CN114836459B - 一种胞嘧啶碱基编辑***及其应用 - Google Patents
一种胞嘧啶碱基编辑***及其应用 Download PDFInfo
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- CN114836459B CN114836459B CN202210265192.3A CN202210265192A CN114836459B CN 114836459 B CN114836459 B CN 114836459B CN 202210265192 A CN202210265192 A CN 202210265192A CN 114836459 B CN114836459 B CN 114836459B
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Classifications
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
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- C12N15/74—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
- C12N15/75—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for Bacillus
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- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
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- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
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- C12Y—ENZYMES
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Abstract
本发明涉及一种胞嘧啶碱基编辑***及其应用,胞嘧啶碱基编辑***包括氨基酸序列如SEQ ID NO.1‑3所示的序列之一的融合蛋白,并基于该融合蛋白设计构建了一种枯草芽孢杆菌胞嘧啶碱基编辑***。本发明构建的胞嘧啶碱基编辑***解决了现有的胞嘧啶碱基编辑器在基因组上的可操作范围受限,且在多位点的碱基编辑(C→T)过程中效率低且操作复杂的问题,实现了快速高效地在基因组上对多个位点进行编辑。
Description
技术领域
本发明涉及生物技术领域,尤其涉及一种胞嘧啶碱基编辑***及其应用。
背景技术
基因编辑是通过在DNA上特定位点引入序列改变,达到基因的敲除、外源DNA片段***或者DNA碱基突变的技术手段。近年来,CRISPR/Cas(clustered regularlyinterspaced short palindromic repeats and CRISPR-associated proteins)***在基因编辑中得到了非常广泛的应用,特别是CRISPR/Cas9***。在该***中,未成熟的前体CRISPR RNA(pre-crRNA)可以与tracrRNA(trans-activating crRNA)相互结合,并在RNA酶III的作用下形成crRNA:tracrRNA复合物,引导Cas9蛋白识别并切割基因组上的特定位点产生双链断裂;然后,通过同源重组修复(HDR)便可将同源模板上的序列改变引入到基因组的目标靶点上;或者,也可以通过非同源末端连接(NHEJ)的方式直接将断裂的DNA片段进行连接,并产生随机的***或缺失(Indel)。由于未被修复的双链断裂是致死的,只有成功修复并引入突变使得Cas9不再识别和切割的细胞才能存活下来,这就是利用CRISPR/Cas9进行基因编辑的基本原理。
为了简化操作过程并提高编辑效率,常常将crRNA和tracrRNA构建成一个嵌合体即sgRNA(small guide RNA)进行表达,这样只需要表达sgRNA和Cas9蛋白便可进行基因编辑。目前除了CRISPR/Cas9***以外,CRISPR/Cpf1***(也被称作CRISPR/Cas12a)也常被应用于基因编辑,与CRISPR/Cas9***不同的是,CRISPR/Cpf1只需要crRNA便可发挥作用;而且Cpf1自身就具有RNA酶的活力,可以对未成熟的包含多个crRNA的mRNA序列进行切割加工;因此,可以设计一段包含多个crRNA的crRNA阵列,当其被Cpf1加工成熟后,可以产生多个具有独立功能的crRNA并引导Cpf1靶向基因组的对应靶点,实现多个位点的同时编辑。
很多物种都缺少NHEJ途径,或者即使有活性也较弱,而HDR途径需要同源模板的参与才能发挥作用,且这两种修复机制之间还会存在竞争关系,这都导致基于上述方式的基因编辑过程致死率较高且编辑效率较低。将Cas9的DNA酶进行失活后,可以得到只能切割一条DNA链的nCas9和无法切割DNA的dCas9。nCas9与dCas9仍然可以在sgRNA的引导下结合到基因组的特定位点,但却不会产生致死的双链断裂。胞嘧啶脱氨酶可以催化胞嘧啶(C)脱氨转化为尿嘧啶(U),并进一步通过DNA修复或复制将U转化为胸腺嘧啶(T),故将胞嘧啶脱氨酶与nCas9或dCas9进行融合,在sgRNA的引导下便可实现不依赖于双链断裂的基因组碱基编辑(C→T)。胞内的尿嘧啶DNA糖基化酶(Uracil DNA N-glycosylase,UNG)可识别错配U·G碱基对,并通过碱基错配修复途径(Base excision repair,BER)将编辑产生的U·G修复恢复为C·G配对,因此还可进一步融合来源于噬菌体PBS的尿嘧啶糖苷酶抑制剂(UGI)来抑制UNG功能,从而进一步提高碱基编辑的效率。上述融合蛋白(nCas9-CBE或dCas9-CBE)在sgRNA的引导下可以实现基因组上特定位点的C→T转化,被称为胞嘧啶碱基编辑***。
Cas9所识别的位点需要具有NGG(N=A,T,C,G)的PAM序列,这限制了基于Cas9的胞嘧啶碱基编辑器在基因组上的可操作范围;此外,引导Cas9的每一个sgRNA都需要完整的转录单元,在进行多位点碱基编辑时(C→T)需要构建多个对应的sgRNA表达框,如此不仅增加了构建过程的复杂性,还会由于启动子等元件的重复使用而降低DNA序列的稳定性。因此,仍需要寻找一种新的胞嘧啶碱基编辑***。
发明内容
为解决上述技术问题,本发明提供了一种基于CRISPR/Cpf1的胞嘧啶碱基编辑***,克服了基于CRISPR/Cas9的胞嘧啶碱基编辑器在基因组上的可操作范围受限,而且在多位点的碱基编辑(C→T)过程中效率低且操作复杂的问题。
本发明的第一个目的是提供一种胞嘧啶碱基编辑***,该胞嘧啶碱基编辑***包括胞嘧啶脱氨酶、尿嘧啶糖苷酶抑制剂和DNA酶失活的Cpf1突变体dCpf1形成的融合蛋白,所述融合蛋白的氨基酸序列为SEQ ID NO.1-3所示的序列之一。其中,SEQ ID NO.1为含有hAPOBEC3A的融合蛋白,SEQ ID NO.2为含有hAID的融合蛋白,SEQ ID NO.3为含有LjCDA1L2_1的融合蛋白。
本发明提供了一种融合蛋白,该融合蛋白可识别TTV作为PAM,其中,V=A,C,G,该融合蛋白的编辑窗口分别在crRNA的第8个碱基到第14个碱基之间、crRNA的第8个碱基到第13个碱基之间或crRNA的第8个碱基到第16个碱基之间将胞嘧啶碱基编辑为胸腺嘧啶,因此该融合蛋白可用于构建胞嘧啶基因编辑器。
进一步地,胞嘧啶碱基编辑***还包括crRNA阵列***区,该crRNA阵列***区用于***与DNA酶失活的Cpf1突变体dCpf1匹配的crRNA,即引导DNA酶失活的Cpf1突变体dCpf1剪切编辑特定靶点的crRNA。
进一步地,crRNA阵列***区的核苷酸序列如SEQ ID NO.4所示。该crRNA阵列***区两端包含两个正向重复的crRNA把手序列,并在中间***了两个反向的Eco31I酶切位点,通过酶切连接便可将所需的识别序列或crRNA阵列放置到两个把手序列之间。
进一步地,上述胞嘧啶脱氨酶、尿嘧啶糖苷酶抑制剂和DNA酶失活的Cpf1突变体dCpf1形成的融合蛋白通过诱导型启动子Pgrac100调控表达。
进一步地,crRNA阵列***区通过组成型启动子Pveg调控表达。
进一步地,上述胞嘧啶碱基编辑***是将编码氨基酸序列为SEQ ID NO.1-3所示之一的基因以及crRNA阵列***区整合到表达载体上得到。
本发明的第二个目的是提供一种枯草芽孢杆菌胞嘧啶碱基编辑***,该胞嘧啶碱基编辑***包括:含有枯草芽孢杆菌的温敏型pE194复制子、编码SEQ ID NO.1-3所示序列之一融合蛋白的基因以及上述crRNA阵列***区的质粒。
进一步地,上述质粒的核苷酸序列如SEQ ID NO.7-9所示序列之一,其中,SEQ IDNO.7为含有hAPOBEC3A的质粒,SEQ ID NO.8为含有hAID的质粒,SEQ ID NO.9为含有LjCDA1L2_1的质粒。
本发明的第三个目的是提供上述胞嘧啶碱基编辑***或枯草芽孢杆菌胞嘧啶碱基编辑***在基因编辑中的应用。
进一步地,上述胞嘧啶碱基编辑***或枯草芽孢杆菌胞嘧啶碱基编辑***用于多位点胞嘧啶碱基编辑。
进一步地,上述基因编辑为crRNA的第8个碱基到第16个碱基之间的多位点C>T碱基转化。
进一步地,上述应用包括采用IPTG进行诱导,优选地,IPTG的终浓度为0.5-5mM,诱导时间为12-36h,更优选为IPTG的终浓度为1mM,诱导时间为12h。
借由上述方案,本发明至少具有以下优点:
本发明为了拓宽胞嘧啶碱基编辑器在基因组上的可操作范围,开发了基于PAM序列为TTV(V=A,C,G)的CRISPR/Cpf1胞嘧啶碱基编辑***,将胞嘧啶脱氨酶、UGI与DNA酶活失活的Cpf1突变体dCpf1进行融合构建得到胞嘧啶碱基编辑器(dCpf1-CBE),同时设计了两端包含有两个相对的Eco31I酶切位点的crRNA***区,用于crRNA阵列的快速组装,由于dCpf1的RNA酶活力得到了保留,因此可以在单个crRNA阵列的引导下实现多个位点的碱基编辑(C→T),实现快速高效地在基因组上对多个位点进行编辑,从而产生极为丰富的突变体组合,这使得本***在蛋白进化与菌株改造中都具有非常大的应用潜力。
上述说明仅是本发明技术方案的概述,为了能够更清楚了解本发明的技术手段,并可依照说明书的内容予以实施,以下以本发明的较佳实施例并配合详细附图说明如后。
附图说明
为了使本发明的内容更容易被清楚的理解,下面根据本发明的具体实施例并结合附图,对本发明作进一步详细的说明。
图1为基于CRISPR/Cpf1的胞嘧啶碱基编辑***示意图;
图2为枯草芽孢杆菌胞嘧啶碱基编辑***质粒图谱;
图3为胞嘧啶脱氨酶选择hAID构建的胞嘧啶碱基编辑***介导的多位点碱基编辑;
图4为多位点胞嘧啶碱基编辑***处理后产生的突变体的具体组成。
具体实施方式
下面结合附图和具体实施例对本发明作进一步说明,以使本领域的技术人员可以更好地理解本发明并能予以实施,但所举实施例不作为对本发明的限定。
实验涉及的材料及试剂:
DNA聚合酶购自Takara公司,限制性内切酶与T4连接酶购买自NEB公司,质粒提取试剂盒购买自生工生物工程(上海)股份有限公司,PCR产物核酸纯化试剂盒购买自ThermoScientific公司。
细胞均使用LB培养基培养,其中含有:胰蛋白胨10g/L,酵母粉5g/L,NaCl 10g/L。培养基中卡那霉素的终浓度为50μg/mL,IPTG的终浓度为1mM。
实施例1基于CRISPR/Cpf1的胞嘧啶碱基编辑***的设计构建
如图1A所示,构建了基于CRISPR/Cpf1的胞嘧啶编辑编辑***,该***由以下两个基本元件组成:
(1)由胞嘧啶脱氨酶、Cpf1的DNA酶失活突变体(D917A)dCpf1和尿嘧啶糖苷酶抑制剂(UGI)构成的融合蛋白,即胞嘧啶碱基编辑器dCpf1-CBE,该融合蛋白可以在dCpf1的帮助下结合到基因组的特定靶点,并在胞嘧啶脱氨酶的作用下将该处的胞嘧啶(C)转化为胸腺嘧啶(T);
具体地,分别将胞嘧啶脱氨酶hAPOBEC3A(GenBank:KM266646.1)、hAID(GenBank:AAM95402.1)及LjCDA1L2_1(GenBank:MG495262.1)通过XTEN短肽linker(SGSETPGTSESATPES)融合到dCpf1的N末端,并将UGI(GenBank:YP_009283008.1)通过短肽linker(GSPKKKRKVSGGS)融合到dCpf1的C末端,得到的融合氨基酸序列分别如序列1-3所示。此外,还设计了DNA序列如4所示的crRNA阵列***区,该区域两端包含两个正向重复(direct repeat)的crRNA把手序列,并在中间***了两个反向的Eco31I酶切位点,通过酶切连接便可将所需的识别序列或crRNA阵列放置到两个把手序列之间。
(2)与Cpf1相匹配的crRNA,该crRNA包含固定的把手序列和与目标靶点互补的识别序列,在dCpf1与crRNA把手序列间的相互作用下dCpf1-CBE会与crRNA结合形成复合物,并在互补序列的作用下识别和靶向基因组的特异性靶点,进而实现碱基编辑(C→T)。由于dCpf1仍然具备RNA酶活力,多个crRNA可通过阵列的形式进行表达,crRNA阵列被dCpf1切割处理以后,便可以将dCpf1-CBE靶向多个位点实现碱基编辑(C→T)过程(图1B)。
实施例2基于CRISPR/Cpf1的胞嘧啶碱基编辑***在多位点碱基编辑中的应用
在枯草芽孢杆菌中进行了基于CRISPR/Cpf1的胞嘧啶碱基编辑***的验证与应用。如图2所示,使用受IPTG诱导的Pgrac100启动子表达不同构造的胞嘧啶碱基编辑器dCpf1-CBE,并将crRNA阵列***区放置到组成型启动子Pveg之后以实现crRNA阵列的表达,将上述两个表达框都放置到了含有温敏型复制子pE194的质粒上用于枯草芽孢杆菌中的碱基编辑(C→T)。
具体包括以下步骤:所使用的质粒骨架来自于pJOE8999(Altenbuchner,J.,2016.Editing of the Bacillus subtilis genome by the CRISPR-Cas9system.Applied and Environmental Microbiology 82,5421–5427),该质粒在大肠杆菌与枯草芽孢杆菌中均为卡那霉素抗性(KanR),且具有多拷贝复制子pBR322可在大肠杆菌中进行质粒的构建与保存,而在枯草芽孢杆菌中则带有温敏型复制子pE194(30℃下可以稳定复制,50℃则会被消除);所使用Pgrac100启动子及其阻遏蛋白基因lacI和dCpf1蛋白均来自于质粒pLCg6-dCpf1(Wu,Y.,Liu,Y.,Lv,X.,Li,J.,Du,G.,Liu,L.,2020.CAMERS-B:CRISPR/Cpf1assisted multiple-genes editing and regulation system for Bacillussubtilis.Biotechnology and Bioengineering 117,1817–1825);crRNA阵列***区及启动子Pveg来自于质粒pcra2(Wu,Y.,Liu,Y.,Lv,X.,Li,J.,Du,G.,Liu,L.,2020.CAMERS-B:CRISPR/Cpf1 assisted multiple-genes editing and regulation system forBacillus subtilis.Biotechnology and Bioengineering 117,1817–1825);胞嘧啶脱氨酶和UGI则通过基因合成的方式获得。上述片段通过PCR扩增之后,使用无缝连接试剂盒(碧云天D7010M)进行连接,并转化到大肠杆菌中进行测序和保存。包含有hAPOBEC3A、hAID及LjCDA1L2_1的质粒序列分别如SEQ ID NO.7-9所示,具体见https://benchling.com/s/seq-3PzEctLun0PjCaMDZGdy?m=slm-sNOplfRLxZJm hoYwp2YQ,https://benchling.com/s/seq-rOIKDKkDCxJnTDZ9PRUE?m=slm-S7dZeQT3eA zJANLyT0Pa,https://benchling.com/s/seq-HcxpTcaW9wzSaVc25a7p?m=slm-4M0HmJNcken NgaPjjCBR。
在大肠杆菌中,通过SOMACA(Synthetic Oligos Mediated Assembly of crRNAArray)方法(参考文献:Wu,Y.,Liu,Y.,Lv,X.,Li,J.,Du,G.,Liu,L.,2020.CAMERS-B:CRISPR/Cpf1 assisted multiple-genes editing and regulation system forBacillus subtilis.Biotechnology and Bioengineering 117,1817–1825)将所需要的crRNA组装到了crRNA阵列***区中,得到可以完成碱基编辑(C→T)所需的质粒。
具体地,当表达单个crRNA时,crRNA可以直接通过带有重叠区的一对引物退火得到(引物浓度为10uM,20uL体系中上下游引物各10uL;反应条件为:98℃2min,0.1℃/S降温至4℃后保温。),退产物稀释10倍后取1uL与使用Eco31I酶切后的载体连接即可。当设计多个crRNA形成crRNA阵列时,首先将多对具有重叠区的引物进行PCR(引物浓度为10uM,20uL体系中含DNA聚合酶10uL以及上下游引物各5uL;使用标准PCR程序,延伸时间5秒,10个循环),结束后稀释10倍便可得到两端具有Eco31I的双链DNA,取上述双链DNA各1μL与质粒进行golden gate组装即可。将上述产物转化到大肠杆菌中之后,在质粒骨架和crRNA上各选一个引物进行菌落PCR,并挑取有条带的单菌落进行测序筛选含有所需crRNA的阳性克隆。
以枯草芽孢杆菌aprE基因和nprE基因中的5个位点作为靶点,进行多位点的碱基编辑,共设计了如表1中所示的5个crRNA,在大肠杆菌中构建并完成测序后即可转化到枯草芽孢杆菌中。
表1用于引导多位点碱基编辑(C→T)的crRNA
多位点碱基编辑(C→T)与分析的操作步骤如下:首先,将编辑质粒转化到枯草芽孢杆菌Bacillus subtilis 168菌株中,涂布含有卡那霉素的平板,30℃培养12h;然后,任意挑取1个单菌落接种到装有2mL LB培养基(含有卡那霉素)的14mL摇菌管中,30℃振荡培养12h;随后,取5μL菌液转接到含装有2mL LB培养基(含有卡那霉素和IPTG)的14mL摇菌管中,30℃振荡培养12h诱导碱基编辑***表达进行多位点碱基编辑(C→T);最后,取1μL诱导后的菌液到50μL PCR反应体系中,扩增aprE和nprE基因并进行sanger测序分析。aprE和nprE扩增使用的引物分别如表2所示,上述DNA片段经过纯化后使用表2中所示引物进行sanger测序,并使用BEAT软件(https://hanlab.cc/beat/)对测序结果进行分析。由于所使用的质粒是温敏的,编辑完成之后,可以将菌液划线到不含抗生素的LB平板上,50℃培养过夜即可将质粒消除。
表2引物序列
使用不同的胞嘧啶脱氨酶构建的胞嘧啶碱基编辑器(dCpf1-CBE)介导的多位点碱基编辑情况如表3所示:当使用胞嘧啶脱氨酶hAID时,5个位点都有C转化为T,且编辑窗口为crRNA的第8个碱基到第13个碱基之间;使用胞嘧啶脱氨酶hAPOBEC3A时,除了位点3以外剩余4个位点也都存在C转化为T,且编辑窗口为crRNA的第8个碱基到第14个碱基之间;使用胞嘧啶脱氨酶LjCDA1L2_1时,仅有位点1和位点2存在C转化为T,但是编辑窗口较大为crRNA的第8个碱基到第16个碱基之间;上述碱基编辑器在不同位点的编辑效率也有所差别,介于1%到46%之间。
表3胞嘧啶碱基编辑器介导的多位点碱基编辑(C→T)及其效率
实施例3诱导时间对多位点碱基编辑效率的影响
由于含有胞嘧啶脱氨酶hAID的胞嘧啶碱基编辑器效率最高,可以实现五个位点的同时编辑。因此,又考察了IPTG的诱导时间对该碱基编辑器的碱基编辑效率的影响。如表4所示,随着诱导时间的增加,各个位点的编辑效率都有一定的提高,但是幅度有限,而且编辑的窗口也并未随着诱导时间的延长而发生变化。上述结果说明诱导12h已经足够实现较为完全的编辑。其中,诱导36h之后的编辑测序结果如图3所示。
