp Pro Ala Gly
85
<210> 65
<211> 84
<212> PRT
<213> 宏基因组(metagenomic)
<400> 65
Met Thr Glu Phe Glu Arg Leu Met Arg Ala Ser Asp Phe Cys His Ser
1 5 10 15
Met Ser Ala Ile Ala Leu Glu Arg Asn Cys Ile Asp Ala Phe His Ala
20 25 30
Trp Arg Ser His Ala Arg Ala Tyr Ser Ser Ala Ala Arg Val Leu His
35 40 45
Arg Ala Gln Trp Lys Arg Ser Val Thr Asp Asp Gly Met Glu Pro Cys
50 55 60
Lys Arg Asn Ala Asp Gly Leu Cys Arg Ser Arg Arg Thr Ala Phe Arg
65 70 75 80
Trp Gly Lys Ser
<210> 66
<211> 61
<212> PRT
<213> 宏基因组(metagenomic)
<400> 66
Met Ser Arg Leu Ser Ala Ile Ala His Leu Arg Ala Tyr Gly Asp Asn
1 5 10 15
Ile Ser Gly Leu Ser Pro Gln Leu Leu Val Leu Ser Leu Ile Gly Val
20 25 30
Ile Thr Arg Asn His Glu Met Asn His Ala Ile Ala Val Phe Ser Ser
35 40 45
Leu Val Ser Trp His Ile Tyr Ser Ser Arg His Ile Asp
50 55 60
<210> 67
<211> 97
<212> PRT
<213> 宏基因组(metagenomic)
<400> 67
Met Leu Val Pro Leu Ser Val Cys Met Leu Ile Ser Pro Gly Ser Gln
1 5 10 15
Val Ser Ser Ser Pro Ala Phe Val Thr Cys Arg Thr Pro Cys Ile Thr
20 25 30
Arg His Pro Ala Leu Arg Ala Leu His Gln Arg Pro Ala Pro Thr Pro
35 40 45
Cys Ile Ile Arg Arg Pro Ala Leu Arg Ala Leu His His Pro Tyr Met
50 55 60
Pro Gln Arg Leu Pro Gly Asp His Lys Ile Met Arg Trp Tyr Ala Pro
65 70 75 80
Cys Ser Cys Leu Ala Gln Trp Arg Val Ala Arg Pro Ser Ser Cys Ile
85 90 95
Phe
<210> 68
<211> 92
<212> PRT
<213> 宏基因组(metagenomic)
<400> 68
Met Pro Cys Ser Lys Arg Leu Leu Ser Asp Ser Gln Tyr Ser Gln Arg
1 5 10 15
Arg His His Pro Gly Tyr Asp Arg Glu Ser Ala Arg Pro Lys Thr Cys
20 25 30
Asn His Pro Ile Pro Arg Arg Arg Arg Pro Gln Lys Phe Lys Lys Phe
35 40 45
Ser Lys Thr Asn Pro Phe Gly Arg Asn Leu Ser Asn His Asn Ala Ile
50 55 60
His Leu Phe Met Asp Asn Thr Ser Pro Arg Cys His Pro Gly Ala Ala
65 70 75 80
Leu Tyr Asn Cys Phe His Ala Asn Gly Ile Pro Ala
85 90
<210> 69
<211> 74
<212> PRT
<213> 宏基因组(metagenomic)
<400> 69
Met Leu Ala Phe Phe Asn Thr Arg Thr Ser Leu Ala Asp Trp Asp Leu
1 5 10 15
Tyr Ser Asp Arg Asn Tyr Arg Leu His Trp Lys Pro Lys His Arg Ser
20 25 30
Leu Ala Asp Trp Asp Leu Tyr Ser Asp Arg Asn Asp Ser Ile Phe Asn
35 40 45
Leu Lys Glu Gly Ile Ser Leu Ala Asp Trp Asp Leu Tyr Ser Asp Arg
50 55 60
Asn Cys Met Gly Gln His Asp Gly Arg Arg
65 70
<210> 70
<211> 90
<212> PRT
<213> 宏基因组(metagenomic)
<400> 70
Met Arg Tyr Tyr Thr Lys His Asp Lys Ser Ile Phe Gly Leu Leu Gly
1 5 10 15
Glu Ile Tyr Asn Gly Phe Met Ile Tyr Asp Thr Lys Pro Lys Lys Ser
20 25 30
Val Ser Phe Ile Thr Gln Asn Ile Leu Glu Arg Tyr Arg Val Val Ala
35 40 45
Asn Ile His Met Glu His Ala Asp Asp Lys Asn Phe Asp Phe Lys Tyr
50 55 60
Thr Ala Glu Leu Ser Ser Asp Arg Tyr Thr Tyr Lys Asp Asp Ser Cys
65 70 75 80
Glu Arg Trp Gly Glu Met Lys Arg Lys Arg
85 90
<210> 71
<211> 58
<212> PRT
<213> 宏基因组(metagenomic)
<400> 71
Met Leu Thr Ala Phe Phe Gln Ile Gly Ser Asn His Phe Phe His Glu
1 5 10 15
Glu Arg Gln Cys Lys Gly Lys Ala His Glu Asn Glu Glu Arg Tyr Glu
20 25 30
Glu Arg Tyr Glu Glu Arg Tyr Arg Asp Pro Phe Ile His Asn Val Ser
35 40 45
Asn His Ile Arg Glu Thr Gly Lys Asn Leu
50 55
<210> 72
<211> 332
<212> PRT
<213> 宏基因组(metagenomic)
<400> 72
Met Arg Ser Leu Pro His Arg Gln Leu Arg Lys Thr Ser Ala Arg Gly
1 5 10 15
Ser Ala Ala Trp Pro Cys Ser Leu Pro His Arg Gln Leu Arg Lys Asp
20 25 30
Ala Pro Pro Ser Ile Ala Met Glu Thr Cys Ser Leu Pro His Arg Gln
35 40 45
Leu Arg Asn Ala Arg Trp Ala Arg Pro Arg Pro Gly Pro Arg Ser Leu
50 55 60
Pro His Arg Gln Leu Arg Lys Asp Gly Ser Thr Phe Thr Leu Pro Ala
65 70 75 80
Ser Met Phe Thr Ala Ala Gln Ala Ala Gln Lys Thr Pro Arg Ala Ala
85 90 95
Trp Leu Ser Leu Ala Leu Phe Thr Ala Ala Gln Ala Ala Gln Lys Ala
100 105 110
Cys Thr Thr Gln Phe Trp Glu Ala Met Val Phe Thr Ala Ala Gln Ala
115 120 125
Ala Gln Lys Ile Arg Pro Ile Ser Asn Ser Ala Gly His Ser Phe Thr
130 135 140
Ala Ala Gln Ala Ala Gln Lys Thr His Phe Gly Ser Pro Val Arg Ala
145 150 155 160
Ala Ser Phe Thr Ala Ala Gln Ala Ala Gln Lys Phe Ser Arg Tyr Leu
165 170 175
Leu Asp Thr Ser Asn Ser Phe Thr Ala Ala Gln Ala Ala Gln Lys Arg
180 185 190
Arg Ser Glu Pro Arg Lys Trp Val Asn Pro Phe Thr Ala Ala Gln Ala
195 200 205
Ala Gln Lys Ser Arg Arg Pro Pro Ser Gln Ala Asp Lys Ser Phe Thr
210 215 220
Ala Ala Gln Ala Ala Gln Lys Thr Gln Glu Gly Tyr Ser Ala Phe Thr
225 230 235 240
Lys Glu Phe Thr Ala Ala Gln Ala Ala Gln Lys Ala Ser Lys Pro Ser
245 250 255
Arg Pro Ser Ser Pro Met Phe Thr Ala Ala Gln Ala Ala Gln Lys Asp
260 265 270
Gly Ala Val Gly Asp Gly Ala Ala Gln Gly Phe Thr Ala Ala Gln Ala
275 280 285
