CN114045302A - 一种单碱基编辑载体及其构建和应用 - Google Patents
一种单碱基编辑载体及其构建和应用 Download PDFInfo
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Abstract
本发明属于生物技术领域，公开了一种包含dFnCpf1序列和crRNA序列的单碱基编辑载体及其构建和应用。与现有的采用包含Cas9序列的编辑载体相比，本发明的单碱基编辑载体在目标基因靶位点的5’端识别富含T的PAM[5’‑(T)TTN‑3’]序列，可以在腺嘌呤/胸腺嘧啶富集区域进行编辑操作。
Description
技术领域
本发明涉及生物技术领域，具体涉及一种单碱基编辑载体及其构建和应用。
背景技术
传统育种中利用的遗传变异主要来自于自然突变、物理或化学诱变，存在变异概率低、周期长、位点不可控等缺点。成簇规律间隔短回文重复序列(clustered regularlyinterspaced short palindromic repeats/CRISPR associated proteins，CRISPR/Cas)系统作为第三代基因编辑技术，由单链引导RNA(single-guide RNA,sgRNA)与切割靶序列的Cas内切核酸酶组成，其主要依赖sgRNA引导核酸内切酶在目标基因组位置产生双链断裂(Double-strand break,DSB)，而DSB可以通过非同源末端连接(non-homologous endjoining,NHEJ)或同源重组(homology recombination,HR)2种方式进行修复，修复过程中会引起靶标位置核苷酸序列的缺失、插入或者替换，从而实现基因编辑。
为满足不同的编辑目的，研究人员以CRISPR/Cas系统为基础，通过融合表达Cas9突变蛋白、胞嘧啶脱氨酶或人工进化的腺嘌呤脱氨酶，开发出能够对靶位点进行精准单碱基编辑的系统。该系统在不引起DNA双链断裂的情况下，实现胞嘧啶(cytosine,C)转化胸腺嘧啶(thymine,T)/鸟嘌呤(guanine,G)转化腺嘌呤(adenine,A)的替换且通过不断改进大大提高单碱基编辑效率，减少插入和删除(Insertion and deletion,indel)和非预期突变。该系统已成功在小麦(Triticumaestivum)、水稻(Oryza sativa)、棉花(Gossypium)、玉米(Zea mays)等物种上实现安全高效的单碱基替换编辑。但目前该系统仅利用nCas9蛋白突变体(Cas9 nicknase,nCas9)或dCas9蛋白突变体(deactivated Cas9,dCas9)作为效应蛋白，所识别的PAM序列为鸟嘌呤/胞嘧啶富集区。因此利用现有的碱基编辑系统无法在腺嘌呤/胸腺嘧啶富集区域进行编辑操作。
发明内容
本发明针对目前亟需一种可以在腺嘌呤/胸腺嘧啶富集区域进行编辑操作的系统，提供了一种单碱基编辑载体及其构建和应用。
为此，本发明一方面提供了一种单碱基编辑载体，其包含dFnCpf1序列和crRNA序列。
在本发明优选的实施方案中，所述的单碱基编辑载体为dFnCpf1-CBE-BT2，载体结构如图2B所示。
本发明另一方面提供了包含本发明所述的单碱基编辑载体的宿主细胞。
在本发明优选的实施方案中，所述的宿主细胞为玉米原生质体。
本发明另一方面提供了一种单碱基编辑载体的构建方法，其包括如下步骤：
1、构建包含dFnCpf1序列的载体；
2、合成包含crRNA序列的表达框；
3、构建包含dFnCpf1序列和crRNA序列的单碱基编辑载体。
在本发明优选的实施方案中，所述的单碱基编辑载体的构建方法，其包括如下步骤：
1、以PB-nCas9-PBE载体为骨架，切除nCas9序列，以pUC57-dFnCpf1载体为模板，扩增dFnCpf1序列，将dFnCpf1序列连入线性化的PB-nCas9-PBE载体，构建dFnCpf1-PBE载体，并切除OsU3-sgRNA-scaffold表达框；
2、选择OsU6启动子，mature-crRNA序列，合成OsU6-crRNA-polyT表达框，并放入pUC57-simple载体；
3、设计包含玉米bt2基因靶序列及同源臂的引物，以OsU6-crRNA-polyT载体为模板进行扩增，连入经酶切线性化的dFnCpf1-PBE载体，构建dFnCpf1-CBE-BT2载体。
本发明另一方面提供了本发明所述的单碱基编辑载体在对靶序列上特定碱基进行定向替换中的应用，其能够识别TTN作为原型间隔序列毗邻基序(PAM)，并对靶位点进行单碱基编辑。
在本发明优选的实施方案中，所述单碱基编辑发生在靶位点5’端8-12bp的编辑框内，单碱基编辑类型是将鸟嘌呤(G)转换为胸腺嘧啶(T)，或者将胞嘧啶(C)转换为腺嘌呤(A)。
在本发明更加优选的实施方案中，所述的单碱基编辑为将靶位点5’端第11位碱基的胞嘧啶(C)转换为腺嘌呤(A)。
本发明最后一方面提供了采用本发明所述的单碱基编辑载体转化的植物体，其中优选为玉米，更加优选为玉米原生质体。
由上述描述可知，与现有技术采用的Cas9突变蛋白相比，本发明采用了FnCpf1蛋白，其属于Cpf1(CRISPR from Prevotella and Francisella 1,Cpf1)。虽然与Cas9同属于Class2蛋白家族，但其具有如下的效果和优势。
1、Cas9需要CRISPR衍生的RNA(CRISPR derived RNA,crRNA)与反式激活RNA(trans-activating RNA,tracrRNA)靶向DNA，而FnCpf1仅需crRNA作为向导，且FnCpf1具有加工crRNA的能力。
2、Cas9及其直系同源蛋白在靶位点3’端识别富含G的PAM(5’-NGG-3’)，而FnCpf1以及其直系同源蛋白在靶位点的5’端识别富含T的PAM[5’-(T)TTN-3’]，可以在腺嘌呤/胸腺嘧啶富集区域进行编辑操作。
3、FnCpf1仅具有保守的RuvC核酸酶结构域，而Cas9具有HNH结构域与RuvC结构域。
4、FnCpf1在目标DNA中产生交错的末端断裂，而Cas9介导的双链断裂切口为平末端。
5、CRISPR/FnCpf1系统的脱靶效率低于CRISPR/Cas9系统。
附图说明
图1：dFnCpf1氨基酸序列。
图2：靶位点表达元件以及dFnCpf1-CBE-BT2载体结构。
2A：OsU6-crRNA表达框序列；
2B：dFnCpf1-CBE-BT2载体结构。
图3：蓝白斑筛选鉴定结果。
3A：三种突变类型以及对应的单菌落测序峰图；
3B：20个单菌落的测序比对序列。
图4：扩增片段二代测序分析结果。
4A：靶标序列单碱基编辑结果热图；
4B：靶序列单碱基突变类型及reads占比情况；
4C：靶序列胞嘧啶变化情况。
图5：脱靶位点测序结果比对。
5A：R2 OT A测序结果；
5B：R2 OT B测序结果；
5C：R2 OT C测序结果。
具体实施方式
下面通过实施例对本发明作进一步的详细说明，旨在用于说明本发明而非限定本发明。应当指出，对于本领域技术人员而言，在不脱离本发明原理的前提下，还可以对本发明进行若干改进和修饰，这些改进和修饰也同样落入本发明的保护范围之内。
实施例1：dFnCpf1-CBE-BT2载体构建
以PB-nCas9-PBE载体为骨架，利用限制性内切酶AflII和MluI切除nCas9。
FnCpf1为Cpf1效应蛋白，为创制dFnCpf1蛋白突变体，获得了FnCpf1氨基酸序列，如序列2所示。并通过序列比对确定dFnCpf1的突变位点，即D917A，E1006A，D1227A，具体见附图1。
根据单子叶植物基因组密码子特点，对其进行优化，创制可在玉米中外源表达的dFnCpf1序列，并放入pUC57中间载体。
以pUC57-dFnCpf1载体为模板PCR扩增已合成的dFnCpf1序列，回收酶切载体以及PCR产物，通过同源重组酶连入线性化的PB-nCas9-PBE载体，替换nCas9序列为dFnCpf1序列，构建dFnCpf1-PBE载体，并用HindIII酶切掉其OsU3-sgRNA-scaffold表达框。
为获得靶位点信息，通过CRISPR RGEN Tools网站(http://www.rgenome.net/cas-designer/)进行靶位点分析，在BT2基因序列的第二外显子区域上挑选打分相对较高，且富含C·G碱基的序列作为CRISPR的靶点。
选择OsU6启动子，mature-crRNA序列，合成OsU6-crRNA-polyT表达框，具体序列如序列3或附图2A所示，并将其放入pUC57-simple载体。
设计包含BT2基因靶序列及同源臂的引物，以OsU6-crRNA-polyT载体为模板进行扩增，通过同源重组酶连入经限制性内切酶HindIII线性化的dFnCpf1-PBE载体，构建成Ubi-rAPOBEC1-dFncpf1-OsU6-crRNA-bt2载体，得到靶向玉米内源基因BT2的胞嘧啶单碱基编辑双元载体，命名为dFnCpf1-CBE-BT2，载体结构见附图2B，载体序列如序列1所示。
载体构建过程中采用的特异性引物见表1。
表1引物名称及序列