表4诱导时间对多位点碱基编辑效率的影响
实施例4多位点胞嘧啶碱基编辑***产生突变体情况分析
为了分析使用胞嘧啶碱基编辑处理后获得的突变体的具体组成,将含有胞嘧啶脱氨酶hAID的胞嘧啶碱基编辑器诱导处理36h后的菌液划线到了不含抗生素LB平板上,并置于37℃过夜培养。然后,挑取了8个单菌落分别扩增aprE和nprE位点进行测序分析。
如图4所示,只需要设计和构建一个特定的crRNA阵列,便可以同时产生丰富的具有不同突变组合的突变体,这对于蛋白进化及菌株改造都是非常有价值的;此外,我们也观察到了使用混合模板测序没有测到的突变位点(如位点1中的C14T、位点3中的C3T和位点5中的C8T),说明某些位点可能存在效率很低的突变,这些突变无法通过实施例2中的方法被检测到,这也说明该碱基编辑器具有更大的编辑窗口。另外,由于挑取的菌落较少,表4中某些低频突变位点也可能在上述突变体中未被检测到。
对比例1其他来源的胞嘧啶脱氨酶的活性对比
基于CRISPR/Cas9的胞嘧啶碱基编辑***中常用的胞嘧啶脱氨酶除上述3种以外,还有rAPOBEC-1(GenBank:KP792760.1)和PmCDA1(GenBank:ABO15149.1)。因此,进一步尝试了将胞嘧啶脱氨酶rAPOBEC-1和PmCDA1分别融合到dCpf1的N末端,并将UGI融合到dCpf1的C末端,构建得到序列5-6所示的胞嘧啶碱基编辑***。
然而同样利用表1中的crRNA进行5个位点的碱基编辑(C→T)时,使用rAPOBEC-1构建的碱基编辑器不具备任何碱基编辑(C→T)的能力,使用PmCDA1构建的碱基编辑器仅在位点1出产生了突变(C8T-40%),说明胞嘧啶脱氨酶rAPOBEC-1和PmCDA1对于dCpf1的活性影响较大,无法基于其构建基于CRISPR/Cpf1的胞嘧啶碱基编辑***。
对比例2尿嘧啶糖苷酶抑制剂(UGI)对编辑效率的影响
为了考察了UGI对于基于dCpf1的胞嘧啶碱基编辑器的编辑效率是否具有促进作用,我们设计了只有胞嘧啶脱氨酶而不含UGI的融合蛋白。在连接了表1中的第一个crRNA后,进行了突变分析。如表5所示,添加UGI后对所有的胞嘧啶碱基编辑器的效率均具有显著的促进作用。
表5 UGI对于胞嘧啶碱基编辑器碱基效率的影响
显然,上述实施例仅仅是为清楚地说明所作的举例,并非对实施方式的限定。对于所属领域的普通技术人员来说,在上述说明的基础上还可以做出其它不同形式变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引申出的显而易见的变化或变动仍处于本发明创造的保护范围之中。
序列表
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Glu Ile Ala Lys Leu Val Ile Glu Tyr Asn Ala Ile Val Val Phe Glu
1205 1210 1215
Asp Leu Asn Phe Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln
1220 1225 1230
Val Tyr Gln Lys Leu Glu Lys Met Leu Ile Glu Lys Leu Asn Tyr Leu
1235 1240 1245
Val Phe Lys Asp Asn Glu Phe Asp Lys Thr Gly Gly Val Leu Arg Ala
1250 1255 1260
Tyr Gln Leu Thr Ala Pro Phe Glu Thr Phe Lys Lys Met Gly Lys Gln
1265 1270 1275 1280
Thr Gly Ile Ile Tyr Tyr Val Pro Ala Gly Phe Thr Ser Lys Ile Cys
1285 1290 1295
Pro Val Thr Gly Phe Val Asn Gln Leu Tyr Pro Lys Tyr Glu Ser Val
1300 1305 1310
Ser Lys Ser Gln Glu Phe Phe Ser Lys Phe Asp Lys Ile Cys Tyr Asn
1315 1320 1325
Leu Asp Lys Gly Tyr Phe Glu Phe Ser Phe Asp Tyr Lys Asn Phe Gly
1330 1335 1340
Asp Lys Ala Ala Lys Gly Lys Trp Thr Ile Ala Ser Phe Gly Ser Arg
1345 1350 1355 1360
Leu Ile Asn Phe Arg Asn Ser Asp Lys Asn His Asn Trp Asp Thr Arg
1365 1370 1375
Glu Val Tyr Pro Thr Lys Glu Leu Glu Lys Leu Leu Lys Asp Tyr Ser
1380 1385 1390
Ile Glu Tyr Gly His Gly Glu Cys Ile Lys Ala Ala Ile Cys Gly Glu
1395 1400 1405
Ser Asp Lys Lys Phe Phe Ala Lys Leu Thr Ser Val Leu Asn Thr Ile
1410 1415 1420
Leu Gln Met Arg Asn Ser Lys Thr Gly Thr Glu Leu Asp Tyr Leu Ile
1425 1430 1435 1440
Ser Pro Val Ala Asp Val Asn Gly Asn Phe Phe Asp Ser Arg Gln Ala
1445 1450 1455
Pro Lys Asn Met Pro Gln Asp Ala Asp Ala Asn Gly Ala Tyr His Ile
1460 1465 1470
Gly Leu Lys Gly Leu Met Leu Leu Gly Arg Ile Lys Asn Asn Gln Glu
1475 1480 1485
Gly Lys Lys Leu Asn Leu Val Ile Lys Asn Glu Glu Tyr Phe Glu Phe
1490 1495 1500
Val Gln Asn Arg Asn Asn Gly Ser Pro Lys Lys Lys Arg Lys Val Ser
1505 1510 1515 1520
Gly Gly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys
1525 1530 1535
Gln Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu
1540 1545 1550
Glu Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala
1555 1560 1565
Tyr Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala
1570 1575 1580
Pro Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu
1585 1590 1595 1600
Asn Lys Ile Lys Met Leu Ser Gly Gly Ser Pro Lys Lys Lys Arg Lys
1605 1610 1615
Val
<210> 3
<211> 1690
<212> PRT
<213> (人工序列)
<400> 3
Met Ser Val Phe Leu Tyr Lys Lys Leu Pro Leu Asp Thr Phe Leu Phe
1 5 10 15
Glu Phe Asn Asn Leu Glu Trp Ala Tyr Gly Arg Asn Ser Cys Tyr Ile
20 25 30
Cys Phe Lys Leu Lys Pro Ile Leu Ala Thr Ser Thr Thr Gly Thr Thr
35 40 45
Glu Ser Glu Leu Trp Gly Tyr Val Thr Asn Lys Gly Glu Val Ala Asp
50 55 60
Gly Ile Pro Cys Glu Ser Pro Glu Lys Arg Gly Met His Ala Glu Glu
65 70 75 80
Leu Leu Leu Glu Glu Met Thr Arg His Val Arg Glu His Gly Gly Ala
85 90 95
Ser Phe Cys Val Glu Trp Phe Ala Ser Trp Ser Pro Cys Asp Arg Cys
100 105 110
Ser Gly Val Leu Leu Arg Trp Leu Arg Asp Val Gly Gly Gly Arg His
115 120 125
Arg Leu Arg Val Trp Phe Ser Arg Ile Tyr Arg Gly Asn Val Gly Ala
130 135 140
Val Arg Ala Gly Leu Arg His Leu Arg Arg Ala Gly Val Gln Leu Gly
145 150 155 160
Val Met Asp Arg Arg Arg His Asp Asn Cys Val His Val Leu Val Asp
165 170 175
Ala Ala Arg Gly Asp Pro Thr Pro Leu Trp Leu Val Gln Trp His Ser
180 185 190
Asn Val Pro Arg Val Gln Arg Glu Phe Asp Lys Ile Met Asp Glu Lys
195 200 205
Val Arg Glu Pro Gly Trp Cys Trp Trp Cys Trp Arg Pro Trp Met Ala
210 215 220
Asp Val Pro Ser Asn Ala Ser Phe Pro Gln Leu Pro Val Asn Asp Gly
225 230 235 240
Ala Ile Val Asp Arg Val Glu Lys Phe Pro Tyr Leu Gly Ser Leu Leu
245 250 255
Met Thr Ser Ser Gly Leu Ala Ala Glu Val Ser Ile Ser Gly Ser Glu
260 265 270
Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser Ile Tyr Gln
275 280 285
Glu Phe Val Asn Lys Tyr Ser Leu Ser Lys Thr Leu Arg Phe Glu Leu
290 295 300
Ile Pro Gln Gly Lys Thr Leu Glu Asn Ile Lys Ala Arg Gly Leu Ile
305 310 315 320
Leu Asp Asp Glu Lys Arg Ala Lys Asp Tyr Lys Lys Ala Lys Gln Ile
325 330 335
Ile Asp Lys Tyr His Gln Phe Phe Ile Glu Glu Ile Leu Ser Ser Val
340 345 350
Cys Ile Ser Glu Asp Leu Leu Gln Asn Tyr Ser Asp Val Tyr Phe Lys
355 360 365
Leu Lys Lys Ser Asp Asp Asp Asn Leu Gln Lys Asp Phe Lys Ser Ala
370 375 380
Lys Asp Thr Ile Lys Lys Gln Ile Ser Glu Tyr Ile Lys Asp Ser Glu
385 390 395 400
Lys Phe Lys Asn Leu Phe Asn Gln Asn Leu Ile Asp Ala Lys Lys Gly
405 410 415
Gln Glu Ser Asp Leu Ile Leu Trp Leu Lys Gln Ser Lys Asp Asn Gly
420 425 430
Ile Glu Leu Phe Lys Ala Asn Ser Asp Ile Thr Asp Ile Asp Glu Ala
435 440 445
Leu Glu Ile Ile Lys Ser Phe Lys Gly Trp Thr Thr Tyr Phe Lys Gly
450 455 460
Phe His Glu Asn Arg Lys Asn Val Tyr Ser Ser Asn Asp Ile Pro Thr
465 470 475 480
Ser Ile Ile Tyr Arg Ile Val Asp Asp Asn Leu Pro Lys Phe Leu Glu
485 490 495
Asn Lys Ala Lys Tyr Glu Ser Leu Lys Asp Lys Ala Pro Glu Ala Ile
500 505 510
Asn Tyr Glu Gln Ile Lys Lys Asp Leu Ala Glu Glu Leu Thr Phe Asp
515 520 525
Ile Asp Tyr Lys Thr Ser Glu Val Asn Gln Arg Val Phe Ser Leu Asp
530 535 540
Glu Val Phe Glu Ile Ala Asn Phe Asn Asn Tyr Leu Asn Gln Ser Gly
545 550 555 560
Ile Thr Lys Phe Asn Thr Ile Ile Gly Gly Lys Phe Val Asn Gly Glu
565 570 575
Asn Thr Lys Arg Lys Gly Ile Asn Glu Tyr Ile Asn Leu Tyr Ser Gln
580 585 590
Gln Ile Asn Asp Lys Thr Leu Lys Lys Tyr Lys Met Ser Val Leu Phe
595 600 605
Lys Gln Ile Leu Ser Asp Thr Glu Ser Lys Ser Phe Val Ile Asp Lys
610 615 620
Leu Glu Asp Asp Ser Asp Val Val Thr Thr Met Gln Ser Phe Tyr Glu
625 630 635 640
Gln Ile Ala Ala Phe Lys Thr Val Glu Glu Lys Ser Ile Lys Glu Thr
645 650 655
Leu Ser Leu Leu Phe Asp Asp Leu Lys Ala Gln Lys Leu Asp Leu Ser
660 665 670
Lys Ile Tyr Phe Lys Asn Asp Lys Ser Leu Thr Asp Leu Ser Gln Gln
675 680 685
Val Phe Asp Asp Tyr Ser Val Ile Gly Thr Ala Val Leu Glu Tyr Ile
690 695 700
Thr Gln Gln Ile Ala Pro Lys Asn Leu Asp Asn Pro Ser Lys Lys Glu
705 710 715 720
Gln Glu Leu Ile Ala Lys Lys Thr Glu Lys Ala Lys Tyr Leu Ser Leu
725 730 735
Glu Thr Ile Lys Leu Ala Leu Glu Glu Phe Asn Lys His Arg Asp Ile
740 745 750
Asp Lys Gln Cys Arg Phe Glu Glu Ile Leu Ala Asn Phe Ala Ala Ile
755 760 765
Pro Met Ile Phe Asp Glu Ile Ala Gln Asn Lys Asp Asn Leu Ala Gln
770 775 780
Ile Ser Ile Lys Tyr Gln Asn Gln Gly Lys Lys Asp Leu Leu Gln Ala
785 790 795 800
Ser Ala Glu Asp Asp Val Lys Ala Ile Lys Asp Leu Leu Asp Gln Thr
805 810 815
Asn Asn Leu Leu His Lys Leu Lys Ile Phe His Ile Ser Gln Ser Glu
820 825 830
Asp Lys Ala Asn Ile Leu Asp Lys Asp Glu His Phe Tyr Leu Val Phe
835 840 845
Glu Glu Cys Tyr Phe Glu Leu Ala Asn Ile Val Pro Leu Tyr Asn Lys
850 855 860
Ile Arg Asn Tyr Ile Thr Gln Lys Pro Tyr Ser Asp Glu Lys Phe Lys
865 870 875 880
Leu Asn Phe Glu Asn Ser Thr Leu Ala Asn Gly Trp Asp Lys Asn Lys
885 890 895
Glu Pro Asp Asn Thr Ala Ile Leu Phe Ile Lys Asp Asp Lys Tyr Tyr
900 905 910
Leu Gly Val Met Asn Lys Lys Asn Asn Lys Ile Phe Asp Asp Lys Ala
915 920 925
Ile Lys Glu Asn Lys Gly Glu Gly Tyr Lys Lys Ile Val Tyr Lys Leu
930 935 940
Leu Pro Gly Ala Asn Lys Met Leu Pro Lys Val Phe Phe Ser Ala Lys
945 950 955 960
Ser Ile Lys Phe Tyr Asn Pro Ser Glu Asp Ile Leu Arg Ile Arg Asn
965 970 975
His Ser Thr His Thr Lys Asn Gly Ser Pro Gln Lys Gly Tyr Glu Lys
980 985 990
Phe Glu Phe Asn Ile Glu Asp Cys Arg Lys Phe Ile Asp Phe Tyr Lys
995 1000 1005
Gln Ser Ile Ser Lys His Pro Glu Trp Lys Asp Phe Gly Phe Arg Phe
1010 1015 1020
Ser Asp Thr Gln Arg Tyr Asn Ser Ile Asp Glu Phe Tyr Arg Glu Val
1025 1030 1035 1040
Glu Asn Gln Gly Tyr Lys Leu Thr Phe Glu Asn Ile Ser Glu Ser Tyr
1045 1050 1055
Ile Asp Ser Val Val Asn Gln Gly Lys Leu Tyr Leu Phe Gln Ile Tyr
1060 1065 1070
Asn Lys Asp Phe Ser Ala Tyr Ser Lys Gly Arg Pro Asn Leu His Thr
1075 1080 1085
Leu Tyr Trp Lys Ala Leu Phe Asp Glu Arg Asn Leu Gln Asp Val Val
1090 1095 1100
Tyr Lys Leu Asn Gly Glu Ala Glu Leu Phe Tyr Arg Lys Gln Ser Ile
1105 1110 1115 1120
Pro Lys Lys Ile Thr His Pro Ala Lys Glu Ala Ile Ala Asn Lys Asn
1125 1130 1135
Lys Asp Asn Pro Lys Lys Glu Ser Val Phe Glu Tyr Asp Leu Ile Lys
1140 1145 1150
Asp Lys Arg Phe Thr Glu Asp Lys Phe Phe Phe His Cys Pro Ile Thr
1155 1160 1165