Ala Gln Lys Ser Lys Ala Arg Ser Pro Lys Lys Thr Ser Pro Val His
290 295 300
Cys Arg Thr Gly Ser Ser Glu Ser His Arg Ile Asp Ala Arg Arg His
305 310 315 320
Ala Val Val His Cys Arg Thr Gly Ser Ser Glu Asn
325 330
<210> 73
<211> 362
<212> PRT
<213> 宏基因组(metagenomic)
<400> 73
Met Glu Gly Asp Ile Arg Phe Asp Glu Ile Leu Asn Pro Lys Trp Ala
1 5 10 15
Phe Gln Ser Thr Pro Ser Ala Trp Arg Glu Thr Glu Ser Arg Gly Val
20 25 30
Leu Ser Val Ser Glu Arg Phe Gln Ser Thr Pro Ser Ala Trp Arg Glu
35 40 45
Thr Phe Arg Leu Arg Thr Ser Leu Asp His Gln His Ile Ser Ile His
50 55 60
Ser Leu Arg Met Glu Gly Asp Cys Arg Lys Ser Cys Gln Asn Lys Tyr
65 70 75 80
Ile Ala Tyr Phe Asn Pro Leu Pro Pro His Gly Gly Arg Leu Val Phe
85 90 95
Ser Leu Pro Gly Ala Asp Leu Gly Ser Ile Ser Ile His Ser Leu Arg
100 105 110
Met Glu Gly Asp Ser Ser Pro Asp Thr Ala Glu Pro Met Leu His Asp
115 120 125
Ile Ser Ile His Ser Leu Arg Met Glu Gly Asp Asn Val Cys Asp Cys
130 135 140
Pro Val Cys Ile His Cys Ile Ser Ile His Ser Leu Arg Met Glu Gly
145 150 155 160
Asp Ile Tyr Arg Met Leu Arg Ile Gln Asn Lys Ile Ile Ser Ile His
165 170 175
Ser Leu Arg Met Glu Gly Asp Ile Tyr Arg Met Leu Arg Ile Gln Asn
180 185 190
Lys Ile Ile Ser Ile His Ser Leu Arg Met Glu Gly Asp Pro Asp Thr
195 200 205
Thr Thr Glu Val Gln Thr Thr Glu Tyr Phe Asn Pro Leu Pro Pro His
210 215 220
Gly Gly Arg Gln Thr Asn His Tyr Ser Glu Thr Leu Thr Ser Pro Phe
225 230 235 240
Gln Ser Thr Pro Ser Ala Trp Arg Glu Thr Ser Ser Asp Trp Met Ser
245 250 255
Arg Arg Arg Pro Asp Ile Ser Ile His Ser Leu Arg Met Glu Gly Asp
260 265 270
Val Val Ser Val Ala Val Ser Gly Lys Pro Leu Tyr Phe Asn Pro Leu
275 280 285
Pro Pro His Gly Gly Arg Gln Lys Thr Ser Arg His Pro Ala Arg Glu
290 295 300
Ser His Phe Asn Pro Leu Pro Pro His Gly Gly Arg Arg Gln Val Leu
305 310 315 320
Ala Ala Val Cys Leu Phe Gln Asp Ile Ser Ile His Ser Leu Arg Met
325 330 335
Glu Gly Asp Ile Val Met Arg Val Gly Phe Gly Tyr Ser Ala Phe Ile
340 345 350
Ser Ile His Ser Leu Arg Met Glu Gly Asp
355 360
<210> 74
<211> 359
<212> PRT
<213> 宏基因组(metagenomic)
<400> 74
Met Tyr Phe Gln Leu Gly Phe His Tyr Leu Cys Leu Lys Ser Asn Cys
1 5 10 15
Glu Phe Pro Glu Gln Tyr Lys Thr Ile Thr Ile Gly Asn Gln Gly Thr
20 25 30
Ile Asn Gly Ala Ile Leu Tyr Gln Leu Val Ala Thr Asn Thr Asn Gly
35 40 45
Leu Pro Val Tyr Thr Leu Asp Asn Asp Gly Gly Asp Val Asp Ala Tyr
50 55 60
Lys Tyr Ala Asn Lys Ser Thr Gly Ala Lys Ile Ser Lys Cys Phe Leu
65 70 75 80
Lys Tyr Ser Glu Lys Ile Gly Tyr Asn Ile Tyr Asp Ala Gly Leu Glu
85 90 95
Leu Phe Gln Asn Met Ser Glu Glu Lys Asn Ile Asn Glu Phe Arg Asp
100 105 110
Tyr Ile Asp His Phe Lys Tyr Phe Val Asp Lys Glu Arg Ser Ile Leu
115 120 125
Glu Leu Tyr Ser Glu Met Phe Asp Arg Phe Met Ala Tyr Asp Gln Lys
130 135 140
Tyr Arg Lys Asn Val Pro Asn Met Leu Thr Asn Ile Leu Lys Arg Asn
145 150 155 160
Tyr Ile Ile Thr His Leu Glu Phe Lys Ser Gly Gln Lys Lys Val Gly
165 170 175
Arg Gly Asn Lys Ser Thr Thr Lys Glu Met Gly Leu Ile Cys Ile Arg
180 185 190
Asp Asn Gly Ile Glu Ser Asp Asp Phe Thr Tyr Lys Leu Ala Asp Lys
195 200 205
Ser Thr Val Glu Leu Pro Ala Arg Gly Thr Asn Tyr Leu Asp Gly Val
210 215 220
Ala Lys Leu Leu Tyr Ser Pro Gln Ser Pro Pro Glu Lys Ala Leu Ala
225 230 235 240
Lys Thr Gln Glu Ala Lys Val Ala Ser Lys Asn Val Asn Pro Lys Ile
245 250 255
Gly Glu Lys Tyr Gln Gly Val Val Lys Asn Ile Asn Lys His Gly Ile
260 265 270
Phe Val Asp Phe Gly Ala Lys Lys Asp Gly Leu Val Arg Tyr Arg Asn
275 280 285
Ile Pro Met Lys Tyr Lys His Asp Ser Thr Ile Val Gly Phe Asp Lys
290 295 300
Gly Asp Thr Ile Asp Val Lys Leu Leu Asp Thr Asn Glu Lys Gly Phe
305 310 315 320
Asp Leu Lys Ile Leu Glu Asp Gly Glu Leu Glu Phe Glu Lys Tyr Lys
325 330 335
Ser Glu Lys Ile Asn Asn Thr Met Gly Ser Val Phe Lys Gly Ile Asp
340 345 350
Leu Ser Phe Leu Pro Lys Gln
355
<210> 75
<211> 362
<212> PRT
<213> 宏基因组(metagenomic)
<400> 75
Met Asp Thr Pro Asn Phe Ser Glu Arg Ile Pro Val Ser Leu Gln Ser
1 5 10 15
His Pro Tyr Tyr Phe Ala His Tyr Leu Asn Met Ala Arg His Asn Ala
20 25 30
Tyr Val Ile Leu Glu Tyr Val Asn Arg Glu Leu Ile Lys Pro Gly Lys
35 40 45
Asn Leu Asp Glu Asp Asn Leu Ile Gln Ser Thr Val Leu Lys Asp Gly
50 55 60
Tyr Phe Asp Arg Lys Pro Asp Glu Leu Ser His Arg Asn Arg Leu Leu
65 70 75 80
Val Gln His Phe Pro Phe Leu Arg Glu Ala Glu Asn Glu Gly Ala Arg
85 90 95
Thr Cys Asn Pro Val Ser Tyr Lys Leu Lys Thr Ala Leu Ala Ala Leu
100 105 110
Asn Gln Trp Arg Asn Asn Ala Ser His Tyr Pro Leu Asn Gln Asn His
115 120 125