PCR体系(50μL)：2×Phanta Master Mix 25μL，模板1μL，正、反向引物(10μmol·L-1)各2μL，Nuclease-free H2O 20μL。反应程序：95℃预变性5min；95℃变性15s，60℃退火15s，72℃延伸(dFnCpf1 4min；OsU6-crRNA表达框20s)，扩增35个循环；72℃再延伸5min。
酶切体系(50μL)：限制性内切酶2μL，模板5μg，CutSmart Buffer 5μL，Nuclease-free H2O补至体系为50μL。37℃孵育2.5h至酶解完全。
实施例2：玉米原生质体制备及转化
准备30株避光土培13d的玉米黄花苗，取第二叶中间较嫩部分并切至1～2mm丝。将其浸泡至20mL酶解液当中(1％纤维素酶R-10，0.2％离析酶R-10，0.4mol·L-1D-甘露醇，20mmol·L-1KCl，20mmol·L-1MES，10mmol·L-1CaCl2，0.1％BSA)，黑暗中室温震荡(40r·min-1)酶解(4-6h)。使用350目尼龙膜过滤酶解产物并置于50mL离心管，4℃，100g·min-1离心3min后弃上清。使用预冷W5 Buffer【2mmol·L-1MES(pH 5.7)，154mmol·L-1NaCl，125mmol·L-1CaCl2，5mmol·L-1KCl】重悬沉淀，洗涤沉淀1次。离心后弃上清，再次加入W5溶液，冰上静置30min。弃掉上清，加入适量MMG buffer【4mmol·L-1MES(pH 5.7)，0.4mmol·L-1D-甘露醇，15mmol·L-1MgCl2】使原生质体浓度达到2×107个·mL-1。
将100μg目的载体与5μg对照载体加入1mL原生质体MMG悬浮液当中，混匀后冰置10min。加入1mL预制PEG-Ca2+溶液(40％PEG-4000，200mmol·L-1D-甘露醇，100mmol·L- 1CaCl2)混匀，室温避光放置15min。加入2倍体积W5 buffer，清洗2次后加入20mL W5Buffer。最后将原生质体培养液置入细胞培养皿(1％BSA孵育0.5h)，避光28℃培养12-16h。
实施例3：靶基因编辑结果初筛
在470nm激发光、525nm发射光条件下，使用全自动荧光倒置显微镜，观察原生质体中绿色荧光信号的表达情况，初步判定转化效率。
采用CTAB法提取转化原生质体基因组DNA，使用2×Phanta Max Master Mix高保真DNA聚合酶，通过两种改良的聚合酶链式反应-限制性核酸内切酶(polymerase chainreaction/Rstriction endonuclease,PCR/RE)方法对编辑位点进行检测：
1、扩增靶位点序列后，使用BglⅡ内切酶酶切扩增产物判断编辑情况。
2、BglⅡ内切酶酶切基因组DNA来富集已编辑序列。以酶切产物为模板，巢式PCR特异扩增目的序列并Sanger测序检测编辑情况。其中巢式PCR反应体系及反应程序、基因组DNA酶切体系均与实施例1中一致，第二轮扩增的反应体系中模板为1μL第一轮PCR产物。
经上述步骤1或步骤2检测，发现编辑现象后，以步骤2中巢式PCR产物为模板进行TA克隆检测。按照北京全式金生物公司的
Cloning Kit的实验步骤转化载体至大肠杆菌感受态细胞中。挑选20个白色单克隆，进行Sanger测序，分析编辑情况。
结果发现靶位点序列存在3种不同突变类型，主要发生在靶位点5’端8-12bp的编辑框内，其中单菌落碱基变化类型5个为G→T，5个为C→A，2个为C→A与G→T，剩余8个则没有发生编辑，具体见附图3，说明该载体对靶位点具有一定编辑能力。
实施例4：靶基因编辑结果二代测序检测
经过初步鉴定后，以原生质体基因组DNA为模板，巢式PCR特异扩增靶位点序列，巢式PCR反应体系及反应程序与实施例3中一致。将扩增产物交由深圳华大基因股份有限公司构建测序文库，文库质控合格后由该公司BGI自主测序平台DNBSEQTM进行文库测序。采用华大Soapnuke软件过滤数据，去除接头污染和低质量reads。将高质量Clean Data根据以下标准使用Python语言进行数据可视化分析：
1、reads数超过1000且质量不低于5；
2、相同变化的reads超过1000条则判定为一种突变类型；
3、位点编辑效率＝(编辑位点reads数/总reads数)×100％；
4、基因插入缺失频率＝(该片段发生indel的reads数/该片段样品中找到完整编辑区域的reads数)×100％。
通过建库测序，共拼接出6903954条质量合格的reads，其中6469637条reads定位到靶序列。超过1000条reads发生相同变化，则会被判定为一种突变类型，以此标准将数据分类汇总后制成热图，具体见附图4A。
其中占比最多的突变类型为胞嘧啶碱基颠换为腺嘌呤碱基(C→A)，共151594条reads，位于靶位点5＇端第11个碱基。
根据位点编辑效率＝(编辑位点reads数/总reads数)×100％，显示该位点的编辑效率为2.5％。
为分析dFnCpf1介导单碱基编辑系统的编辑偏好位点，将各碱基的变化情况及对应编辑效率汇总，具体见附图4B。发现除了C→A外，位点编辑效率相对较高的为0.9％的G→T、0.3％的G→C以及0.2％的C→G，分别发生在靶位点5’端第8个碱基、第23与24个碱基。
由此可见，本发明所用单碱基编辑载体主要作用为产生胞嘧啶到胸腺嘧啶的改变，因此统计了靶序列中胞嘧啶变化的种类，具体见附图4C。结果表明，靶位点5’末端出现胞嘧啶转换为胸腺嘧啶(C→T)，位点的编辑效率为0.1％，具体见附图4B；位于靶序列5’端第11个碱基也出现C→T现象，但该点发生变化的reads数仅为1073，位点编辑效率不足0.02％。
实施例5：脱靶效率分析
为分析dFnCpf1-CBE-BT2编辑载体是否存在脱靶情况，根据已设计靶位点序列，使用CRISPR RGEN Tools网站(http://www.rgenome.net/cas-designer/)进行脱靶位点预测，设置Mismatch number为5，并选择打分较高的3个靶位点。选择3个排名靠前的靶序列，具体见表2，命名为R2-OT-A，R2-OT-B，R2-OT-C。
表2 R2脱靶位点预测
根据筛选结果从Zm-B73-REFERENCE-GRAMENE-4.0数据库查询基因序列，设计特异引物并以原生质体瞬时转化基因组DNA为模板扩增，Sanger测序鉴定编辑情况，其中PCR反应体系及反应程序与实施例1中一致。
Sanger测序结果显示未发现碱基突变，具体见附图5，说明dFnCpf1-CBE-BT2编辑载体不存在脱靶情况。
序列表
<110> 三亚中国农业科学院国家南繁研究院
<120> 一种单碱基编辑载体及其构建和应用
<160> 22
<170> PatentIn version 3.3
<210> 1
<211> 15971
<212> DNA
<213> 人工合成
<400> 1
taaacgctct tttctcttag gtttacccgc caatatatcc tgtcaaacac tgatagttta 60
aactgaaggc gggaaacgac aatctgatcc aagctcaagc tgctctagca ttcgccattc 120
aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg 180
gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca 240
cgacgttgta aaacgacggc cagtgccaag cttggatcat gaaccaacgg cctggctgta 300
tttggtggtt gtgtagggag atggggagaa gaaaagcccg attctcttcg ctgtgatggg 360
ctggatgcat gcgggggagc gggaggccca agtacgtgca cggtgagcgg cccacagggc 420
gagtgtgagc gcgagaggcg ggaggaacag tttagtacca cattgcccag ctaactcgaa 480
cgcgaccaac ttataaaccc gcgcgctgtc gcttgtgtgt aatttctact gttgtagatg 540
cacatagatc ttggatatgt tgcttttttt taagcttgca tgcctgcagt gcagcgtgac 600
ccggtcgtgc ccctctctag agataatgag cattgcatgt ctaagttata aaaaattacc 660
acatattttt tttgtcacac ttgtttgaag tgcagtttat ctatctttat acatatattt 720
aaactttact ctacgaataa tataatctat agtactacaa taatatcagt gttttagaga 780
atcatataaa tgaacagtta gacatggtct aaaggacaat tgagtatttt gacaacagga 840
ctctacagtt ttatcttttt agtgtgcatg tgttctcctt tttttttgca aatagcttca 900
cctatataat acttcatcca ttttattagt acatccattt agggtttagg gttaatggtt 960