Ile Asn Phe Lys Ser Ser Gly Ala Asn Lys Phe Asn Asp Glu Ile Asn
1170 1175 1180
Leu Leu Leu Lys Glu Lys Ala Asn Asp Val His Ile Leu Ser Ile Ala
1185 1190 1195 1200
Arg Gly Glu Arg His Leu Ala Tyr Tyr Thr Leu Val Asp Gly Lys Gly
1205 1210 1215
Asn Ile Ile Lys Gln Asp Thr Phe Asn Ile Ile Gly Asn Asp Arg Met
1220 1225 1230
Lys Thr Asn Tyr His Asp Lys Leu Ala Ala Ile Glu Lys Asp Arg Asp
1235 1240 1245
Ser Ala Arg Lys Asp Trp Lys Lys Ile Asn Asn Ile Lys Glu Met Lys
1250 1255 1260
Glu Gly Tyr Leu Ser Gln Val Val His Glu Ile Ala Lys Leu Val Ile
1265 1270 1275 1280
Glu Tyr Asn Ala Ile Val Val Phe Glu Asp Leu Asn Phe Gly Phe Lys
1285 1290 1295
Arg Gly Arg Phe Lys Val Glu Lys Gln Val Tyr Gln Lys Leu Glu Lys
1300 1305 1310
Met Leu Ile Glu Lys Leu Asn Tyr Leu Val Phe Lys Asp Asn Glu Phe
1315 1320 1325
Asp Lys Thr Gly Gly Val Leu Arg Ala Tyr Gln Leu Thr Ala Pro Phe
1330 1335 1340
Glu Thr Phe Lys Lys Met Gly Lys Gln Thr Gly Ile Ile Tyr Tyr Val
1345 1350 1355 1360
Pro Ala Gly Phe Thr Ser Lys Ile Cys Pro Val Thr Gly Phe Val Asn
1365 1370 1375
Gln Leu Tyr Pro Lys Tyr Glu Ser Val Ser Lys Ser Gln Glu Phe Phe
1380 1385 1390
Ser Lys Phe Asp Lys Ile Cys Tyr Asn Leu Asp Lys Gly Tyr Phe Glu
1395 1400 1405
Phe Ser Phe Asp Tyr Lys Asn Phe Gly Asp Lys Ala Ala Lys Gly Lys
1410 1415 1420
Trp Thr Ile Ala Ser Phe Gly Ser Arg Leu Ile Asn Phe Arg Asn Ser
1425 1430 1435 1440
Asp Lys Asn His Asn Trp Asp Thr Arg Glu Val Tyr Pro Thr Lys Glu
1445 1450 1455
Leu Glu Lys Leu Leu Lys Asp Tyr Ser Ile Glu Tyr Gly His Gly Glu
1460 1465 1470
Cys Ile Lys Ala Ala Ile Cys Gly Glu Ser Asp Lys Lys Phe Phe Ala
1475 1480 1485
Lys Leu Thr Ser Val Leu Asn Thr Ile Leu Gln Met Arg Asn Ser Lys
1490 1495 1500
Thr Gly Thr Glu Leu Asp Tyr Leu Ile Ser Pro Val Ala Asp Val Asn
1505 1510 1515 1520
Gly Asn Phe Phe Asp Ser Arg Gln Ala Pro Lys Asn Met Pro Gln Asp
1525 1530 1535
Ala Asp Ala Asn Gly Ala Tyr His Ile Gly Leu Lys Gly Leu Met Leu
1540 1545 1550
Leu Gly Arg Ile Lys Asn Asn Gln Glu Gly Lys Lys Leu Asn Leu Val
1555 1560 1565
Ile Lys Asn Glu Glu Tyr Phe Glu Phe Val Gln Asn Arg Asn Asn Gly
1570 1575 1580
Ser Pro Lys Lys Lys Arg Lys Val Ser Gly Gly Ser Thr Asn Leu Ser
1585 1590 1595 1600
Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val Ile Gln Glu Ser
1605 1610 1615
Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile Gly Asn Lys Pro
1620 1625 1630
Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu Ser Thr Asp Glu
1635 1640 1645
Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr Lys Pro Trp Ala
1650 1655 1660
Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile Lys Met Leu Ser
1665 1670 1675 1680
Gly Gly Ser Pro Lys Lys Lys Arg Lys Val
1685 1690
<210> 4
<211> 82
<212> DNA
<213> (人工序列)
<400> 4
gtctaagaac tttaaataat ttctactgtt gtagatagag accgtgaagt taataaggtc 60
tcaaatttct actgttgtag at 82
<210> 5
<211> 1651
<212> PRT
<213> (人工序列)
<400> 5
Met Ser Ser Glu Thr Gly Pro Val Ala Val Asp Pro Thr Leu Arg Arg
1 5 10 15
Arg Ile Glu Pro His Glu Phe Glu Val Phe Phe Asp Pro Arg Glu Leu
20 25 30
Arg Lys Glu Thr Cys Leu Leu Tyr Glu Ile Asn Trp Gly Gly Arg His
35 40 45
Ser Ile Trp Arg His Thr Ser Gln Asn Thr Asn Lys His Val Glu Val
50 55 60
Asn Phe Ile Glu Lys Phe Thr Thr Glu Arg Tyr Phe Cys Pro Asn Thr
65 70 75 80
Arg Cys Ser Ile Thr Trp Phe Leu Ser Trp Ser Pro Cys Gly Glu Cys
85 90 95
Ser Arg Ala Ile Thr Glu Phe Leu Ser Arg Tyr Pro His Val Thr Leu
100 105 110
Phe Ile Tyr Ile Ala Arg Leu Tyr His His Ala Asp Pro Arg Asn Arg
115 120 125
Gln Gly Leu Arg Asp Leu Ile Ser Ser Gly Val Thr Ile Gln Ile Met
130 135 140
Thr Glu Gln Glu Ser Gly Tyr Cys Trp Arg Asn Phe Val Asn Tyr Ser
145 150 155 160
Pro Ser Asn Glu Ala His Trp Pro Arg Tyr Pro His Leu Trp Val Arg
165 170 175
Leu Tyr Val Leu Glu Leu Tyr Cys Ile Ile Leu Gly Leu Pro Pro Cys
180 185 190
Leu Asn Ile Leu Arg Arg Lys Gln Pro Gln Leu Thr Phe Phe Thr Ile
195 200 205
Ala Leu Gln Ser Cys His Tyr Gln Arg Leu Pro Pro His Ile Leu Trp
210 215 220
Ala Thr Gly Leu Lys Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser
225 230 235 240
Ala Thr Pro Glu Ser Ser Ile Tyr Gln Glu Phe Val Asn Lys Tyr Ser
245 250 255
Leu Ser Lys Thr Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu
260 265 270
Glu Asn Ile Lys Ala Arg Gly Leu Ile Leu Asp Asp Glu Lys Arg Ala
275 280 285
Lys Asp Tyr Lys Lys Ala Lys Gln Ile Ile Asp Lys Tyr His Gln Phe
290 295 300
Phe Ile Glu Glu Ile Leu Ser Ser Val Cys Ile Ser Glu Asp Leu Leu
305 310 315 320
Gln Asn Tyr Ser Asp Val Tyr Phe Lys Leu Lys Lys Ser Asp Asp Asp
325 330 335
Asn Leu Gln Lys Asp Phe Lys Ser Ala Lys Asp Thr Ile Lys Lys Gln
340 345 350
Ile Ser Glu Tyr Ile Lys Asp Ser Glu Lys Phe Lys Asn Leu Phe Asn
355 360 365
Gln Asn Leu Ile Asp Ala Lys Lys Gly Gln Glu Ser Asp Leu Ile Leu
370 375 380
Trp Leu Lys Gln Ser Lys Asp Asn Gly Ile Glu Leu Phe Lys Ala Asn
385 390 395 400
Ser Asp Ile Thr Asp Ile Asp Glu Ala Leu Glu Ile Ile Lys Ser Phe
405 410 415
Lys Gly Trp Thr Thr Tyr Phe Lys Gly Phe His Glu Asn Arg Lys Asn
420 425 430
Val Tyr Ser Ser Asn Asp Ile Pro Thr Ser Ile Ile Tyr Arg Ile Val
435 440 445
Asp Asp Asn Leu Pro Lys Phe Leu Glu Asn Lys Ala Lys Tyr Glu Ser
450 455 460
Leu Lys Asp Lys Ala Pro Glu Ala Ile Asn Tyr Glu Gln Ile Lys Lys
465 470 475 480
Asp Leu Ala Glu Glu Leu Thr Phe Asp Ile Asp Tyr Lys Thr Ser Glu
485 490 495
Val Asn Gln Arg Val Phe Ser Leu Asp Glu Val Phe Glu Ile Ala Asn
500 505 510
Phe Asn Asn Tyr Leu Asn Gln Ser Gly Ile Thr Lys Phe Asn Thr Ile
515 520 525
Ile Gly Gly Lys Phe Val Asn Gly Glu Asn Thr Lys Arg Lys Gly Ile
530 535 540
Asn Glu Tyr Ile Asn Leu Tyr Ser Gln Gln Ile Asn Asp Lys Thr Leu
545 550 555 560
Lys Lys Tyr Lys Met Ser Val Leu Phe Lys Gln Ile Leu Ser Asp Thr
565 570 575
Glu Ser Lys Ser Phe Val Ile Asp Lys Leu Glu Asp Asp Ser Asp Val
580 585 590
Val Thr Thr Met Gln Ser Phe Tyr Glu Gln Ile Ala Ala Phe Lys Thr
595 600 605
Val Glu Glu Lys Ser Ile Lys Glu Thr Leu Ser Leu Leu Phe Asp Asp
610 615 620
Leu Lys Ala Gln Lys Leu Asp Leu Ser Lys Ile Tyr Phe Lys Asn Asp
625 630 635 640
Lys Ser Leu Thr Asp Leu Ser Gln Gln Val Phe Asp Asp Tyr Ser Val
645 650 655
Ile Gly Thr Ala Val Leu Glu Tyr Ile Thr Gln Gln Ile Ala Pro Lys
660 665 670
Asn Leu Asp Asn Pro Ser Lys Lys Glu Gln Glu Leu Ile Ala Lys Lys
675 680 685
Thr Glu Lys Ala Lys Tyr Leu Ser Leu Glu Thr Ile Lys Leu Ala Leu
690 695 700
Glu Glu Phe Asn Lys His Arg Asp Ile Asp Lys Gln Cys Arg Phe Glu
705 710 715 720
Glu Ile Leu Ala Asn Phe Ala Ala Ile Pro Met Ile Phe Asp Glu Ile
725 730 735
Ala Gln Asn Lys Asp Asn Leu Ala Gln Ile Ser Ile Lys Tyr Gln Asn
740 745 750
Gln Gly Lys Lys Asp Leu Leu Gln Ala Ser Ala Glu Asp Asp Val Lys
755 760 765
Ala Ile Lys Asp Leu Leu Asp Gln Thr Asn Asn Leu Leu His Lys Leu
770 775 780
Lys Ile Phe His Ile Ser Gln Ser Glu Asp Lys Ala Asn Ile Leu Asp
785 790 795 800
Lys Asp Glu His Phe Tyr Leu Val Phe Glu Glu Cys Tyr Phe Glu Leu
805 810 815
Ala Asn Ile Val Pro Leu Tyr Asn Lys Ile Arg Asn Tyr Ile Thr Gln
820 825 830
Lys Pro Tyr Ser Asp Glu Lys Phe Lys Leu Asn Phe Glu Asn Ser Thr
835 840 845
Leu Ala Asn Gly Trp Asp Lys Asn Lys Glu Pro Asp Asn Thr Ala Ile
850 855 860
Leu Phe Ile Lys Asp Asp Lys Tyr Tyr Leu Gly Val Met Asn Lys Lys
865 870 875 880
Asn Asn Lys Ile Phe Asp Asp Lys Ala Ile Lys Glu Asn Lys Gly Glu
885 890 895
Gly Tyr Lys Lys Ile Val Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met
900 905 910
Leu Pro Lys Val Phe Phe Ser Ala Lys Ser Ile Lys Phe Tyr Asn Pro
915 920 925
Ser Glu Asp Ile Leu Arg Ile Arg Asn His Ser Thr His Thr Lys Asn
930 935 940
Gly Ser Pro Gln Lys Gly Tyr Glu Lys Phe Glu Phe Asn Ile Glu Asp
945 950 955 960
Cys Arg Lys Phe Ile Asp Phe Tyr Lys Gln Ser Ile Ser Lys His Pro
965 970 975
Glu Trp Lys Asp Phe Gly Phe Arg Phe Ser Asp Thr Gln Arg Tyr Asn
980 985 990
Ser Ile Asp Glu Phe Tyr Arg Glu Val Glu Asn Gln Gly Tyr Lys Leu
995 1000 1005
Thr Phe Glu Asn Ile Ser Glu Ser Tyr Ile Asp Ser Val Val Asn Gln
1010 1015 1020
Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Ala Tyr
1025 1030 1035 1040
Ser Lys Gly Arg Pro Asn Leu His Thr Leu Tyr Trp Lys Ala Leu Phe
1045 1050 1055
Asp Glu Arg Asn Leu Gln Asp Val Val Tyr Lys Leu Asn Gly Glu Ala
1060 1065 1070
Glu Leu Phe Tyr Arg Lys Gln Ser Ile Pro Lys Lys Ile Thr His Pro
1075 1080 1085
Ala Lys Glu Ala Ile Ala Asn Lys Asn Lys Asp Asn Pro Lys Lys Glu
1090 1095 1100
Ser Val Phe Glu Tyr Asp Leu Ile Lys Asp Lys Arg Phe Thr Glu Asp
1105 1110 1115 1120
Lys Phe Phe Phe His Cys Pro Ile Thr Ile Asn Phe Lys Ser Ser Gly
1125 1130 1135
Ala Asn Lys Phe Asn Asp Glu Ile Asn Leu Leu Leu Lys Glu Lys Ala
1140 1145 1150
Asn Asp Val His Ile Leu Ser Ile Ala Arg Gly Glu Arg His Leu Ala
1155 1160 1165
Tyr Tyr Thr Leu Val Asp Gly Lys Gly Asn Ile Ile Lys Gln Asp Thr
1170 1175 1180
Phe Asn Ile Ile Gly Asn Asp Arg Met Lys Thr Asn Tyr His Asp Lys
1185 1190 1195 1200
Leu Ala Ala Ile Glu Lys Asp Arg Asp Ser Ala Arg Lys Asp Trp Lys
1205 1210 1215
Lys Ile Asn Asn Ile Lys Glu Met Lys Glu Gly Tyr Leu Ser Gln Val
1220 1225 1230
Val His Glu Ile Ala Lys Leu Val Ile Glu Tyr Asn Ala Ile Val Val
1235 1240 1245
Phe Glu Asp Leu Asn Phe Gly Phe Lys Arg Gly Arg Phe Lys Val Glu