Glu Lys Asp Phe Asp Leu Gln Pro Phe Phe Ser Phe Ala Ile Glu Ala
130 135 140
Cys Lys Lys Arg Met Arg Glu Val Phe Gln Pro Asp Asp Phe Tyr Leu
145 150 155 160
Leu Glu Thr Asn Glu Lys Gln Phe Tyr Thr Leu His Asn Glu Asn Gly
165 170 175
Phe Thr Glu Lys Gly Leu Tyr Cys Phe Ile Cys Phe Phe Leu Glu Lys
180 185 190
Lys Tyr Ala Phe Gln Phe Leu Ala Gly Ile Lys Gly Phe Lys Asn Thr
195 200 205
Thr Asp Asn Lys Phe Arg Ala Thr Leu Glu Thr Phe Thr Glu His Cys
210 215 220
Cys Arg Leu Pro Lys Pro Lys Leu Asp Ser Ser Asp Ile Lys Leu Asp
225 230 235 240
Met Leu Gly Glu Leu Ser Arg Cys Pro Ala Pro Leu Phe Asp Leu Leu
245 250 255
Asp Ile Glu Glu Arg Lys Lys Phe Ile Arg Glu Pro Glu Glu Val Lys
260 265 270
Pro Asp Glu Ser Gly Asp Arg Glu Glu Val Gln Gln Val Leu Met Lys
275 280 285
Arg Tyr Asp Asp Arg Phe Pro Tyr Phe Ala Leu Arg Tyr Phe Glu Glu
290 295 300
Lys Asn Leu Leu Lys Gly Ile Ser Phe His Ile His Ile Gly Arg Trp
305 310 315 320
Ile Lys Ser Glu His Thr Lys Lys Ile Met Gly Ala Glu Arg Asp Arg
325 330 335
Arg Leu Leu Lys Asp Ile Arg Thr Phe Gly Glu Leu Lys Glu Phe Ser
340 345 350
Pro Glu His Glu Pro Tyr Arg Tyr Lys Thr
355 360
<210> 76
<211> 377
<212> PRT
<213> 宏基因组(metagenomic)
<400> 76
Met Gly Pro Trp Leu Cys Ser His Gly Arg Pro Ala Gly Met Ser Arg
1 5 10 15
Lys Arg Lys Gln Ser Leu Leu Gln Trp Gly Arg Gly Phe Ala Ala Thr
20 25 30
Glu Gly Arg Cys Arg Leu Lys Cys Trp Ser Cys Cys Ser Gln Leu Gln
35 40 45
Trp Gly Arg Gly Phe Ala Ala Thr Glu Gly Pro Gly Ile Pro Ser Thr
50 55 60
Ala Gly Pro Leu Pro Arg Ala Ser Met Gly Pro Trp Leu Cys Ser His
65 70 75 80
Gly Arg Leu Pro Ala Ser Asn Gly Ser Ala Met Pro Thr Pro Ser Phe
85 90 95
Asn Gly Ala Val Ala Leu Gln Pro Arg Lys Glu Met Pro Ile Ala Pro
100 105 110
Ser Gln Arg His Cys Pro Thr Leu Gln Trp Gly Arg Gly Phe Ala Ala
115 120 125
Thr Glu Gly Ser Ser Gly Val Pro Leu Cys Val Gly His Arg Ala Leu
130 135 140
Gln Trp Gly Arg Gly Phe Ala Ala Thr Glu Gly Ala Gly Arg Arg Trp
145 150 155 160
Asp Thr Arg Gly Pro Ala Gly Phe Asn Gly Ala Val Ala Leu Gln Pro
165 170 175
Arg Lys Gly Pro Thr Thr Ser Ala Val Ala Trp Pro Pro Arg Ser Phe
180 185 190
Asn Gly Ala Val Ala Leu Gln Pro Arg Lys Gly Ala Glu Arg Leu His
195 200 205
Val Gly Pro His Arg Gly Ala Ser Met Gly Pro Trp Leu Cys Ser His
210 215 220
Gly Arg Val Ser Arg Cys Arg Arg Met Gly Arg Ser Cys Met Ala Leu
225 230 235 240
Gln Trp Gly Arg Gly Leu Ala Ala Thr Glu Gly Arg Pro Ser Arg Glu
245 250 255
Gln Phe Ala Ala Ala Tyr Lys Leu Gln Trp Gly Arg Gly Phe Ala Ala
260 265 270
Thr Glu Gly Asn Arg Leu Pro Ala Ala Ala Ile Thr Ala Ala Leu Leu
275 280 285
Gln Trp Gly Arg Gly Phe Ala Ala Thr Glu Gly Arg Pro Ala Arg Pro
290 295 300
Cys Cys Gly Ala Arg Ser Ser Arg Phe Asn Gly Ala Val Ala Leu Gln
305 310 315 320
Pro Arg Lys Val Pro Ala Asn Arg Ser Asp Val Pro Asn Ser Ser Gln
325 330 335
Leu Gln Trp Gly Arg Gly Phe Ala Ala Thr Glu Gly Ala Pro Gly Arg
340 345 350
Trp Ala Ala Ser Ala Thr Ala Gly Ala Ser Met Gly Pro Trp Leu Cys
355 360 365
Ser His Gly Arg Lys Ser Gln Arg Ser
370 375
<210> 77
<211> 369
<212> PRT
<213> 宏基因组(metagenomic)
<400> 77
Met Lys Asn Lys Ala Asn Asp Ser Arg Phe Gly Ala Gly Gln Ser Lys
1 5 10 15
Lys Gln Lys Pro Ile Thr Arg Ala Gln Gln Phe Met Gly Gly Lys His
20 25 30
Pro Glu Phe Ala Ile Phe Tyr Asn Ile Ala Phe Phe Arg Leu Ala Gly
35 40 45
Tyr Ile Asn Phe Leu Gln Gly Asn Ser Tyr Glu Gly Asn Lys Lys Glu
50 55 60
Leu Glu Asp Ile Phe Ile Lys Trp Lys Ser Lys Val Asp Asp Gly Val
65 70 75 80
Lys Val Leu Gly Gln Thr Ala Tyr Ser Lys Leu Arg Asn Tyr Leu Trp
85 90 95
Lys Ala Tyr Met Phe Glu Lys Asn Ser Ser Gly Phe Ala Leu Glu Glu
100 105 110
Lys Asp Ile Asn Leu Ile Asn Glu Met Ile Phe Lys Leu Asn His Ile
115 120 125
Arg Asn Phe Gln Ser His Ile Trp His Asp Asn Gln Pro Leu Val Phe
130 135 140
Ser Arg Glu Leu Ser Ala Phe Ile Glu Glu Phe His Glu Asn Ala Val
145 150 155 160
Met Ala Gln Arg Lys Asp Asn Lys Leu Asn Tyr Glu Val Asp Asp Tyr
165 170 175
Leu Lys Glu Tyr Cys Asp Ser Arg Lys Ser Arg Lys Pro Gly Arg Asn
180 185 190
Asp Met Arg Phe Phe Asp Ile His Asp Gly Lys Ile Met Met Thr Gln
195 200 205
Asp Gly Arg Val Phe Phe Leu Ser Leu Phe Leu Thr Arg Gly Glu Met
210 215 220
Asn Arg Phe Leu Gln Leu Lys Lys Gly Phe Lys Arg Asn Asp Leu Pro
225 230 235 240
Lys Tyr Lys Ile Lys Arg Arg Ile Phe Gln His Leu Thr His Arg Asp
245 250 255
Gly Ala Ser Arg Asn Tyr Tyr Gly Phe Glu Asp Asn Ile Leu Asp Ser
260 265 270
Leu Pro Lys Glu Gln Lys Ser Glu Thr Leu Asn Ile Arg Gln Thr Phe
275 280 285
Lys Leu Ile Ser Tyr Leu Asn Asp Leu Pro Ile Glu Val Asn Asn Pro