tttatagact aattttttta gtacatctat tttattctat tttagcctct aaattaagaa 1020
aactaaaact ctattttagt ttttttattt aataatttag atataaaata gaataaaata 1080
aagtgactaa aaattaaaca aatacccttt aagaaattaa aaaaactaag gaaacatttt 1140
tcttgtttcg agtagataat gccagcctgt taaacgccgt cgacgagtct aacggacacc 1200
aaccagcgaa ccagcagcgt cgcgtcgggc caagcgaagc agacggcacg gcatctctgt 1260
cgctgcctct ggacccctct cgagagttcc gctccaccgt tggacttgct ccgctgtcgg 1320
catccagaaa ttgcgtggcg gagcggcaga cgtgagccgg cacggcaggc ggcctcctcc 1380
tcctctcacg gcacggcagc tacgggggat tcctttccca ccgctccttc gctttccctt 1440
cctcgcccgc cgtaataaat agacaccccc tccacaccct ctttccccaa cctcgtgttg 1500
ttcggagcgc acacacacac aaccagatct cccccaaatc cacccgtcgg cacctccgct 1560
tcaaggtacg ccgctcgtcc tccccccccc cccctctcta ccttctctag atcggcgttc 1620
cggtccatgg ttagggcccg gtagttctac ttctgttcat gtttgtgtta gatccgtgtt 1680
tgtgttagat ccgtgctgct agcgttcgta cacggatgcg acctgtacgt cagacacgtt 1740
ctgattgcta acttgccagt gtttctcttt ggggaatcct gggatggctc tagccgttcc 1800
gcagacggga tcgatttcat gatttttttt gtttcgttgc atagggtttg gtttgccctt 1860
ttcctttatt tcaatatatg ccgtgcactt gtttgtcggg tcatcttttc atgctttttt 1920
ttgtcttggt tgtgatgatg tggtctggtt gggcggtcgt tctagatcgg agtagaattc 1980
tgtttcaaac tacctggtgg atttattaat tttggatctg tatgtgtgtg ccatacatat 2040
tcatagttac gaattgaaga tgatggatgg aaatatcgat ctaggatagg tatacatgtt 2100
gatgcgggtt ttactgatgc atatacagag atgctttttg ttcgcttggt tgtgatgatg 2160
tggtgtggtt gggcggtcgt tcattcgttc tagatcggag tagaatactg tttcaaacta 2220
cctggtgtat ttattaattt tggaactgta tgtgtgtgtc atacatcttc atagttacga 2280
gtttaagatg gatggaaata tcgatctagg ataggtatac atgttgatgt gggttttact 2340
gatgcatata catgatggca tatgcagcat ctattcatat gctctaacct tgagtaccta 2400
tctattataa taaacaagta tgttttataa ttattttgat cttgatatac ttggatgatg 2460
gcatatgcag cagctatatg tggatttttt tagccctgcc ttcatacgct atttatttgc 2520
ttggtactgt ttcttttgtc gatgctcacc ctgttgtttg gtgttacttc tgcagcccta 2580
ggatgccaaa gaagaagagg aaggtttcat cggagaccgg ccctgttgct gttgacccca 2640
ccctgcggcg gagaatcgag ccacacgagt tcgaggtgtt cttcgaccca agggagctcc 2700
gcaaggagac gtgcctcctg tacgagatca actggggcgg caggcactcc atctggaggc 2760
acaccagcca aaacaccaac aagcacgtgg aggtcaactt catcgagaag ttcaccaccg 2820
agaggtactt ctgcccaaac acccgctgct ccatcacctg gttcctgtcc tggagcccat 2880
gcggcgagtg ctccagggcc atcaccgagt tcctcagccg ctacccacac gtcaccctgt 2940
tcatctacat cgccaggctc taccaccacg ccgacccaag gaacaggcag ggcctccgcg 3000
acctgatctc cagcggcgtg accatccaaa tcatgaccga gcaggagtcc ggctactgct 3060
ggaggaactt cgtcaactac tccccaagca acgaggccca ctggccaagg tacccacacc 3120
tctgggtgcg cctctacgtg ctcgagctgt actgcatcat cctcggcctg ccaccatgcc 3180
tcaacatcct gaggcgcaag caaccacagc tgaccttctt caccatcgcc ctccaaagct 3240
gccactacca gaggctccca ccacacatcc tgtgggctac cggcctcaag tccggcagcg 3300
agacgccagg cacctccgag agcgctacgc ctgaacttaa gatgtccatc taccaggagt 3360
tcgtgaacaa gtactccctc agcaagaccc tgaggttcga gctgatcccg cagggcaaga 3420
ccctggagaa catcaaggcc cgcggcctga tcctggacga cgagaagagg gccaaggact 3480
acaagaaggc caagcagatc atcgacaagt accaccagtt cttcatcgag gagatcctga 3540
gcagcgtgtg catctccgag gacctgctcc agaactactc cgacgtctac ttcaagctga 3600
agaagagcga cgacgacaac ctccagaagg acttcaagag cgccaaggac accatcaaga 3660
agcagatctc cgagtacatc aaggactccg agaagttcaa gaacctcttc aaccagaacc 3720
tgatcgacgc caagaagggc caggagagcg acctcatcct ctggctcaag cagagcaagg 3780
acaacggcat cgagctgttc aaggccaaca gcgacatcac cgacatcgac gaggccctcg 3840
agatcatcaa gagcttcaag ggctggacca cctacttcaa gggcttccac gagaacagga 3900
agaacgtcta ctcctccaac gacatcccaa cctccatcat ctaccgcatc gtcgacgaca 3960
acctgccgaa gttcctcgag aacaaggcca agtacgagtc cctgaaggac aaggccccag 4020
aggccatcaa ctacgagcag atcaagaagg acctcgccga ggagctgacc ttcgacatcg 4080
actacaagac cagcgaggtc aaccagaggg tgttctccct ggacgaggtg ttcgagatcg 4140
ccaacttcaa caactacctg aaccagtccg gcatcaccaa gttcaacacc atcatcggcg 4200
gcaagttcgt gaacggcgag aacaccaaga ggaagggcat caacgagtac atcaacctgt 4260
actcccagca gatcaacgac aagaccctga agaagtacaa gatgtccgtg ctgttcaagc 4320