1250 1255 1260
Lys Gln Val Tyr Gln Lys Leu Glu Lys Met Leu Ile Glu Lys Leu Asn
1265 1270 1275 1280
Tyr Leu Val Phe Lys Asp Asn Glu Phe Asp Lys Thr Gly Gly Val Leu
1285 1290 1295
Arg Ala Tyr Gln Leu Thr Ala Pro Phe Glu Thr Phe Lys Lys Met Gly
1300 1305 1310
Lys Gln Thr Gly Ile Ile Tyr Tyr Val Pro Ala Gly Phe Thr Ser Lys
1315 1320 1325
Ile Cys Pro Val Thr Gly Phe Val Asn Gln Leu Tyr Pro Lys Tyr Glu
1330 1335 1340
Ser Val Ser Lys Ser Gln Glu Phe Phe Ser Lys Phe Asp Lys Ile Cys
1345 1350 1355 1360
Tyr Asn Leu Asp Lys Gly Tyr Phe Glu Phe Ser Phe Asp Tyr Lys Asn
1365 1370 1375
Phe Gly Asp Lys Ala Ala Lys Gly Lys Trp Thr Ile Ala Ser Phe Gly
1380 1385 1390
Ser Arg Leu Ile Asn Phe Arg Asn Ser Asp Lys Asn His Asn Trp Asp
1395 1400 1405
Thr Arg Glu Val Tyr Pro Thr Lys Glu Leu Glu Lys Leu Leu Lys Asp
1410 1415 1420
Tyr Ser Ile Glu Tyr Gly His Gly Glu Cys Ile Lys Ala Ala Ile Cys
1425 1430 1435 1440
Gly Glu Ser Asp Lys Lys Phe Phe Ala Lys Leu Thr Ser Val Leu Asn
1445 1450 1455
Thr Ile Leu Gln Met Arg Asn Ser Lys Thr Gly Thr Glu Leu Asp Tyr
1460 1465 1470
Leu Ile Ser Pro Val Ala Asp Val Asn Gly Asn Phe Phe Asp Ser Arg
1475 1480 1485
Gln Ala Pro Lys Asn Met Pro Gln Asp Ala Asp Ala Asn Gly Ala Tyr
1490 1495 1500
His Ile Gly Leu Lys Gly Leu Met Leu Leu Gly Arg Ile Lys Asn Asn
1505 1510 1515 1520
Gln Glu Gly Lys Lys Leu Asn Leu Val Ile Lys Asn Glu Glu Tyr Phe
1525 1530 1535
Glu Phe Val Gln Asn Arg Asn Asn Gly Ser Pro Lys Lys Lys Arg Lys
1540 1545 1550
Val Ser Gly Gly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr
1555 1560 1565
Gly Lys Gln Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu
1570 1575 1580
Val Glu Glu Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His
1585 1590 1595 1600
Thr Ala Tyr Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser
1605 1610 1615
Asp Ala Pro Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn
1620 1625 1630
Gly Glu Asn Lys Ile Lys Met Leu Ser Gly Gly Ser Pro Lys Lys Lys
1635 1640 1645
Arg Lys Val
1650
<210> 6
<211> 1630
<212> PRT
<213> (人工序列)
<400> 6
Met Thr Asp Ala Glu Tyr Val Arg Ile His Glu Lys Leu Asp Ile Tyr
1 5 10 15
Thr Phe Lys Lys Gln Phe Phe Asn Asn Lys Lys Ser Val Ser His Arg
20 25 30
Cys Tyr Val Leu Phe Glu Leu Lys Arg Arg Gly Glu Arg Arg Ala Cys
35 40 45
Phe Trp Gly Tyr Ala Val Asn Lys Pro Gln Ser Gly Thr Glu Arg Gly
50 55 60
Ile His Ala Glu Ile Phe Ser Ile Arg Lys Val Glu Glu Tyr Leu Arg
65 70 75 80
Asp Asn Pro Gly Gln Phe Thr Ile Asn Trp Tyr Ser Ser Trp Ser Pro
85 90 95
Cys Ala Asp Cys Ala Glu Lys Ile Leu Glu Trp Tyr Asn Gln Glu Leu
100 105 110
Arg Gly Asn Gly His Thr Leu Lys Ile Trp Ala Cys Lys Leu Tyr Tyr
115 120 125
Glu Lys Asn Ala Arg Asn Gln Ile Gly Leu Trp Asn Leu Arg Asp Asn
130 135 140
Gly Val Gly Leu Asn Val Met Val Ser Glu His Tyr Gln Cys Cys Arg
145 150 155 160
Lys Ile Phe Ile Gln Ser Ser His Asn Gln Leu Asn Glu Asn Arg Trp
165 170 175
Leu Glu Lys Thr Leu Lys Arg Ala Glu Lys Arg Arg Ser Glu Leu Ser
180 185 190
Ile Met Ile Gln Val Lys Ile Leu His Thr Thr Lys Ser Pro Ala Val
195 200 205
Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser
210 215 220
Ser Ile Tyr Gln Glu Phe Val Asn Lys Tyr Ser Leu Ser Lys Thr Leu
225 230 235 240
Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Glu Asn Ile Lys Ala
245 250 255
Arg Gly Leu Ile Leu Asp Asp Glu Lys Arg Ala Lys Asp Tyr Lys Lys
260 265 270
Ala Lys Gln Ile Ile Asp Lys Tyr His Gln Phe Phe Ile Glu Glu Ile
275 280 285
Leu Ser Ser Val Cys Ile Ser Glu Asp Leu Leu Gln Asn Tyr Ser Asp
290 295 300
Val Tyr Phe Lys Leu Lys Lys Ser Asp Asp Asp Asn Leu Gln Lys Asp
305 310 315 320
Phe Lys Ser Ala Lys Asp Thr Ile Lys Lys Gln Ile Ser Glu Tyr Ile
325 330 335
Lys Asp Ser Glu Lys Phe Lys Asn Leu Phe Asn Gln Asn Leu Ile Asp
340 345 350
Ala Lys Lys Gly Gln Glu Ser Asp Leu Ile Leu Trp Leu Lys Gln Ser
355 360 365
Lys Asp Asn Gly Ile Glu Leu Phe Lys Ala Asn Ser Asp Ile Thr Asp
370 375 380
Ile Asp Glu Ala Leu Glu Ile Ile Lys Ser Phe Lys Gly Trp Thr Thr
385 390 395 400
Tyr Phe Lys Gly Phe His Glu Asn Arg Lys Asn Val Tyr Ser Ser Asn
405 410 415
Asp Ile Pro Thr Ser Ile Ile Tyr Arg Ile Val Asp Asp Asn Leu Pro
420 425 430
Lys Phe Leu Glu Asn Lys Ala Lys Tyr Glu Ser Leu Lys Asp Lys Ala
435 440 445
Pro Glu Ala Ile Asn Tyr Glu Gln Ile Lys Lys Asp Leu Ala Glu Glu
450 455 460
Leu Thr Phe Asp Ile Asp Tyr Lys Thr Ser Glu Val Asn Gln Arg Val
465 470 475 480
Phe Ser Leu Asp Glu Val Phe Glu Ile Ala Asn Phe Asn Asn Tyr Leu
485 490 495
Asn Gln Ser Gly Ile Thr Lys Phe Asn Thr Ile Ile Gly Gly Lys Phe
500 505 510
Val Asn Gly Glu Asn Thr Lys Arg Lys Gly Ile Asn Glu Tyr Ile Asn
515 520 525
Leu Tyr Ser Gln Gln Ile Asn Asp Lys Thr Leu Lys Lys Tyr Lys Met
530 535 540
Ser Val Leu Phe Lys Gln Ile Leu Ser Asp Thr Glu Ser Lys Ser Phe
545 550 555 560
Val Ile Asp Lys Leu Glu Asp Asp Ser Asp Val Val Thr Thr Met Gln
565 570 575
Ser Phe Tyr Glu Gln Ile Ala Ala Phe Lys Thr Val Glu Glu Lys Ser
580 585 590
Ile Lys Glu Thr Leu Ser Leu Leu Phe Asp Asp Leu Lys Ala Gln Lys
595 600 605
Leu Asp Leu Ser Lys Ile Tyr Phe Lys Asn Asp Lys Ser Leu Thr Asp
610 615 620
Leu Ser Gln Gln Val Phe Asp Asp Tyr Ser Val Ile Gly Thr Ala Val
625 630 635 640
Leu Glu Tyr Ile Thr Gln Gln Ile Ala Pro Lys Asn Leu Asp Asn Pro
645 650 655
Ser Lys Lys Glu Gln Glu Leu Ile Ala Lys Lys Thr Glu Lys Ala Lys
660 665 670
Tyr Leu Ser Leu Glu Thr Ile Lys Leu Ala Leu Glu Glu Phe Asn Lys
675 680 685
His Arg Asp Ile Asp Lys Gln Cys Arg Phe Glu Glu Ile Leu Ala Asn
690 695 700
Phe Ala Ala Ile Pro Met Ile Phe Asp Glu Ile Ala Gln Asn Lys Asp
705 710 715 720
Asn Leu Ala Gln Ile Ser Ile Lys Tyr Gln Asn Gln Gly Lys Lys Asp
725 730 735
Leu Leu Gln Ala Ser Ala Glu Asp Asp Val Lys Ala Ile Lys Asp Leu
740 745 750
Leu Asp Gln Thr Asn Asn Leu Leu His Lys Leu Lys Ile Phe His Ile
755 760 765
Ser Gln Ser Glu Asp Lys Ala Asn Ile Leu Asp Lys Asp Glu His Phe
770 775 780
Tyr Leu Val Phe Glu Glu Cys Tyr Phe Glu Leu Ala Asn Ile Val Pro
785 790 795 800
Leu Tyr Asn Lys Ile Arg Asn Tyr Ile Thr Gln Lys Pro Tyr Ser Asp
805 810 815
Glu Lys Phe Lys Leu Asn Phe Glu Asn Ser Thr Leu Ala Asn Gly Trp
820 825 830
Asp Lys Asn Lys Glu Pro Asp Asn Thr Ala Ile Leu Phe Ile Lys Asp
835 840 845
Asp Lys Tyr Tyr Leu Gly Val Met Asn Lys Lys Asn Asn Lys Ile Phe
850 855 860
Asp Asp Lys Ala Ile Lys Glu Asn Lys Gly Glu Gly Tyr Lys Lys Ile
865 870 875 880
Val Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro Lys Val Phe
885 890 895
Phe Ser Ala Lys Ser Ile Lys Phe Tyr Asn Pro Ser Glu Asp Ile Leu
900 905 910
Arg Ile Arg Asn His Ser Thr His Thr Lys Asn Gly Ser Pro Gln Lys
915 920 925
Gly Tyr Glu Lys Phe Glu Phe Asn Ile Glu Asp Cys Arg Lys Phe Ile
930 935 940
Asp Phe Tyr Lys Gln Ser Ile Ser Lys His Pro Glu Trp Lys Asp Phe
945 950 955 960
Gly Phe Arg Phe Ser Asp Thr Gln Arg Tyr Asn Ser Ile Asp Glu Phe
965 970 975
Tyr Arg Glu Val Glu Asn Gln Gly Tyr Lys Leu Thr Phe Glu Asn Ile
980 985 990
Ser Glu Ser Tyr Ile Asp Ser Val Val Asn Gln Gly Lys Leu Tyr Leu
995 1000 1005
Phe Gln Ile Tyr Asn Lys Asp Phe Ser Ala Tyr Ser Lys Gly Arg Pro
1010 1015 1020
Asn Leu His Thr Leu Tyr Trp Lys Ala Leu Phe Asp Glu Arg Asn Leu
1025 1030 1035 1040
Gln Asp Val Val Tyr Lys Leu Asn Gly Glu Ala Glu Leu Phe Tyr Arg
1045 1050 1055
Lys Gln Ser Ile Pro Lys Lys Ile Thr His Pro Ala Lys Glu Ala Ile
1060 1065 1070
Ala Asn Lys Asn Lys Asp Asn Pro Lys Lys Glu Ser Val Phe Glu Tyr
1075 1080 1085
Asp Leu Ile Lys Asp Lys Arg Phe Thr Glu Asp Lys Phe Phe Phe His
1090 1095 1100
Cys Pro Ile Thr Ile Asn Phe Lys Ser Ser Gly Ala Asn Lys Phe Asn
1105 1110 1115 1120
Asp Glu Ile Asn Leu Leu Leu Lys Glu Lys Ala Asn Asp Val His Ile
1125 1130 1135
Leu Ser Ile Ala Arg Gly Glu Arg His Leu Ala Tyr Tyr Thr Leu Val
1140 1145 1150
Asp Gly Lys Gly Asn Ile Ile Lys Gln Asp Thr Phe Asn Ile Ile Gly
1155 1160 1165
Asn Asp Arg Met Lys Thr Asn Tyr His Asp Lys Leu Ala Ala Ile Glu
1170 1175 1180
Lys Asp Arg Asp Ser Ala Arg Lys Asp Trp Lys Lys Ile Asn Asn Ile
1185 1190 1195 1200
Lys Glu Met Lys Glu Gly Tyr Leu Ser Gln Val Val His Glu Ile Ala
1205 1210 1215
Lys Leu Val Ile Glu Tyr Asn Ala Ile Val Val Phe Glu Asp Leu Asn
1220 1225 1230
Phe Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Val Tyr Gln
1235 1240 1245
Lys Leu Glu Lys Met Leu Ile Glu Lys Leu Asn Tyr Leu Val Phe Lys
1250 1255 1260
Asp Asn Glu Phe Asp Lys Thr Gly Gly Val Leu Arg Ala Tyr Gln Leu
1265 1270 1275 1280
Thr Ala Pro Phe Glu Thr Phe Lys Lys Met Gly Lys Gln Thr Gly Ile
1285 1290 1295
Ile Tyr Tyr Val Pro Ala Gly Phe Thr Ser Lys Ile Cys Pro Val Thr
1300 1305 1310
Gly Phe Val Asn Gln Leu Tyr Pro Lys Tyr Glu Ser Val Ser Lys Ser
1315 1320 1325
Gln Glu Phe Phe Ser Lys Phe Asp Lys Ile Cys Tyr Asn Leu Asp Lys
1330 1335 1340
Gly Tyr Phe Glu Phe Ser Phe Asp Tyr Lys Asn Phe Gly Asp Lys Ala
1345 1350 1355 1360
Ala Lys Gly Lys Trp Thr Ile Ala Ser Phe Gly Ser Arg Leu Ile Asn
1365 1370 1375
Phe Arg Asn Ser Asp Lys Asn His Asn Trp Asp Thr Arg Glu Val Tyr
1380 1385 1390
Pro Thr Lys Glu Leu Glu Lys Leu Leu Lys Asp Tyr Ser Ile Glu Tyr
1395 1400 1405
Gly His Gly Glu Cys Ile Lys Ala Ala Ile Cys Gly Glu Ser Asp Lys
1410 1415 1420