290 295 300
Glu Gln Phe Pro Ile Phe Leu Ser Asn Gly Lys Asn Val Glu Ser Ser
305 310 315 320
Gln Glu Val Ile Glu Trp Cys Lys Glu Asn Asp Phe Leu Lys Glu Leu
325 330 335
Gln Phe Asn Leu Ile Lys Lys Glu Pro Lys Glu Gly Glu Ala Tyr Leu
340 345 350
Leu Asp Gly Ser Ile Glu Cys Gln Leu Ile Ser Gln Pro Asp Tyr Ser
355 360 365
Phe
<210> 78
<211> 313
<212> PRT
<213> 宏基因组(metagenomic)
<400> 78
Met Ala Val Ala Pro Leu Pro Leu Ser Cys Ser Ala Asp Ala Ala Lys
1 5 10 15
Cys Ser Asp Pro Met Lys Asp Val Val Asp Ala Ala Leu Arg Ile Ser
20 25 30
Ser Leu Val Asp Arg Val Leu Ala Ala His Arg Leu Lys Leu Glu Ala
35 40 45
Ala Glu Arg Tyr Pro His Leu His Val Pro Thr Asn Asp Asp Leu Leu
50 55 60
Gly Ala Ile Leu Val Gly Ile Ala His Ala Ile Ser Asp Thr Asp Gly
65 70 75 80
Leu Phe Glu Ala Asp Met Tyr Asp Thr Ile Glu Asp Leu Tyr Gly Gly
85 90 95
Ser Gly Ala Thr Asp Ser Ala Thr Asp Asp Glu Asp Asp Thr Val Trp
100 105 110
Glu Glu Tyr Glu Arg Ile Gln Asp Val Ala Tyr Gln Gln Thr Cys Ala
115 120 125
Trp Phe Lys Gly Val Ala Pro Trp Ile Asp Pro Cys Asp Pro Val Asn
130 135 140
Ala Asp Tyr Arg Leu Val Pro Val Leu Val Leu Ser Ala Ile Gly Pro
145 150 155 160
Ala Leu Gly Ala Trp Leu Ser Arg Glu Ile Ala Arg Leu Leu Glu Ser
165 170 175
Glu Asp Asp Ala Ile Phe Pro Leu Tyr Gly Pro Leu Tyr Gly Ala Leu
180 185 190
Ile Cys Ser Gly Ile Gly Leu Asp Asp Ala Gly Ala Val Gly Ala Phe
195 200 205
Ile Arg Glu Leu Lys Pro Arg Val Asp Pro Trp Ser Ala Val Gly Cys
210 215 220
Asp His Arg Leu Phe Glu Arg His Ser Arg Val Val Ser Arg Arg Leu
225 230 235 240
Gly Glu Ile Leu Gly Val Ser His Ala Gln Leu Asn Val Glu Arg Ile
245 250 255
Lys Ala Val Ala Asp Leu Asp Gln Met Arg Ser Ala Ile Trp His Gln
260 265 270
Arg Gln Gln Ser Asp Arg Lys Leu Lys Met Gly Gln Leu Val Ser Lys
275 280 285
Val Gly Ser Glu Arg Ala Arg Trp Leu Val Glu Lys Ala Arg Phe Lys
290 295 300
Ala Arg Ala Lys Ala Cys Val Gly Arg
305 310
<210> 79
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 79
tcttcaaatt gtgatacgtc ccaa 24
<210> 80
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 80
ttgggacgta tcacaatttg aaga 24
<210> 81
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 81
gtttccattc aattaattgc ctctattaaa agagac 36
<210> 82
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 82
gtcgtccccg cgcccgcggg ggttgctc 28
<210> 83
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 83
gtcgtccccg cgcccgcggg ggttgctcc 29
<210> 84
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 84
ggagcaaccc ccgcgggcgc ggggacgac 29
<210> 85
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 85
gtttgccatc gcccagatgg tttagaag 28
<210> 86
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 86
cttctaaacc atctgggcga tggcaaac 28
<210> 87
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 87
gtcctcatcg cccctacgag gggtcgcaac 30
<210> 88
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 88
gttgcgaccc ctcgtagggg cgatgaggac 30
<210> 89
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 89
gttcactgcc gcgtaggcag ctcagaaa 28
<210> 90
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 90
tttctgagct gcctacgcgg cagtgaac 28
<210> 91
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 91
gtggcggtcg cccctcgggg ggaccgagga tcgcaac 37
<210> 92
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 92
gttgcgatcc tcggtccccc cgaggggcga ccgccac 37
<210> 93
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 93
gttctctccg cgcgagcgga ggtggtccg 29
<210> 94
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 94
cggaccacct ccgctcgcgc ggagagaac 29
<210> 95
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 95
gttgtaattg ctcttatttt gaagggtata cacaac 36
<210> 96
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 96
gttgtgtata cccttcaaaa taagagcaat tacaac 36
<210> 97
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 97
gctgtactca cccttcaaat aaagggcttt tacagc 36
<210> 98
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 98
gctgtaaaag ccctttattt gaagggtgag tacagc 36
<210> 99
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 99
gttgggaata cccttagtta gaagggtgga gacaac 36
<210> 100
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 100
gttgggaata cccttagtta gaagggtgga gacaact 37
<210> 101
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 101
gttgggaata cccttagtta gaagggtgga gacaac 36
<210> 102
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 102
gttgtctcca cccttctaac taagggtatt cccaac 36
<210> 103
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 103