agatcctgag cgacaccgag tccaagagct tcgtgatcga caagctggag gacgacagcg 4380
acgtcgtcac caccatgcag agcttctacg agcagattgc cgccttcaag accgtcgagg 4440
agaagagcat caaggagacc ctgagcctgc tcttcgacga cctcaaggcc cagaagctcg 4500
acctcagcaa gatctacttc aagaacgaca agagcctgac cgacctctcc cagcaggtgt 4560
tcgacgacta cagcgtgatc ggcaccgccg tgctggagta catcacccag cagatcgccc 4620
cgaagaacct cgacaacccg agcaagaagg agcaggagct gatcgccaag aagaccgaga 4680
aggccaagta tctctccctg gagaccatca agctggccct cgaggagttc aacaagcaca 4740
gggacatcga caagcagtgc aggttcgagg agatcctcgc caacttcgcc gccatcccga 4800
tgatcttcga cgagatcgcc cagaacaagg acaacctcgc ccagatctcc atcaagtacc 4860
agaaccaggg caagaaggac ctgctccaag ccagcgccga ggacgacgtg aaggccatca 4920
aggacctgct ggaccagacc aacaacctgc tgcacaagct caagatcttc cacatctccc 4980
agagcgagga caaggccaac atcctcgaca aggacgagca cttctacctg gtcttcgagg 5040
agtgctactt cgagctggcc aacatcgtgc cgctctacaa caagatcagg aactacatca 5100
cccaaaagcc atactccgac gagaagttca aactcaactt cgagaactcc accctcgcca 5160
acggctggga caagaacaag gagccggaca acaccgccat cctcttcatc aaggacgaca 5220
agtactacct cggcgtgatg aacaagaaga acaacaagat cttcgacgac aaggccatca 5280
aagagaacaa gggcgagggc tacaagaaga tcgtctacaa gctcctgccg ggcgccaaca 5340
agatgctccc aaaggtcttc ttctccgcca agagcatcaa attctacaac ccaagcgagg 5400
acatcctccg catccgcaac cactccaccc acaccaagaa cggctcccca cagaagggct 5460
acgagaagtt cgagttcaac atcgaggact gcaggaagtt catcgacttc tacaagcagt 5520
ccatcagcaa gcacccagag tggaaggact tcggcttccg cttcagcgac acccagaggt 5580
acaactccat cgacgagttc taccgcgagg tggagaacca gggctacaag ctcaccttcg 5640
agaacatctc cgagtcctac atcgactccg tcgtcaacca gggcaaactg tacctgttcc 5700
agatctacaa caaggacttc tccgcctaca gcaagggccg cccaaacctg cacaccctct 5760
actggaaggc cctcttcgac gagcgcaacc tccaggacgt ggtctacaag ctgaacggcg 5820
aggccgagct gttctaccgc aagcagtcca ttccgaagaa gatcacccac ccagccaagg 5880
aggccatcgc caacaagaac aaggacaatc cgaagaagga gtccgtgttc gagtacgacc 5940
tgatcaagga caagcgcttc accgaggaca agttcttctt ccactgcccg atcaccatca 6000
acttcaagag cagcggcgcc aacaaattca acgacgagat caacctgctc ctcaaggaga 6060
aggccaacga cgtgcacatc ctgagcatcg ccaggggcga gaggcacctg gcctactaca 6120
ccctcgtgga cggcaagggc aacatcatca agcaggacac cttcaacatc atcggcaacg 6180
acaggatgaa gaccaactac cacgacaagc tcgccgccat cgagaaggac agggacagcg 6240
ccaggaagga ctggaagaag atcaacaaca tcaaggagat gaaggagggc tacctgtccc 6300
aggtggtcca cgagatcgcc aagctggtca ttgagtacaa cgccatcgtc gtcttcgccg 6360
acctcaactt cggcttcaag aggggccgct tcaaggtcga gaagcaggtg taccagaagc 6420
tggagaagat gctcatcgag aagctgaact acctcgtctt caaggacaac gagttcgaca 6480
agaccggcgg cgtgctgcgc gcctaccagc ttaccgcccc gttcgagacc ttcaagaaga 6540
tgggcaagca gaccggcatc atctactacg tgccagccgg cttcaccagc aagatctgcc 6600
cggtcaccgg cttcgtgaac cagctgtacc caaagtacga gagcgtctcc aagtcccagg 6660
agttcttctc caagttcgac aagatctgct acaacctcga caagggctac ttcgagttca 6720
gcttcgacta caagaacttc ggcgacaagg ccgccaaggg caagtggacc atcgcctcct 6780
tcggcagccg cctcatcaac ttcaggaaca gcgacaagaa ccacaactgg gacaccaggg 6840
aggtctaccc gaccaaggag ctggagaagc tcctcaagga ctacagcatc gagtacggcc 6900
acggcgagtg catcaaggcc gccatctgcg gcgagtccga caagaagttc ttcgccaagc 6960
tcaccagcgt cctcaacacc atcctgcaga tgcgcaacag caagaccggc accgagctgg 7020
cctacctgat cagcccagtg gccgacgtca acggcaactt cttcgacagc cgccaggccc 7080
caaagaacat gccacaggac gccgacgcca acggcgccta ccacatcggc ctcaagggcc 7140
tgatgctcct cggcaggatc aagaacaacc aggagggcaa gaagctgaac ctggtcatca 7200
agaacgagga gtacttcgag tttgtgcaga acaggaacaa cacgcgtgac tccggcggca 7260
gcaccaacct gtccgacatc atcgagaagg agacgggcaa gcaactcgtg atccaggaga 7320
gcatcctcat gctgccagag gaggtggagg aggtcatcgg caacaagcca gagtccgaca 7380
tcctggtgca caccgcctac gacgagtcca ccgacgagaa cgtcatgctc ctgaccagcg 7440
acgccccaga gtacaagcca tgggccctcg tcatccagga cagcaacggg gagaacaaga 7500
tcaagatgct gtcggggggg agcccaaaga agaagcggaa ggtgtagagc tcagagcttt 7560
cgttcgtatc atcggtttcg acaacgttcg tcaagttcaa tgcatcagtt tcattgcgca 7620
cacaccagaa tcctactgag tttgagtatt atggcattgg gaaaactgtt tttcttgtac 7680