Lys Phe Phe Ala Lys Leu Thr Ser Val Leu Asn Thr Ile Leu Gln Met
1425 1430 1435 1440
Arg Asn Ser Lys Thr Gly Thr Glu Leu Asp Tyr Leu Ile Ser Pro Val
1445 1450 1455
Ala Asp Val Asn Gly Asn Phe Phe Asp Ser Arg Gln Ala Pro Lys Asn
1460 1465 1470
Met Pro Gln Asp Ala Asp Ala Asn Gly Ala Tyr His Ile Gly Leu Lys
1475 1480 1485
Gly Leu Met Leu Leu Gly Arg Ile Lys Asn Asn Gln Glu Gly Lys Lys
1490 1495 1500
Leu Asn Leu Val Ile Lys Asn Glu Glu Tyr Phe Glu Phe Val Gln Asn
1505 1510 1515 1520
Arg Asn Asn Gly Ser Pro Lys Lys Lys Arg Lys Val Ser Gly Gly Ser
1525 1530 1535
Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val
1540 1545 1550
Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile
1555 1560 1565
Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu
1570 1575 1580
Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr
1585 1590 1595 1600
Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile
1605 1610 1615
Lys Met Leu Ser Gly Gly Ser Pro Lys Lys Lys Arg Lys Val
1620 1625 1630
<210> 7
<211> 9780
<212> DNA
<213> (人工序列)
<400> 7
atgtcatgac attggtgtac agaaatggcg cagcaatggc aagaacgtcc cgggcggagc 60
tcaggcctta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 120
tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 180
ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc 240
gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa 300
tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat 360
cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg 420
cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc 480
atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc 540
ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag 600
acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc 660
tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg 720
tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca 780
tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg 840
tgcaccgccg ttttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg 900
gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg 960
gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc 1020
acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc 1080
gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca 1140
tactctgcga catcgtataa cgttactggt ttcatcaaaa tcgtctccct ccgtttgaat 1200
atttgattga tcgtaaccag atgaagcact ctttccacta tccctacagt gttatggctt 1260
gaacaatcac gaaacaataa ttggtacgta cgatctttca gccgactcaa acatcaaatc 1320
ttacaaatgt agtctttgaa agtattacat atgtaagatt taaatgcaac cgttttttcg 1380
gaaggaaatg atgacctcgt ttccaccgga attagcttgg taccagctat tgtaacataa 1440
tcggtacggg ggtgaaaaag ctaacggaaa agggagcgga aaagaatgat gtaagcgtga 1500
aaaatttttt aaaaaatctc ttgacattgg aagggagata tgttattata agaattgcgg 1560
aattgtgagc ggataacaat tcataattgt gagcggataa caattcaacc ccaaaggagg 1620
tgggatccat ggaagcatca cctgcatcag gaccacgcca cctaatggac ccacacatct 1680
tcacgagtaa tttcaataat ggtatcggtc ggcacaagac ttacctttgt tatgaagttg 1740
aaagacttga taacggaaca tcagttaaga tggaccagca tagaggattt cttcataacc 1800
aggcaaagaa tctcctttgt ggattctacg gtagacatgc agaacttaga tttcttgatc 1860
ttgttccttc acttcagctt gatcctgcac agatatatag agttacatgg tttatatcat 1920
ggtcaccttg tttcagttgg ggatgcgctg gagaagttag agcatttctt caggagaata 1980
ctcacgtgcg cctgcggatc tttgcagcgc ggatttacga ctatgatcct ctttataaag 2040
aagcacttca gatgcttaga gatgcaggag cacaggtttc aataatgaca tatgatgaat 2100
ttaaacattg ttgggataca tttgttgatc atcagggatg tcctttccaa ccatgggatg 2160
gacttgatga acattcacag gcactttcag gaagacttag agcaatactt cagaaccagg 2220
gaaactctgg ctctgaaaca cctggcacat ctgaatctgc tacacctgaa tcttcaattt 2280
atcaagaatt tgttaataaa tatagtttaa gtaaaactct aagatttgag ttaatcccac 2340
agggtaaaac acttgaaaac ataaaagcaa gaggtttgat tttagatgat gagaaaagag 2400
ctaaagacta caaaaaggct aaacaaataa ttgataaata tcatcagttt tttatagagg 2460
agatattaag ttcggtttgt attagcgaag atttattaca aaactattct gatgtttatt 2520
ttaaacttaa aaagagtgat gatgataatc tacaaaaaga ttttaaaagt gcaaaagata 2580
cgataaagaa acaaatatct gaatatataa aggactcaga gaaatttaag aatttgttta 2640
atcaaaacct tatcgatgct aaaaaagggc aagagtcaga tttaattcta tggctaaagc 2700
aatctaagga taatggtata gaactattta aagccaatag tgatatcaca gatatagatg 2760
aggcgttaga aataatcaaa tcttttaaag gttggacaac ttattttaag ggttttcatg 2820
aaaatagaaa aaatgtttat agtagcaatg atattcctac atctattatt tataggatag 2880
tagatgataa tttgcctaaa tttctagaaa ataaagctaa gtatgagagt ttaaaagaca 2940
aagctccaga agctataaac tatgaacaaa ttaaaaaaga tttggcagaa gagctaacct 3000
ttgatattga ctacaaaaca tctgaagtta atcaaagagt tttttcactt gatgaagttt 3060
ttgagatagc aaactttaat aattatctaa atcaaagtgg tattactaaa tttaatacta 3120
ttattggtgg taaatttgta aatggtgaaa atacaaagag aaaaggtata aatgaatata 3180
taaatctata ctcacagcaa ataaatgata aaacactcaa aaaatataaa atgagtgttt 3240
tatttaagca aattttaagt gatacagaat ctaaatcttt tgtaattgat aagttagaag 3300
atgatagtga tgtagttaca acgatgcaaa gtttttatga gcaaatagca gcttttaaaa 3360
cagtagaaga aaaatctatt aaagaaacac tatctttatt atttgatgat ttaaaagctc 3420
aaaaacttga tttgagtaaa atttatttta aaaatgataa atctcttact gatctatcac 3480
aacaagtttt tgatgattat agtgttattg gtacagcggt actagaatat ataactcaac 3540
aaatagcacc taaaaatctt gataacccta gtaagaaaga gcaagaatta atagccaaaa 3600
aaactgaaaa agcaaaatac ttatctctag aaactataaa gcttgcctta gaagaattta 3660
ataagcatag agatatagat aaacagtgta ggtttgaaga aatacttgca aactttgcgg 3720
ctattccgat gatatttgat gaaatagctc aaaacaaaga caatttggca cagatatcta 3780
tcaaatatca aaatcaaggt aaaaaagacc tacttcaagc tagtgcggaa gatgatgtta 3840
aagctatcaa ggatctttta gatcaaacta ataatctctt acataaacta aaaatatttc 3900
atattagtca gtcagaagat aaggcaaata ttttagacaa ggatgagcat ttttatctag 3960
tatttgagga gtgctacttt gagctagcga atatagtgcc tctttataac aaaattagaa 4020
actatataac tcaaaagcca tatagtgatg agaaatttaa gctcaatttt gagaactcga 4080
ctttggctaa tggttgggat aaaaataaag agcctgacaa tacggcaatt ttatttatca 4140
aagatgataa atattatctg ggtgtgatga ataagaaaaa taacaaaata tttgatgata 4200
aagctatcaa agaaaataaa ggcgagggtt ataaaaaaat tgtttataaa cttttacctg 4260
gcgcaaataa aatgttacct aaggttttct tttctgctaa atctataaaa ttttataatc 4320
ctagtgaaga tatacttaga ataagaaatc attccacaca tacaaaaaat ggtagtcctc 4380
aaaaaggata tgaaaaattt gagtttaata ttgaagattg ccgaaaattt atagattttt 4440
ataaacagtc tataagtaag catccggagt ggaaagattt tggatttaga ttttctgata 4500
ctcaaagata taattctata gatgaatttt atagagaagt tgaaaatcaa ggctacaaac 4560
taacttttga aaatatatca gagagctata ttgatagcgt agttaatcag ggtaaattgt 4620
acctattcca aatctataat aaagattttt cagcttatag caaagggcga ccaaatctac 4680
atactttata ttggaaagcg ctgtttgatg agagaaatct tcaagatgtg gtttataagc 4740
taaatggtga ggcagagctt ttttatcgta aacaatcaat acctaaaaaa atcactcacc 4800
cagctaaaga ggcaatagct aataaaaaca aagataatcc taaaaaagag agtgtttttg 4860
aatatgattt aatcaaagat aaacgcttta ctgaagataa gtttttcttt cactgtccta 4920
ttacaatcaa ttttaaatct agtggagcta ataagtttaa tgatgaaatc aatttattgc 4980
taaaagaaaa agcaaatgat gttcatatat taagtatagc tagaggtgaa agacatttag 5040
cttactatac tttggtagat ggtaaaggca atatcatcaa acaagatact ttcaacatca 5100
ttggtaatga tagaatgaaa acaaactacc atgataagct tgctgcaata gagaaagata 5160
gggattcagc taggaaagac tggaaaaaga taaataacat caaagagatg aaagagggct 5220
atctatctca ggtagttcat gaaatagcta agctagttat agagtataat gctattgtgg 5280
tttttgagga tttaaatttt ggatttaaaa gagggcgttt caaggtagag aagcaggtct 5340
atcaaaagtt agaaaaaatg ctaattgaga aactaaacta tctagttttc aaagataatg 5400
agtttgataa aactggggga gtgcttagag cttatcagct aacagcacct tttgagactt 5460
ttaaaaagat gggtaaacaa acaggtatta tctactatgt accagctggt tttacttcaa 5520
aaatttgtcc tgtaactggt tttgtaaatc agttatatcc taagtatgaa agtgtcagca 5580
aatctcaaga gttctttagt aagtttgaca agatttgtta taaccttgat aagggctatt 5640
ttgagtttag ttttgattat aaaaactttg gtgacaaggc tgccaaaggc aagtggacta 5700
tagctagctt tgggagtaga ttgattaact ttagaaattc agataaaaat cataattggg 5760
atactcgaga agtttatcca actaaagagt tggagaaatt gctaaaagat tattctatcg 5820
aatatgggca tggcgaatgt atcaaagcag ctatttgcgg tgagagcgac aaaaagtttt 5880
ttgctaagct aactagtgtc ctaaatacta tcttacaaat gcgtaactca aaaacaggta 5940
ctgagttaga ttatctaatt tcaccagtag cagatgtaaa tggcaatttc tttgattcgc 6000
gacaggcgcc aaaaaatatg cctcaagatg ctgatgccaa tggtgcttat catattgggc 6060
taaaaggtct gatgctacta ggtaggatca aaaataatca agagggcaaa aaactcaatt 6120
tggttatcaa aaatgaagag tattttgagt tcgtgcagaa taggaataac ggctctccta 6180
aaaagaaacg taaagtttct ggcggctcta cgaatcttag cgacatcatc gagaaggaaa 6240
cgggcaaaca gcttgtcatc caagaaagca ttcttatgct gccggaagag gtcgaagagg 6300
tcatcggcaa caaaccggaa agcgacattc ttgtccatac agcctacgat gagagcacgg 6360
atgagaacgt catgcttctg acgagcgatg ccccggaata taaaccgtgg gctcttgtca 6420
ttcaagatag caacggcgag aataagatca aaatgctttc tggtggttct cccaagaaga 6480
agaggaaagt ctaactgcag tataatcaga aacagcccgc ggatgttgat ctgcgggctg 6540
ttttttattg atcgaatggc catgaccaaa atcccttaac gtgagttttc gttccactga 6600
gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 6660
atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 6720
gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 6780
gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 6840
tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 6900
accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 6960
ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 7020
cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 7080
agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 7140
ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 7200
tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc 7260
ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac 7320
cgtattaccg cctttgagtg agctgagaca ggtcattcag actggctaat gcacccagta 7380
aggcagcggt atcatcaact caaaatggta tgcgttttga cacatccact atatatccgt 7440
gtcgttctgt ccactcctga atcccattcc agaaattctc tagcgattcc agaagtttct 7500
cagagtcgga aagttgacca gacattacga actggcacag atggtcataa cctgaaggaa 7560
gatctgattg cttaactgct tcagttaaga ccgaagcgct cgtcgtataa cagatgcgat 7620
gatgcagacc aatcaacatg gcacctgcca ttgctacctg cacagtcaag gatggtagaa 7680
atgttgtcgg tccttgcaca cgaatattac gccatttgcc tgcatattca aacagctctt 7740
ctacgataag ggcacaaatc gcatcgtgga acgtttgggc ttctaccgat ttagcagttt 7800
gatacacttt ctctaagtat ccacctgaat cataaatcgg caaaatagag aaaaattgac 7860
catgtgtaag cggccaatct gattccacct gagatgcata atctagtaga atctcttcgc 7920
tatcaaaatt cacttccacc ttccactcac cggttgtcca ttcatggctg aactctgctt 7980
cctctgttga catgacacac atcatctcaa tatccgaata gggcccatca gtctgacgac 8040
caagagagcc ataaacacca atagccttaa catcatcccc atatttatcc aatattcgtt 8100
ccttaatttc atgaacaatc ttcattcttt cttctctagt cattattatt ggtccattca 8160
ctattctcat tcccctttca gataatttta gatttgcttt tctaaataag aatatttgga 8220
gagcaccgtt cttattcagc tattaaaccc attatatcgg gtttttgagg ggatttcaac 8280
tgcagacacc taaattcaaa atctatcggt cagatttata ccgatttgat tttatatatt 8340
cttgaataac atacgccgag ttatcacata aaagcgggaa ccaatcatca aatttaaact 8400
tcattgcata atccattaaa ctcttaaatt ctacgattcc ttgttcatca ataaactcaa 8460
tcatttcttt aattaattta tatctatctg ttgttgtttt ctttaataat tcatcaacat 8520
ctacaccgcc ataaactatc atatcttctt tttgatattt aaatttatta ggatcgtcca 8580
tgtgaagcat atatctcaca agacctttca cacttcctgc aatctgcgga atagtcgcat 8640
tcaattcttc tgtaattatt tttatctgtt cataagattt attaccctca tacatcacta 8700
gaatatgata atgctctttt ttcatcctat cttctgtatc agtatcccta tcatgtaatg 8760
gagacactac aaattgaatg tgtaactctt ttaaatactc taaccactcg gcttttgctg 8820
attctggata taaaacaaat gtccaattac gtcctcttga atttttcttg ttttcagttt 8880
cttttattac attttcgctc atgatataat aacggtgcta atacatttaa caaaatttag 8940
tcatagatag gcagcatgcc agtgctgtct atcttttttt gtttaaaatg caccgtattc 9000
ctcctttgca tattttttta ttagaatacc ggttgcatct gatttgctaa tattatattt 9060
ttctttgatt ctatttaata tctcattttc ttctgttgta agtcttaaag taacagcaac 9120
ttttttctct tcttttctat ctacaaccat cactgtacct cccaacatct gtttttttca 9180
ctttaacata aaaaacaacc ttttaacatt aaaaacccaa tatttattta tttgtttgga 9240
caatggacaa tggacaccta ggggggaggt cgtagtaccc ccctatgttt tctcccctaa 9300
ataaccccaa aaatctaaga aaaaaagacc tcaaaaaggt ctttaattaa catctcaaat 9360
ttcgcattta ttccaatttc ctttttgcgt gtgatgcgtt attaacgttg atataattta 9420
aattttattt gacaaaaatg ggctcgtgtt gtacaataaa tgtagaggta gagacgcgag 9480
gtctaagaac tttaaataat ttctactgtt gtagatagag accgtgaagt taataaggtc 9540
tcaaatttct actgttgtag atcgtctctg aactgattca agcaagctta aacccagctc 9600
aatgagctgg gttttttgtt tgttttttca aacttagtta gcttggccag tgcctctaga 9660
gtcaagtaaa gagtcgacct gttacgaacg gcagatcaga attttgtaat aaaaaaagag 9720
cctgcttatt acactgcggg ctctttttca tggtcagaag acgggtaacc aagataacaa 9780
<210> 8
<211> 9768
<212> DNA
<213> (人工序列)
<400> 8
atgtcatgac attggtgtac agaaatggcg cagcaatggc aagaacgtcc cgggcggagc 60
tcaggcctta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 120
tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 180
ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc 240
gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa 300
tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat 360
cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg 420
cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc 480
atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc 540
ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag 600
acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc 660
tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg 720
tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca 780
tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg 840
tgcaccgccg ttttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg 900
gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg 960
gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc 1020
acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc 1080
gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca 1140
tactctgcga catcgtataa cgttactggt ttcatcaaaa tcgtctccct ccgtttgaat 1200
atttgattga tcgtaaccag atgaagcact ctttccacta tccctacagt gttatggctt 1260
gaacaatcac gaaacaataa ttggtacgta cgatctttca gccgactcaa acatcaaatc 1320
ttacaaatgt agtctttgaa agtattacat atgtaagatt taaatgcaac cgttttttcg 1380
gaaggaaatg atgacctcgt ttccaccgga attagcttgg taccagctat tgtaacataa 1440
tcggtacggg ggtgaaaaag ctaacggaaa agggagcgga aaagaatgat gtaagcgtga 1500
aaaatttttt aaaaaatctc ttgacattgg aagggagata tgttattata agaattgcgg 1560
aattgtgagc ggataacaat tcataattgt gagcggataa caattcaacc ccaaaggagg 1620
tgggatccat ggattcactt cttatgaaca gaagagaatt tctttatcag tttaagaatg 1680
tgagatgggc aaagggtcgg cgagaaacat atctttgtta tgttgttaag cgccgcgatt 1740
cagcaacatc attcagcttg gatttcggct accttagaaa caagaatggt tgtcatgttg 1800
aacttctgtt cctaaggtat atatcagatt gggatcttga tcctggaaga tgttatagag 1860
ttacatggtt tatatcatgg tcaccttgtt atgattgtgc aagacatgtt gcagatttcc 1920
tgcgcggaaa ccctaacctt tcacttagaa tatttacagc aagactttat ttctgcgagg 1980
atagaaaggc ggagcctgaa ggacttagaa gacttcatag agctggcgtt cagattgcta 2040
tcatgacgtt caaggactac ttctactgtt ggaacacatt tgttgagaat cacggaagaa 2100
catttaaagc atgggaagga cttcatgaga attccgttag actttcaaga cagcttagaa 2160
gaatacttct tcctctttat gaagttgatg atcttagaga tgcatttaga acatctggct 2220
ctgaaacacc tggcacatct gaatctgcta cacctgaatc ttcaatttat caagaatttg 2280
ttaataaata tagtttaagt aaaactctaa gatttgagtt aatcccacag ggtaaaacac 2340
ttgaaaacat aaaagcaaga ggtttgattt tagatgatga gaaaagagct aaagactaca 2400
aaaaggctaa acaaataatt gataaatatc atcagttttt tatagaggag atattaagtt 2460
cggtttgtat tagcgaagat ttattacaaa actattctga tgtttatttt aaacttaaaa 2520
agagtgatga tgataatcta caaaaagatt ttaaaagtgc aaaagatacg ataaagaaac 2580
aaatatctga atatataaag gactcagaga aatttaagaa tttgtttaat caaaacctta 2640
tcgatgctaa aaaagggcaa gagtcagatt taattctatg gctaaagcaa tctaaggata 2700
atggtataga actatttaaa gccaatagtg atatcacaga tatagatgag gcgttagaaa 2760
taatcaaatc ttttaaaggt tggacaactt attttaaggg ttttcatgaa aatagaaaaa 2820
atgtttatag tagcaatgat attcctacat ctattattta taggatagta gatgataatt 2880
tgcctaaatt tctagaaaat aaagctaagt atgagagttt aaaagacaaa gctccagaag 2940
ctataaacta tgaacaaatt aaaaaagatt tggcagaaga gctaaccttt gatattgact 3000
acaaaacatc tgaagttaat caaagagttt tttcacttga tgaagttttt gagatagcaa 3060
actttaataa ttatctaaat caaagtggta ttactaaatt taatactatt attggtggta 3120
aatttgtaaa tggtgaaaat acaaagagaa aaggtataaa tgaatatata aatctatact 3180
cacagcaaat aaatgataaa acactcaaaa aatataaaat gagtgtttta tttaagcaaa 3240
ttttaagtga tacagaatct aaatcttttg taattgataa gttagaagat gatagtgatg 3300
tagttacaac gatgcaaagt ttttatgagc aaatagcagc ttttaaaaca gtagaagaaa 3360
aatctattaa agaaacacta tctttattat ttgatgattt aaaagctcaa aaacttgatt 3420
tgagtaaaat ttattttaaa aatgataaat ctcttactga tctatcacaa caagtttttg 3480
atgattatag tgttattggt acagcggtac tagaatatat aactcaacaa atagcaccta 3540
aaaatcttga taaccctagt aagaaagagc aagaattaat agccaaaaaa actgaaaaag 3600
caaaatactt atctctagaa actataaagc ttgccttaga agaatttaat aagcatagag 3660
atatagataa acagtgtagg tttgaagaaa tacttgcaaa ctttgcggct attccgatga 3720
tatttgatga aatagctcaa aacaaagaca atttggcaca gatatctatc aaatatcaaa 3780
atcaaggtaa aaaagaccta cttcaagcta gtgcggaaga tgatgttaaa gctatcaagg 3840
atcttttaga tcaaactaat aatctcttac ataaactaaa aatatttcat attagtcagt 3900
cagaagataa ggcaaatatt ttagacaagg atgagcattt ttatctagta tttgaggagt 3960
gctactttga gctagcgaat atagtgcctc tttataacaa aattagaaac tatataactc 4020
aaaagccata tagtgatgag aaatttaagc tcaattttga gaactcgact ttggctaatg 4080
gttgggataa aaataaagag cctgacaata cggcaatttt atttatcaaa gatgataaat 4140
attatctggg tgtgatgaat aagaaaaata acaaaatatt tgatgataaa gctatcaaag 4200
aaaataaagg cgagggttat aaaaaaattg tttataaact tttacctggc gcaaataaaa 4260
tgttacctaa ggttttcttt tctgctaaat ctataaaatt ttataatcct agtgaagata 4320
tacttagaat aagaaatcat tccacacata caaaaaatgg tagtcctcaa aaaggatatg 4380
aaaaatttga gtttaatatt gaagattgcc gaaaatttat agatttttat aaacagtcta 4440
taagtaagca tccggagtgg aaagattttg gatttagatt ttctgatact caaagatata 4500
attctataga tgaattttat agagaagttg aaaatcaagg ctacaaacta acttttgaaa 4560
atatatcaga gagctatatt gatagcgtag ttaatcaggg taaattgtac ctattccaaa 4620
tctataataa agatttttca gcttatagca aagggcgacc aaatctacat actttatatt 4680
ggaaagcgct gtttgatgag agaaatcttc aagatgtggt ttataagcta aatggtgagg 4740
cagagctttt ttatcgtaaa caatcaatac ctaaaaaaat cactcaccca gctaaagagg 4800
caatagctaa taaaaacaaa gataatccta aaaaagagag tgtttttgaa tatgatttaa 4860