gttgtagttc cctgatcgtt cttggtatgg tataat 36
<210> 104
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 104
attataccat accaagaacg atcagggaac tacaac 36
<210> 105
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 105
aggatagcag ttcagaaatc gcggtccagc tgcaac 36
<210> 106
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 106
gttgcagctg gaccgcgatt tctgaactgc tatcct 36
<210> 107
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 107
gggctcatcc ccgcacgcgc ggggagcac 29
<210> 108
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 108
gtgctccccg cgcgtgcggg gatgagccc 29
<210> 109
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 109
agtcttcccc acatgggtgg gggtgtttct a 31
<210> 110
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 110
gtcttcccca catgggtggg ggtgtttcta 30
<210> 111
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 111
gttgcaaagg ctgtccctcg gtagagggat tgaaac 36
<210> 112
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 112
gttgcaaagg ctgtccctcg gtagagggat tgaaacac 38
<210> 113
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 113
cggaccatcc ccacgcacgt ggggagaac 29
<210> 114
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 114
gttctcccca cgtgcgtggg gatggtccg 29
<210> 115
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 115
gtccgctttc atctaggaag tggaattaat ggaaac 36
<210> 116
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 116
gtttccatta attccacttc ctagatgaaa gcggac 36
<210> 117
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 117
gtcgcagctc cttcgggagc tgctcttcat tgaggc 36
<210> 118
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 118
gcctcaatga agagcagctc ccgaaggagc tgcgac 36
<210> 119
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 119
gtcgcgcccc gcacggggcg cgtggattga aac 33
<210> 120
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 120
gtttcaatcc acgcgccccg tgcggggcgc g 31
<210> 121
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 121
ggtgtgaaag ccatcttttt gtatggtagg gacacc 36
<210> 122
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 122
ggtgtcccta ccatacaaaa agatggcttt cacacc 36
<210> 123
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 123
gtcgctcccc tcgcgggagc gtggattgaa ata 33
<210> 124
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 124
tatttcaatc cacgctcccg cgaggggagc gac 33
<210> 125
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 125
aaataccacc caagaatgag ggggttctat aacc 34
<210> 126
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 126
ggttatagaa ccccctcatt cttgggtggt attt 34
<210> 127
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 127
agtttatccg atgggagatc ggggaggaac cgcaac 36
<210> 128
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 128
gttgcggttc ctccccgatc tcccatcgga taaact 36
<210> 129
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 129
cttccaattt gcgcgtgggc gtgagttggg ggcac 35
<210> 130
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 130
gtgcccccaa ctcacgccca cgcgcaaatt ggaag 35
<210> 131
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 131
gtcgcagtcc tcactaaaat tggacatgac 30
<210> 132
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 132
gtcatgtcca attttagtga ggactgcgac 30
<210> 133
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 133
gctgtgatag acctcgattt gtggggtagt aacagc 36
<210> 134
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 134
gctgttacta ccccacaaat cgaggtctat cacagc 36
<210> 135
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 135
tgaatacagc tcgatatagt gagcaataac t 31
<210> 136
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 136
agttattgct cactatatcg agctgtattc a 31
<210> 137
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 137
gtttcaccag ccgatttttt aaacggtaac tgaaac 36
<210> 138
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 138
gtttcagtta ccgtttaaaa aatcggctgg tgaaac 36
<210> 139
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 139
gttgtagaag ccacttgttt gaaatggcat gacaac 36
<210> 140
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 140
gttgtcatgc catttcaaac aagtggcttc tacaac 36
<210> 141
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 141
gttggagatc acccccaaat cgagggggac tgcacc 36
<210> 142
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 142
ggtgcagtcc ccctcgattt gggggtgatc tccaac 36
<210> 143
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 143
gaatcgcccg gcttcccagc cgggcgcgga ttgaaac 37
<210> 144