catttgttgt gcttgtaatt tactgtgttt tttattcggt tttcgctatc gaactgtgaa 7740
atggaaatgg atggagaaga gttaatgaat gatatggtcc ttttgttcat tctcaaatta 7800
atattatttg ttttttctct tatttgttgt gtgttgaatt tgaaattata agagatatgc 7860
aaacattttg ttttgagtaa aaatgtgtca aatcgtggcc tctaatgacc gaagttaata 7920
tgaggagtaa aacacttgta gttgtaccat tatgcttatt cactaggcaa caaatatatt 7980
ttcagaccta gaaaagctgc aaatgttact gaatacaagt atgtcctctt gtgttttaga 8040
catttatgaa ctttccttta tgtaattttc cagaatcctt gtcagattct aatcattgct 8100
ttataattat agttatactc atggatttgt agttgagtat gaaaatattt tttaatgcat 8160
tttatgactt gccaattgat tgacaacgaa ttcgtaatca tggtcatagc tgtttcctgt 8220
gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 8280
agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 8340
tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 8400
aggcggtttg cgtattggct agagcagctt gccaacatgg tggagcacga cactctcgtc 8460
tactccaaga atatcaaaga tacagtctca gaagaccaaa gggctattga gacttttcaa 8520
caaagggtaa tatcgggaaa cctcctcgga ttccattgcc cagctatctg tcacttcatc 8580
aaaaggacag tagaaaagga aggtggcacc tacaaatgcc atcattgcga taaaggaaag 8640
gctatcgttc aagatgcctc tgccgacagt ggtcccaaag atggaccccc acccacgagg 8700
agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat 8760
atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga ccttcctcta 8820
tataaggaag ttcatttcat ttggagagga cacgctgaaa tcaccagtct ctctctacaa 8880
atctatctct ctcgagtcta ccatgagccc agaacgacgc ccggccgaca tccgccgtgc 8940
caccgaggcg gacatgccgg cggtctgcac catcgtcaac cactacatcg agacaagcac 9000
ggtcaacttc cgtaccgagc cgcaggaacc gcaggagtgg acggacgacc tcgtccgtct 9060
gcgggagcgc tatccctggc tcgtcgccga ggtggacggc gaggtcgccg gcatcgccta 9120
cgcgggcccc tggaaggcac gcaacgccta cgactggacg gccgagtcga ccgtgtacgt 9180
ctccccccgc caccagcgga cgggactggg ctccacgctc tacacccacc tgctgaagtc 9240
cctggaggca cagggcttca agagcgtggt cgctgtcatc gggctgccca acgacccgag 9300
cgtgcgcatg cacgaggcgc tcggatatgc cccccgcggc atgctgcggg cggccggctt 9360
caagcacggg aactggcatg acgtgggttt ctggcagctg gacttcagcc tgccggtacc 9420
gccccgtccg gtcctgcccg tcaccgagat ttgactcgag tttctccata ataatgtgtg 9480
agtagttccc agataaggga attagggttc ctatagggtt tcgctcatgt gttgagcata 9540
taagaaaccc ttagtatgta tttgtatttg taaaatactt ctatcaataa aatttctaat 9600
tcctaaaacc aaaatccagt actaaaatcc agatcccccg aattaattcg gcgttaattc 9660
agtacattaa aaacgtccgc aatgtgttat taagttgtct aagcgtcaat ttgtttacac 9720
cacaatatat cctgccacca gccagccaac agctccccga ccggcagctc ggcacaaaat 9780
caccactcga tacaggcagc ccatcagtcc gggacggcgt cagcgggaga gccgttgtaa 9840
ggcggcagac tttgctcatg ttaccgatgc tattcggaag aacggcaact aagctgccgg 9900
gtttgaaaca cggatgatct cgcggagggt agcatgttga ttgtaacgat gacagagcgt 9960
tgctgcctgt gatcaccgcg gtttcaaaat cggctccgtc gatactatgt tatacgccaa 10020
ctttgaaaac aactttgaaa aagctgtttt ctggtattta aggttttaga atgcaaggaa 10080
cagtgaattg gagttcgtct tgttataatt agcttcttgg ggtatcttta aatactgtag 10140
aaaagaggaa ggaaataata aatggctaaa atgagaatat caccggaatt gaaaaaactg 10200
atcgaaaaat accgctgcgt aaaagatacg gaaggaatgt ctcctgctaa ggtatataag 10260
ctggtgggag aaaatgaaaa cctatattta aaaatgacgg acagccggta taaagggacc 10320
acctatgatg tggaacggga aaaggacatg atgctatggc tggaaggaaa gctgcctgtt 10380
ccaaaggtcc tgcactttga acggcatgat ggctggagca atctgctcat gagtgaggcc 10440
gatggcgtcc tttgctcgga agagtatgaa gatgaacaaa gccctgaaaa gattatcgag 10500
ctgtatgcgg agtgcatcag gctctttcac tccatcgaca tatcggattg tccctatacg 10560
aatagcttag acagccgctt agccgaattg gattacttac tgaataacga tctggccgat 10620
gtggattgcg aaaactggga agaagacact ccatttaaag atccgcgcga gctgtatgat 10680
tttttaaaga cggaaaagcc cgaagaggaa cttgtctttt cccacggcga cctgggagac 10740
agcaacatct ttgtgaaaga tggcaaagta agtggcttta ttgatcttgg gagaagcggc 10800
agggcggaca agtggtatga cattgccttc tgcgtccggt cgatcaggga ggatatcggg 10860
gaagaacagt atgtcgagct attttttgac ttactgggga tcaagcctga ttgggagaaa 10920
ataaaatatt atattttact ggatgaattg ttttagtacc tagaatgcat gaccaaaatc 10980
ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 11040
tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 11100
ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 11160
ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac 11220
ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 11280
gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 11340
aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 11400
acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 11460
gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 11520
gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 11580
cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 11640
aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct 11700
gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct 11760
cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg 11820
atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc 11880
agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg 11940
actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt 12000
gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc 12060
agaggttttc accgtcatca ccgaaacgcg cgaggcaggg tgccttgatg tgggcgccgg 12120
cggtcgagtg gcgacggcgc ggcttgtccg cgccctggta gattgcctgg ccgtaggcca 12180
gccatttttg agcggccagc ggccgcgata ggccgacgcg aagcggcggg gcgtagggag 12240
cgcagcgacc gaagggtagg cgctttttgc agctcttcgg ctgtgcgctg gccagacagt 12300
tatgcacagg ccaggcgggt tttaagagtt ttaataagtt ttaaagagtt ttaggcggaa 12360
aaatcgcctt ttttctcttt tatatcagtc acttacatgt gtgaccggtt cccaatgtac 12420
ggctttgggt tcccaatgta cgggttccgg ttcccaatgt acggctttgg gttcccaatg 12480
tacgtgctat ccacaggaaa cagacctttt cgaccttttt cccctgctag ggcaatttgc 12540
cctagcatct gctccgtaca ttaggaaccg gcggatgctt cgccctcgat caggttgcgg 12600
tagcgcatga ctaggatcgg gccagcctgc cccgcctcct ccttcaaatc gtactccggc 12660
aggtcatttg acccgatcag cttgcgcacg gtgaaacaga acttcttgaa ctctccggcg 12720
ctgccactgc gttcgtagat cgtcttgaac aaccatctgg cttctgcctt gcctgcggcg 12780
cggcgtgcca ggcggtagag aaaacggccg atgccgggat cgatcaaaaa gtaatcgggg 12840
tgaaccgtca gcacgtccgg gttcttgcct tctgtgatct cgcggtacat ccaatcagct 12900
agctcgatct cgatgtactc cggccgcccg gtttcgctct ttacgatctt gtagcggcta 12960
atcaaggctt caccctcgga taccgtcacc aggcggccgt tcttggcctt cttcgtacgc 13020
tgcatggcaa cgtgcgtggt gtttaaccga atgcaggttt ctaccaggtc gtctttctgc 13080
tttccgccat cggctcgccg gcagaacttg agtacgtccg caacgtgtgg acggaacacg 13140
cggccgggct tgtctccctt cccttcccgg tatcggttca tggattcggt tagatgggaa 13200
accgccatca gtaccaggtc gtaatcccac acactggcca tgccggccgg ccctgcggaa 13260
acctctacgt gcccgtctgg aagctcgtag cggatcacct cgccagctcg tcggtcacgc 13320
ttcgacagac ggaaaacggc cacgtccatg atgctgcgac tatcgcgggt gcccacgtca 13380
tagagcatcg gaacgaaaaa atctggttgc tcgtcgccct tgggcggctt cctaatcgac 13440
ggcgcaccgg ctgccggcgg ttgccgggat tctttgcgga ttcgatcagc ggccgcttgc 13500
cacgattcac cggggcgtgc ttctgcctcg atgcgttgcc gctgggcggc ctgcgcggcc 13560
ttcaacttct ccaccaggtc atcacccagc gccgcgccga tttgtaccgg gccggatggt 13620
ttgcgaccgc tcacgccgat tcctcgggct tgggggttcc agtgccattg cagggccggc 13680
agacaaccca gccgcttacg cctggccaac cgcccgttcc tccacacatg gggcattcca 13740
cggcgtcggt gcctggttgt tcttgatttt ccatgccgcc tcctttagcc gctaaaattc 13800
atctactcat ttattcattt gctcatttac tctggtagct gcgcgatgta ttcagatagc 13860
agctcggtaa tggtcttgcc ttggcgtacc gcgtacatct tcagcttggt gtgatcctcc 13920
gccggcaact gaaagttgac ccgcttcatg gctggcgtgt ctgccaggct ggccaacgtt 13980
gcagccttgc tgctgcgtgc gctcggacgg ccggcactta gcgtgtttgt gcttttgctc 14040
attttctctt tacctcatta actcaaatga gttttgattt aatttcagcg gccagcgcct 14100
ggacctcgcg ggcagcgtcg ccctcgggtt ctgattcaag aacggttgtg ccggcggcgg 14160
cagtgcctgg gtagctcacg cgctgcgtga tacgggactc aagaatgggc agctcgtacc 14220
cggccagcgc ctcggcaacc tcaccgccga tgcgcgtgcc tttgatcgcc cgcgacacga 14280
caaaggccgc ttgtagcctt ccatccgtga cctcaatgcg ctgcttaacc agctccacca 14340
ggtcggcggt ggcccatatg tcgtaagggc ttggctgcac cggaatcagc acgaagtcgg 14400
ctgccttgat cgcggacaca gccaagtccg ccgcctgggg cgctccgtcg atcactacga 14460
agtcgcgccg gccgatggcc ttcacgtcgc ggtcaatcgt cgggcggtcg atgccgacaa 14520
cggttagcgg ttgatcttcc cgcacggccg cccaatcgcg ggcactgccc tggggatcgg 14580