tcaaagataa acgctttact gaagataagt ttttctttca ctgtcctatt acaatcaatt 4920
ttaaatctag tggagctaat aagtttaatg atgaaatcaa tttattgcta aaagaaaaag 4980
caaatgatgt tcatatatta agtatagcta gaggtgaaag acatttagct tactatactt 5040
tggtagatgg taaaggcaat atcatcaaac aagatacttt caacatcatt ggtaatgata 5100
gaatgaaaac aaactaccat gataagcttg ctgcaataga gaaagatagg gattcagcta 5160
ggaaagactg gaaaaagata aataacatca aagagatgaa agagggctat ctatctcagg 5220
tagttcatga aatagctaag ctagttatag agtataatgc tattgtggtt tttgaggatt 5280
taaattttgg atttaaaaga gggcgtttca aggtagagaa gcaggtctat caaaagttag 5340
aaaaaatgct aattgagaaa ctaaactatc tagttttcaa agataatgag tttgataaaa 5400
ctgggggagt gcttagagct tatcagctaa cagcaccttt tgagactttt aaaaagatgg 5460
gtaaacaaac aggtattatc tactatgtac cagctggttt tacttcaaaa atttgtcctg 5520
taactggttt tgtaaatcag ttatatccta agtatgaaag tgtcagcaaa tctcaagagt 5580
tctttagtaa gtttgacaag atttgttata accttgataa gggctatttt gagtttagtt 5640
ttgattataa aaactttggt gacaaggctg ccaaaggcaa gtggactata gctagctttg 5700
ggagtagatt gattaacttt agaaattcag ataaaaatca taattgggat actcgagaag 5760
tttatccaac taaagagttg gagaaattgc taaaagatta ttctatcgaa tatgggcatg 5820
gcgaatgtat caaagcagct atttgcggtg agagcgacaa aaagtttttt gctaagctaa 5880
ctagtgtcct aaatactatc ttacaaatgc gtaactcaaa aacaggtact gagttagatt 5940
atctaatttc accagtagca gatgtaaatg gcaatttctt tgattcgcga caggcgccaa 6000
aaaatatgcc tcaagatgct gatgccaatg gtgcttatca tattgggcta aaaggtctga 6060
tgctactagg taggatcaaa aataatcaag agggcaaaaa actcaatttg gttatcaaaa 6120
atgaagagta ttttgagttc gtgcagaata ggaataacgg ctctcctaaa aagaaacgta 6180
aagtttctgg cggctctacg aatcttagcg acatcatcga gaaggaaacg ggcaaacagc 6240
ttgtcatcca agaaagcatt cttatgctgc cggaagaggt cgaagaggtc atcggcaaca 6300
aaccggaaag cgacattctt gtccatacag cctacgatga gagcacggat gagaacgtca 6360
tgcttctgac gagcgatgcc ccggaatata aaccgtgggc tcttgtcatt caagatagca 6420
acggcgagaa taagatcaaa atgctttctg gtggttctcc caagaagaag aggaaagtct 6480
aactgcagta taatcagaaa cagcccgcgg atgttgatct gcgggctgtt ttttattgat 6540
cgaatggcca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc 6600
gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 6660
caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 6720
ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg 6780
tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg 6840
ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac 6900
tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca 6960
cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga 7020
gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc 7080
ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct 7140
gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg 7200
agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct 7260
tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc 7320
tttgagtgag ctgagacagg tcattcagac tggctaatgc acccagtaag gcagcggtat 7380
catcaactca aaatggtatg cgttttgaca catccactat atatccgtgt cgttctgtcc 7440
actcctgaat cccattccag aaattctcta gcgattccag aagtttctca gagtcggaaa 7500
gttgaccaga cattacgaac tggcacagat ggtcataacc tgaaggaaga tctgattgct 7560
taactgcttc agttaagacc gaagcgctcg tcgtataaca gatgcgatga tgcagaccaa 7620
tcaacatggc acctgccatt gctacctgca cagtcaagga tggtagaaat gttgtcggtc 7680
cttgcacacg aatattacgc catttgcctg catattcaaa cagctcttct acgataaggg 7740
cacaaatcgc atcgtggaac gtttgggctt ctaccgattt agcagtttga tacactttct 7800
ctaagtatcc acctgaatca taaatcggca aaatagagaa aaattgacca tgtgtaagcg 7860
gccaatctga ttccacctga gatgcataat ctagtagaat ctcttcgcta tcaaaattca 7920
cttccacctt ccactcaccg gttgtccatt catggctgaa ctctgcttcc tctgttgaca 7980
tgacacacat catctcaata tccgaatagg gcccatcagt ctgacgacca agagagccat 8040
aaacaccaat agccttaaca tcatccccat atttatccaa tattcgttcc ttaatttcat 8100
gaacaatctt cattctttct tctctagtca ttattattgg tccattcact attctcattc 8160
ccctttcaga taattttaga tttgcttttc taaataagaa tatttggaga gcaccgttct 8220
tattcagcta ttaaacccat tatatcgggt ttttgagggg atttcaactg cagacaccta 8280
aattcaaaat ctatcggtca gatttatacc gatttgattt tatatattct tgaataacat 8340
acgccgagtt atcacataaa agcgggaacc aatcatcaaa tttaaacttc attgcataat 8400
ccattaaact cttaaattct acgattcctt gttcatcaat aaactcaatc atttctttaa 8460
ttaatttata tctatctgtt gttgttttct ttaataattc atcaacatct acaccgccat 8520
aaactatcat atcttctttt tgatatttaa atttattagg atcgtccatg tgaagcatat 8580
atctcacaag acctttcaca cttcctgcaa tctgcggaat agtcgcattc aattcttctg 8640
taattatttt tatctgttca taagatttat taccctcata catcactaga atatgataat 8700
gctctttttt catcctatct tctgtatcag tatccctatc atgtaatgga gacactacaa 8760
attgaatgtg taactctttt aaatactcta accactcggc ttttgctgat tctggatata 8820
aaacaaatgt ccaattacgt cctcttgaat ttttcttgtt ttcagtttct tttattacat 8880
tttcgctcat gatataataa cggtgctaat acatttaaca aaatttagtc atagataggc 8940
agcatgccag tgctgtctat ctttttttgt ttaaaatgca ccgtattcct cctttgcata 9000
tttttttatt agaataccgg ttgcatctga tttgctaata ttatattttt ctttgattct 9060
atttaatatc tcattttctt ctgttgtaag tcttaaagta acagcaactt ttttctcttc 9120
ttttctatct acaaccatca ctgtacctcc caacatctgt ttttttcact ttaacataaa 9180
aaacaacctt ttaacattaa aaacccaata tttatttatt tgtttggaca atggacaatg 9240
gacacctagg ggggaggtcg tagtaccccc ctatgttttc tcccctaaat aaccccaaaa 9300
atctaagaaa aaaagacctc aaaaaggtct ttaattaaca tctcaaattt cgcatttatt 9360
ccaatttcct ttttgcgtgt gatgcgttat taacgttgat ataatttaaa ttttatttga 9420
caaaaatggg ctcgtgttgt acaataaatg tagaggtaga gacgcgaggt ctaagaactt 9480
taaataattt ctactgttgt agatagagac cgtgaagtta ataaggtctc aaatttctac 9540
tgttgtagat cgtctctgaa ctgattcaag caagcttaaa cccagctcaa tgagctgggt 9600
tttttgtttg ttttttcaaa cttagttagc ttggccagtg cctctagagt caagtaaaga 9660
gtcgacctgt tacgaacggc agatcagaat tttgtaataa aaaaagagcc tgctcattac 9720
actgcgggct ctttttcatg gtcagaagac gggtaaccaa gataacaa 9768
<210> 9
<211> 9987
<212> DNA
<213> (人工序列)
<400> 9
atgtcatgac attggtgtac agaaatggcg cagcaatggc aagaacgtcc cgggcggagc 60
tcaggcctta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 120
tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 180
ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc 240
gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa 300
tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat 360
cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg 420
cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc 480
atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc 540
ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag 600
acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc 660
tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg 720
tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca 780
tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg 840
tgcaccgccg ttttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg 900
gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg 960
gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc 1020
acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc 1080
gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca 1140
tactctgcga catcgtataa cgttactggt ttcatcaaaa tcgtctccct ccgtttgaat 1200
atttgattga tcgtaaccag atgaagcact ctttccacta tccctacagt gttatggctt 1260
gaacaatcac gaaacaataa ttggtacgta cgatctttca gccgactcaa acatcaaatc 1320
ttacaaatgt agtctttgaa agtattacat atgtaagatt taaatgcaac cgttttttcg 1380
gaaggaaatg atgacctcgt ttccaccgga attagcttgg taccagctat tgtaacataa 1440
tcggtacggg ggtgaaaaag ctaacggaaa agggagcgga aaagaatgat gtaagcgtga 1500
aaaatttttt aaaaaatctc ttgacattgg aagggagata tgttattata agaattgcgg 1560
aattgtgagc ggataacaat tcataattgt gagcggataa caattcaacc ccaaaggagg 1620
tgggatccat gtcagttttt ctgtataaaa aactgccgct ggatacattt ctgtttgaat 1680
ttaataatct ggaatgggca tatggcagaa attcatgcta tatttgcttt aaactgaaac 1740
cgattctggc aacatcaaca acgggcacaa cagaatcaga actgtggggc tatgttacaa 1800
ataaaggcga agttgcagat ggcattccgt gcgaatcacc ggaaaaaaga ggcatgcatg 1860
cagaagaact gcttctggaa gaaatgacaa gacatgttag agaacatggc ggcgcatcat 1920
tttgcgttga atggtttgca tcatggtcac cgtgcgatag atgcagcggc gttctgctga 1980
gatggctgag agatgttggc ggaggcagac atagactgag agtttggttt tcaagaattt 2040
atagaggcaa tgttggcgca gttagagcgg gcctgagaca tctgagaaga gcgggcgttc 2100
aactgggcgt tatggataga cgcagacatg ataattgcgt tcatgttctg gttgatgcag 2160
caagaggcga tccgacaccg ctgtggctgg ttcaatggca ttcaaatgtt ccgagagttc 2220
aaagagaatt tgataaaatt atggatgaaa aagttagaga accgggctgg tgctggtggt 2280
gctggcgccc gtggatggca gatgttccgt caaatgcatc atttccgcaa ctgccggtta 2340
atgatggcgc aattgttgat agagttgaaa aatttccgta tctgggctca ctgctgatga 2400
catcaagcgg cctggcagca gaagtttcaa tttctggctc tgaaacacct ggcacatctg 2460
aatctgctac acctgaatct tcaatttatc aagaatttgt taataaatat agtttaagta 2520
aaactctaag atttgagtta atcccacagg gtaaaacact tgaaaacata aaagcaagag 2580
gtttgatttt agatgatgag aaaagagcta aagactacaa aaaggctaaa caaataattg 2640
ataaatatca tcagtttttt atagaggaga tattaagttc ggtttgtatt agcgaagatt 2700
tattacaaaa ctattctgat gtttatttta aacttaaaaa gagtgatgat gataatctac 2760
aaaaagattt taaaagtgca aaagatacga taaagaaaca aatatctgaa tatataaagg 2820