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 144
gtttcaatcc gcgcccggct gggaagccgg gcgattc 37
<210> 145
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 145
gttcaatttt gagtactata 20
<210> 146
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 146
tatagtactc aaaattgaac 20
<210> 147
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 147
gagcatacgc acaaagtcca cagt 24
<210> 148
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 148
actgtggact ttgtgcgtat gctc 24
<210> 149
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 149
gttttagagc tgtgctgttt cgaatggttc caaaac 36
<210> 150
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 150
gttttggaac cattcgaaac agcacagctc taaaac 36
<210> 151
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 151
gttttagagc tgtgctgttt cgaatggttc caaaac 36
<210> 152
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 152
gttttggaac cattcgaaac agcacagctc taaaac 36
<210> 153
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 153
ggttcacccg cgcacgcgcg tgtaagg 27
<210> 154
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 154
ccttacacgc gcgtgcgcgg gtgaacc 27
<210> 155
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 155
gtctccctcc atgcggaggg agtggattga aat 33
<210> 156
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 156
atttcaatcc actccctccg catggaggga gac 33
<210> 157
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 157
gttgtagttc cctttcattt tgggatcatt cacacc 36
<210> 158
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 158
ggtgtgaatg atcccaaaat gaaagggaac tacaac 36
<210> 159
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 159
gttgtagaag cctatcgttt ggataggtat gacaac 36
<210> 160
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 160
gttgtcatac ctatccaaac gataggcttc tacaac 36
<210> 161
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 161
gttcgctgcc gcgcaggcag ctcagaaa 28
<210> 162
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 162
tttctgagct gcctgcgcgg cagcgaac 28
<210> 163
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 163
gttgcaccga ccacgcccac tgaagggcga ctgcacc 37
<210> 164
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 164
ggtgcagtcg cccttcagtg ggcgtggtcg gtgcaac 37
<210> 165
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 165
gtcgctcccc attcggggag cgtggattga aat 33
<210> 166
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 166
atttcaatcc acgctccccg aatggggagc gac 33
<210> 167
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 167
gttgtagaag ccctcatttt gagagggtat aacaac 36
<210> 168
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 168
gttgttatac cctctcaaaa tgagggcttc tacaac 36
<210> 169
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 169
gttttagata taagtcattt taagtacata gaaccc 36
<210> 170
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 170
gggttctatg tacttaaaat gacttatatc taaaac 36
<210> 171
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 171
gtggcgacgg gtgaggaggc cggatcgggt tggagg 36
<210> 172
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 172
cctccaaccc gatccggcct cctcacccgt cgccac 36
<210> 173
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 173
gtttttatcg tccctataag gggttgaaac 30
<210> 174
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 174
gtttcaaccc cttataggga cgataaaaac 30
<210> 175
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 175
gttgtagttc cctttcattt tgggatcatt cacacc 36
<210> 176
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 176
ggtgtgaatg atcccaaaat gaaagggaac tacaac 36
<210> 177
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 177
gcccccaaca aaccatcagc cgaaaggcga ttgagac 37
<210> 178
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 178
gtctcaatcg cctttcggct gatggtttgt tgggggc 37
<210> 179
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 179
gttggagatc acccccaaat cgagggggac tgcacc 36
<210> 180
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 180
ggtgcagtcc ccctcgattt gggggtgatc tccaac 36
<210> 181
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 181
gtcgaggctc gcgagagcct tgtggattga aat 33
<210> 182
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 182
atttcaatcc acaaggctct cgcgagcctc gac 33
<210> 183
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 183
gtcgccttcc ccccggaagg cgtggattga aac 33
<210> 184
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 184
gtttcaatcc acgccttccg gggggaaggc gac 33
<210> 185