aatcgactaa cagaacatcg gccccggcga gttgcagggc gcgggctaga tgggttgcga 14640
tggtcgtctt gcctgacccg cctttctggt taagtacagc gataaccttc atgcgttccc 14700
cttgcgtatt tgtttattta ctcatcgcat catatacgca gcgaccgcat gacgcaagct 14760
gttttactca aatacacatc acctttttag acggcggcgc tcggtttctt cagcggccaa 14820
gctggccggc caggccgcca gcttggcatc agacaaaccg gccaggattt catgcagccg 14880
cacggttgag acgtgcgcgg gcggctcgaa cacgtacccg gccgcgatca tctccgcctc 14940
gatctcttcg gtaatgaaaa acggttcgtc ctggccgtcc tggtgcggtt tcatgcttgt 15000
tcctcttggc gttcattctc ggcggccgcc agggcgtcgg cctcggtcaa tgcgtcctca 15060
cggaaggcac cgcgccgcct ggcctcggtg ggcgtcactt cctcgctgcg ctcaagtgcg 15120
cggtacaggg tcgagcgatg cacgccaagc agtgcagccg cctctttcac ggtgcggcct 15180
tcctggtcga tcagctcgcg ggcgtgcgcg atctgtgccg gggtgagggt agggcggggg 15240
ccaaacttca cgcctcgggc cttggcggcc tcgcgcccgc tccgggtgcg gtcgatgatt 15300
agggaacgct cgaactcggc aatgccggcg aacacggtca acaccatgcg gccggccggc 15360
gtggtggtgt cggcccacgg ctctgccagg ctacgcaggc ccgcgccggc ctcctggatg 15420
cgctcggcaa tgtccagtag gtcgcgggtg ctgcgggcca ggcggtctag cctggtcact 15480
gtcacaacgt cgccagggcg taggtggtca agcatcctgg ccagctccgg gcggtcgcgc 15540
ctggtgccgg tgatcttctc ggaaaacagc ttggtgcagc cggccgcgtg cagttcggcc 15600
cgttggttgg tcaagtcctg gtcgtcggtg ctgacgcggg catagcccag caggccagcg 15660
gcggcgctct tgttcatggc gtaatgtctc cggttctagt cgcaagtatt ctactttatg 15720
cgactaaaac acgcgacaag aaaacgccag gaaaagggca gggcggcagc ctgtcgcgta 15780
acttaggact tgtgcgacat gtcgttttca gaagacggct gcactgaacg tcagaagccg 15840
actgcactat agcagcggag gggttggatc aaagtacttt gatcccgagg ggaaccctgt 15900
ggttggcatg cacatacaaa tggacgaacg gataaacctt ttcacgccct tttaaatatc 15960
cgttattcta a 15971
<210> 2
<211> 1300
<212> PRT
<213> 人工合成
<400> 2
Met Ser Ile Tyr Gln Glu Phe Val Asn Lys Tyr Ser Leu Ser Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Glu Asn Ile Lys
20 25 30
Ala Arg Gly Leu Ile Leu Asp Asp Glu Lys Arg Ala Lys Asp Tyr Lys
35 40 45
Lys Ala Lys Gln Ile Ile Asp Lys Tyr His Gln Phe Phe Ile Glu Glu
50 55 60
Ile Leu Ser Ser Val Cys Ile Ser Glu Asp Leu Leu Gln Asn Tyr Ser
65 70 75 80
Asp Val Tyr Phe Lys Leu Lys Lys Ser Asp Asp Asp Asn Leu Gln Lys
85 90 95
Asp Phe Lys Ser Ala Lys Asp Thr Ile Lys Lys Gln Ile Ser Glu Tyr
100 105 110
Ile Lys Asp Ser Glu Lys Phe Lys Asn Leu Phe Asn Gln Asn Leu Ile
115 120 125
Asp Ala Lys Lys Gly Gln Glu Ser Asp Leu Ile Leu Trp Leu Lys Gln
130 135 140
Ser Lys Asp Asn Gly Ile Glu Leu Phe Lys Ala Asn Ser Asp Ile Thr
145 150 155 160
Asp Ile Asp Glu Ala Leu Glu Ile Ile Lys Ser Phe Lys Gly Trp Thr
165 170 175
Thr Tyr Phe Lys Gly Phe His Glu Asn Arg Lys Asn Val Tyr Ser Ser
180 185 190
Asn Asp Ile Pro Thr Ser Ile Ile Tyr Arg Ile Val Asp Asp Asn Leu
195 200 205
Pro Lys Phe Leu Glu Asn Lys Ala Lys Tyr Glu Ser Leu Lys Asp Lys
210 215 220
Ala Pro Glu Ala Ile Asn Tyr Glu Gln Ile Lys Lys Asp Leu Ala Glu
225 230 235 240
Glu Leu Thr Phe Asp Ile Asp Tyr Lys Thr Ser Glu Val Asn Gln Arg
245 250 255
Val Phe Ser Leu Asp Glu Val Phe Glu Ile Ala Asn Phe Asn Asn Tyr
260 265 270
Leu Asn Gln Ser Gly Ile Thr Lys Phe Asn Thr Ile Ile Gly Gly Lys
275 280 285
Phe Val Asn Gly Glu Asn Thr Lys Arg Lys Gly Ile Asn Glu Tyr Ile
290 295 300
Asn Leu Tyr Ser Gln Gln Ile Asn Asp Lys Thr Leu Lys Lys Tyr Lys
305 310 315 320
Met Ser Val Leu Phe Lys Gln Ile Leu Ser Asp Thr Glu Ser Lys Ser
325 330 335
Phe Val Ile Asp Lys Leu Glu Asp Asp Ser Asp Val Val Thr Thr Met
340 345 350
Gln Ser Phe Tyr Glu Gln Ile Ala Ala Phe Lys Thr Val Glu Glu Lys
355 360 365
Ser Ile Lys Glu Thr Leu Ser Leu Leu Phe Asp Asp Leu Lys Ala Gln
370 375 380
Lys Leu Asp Leu Ser Lys Ile Tyr Phe Lys Asn Asp Lys Ser Leu Thr
385 390 395 400
Asp Leu Ser Gln Gln Val Phe Asp Asp Tyr Ser Val Ile Gly Thr Ala
405 410 415
Val Leu Glu Tyr Ile Thr Gln Gln Ile Ala Pro Lys Asn Leu Asp Asn
420 425 430
Pro Ser Lys Lys Glu Gln Glu Leu Ile Ala Lys Lys Thr Glu Lys Ala
435 440 445
Lys Tyr Leu Ser Leu Glu Thr Ile Lys Leu Ala Leu Glu Glu Phe Asn
450 455 460
Lys His Arg Asp Ile Asp Lys Gln Cys Arg Phe Glu Glu Ile Leu Ala