actcagagaa atttaagaat ttgtttaatc aaaaccttat cgatgctaaa aaagggcaag 2880
agtcagattt aattctatgg ctaaagcaat ctaaggataa tggtatagaa ctatttaaag 2940
ccaatagtga tatcacagat atagatgagg cgttagaaat aatcaaatct tttaaaggtt 3000
ggacaactta ttttaagggt tttcatgaaa atagaaaaaa tgtttatagt agcaatgata 3060
ttcctacatc tattatttat aggatagtag atgataattt gcctaaattt ctagaaaata 3120
aagctaagta tgagagttta aaagacaaag ctccagaagc tataaactat gaacaaatta 3180
aaaaagattt ggcagaagag ctaacctttg atattgacta caaaacatct gaagttaatc 3240
aaagagtttt ttcacttgat gaagtttttg agatagcaaa ctttaataat tatctaaatc 3300
aaagtggtat tactaaattt aatactatta ttggtggtaa atttgtaaat ggtgaaaata 3360
caaagagaaa aggtataaat gaatatataa atctatactc acagcaaata aatgataaaa 3420
cactcaaaaa atataaaatg agtgttttat ttaagcaaat tttaagtgat acagaatcta 3480
aatcttttgt aattgataag ttagaagatg atagtgatgt agttacaacg atgcaaagtt 3540
tttatgagca aatagcagct tttaaaacag tagaagaaaa atctattaaa gaaacactat 3600
ctttattatt tgatgattta aaagctcaaa aacttgattt gagtaaaatt tattttaaaa 3660
atgataaatc tcttactgat ctatcacaac aagtttttga tgattatagt gttattggta 3720
cagcggtact agaatatata actcaacaaa tagcacctaa aaatcttgat aaccctagta 3780
agaaagagca agaattaata gccaaaaaaa ctgaaaaagc aaaatactta tctctagaaa 3840
ctataaagct tgccttagaa gaatttaata agcatagaga tatagataaa cagtgtaggt 3900
ttgaagaaat acttgcaaac tttgcggcta ttccgatgat atttgatgaa atagctcaaa 3960
acaaagacaa tttggcacag atatctatca aatatcaaaa tcaaggtaaa aaagacctac 4020
ttcaagctag tgcggaagat gatgttaaag ctatcaagga tcttttagat caaactaata 4080
atctcttaca taaactaaaa atatttcata ttagtcagtc agaagataag gcaaatattt 4140
tagacaagga tgagcatttt tatctagtat ttgaggagtg ctactttgag ctagcgaata 4200
tagtgcctct ttataacaaa attagaaact atataactca aaagccatat agtgatgaga 4260
aatttaagct caattttgag aactcgactt tggctaatgg ttgggataaa aataaagagc 4320
ctgacaatac ggcaatttta tttatcaaag atgataaata ttatctgggt gtgatgaata 4380
agaaaaataa caaaatattt gatgataaag ctatcaaaga aaataaaggc gagggttata 4440
aaaaaattgt ttataaactt ttacctggcg caaataaaat gttacctaag gttttctttt 4500
ctgctaaatc tataaaattt tataatccta gtgaagatat acttagaata agaaatcatt 4560
ccacacatac aaaaaatggt agtcctcaaa aaggatatga aaaatttgag tttaatattg 4620
aagattgccg aaaatttata gatttttata aacagtctat aagtaagcat ccggagtgga 4680
aagattttgg atttagattt tctgatactc aaagatataa ttctatagat gaattttata 4740
gagaagttga aaatcaaggc tacaaactaa cttttgaaaa tatatcagag agctatattg 4800
atagcgtagt taatcagggt aaattgtacc tattccaaat ctataataaa gatttttcag 4860
cttatagcaa agggcgacca aatctacata ctttatattg gaaagcgctg tttgatgaga 4920
gaaatcttca agatgtggtt tataagctaa atggtgaggc agagcttttt tatcgtaaac 4980
aatcaatacc taaaaaaatc actcacccag ctaaagaggc aatagctaat aaaaacaaag 5040
ataatcctaa aaaagagagt gtttttgaat atgatttaat caaagataaa cgctttactg 5100
aagataagtt tttctttcac tgtcctatta caatcaattt taaatctagt ggagctaata 5160
agtttaatga tgaaatcaat ttattgctaa aagaaaaagc aaatgatgtt catatattaa 5220
gtatagctag aggtgaaaga catttagctt actatacttt ggtagatggt aaaggcaata 5280
tcatcaaaca agatactttc aacatcattg gtaatgatag aatgaaaaca aactaccatg 5340
ataagcttgc tgcaatagag aaagataggg attcagctag gaaagactgg aaaaagataa 5400
ataacatcaa agagatgaaa gagggctatc tatctcaggt agttcatgaa atagctaagc 5460
tagttataga gtataatgct attgtggttt ttgaggattt aaattttgga tttaaaagag 5520
ggcgtttcaa ggtagagaag caggtctatc aaaagttaga aaaaatgcta attgagaaac 5580
taaactatct agttttcaaa gataatgagt ttgataaaac tgggggagtg cttagagctt 5640
atcagctaac agcacctttt gagactttta aaaagatggg taaacaaaca ggtattatct 5700
actatgtacc agctggtttt acttcaaaaa tttgtcctgt aactggtttt gtaaatcagt 5760
tatatcctaa gtatgaaagt gtcagcaaat ctcaagagtt ctttagtaag tttgacaaga 5820
tttgttataa ccttgataag ggctattttg agtttagttt tgattataaa aactttggtg 5880
acaaggctgc caaaggcaag tggactatag ctagctttgg gagtagattg attaacttta 5940
gaaattcaga taaaaatcat aattgggata ctcgagaagt ttatccaact aaagagttgg 6000
agaaattgct aaaagattat tctatcgaat atgggcatgg cgaatgtatc aaagcagcta 6060
tttgcggtga gagcgacaaa aagttttttg ctaagctaac tagtgtccta aatactatct 6120
tacaaatgcg taactcaaaa acaggtactg agttagatta tctaatttca ccagtagcag 6180
atgtaaatgg caatttcttt gattcgcgac aggcgccaaa aaatatgcct caagatgctg 6240
atgccaatgg tgcttatcat attgggctaa aaggtctgat gctactaggt aggatcaaaa 6300
ataatcaaga gggcaaaaaa ctcaatttgg ttatcaaaaa tgaagagtat tttgagttcg 6360
tgcagaatag gaataacggc tctcctaaaa agaaacgtaa agtttctggc ggctctacga 6420
atcttagcga catcatcgag aaggaaacgg gcaaacagct tgtcatccaa gaaagcattc 6480
ttatgctgcc ggaagaggtc gaagaggtca tcggcaacaa accggaaagc gacattcttg 6540
tccatacagc ctacgatgag agcacggatg agaacgtcat gcttctgacg agcgatgccc 6600
cggaatataa accgtgggct cttgtcattc aagatagcaa cggcgagaat aagatcaaaa 6660
tgctttctgg tggttctccc aagaagaaga ggaaagtcta actgcagtat aatcagaaac 6720
agcccgcgga tgttgatctg cgggctgttt tttattgatc gaatggccat gaccaaaatc 6780
ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 6840
tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 6900
ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 6960
ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac 7020
ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 7080
gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 7140
aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 7200
acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 7260
gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 7320
gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 7380
cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 7440
aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct 7500
gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgagacaggt 7560
cattcagact ggctaatgca cccagtaagg cagcggtatc atcaactcaa aatggtatgc 7620
gttttgacac atccactata tatccgtgtc gttctgtcca ctcctgaatc ccattccaga 7680
aattctctag cgattccaga agtttctcag agtcggaaag ttgaccagac attacgaact 7740
ggcacagatg gtcataacct gaaggaagat ctgattgctt aactgcttca gttaagaccg 7800
aagcgctcgt cgtataacag atgcgatgat gcagaccaat caacatggca cctgccattg 7860
ctacctgcac agtcaaggat ggtagaaatg ttgtcggtcc ttgcacacga atattacgcc 7920
atttgcctgc atattcaaac agctcttcta cgataagggc acaaatcgca tcgtggaacg 7980
tttgggcttc taccgattta gcagtttgat acactttctc taagtatcca cctgaatcat 8040
aaatcggcaa aatagagaaa aattgaccat gtgtaagcgg ccaatctgat tccacctgag 8100
atgcataatc tagtagaatc tcttcgctat caaaattcac ttccaccttc cactcaccgg 8160
ttgtccattc atggctgaac tctgcttcct ctgttgacat gacacacatc atctcaatat 8220
ccgaataggg cccatcagtc tgacgaccaa gagagccata aacaccaata gccttaacat 8280
catccccata tttatccaat attcgttcct taatttcatg aacaatcttc attctttctt 8340
ctctagtcat tattattggt ccattcacta ttctcattcc cctttcagat aattttagat 8400
ttgcttttct aaataagaat atttggagag caccgttctt attcagctat taaacccatt 8460
atatcgggtt tttgagggga tttcaactgc agacacctaa attcaaaatc tatcggtcag 8520
atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa 8580
gcgggaacca atcatcaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta 8640
cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg 8700
ttgttttctt taataattca tcaacatcta caccgccata aactatcata tcttcttttt 8760
gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac 8820
ttcctgcaat ctgcggaata gtcgcattca attcttctgt aattattttt atctgttcat 8880
aagatttatt accctcatac atcactagaa tatgataatg ctcttttttc atcctatctt 8940
ctgtatcagt atccctatca tgtaatggag acactacaaa ttgaatgtgt aactctttta 9000
aatactctaa ccactcggct tttgctgatt ctggatataa aacaaatgtc caattacgtc 9060
ctcttgaatt tttcttgttt tcagtttctt ttattacatt ttcgctcatg atataataac 9120
ggtgctaata catttaacaa aatttagtca tagataggca gcatgccagt gctgtctatc 9180
tttttttgtt taaaatgcac cgtattcctc ctttgcatat ttttttatta gaataccggt 9240
tgcatctgat ttgctaatat tatatttttc tttgattcta tttaatatct cattttcttc 9300
tgttgtaagt cttaaagtaa cagcaacttt tttctcttct tttctatcta caaccatcac 9360
tgtacctccc aacatctgtt tttttcactt taacataaaa aacaaccttt taacattaaa 9420
aacccaatat ttatttattt gtttggacaa tggacaatgg acacctaggg gggaggtcgt 9480
agtacccccc tatgttttct cccctaaata accccaaaaa tctaagaaaa aaagacctca 9540
aaaaggtctt taattaacat ctcaaatttc gcatttattc caatttcctt tttgcgtgtg 9600
atgcgttatt aacgttgata taatttaaat tttatttgac aaaaatgggc tcgtgttgta 9660
caataaatgt agaggtagag acgcgaggtc taagaacttt aaataatttc tactgttgta 9720
gatagagacc gtgaagttaa taaggtctca aatttctact gttgtagatc gtctctgaac 9780
tgattcaagc aagcttaaac ccagctcaat gagctgggtt ttttgtttgt tttttcaaac 9840
ttagttagct tggccagtgc ctctagagtc aagtaaagag tcgacctgtt acgaacggca 9900
gatcagaatt ttgtaataaa aaaagagcct gctcattaca ctgcgggctc tttttcatgg 9960
tcagaagacg ggtaaccaag ataacaa 9987
Claims (8)
1.一种胞嘧啶碱基编辑***,其特征在于,所述胞嘧啶碱基编辑***包括氨基酸序列如SEQ ID NO.2所示的融合蛋白,以及核苷酸序列如SEQ ID NO.4所示的crRNA阵列***区。
2.根据权利要求1所述的胞嘧啶碱基编辑***,其特征在于:所述crRNA阵列***区通过组成型启动子P veg 调控表达。
3.根据权利要求1所述的胞嘧啶碱基编辑***,其特征在于:所述融合蛋白通过诱导型启动子P grac100 调控表达。
4.根据权利要求1所述的胞嘧啶碱基编辑***,其特征在于:所述胞嘧啶碱基编辑***通过将编码SEQ ID NO.2所示序列的基因以及crRNA阵列***区整合到表达载体上得到。
5.一种枯草芽孢杆菌胞嘧啶碱基编辑***,其特征在于,所述枯草芽孢杆菌胞嘧啶碱基编辑***包括:含有pE194复制子、编码权利要求1所述融合蛋白的基因以及crRNA阵列***区的质粒。
6.根据权利要求5所述的枯草芽孢杆菌胞嘧啶碱基编辑***,其特征在于:所述质粒的核苷酸序列如SEQ ID NO.8所示。
7.权利要求1-4任一项所述的胞嘧啶碱基编辑***或权利要求5或6所述的枯草芽孢杆菌胞嘧啶碱基编辑***在枯草芽孢杆菌基因编辑中的应用。
8.根据权利要求7所述的应用,其特征在于:所述的基因编辑为将多位点的胞嘧啶转化为胸腺嘧啶。
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