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 185
gtttgccccg catgtgcggg gatgatccg 29
<210> 186
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 186
cggatcatcc ccgcacatgc ggggcaaac 29
<210> 187
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 187
ctccttctgc tcaggcgtgg ctt 23
<210> 188
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 188
aagccacgcc tgagcagaag gag 23
<210> 189
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 189
cgtttccacg gcatcacagc cgtggccgaa ttgaagc 37
<210> 190
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 190
gcttcaattc ggccacggct gtgatgccgt ggaaacg 37
<210> 191
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 191
gtaagaatca aataatcccg atacgcggga ttaagac 37
<210> 192
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 192
gtcttaatcc cgcgtatcgg gattatttga ttcttac 37
<210> 193
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 193
gctgcattcc ccgcgcgaga ggggattgag ac 32
<210> 194
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 194
gtctcaatcc cctctcgcgc ggggaatgca gc 32
<210> 195
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 195
gttgtgtgta cccttcgaat agagggtaga tccaac 36
<210> 196
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 196
gttggatcta ccctctattc gaagggtaca cacaac 36
<210> 197
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 197
gtcgcgcctt cgcgggcgcg tgagttgaaa c 31
<210> 198
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 198
gtttcaactc acgcgcccgc gaaggcgcga c 31
<210> 199
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 199
ggttcccccg tacacgcggg gatagacc 28
<210> 200
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 200
ggtctatccc cgcgtgtacg ggggaacc 28
<210> 201
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 201
gtgctccccg cacacgcggg gatgatccc 29
<210> 202
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 202
gggatcatcc ccgcgtgtgc ggggagcac 29
<210> 203
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 203
ggtggagaca cgcggattta ggggtgtgat gacagg 36
<210> 204
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 204
cctgtcatca cacccctaaa tccgcgtgtc tccacc 36
<210> 205
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 205
attcctaagc ttttacgctt aggacttcat tgagg 35
<210> 206
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 206
cctcaatgaa gtcctaagcg taaaagctta ggaat 35
<210> 207
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 207
ccctcaacta ttgaaacgtg tttcagtcgt ttcagg 36
<210> 208
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 208
cctgaaacga ctgaaacacg tttcaatagt tgaggg 36
<210> 209
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 209
ggtttccgtc cccgtgaagg ggaagttgta tgaaac 36
<210> 210
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 210
gtttcataca acttcccctt cacggggacg gaaacc 36
<210> 211
<211> 41
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 211
ttatgtgctc agggccactg catggtgctg atggaggcca c 41
<210> 212
<211> 41
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 212
gtggcctcca tcagcaccat gcagtggccc tgagcacata a 41
<210> 213
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 213
ggtgtcggaa accgctaatt caggggccgc tacaac 36
<210> 214
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 214
gttgtagcgg cccctgaatt agcggtttcc gacacc 36
<210> 215
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 215
agtttagcag attgggattt gtactctgac cggaac 36
<210> 216
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 216
gttccggtca gagtacaaat cccaatctgc taaact 36
<210> 217
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 217
gtagaaatga gtacaaagcg atagagagct taataac 37
<210> 218
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 218
gttattaagc tctctatcgc tttgtactca tttctac 37
<210> 219
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 219
aactcggaag gattcagaag aagctttcat ct 32
<210> 220
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 220
agatgaaagc ttcttctgaa tccttccgag tt 32
<210> 221
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 221
gttcactgcc gcacaggcag ctcagaaa 28
<210> 222
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 222
tttctgagct gcctgtgcgg cagtgaac 28
<210> 223
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 223
gtctccctcc atgcggaggg agtggattga aat 33
<210> 224
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 224
atttcaatcc actccctccg catggaggga gac 33
<210> 225
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 225
gttgaaagag aatagcccga catagtgggc aatcaa 36
<210> 226
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 226
ttgattgccc actatgtcgg gctattctct ttcaac 36
<210> 227
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 227
gttgttctca ccttccaaaa ttaaggcat 29
<210> 228
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 228
atgccttaat tttggaaggt gagaacaac 29
<210> 229
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 229
ccttccgtgg ctgcaaagcc acggccccat tgaagc 36
<210> 230
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 230
gcttcaatgg ggccgtggct ttgcagccac ggaagg 36
<210> 231
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 231
ggtgtggatg cctctatttt gagaggtaga atcacc 36
<210> 232
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 232
ggtgattcta cctctcaaaa tagaggcatc cacacc 36
<210> 233
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 233
gtcgcagcta caaggccgcc gcaatggcca ttggaacat 39
<210> 234
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 234
atgttccaat ggccattgcg gcggccttgt agctgcgac 39
Claims (27)
1.Cas13蛋白，其包含如SEQ ID NO:1至78中任一项所述的氨基酸序列，或具有一个或更多个残基的保守氨基酸取代的SEQ ID NO:1至78中任一项所述的氨基酸序列。
2.根据权利要求1所述Cas13蛋白，其RNA切割活性被保留。
3.根据权利要求1所述Cas13蛋白，其HEPN结构域或RNA切割结构域经进一步修饰或改造，而使其RNA切割活性降低或消除，成为RNA切割活性降低或消除的dCas13。
4.根据权利要求1至3中任一项所述的Cas13蛋白，其中所述Cas13蛋白与一个或更多个异源功能性结构域融合，其中所述融合在所述Cas13蛋白的N端、C端或者内部。
5.根据权利要求4所述的Cas13蛋白，其中所述一个或更多个异源功能性结构域具有以下活性：脱氨酶如胞苷脱氨基酶和脱氧腺苷脱氨基酶、甲基化酶、去甲基化酶、转录激活、转录抑制、核酸酶、单链RNA裂解、双链RNA裂解、单链DNA裂解、双链DNA裂解、DNA或RNA连接酶、报告蛋白、检测蛋白、定位信号、或其任意组合。
6.核酸分子，其包含编码权利要求1至5中任一项所述Cas13蛋白的核苷酸序列。
7.根据权利要求6所述的核酸分子，其针对在特定宿主细胞中的表达而进行了密码子优化。
8.根据权利要求7所述的核酸分子，其中所述宿主细胞是原核或真核生物细胞，优选人细胞。
9.根据权利要求6至8中任一项所述的核酸分子，其包含与编码Cas13的核苷酸序列有效链接的启动子，其为组成型启动子、诱导型启动子、组织特异性启动子、嵌合型启动子或发育特异性启动子。
10.表达载体，其包含权利要求6至9中任一项所述核酸分子，以DNA或RNA或蛋白等形式表达权利要求1的氨基酸序列或权利要求6至9中任一项的核苷酸序列。
11.根据权利要求10所述的表达载体，其为腺相关病毒(AAV)、腺病毒、慢病毒、逆转录病毒、单纯孢疹病毒、溶瘤病毒。
12.递送系统，包含(1)权利要求10所述的表达载体，或权利要求6至11中任一项所述的Cas13蛋白；以及(2)递送载体。
13.根据权利要求12所述的递送系统，其中所述递送载体是纳米颗粒、脂质体、外泌体、微囊泡或基因枪。
14.CRISPR-Cas系统，其包含：(1)根据权利要求1至5中任一项所述的Cas13蛋白或者其衍生物或功能片段，或权利要求6至9中任一项所述核酸分子；(2)用于靶向目标RNA的gRNA序列。
15.根据权利要求14所述的CRISPR-Cas系统，Cas13蛋白的功能片段应指含有一个或多个SEQ ID NO:1至78中任何一个氨基酸(例如1、2、3、4、5、6、7、8、9或10个残基)的添加、缺失和/或取代(例如保守取代)。
16.根据权利要求15所述的CRISPR-Cas系统，Cas13蛋白的衍生物应指至少具有与SEQID NO:1至78中任意一个蛋白片段达到≥70％氨基酸序列同一性(如70％、80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％的一致性)。
17.根据权利要求14所述的CRISPR-Cas系统，其中所述gRNA序列包含同向重复(DR)序列和靶向靶RNA部分的间隔区域的序列。
18.根据权利要求17所述的CRISPR-Cas系统，其中所述DR序列为表1中所示序列；其中所述间隔区序列为15-60个核苷酸，优选25-50个核苷酸，更优选30个核苷酸。
19.根据权利要求18所述CRISPR-Cas系统，其中所述DR序列可以是对应以下任一项的衍生物，其中所述衍生物(i)与表1中所示序列中的任一个相比，具有一个或多个(例如1、2、3、4、5、6、7、8、9或10)个核苷酸的添加、缺失、或取代；(ii)与表1中所示序列中任何一个具有至少20％、30％、40％、50％、60％、70％、80％、90％、95％或97％的序列同一性；(iii)在严格条件下与表1中所示序列任意一个，或与(i)和(ii)中的任意一个杂交；或(iv)是(i)-(iii)中任何一个的互补物，条件是所述衍生物非表1中所示序列中的任何一个，并且所述衍生物编码一个RNA，或本身即是一个RNA，所述RNA与SEQ ID NO：79-234编码的任意RNA基本保持相同的二级结构。
20.根据权利要求14所述的CRISPR-Cas系统，其还包含：(3)靶RNA。
21.根据权利要求14所述的CRISPR-Cas系统，其引起靶RNA序列的降解、切割或序列的改变。
22.根据权利要求20所述的CRISPR-Cas系统，其中所述靶RNA是mRNA或ncRNA，包括选自lncRNA、miRNA、misc_RNA、Mt_rRNA、Mt_tRNA、rRNA、scaRNA、scRNA、snoRNA、snRNA、sRNA的非编码RNA。
23.细胞，其包含权利要求1至5中任一项所述Cas13蛋白、权利要求6至9中任一项所述核酸分子、权利要求10或11所述表达载体、权利要求12或13所述递送系统、或权利要求14至22中任一项所述CRISPR-Cas系统。
24.根据权利要求23所述的细胞，其为原核细胞或真核细胞，优选人细胞。
25.降解或切割目的细胞中靶RNA、修饰目的细胞中靶RNA的序列的方法，其包括使用权利要求1至5中任一项所述Cas13蛋白、权利要求6至9中任一项所述核酸分子、权利要求10或11所述表达载体、权利要求12或13所述递送系统、或权利要求14至22中任一项所述CRISPR-Cas系统。
26.根据权利要求25所述的方法，所述目的细胞为原核细胞或真核细胞，优选人细胞。
27.根据权利要求25所述的方法，其中所述目的细胞为离体细胞、体外细胞或体内细胞。
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| CN117230043A (zh) * | 2023-11-14 | 2023-12-15 | 广州瑞风生物科技有限公司 | Cas13蛋白及其应用 |
| CN117384884A (zh) * | 2023-11-30 | 2024-01-12 | 辉大(上海)生物科技有限公司 | IscB多肽及其用途 |
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| KR20210053898A (ko) * | 2018-07-31 | 2021-05-12 | 더 브로드 인스티튜트, 인코퍼레이티드 | 신규 crispr 효소 및 시스템 |
| CN113544267A (zh) * | 2019-01-14 | 2021-10-22 | 罗切斯特大学 | 使用CRISPR-Cas进行靶向核RNA裂解和聚腺苷酸化 |
| CN115315519A (zh) * | 2020-02-28 | 2022-11-08 | 辉大(上海)生物科技有限公司 | VI-E型和VI-F型CRISPR-Cas系统及其用途 |
| CN112430586B (zh) * | 2020-11-16 | 2021-09-07 | 珠海舒桐医疗科技有限公司 | 一种VI-B型CRISPR/Cas13基因编辑系统及其应用 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117230043A (zh) * | 2023-11-14 | 2023-12-15 | 广州瑞风生物科技有限公司 | Cas13蛋白及其应用 |
| CN117230043B (zh) * | 2023-11-14 | 2024-04-12 | 广州瑞风生物科技有限公司 | Cas13蛋白及其应用 |
| CN117384884A (zh) * | 2023-11-30 | 2024-01-12 | 辉大(上海)生物科技有限公司 | IscB多肽及其用途 |
| CN117384884B (zh) * | 2023-11-30 | 2024-03-08 | 辉大(上海)生物科技有限公司 | IscB多肽及其用途 |
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