465 470 475 480
Asn Phe Ala Ala Ile Pro Met Ile Phe Asp Glu Ile Ala Gln Asn Lys
485 490 495
Asp Asn Leu Ala Gln Ile Ser Ile Lys Tyr Gln Asn Gln Gly Lys Lys
500 505 510
Asp Leu Leu Gln Ala Ser Ala Glu Asp Asp Val Lys Ala Ile Lys Asp
515 520 525
Leu Leu Asp Gln Thr Asn Asn Leu Leu His Lys Leu Lys Ile Phe His
530 535 540
Ile Ser Gln Ser Glu Asp Lys Ala Asn Ile Leu Asp Lys Asp Glu His
545 550 555 560
Phe Tyr Leu Val Phe Glu Glu Cys Tyr Phe Glu Leu Ala Asn Ile Val
565 570 575
Pro Leu Tyr Asn Lys Ile Arg Asn Tyr Ile Thr Gln Lys Pro Tyr Ser
580 585 590
Asp Glu Lys Phe Lys Leu Asn Phe Glu Asn Ser Thr Leu Ala Asn Gly
595 600 605
Trp Asp Lys Asn Lys Glu Pro Asp Asn Thr Ala Ile Leu Phe Ile Lys
610 615 620
Asp Asp Lys Tyr Tyr Leu Gly Val Met Asn Lys Lys Asn Asn Lys Ile
625 630 635 640
Phe Asp Asp Lys Ala Ile Lys Glu Asn Lys Gly Glu Gly Tyr Lys Lys
645 650 655
Ile Val Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro Lys Val
660 665 670
Phe Phe Ser Ala Lys Ser Ile Lys Phe Tyr Asn Pro Ser Glu Asp Ile
675 680 685
Leu Arg Ile Arg Asn His Ser Thr His Thr Lys Asn Gly Ser Pro Gln
690 695 700
Lys Gly Tyr Glu Lys Phe Glu Phe Asn Ile Glu Asp Cys Arg Lys Phe
705 710 715 720
Ile Asp Phe Tyr Lys Gln Ser Ile Ser Lys His Pro Glu Trp Lys Asp
725 730 735
Phe Gly Phe Arg Phe Ser Asp Thr Gln Arg Tyr Asn Ser Ile Asp Glu
740 745 750
Phe Tyr Arg Glu Val Glu Asn Gln Gly Tyr Lys Leu Thr Phe Glu Asn
755 760 765
Ile Ser Glu Ser Tyr Ile Asp Ser Val Val Asn Gln Gly Lys Leu Tyr
770 775 780
Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Ala Tyr Ser Lys Gly Arg
785 790 795 800
Pro Asn Leu His Thr Leu Tyr Trp Lys Ala Leu Phe Asp Glu Arg Asn
805 810 815
Leu Gln Asp Val Val Tyr Lys Leu Asn Gly Glu Ala Glu Leu Phe Tyr
820 825 830
Arg Lys Gln Ser Ile Pro Lys Lys Ile Thr His Pro Ala Lys Glu Ala
835 840 845
Ile Ala Asn Lys Asn Lys Asp Asn Pro Lys Lys Glu Ser Val Phe Glu
850 855 860
Tyr Asp Leu Ile Lys Asp Lys Arg Phe Thr Glu Asp Lys Phe Phe Phe
865 870 875 880
His Cys Pro Ile Thr Ile Asn Phe Lys Ser Ser Gly Ala Asn Lys Phe
885 890 895
Asn Asp Glu Ile Asn Leu Leu Leu Lys Glu Lys Ala Asn Asp Val His
900 905 910
Ile Leu Ser Ile Ala Arg Gly Glu Arg His Leu Ala Tyr Tyr Thr Leu
915 920 925
Val Asp Gly Lys Gly Asn Ile Ile Lys Gln Asp Thr Phe Asn Ile Ile
930 935 940
Gly Asn Asp Arg Met Lys Thr Asn Tyr His Asp Lys Leu Ala Ala Ile
945 950 955 960
Glu Lys Asp Arg Asp Ser Ala Arg Lys Asp Trp Lys Lys Ile Asn Asn
965 970 975
Ile Lys Glu Met Lys Glu Gly Tyr Leu Ser Gln Val Val His Glu Ile
980 985 990
Ala Lys Leu Val Ile Glu Tyr Asn Ala Ile Val Val Phe Ala Asp Leu
995 1000 1005
Asn Phe Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Val
1010 1015 1020
Tyr Gln Lys Leu Glu Lys Met Leu Ile Glu Lys Leu Asn Tyr Leu
1025 1030 1035
Val Phe Lys Asp Asn Glu Phe Asp Lys Thr Gly Gly Val Leu Arg
1040 1045 1050
Ala Tyr Gln Leu Thr Ala Pro Phe Glu Thr Phe Lys Lys Met Gly
1055 1060 1065
Lys Gln Thr Gly Ile Ile Tyr Tyr Val Pro Ala Gly Phe Thr Ser
1070 1075 1080
Lys Ile Cys Pro Val Thr Gly Phe Val Asn Gln Leu Tyr Pro Lys
1085 1090 1095
Tyr Glu Ser Val Ser Lys Ser Gln Glu Phe Phe Ser Lys Phe Asp
1100 1105 1110
Lys Ile Cys Tyr Asn Leu Asp Lys Gly Tyr Phe Glu Phe Ser Phe
1115 1120 1125
Asp Tyr Lys Asn Phe Gly Asp Lys Ala Ala Lys Gly Lys Trp Thr
1130 1135 1140
Ile Ala Ser Phe Gly Ser Arg Leu Ile Asn Phe Arg Asn Ser Asp
1145 1150 1155
Lys Asn His Asn Trp Asp Thr Arg Glu Val Tyr Pro Thr Lys Glu
1160 1165 1170
Leu Glu Lys Leu Leu Lys Asp Tyr Ser Ile Glu Tyr Gly His Gly
1175 1180 1185
Glu Cys Ile Lys Ala Ala Ile Cys Gly Glu Ser Asp Lys Lys Phe
1190 1195 1200
Phe Ala Lys Leu Thr Ser Val Leu Asn Thr Ile Leu Gln Met Arg
1205 1210 1215
Asn Ser Lys Thr Gly Thr Glu Leu Ala Tyr Leu Ile Ser Pro Val
1220 1225 1230
Ala Asp Val Asn Gly Asn Phe Phe Asp Ser Arg Gln Ala Pro Lys
1235 1240 1245
Asn Met Pro Gln Asp Ala Asp Ala Asn Gly Ala Tyr His Ile Gly
1250 1255 1260
Leu Lys Gly Leu Met Leu Leu Gly Arg Ile Lys Asn Asn Gln Glu
1265 1270 1275
Gly Lys Lys Leu Asn Leu Val Ile Lys Asn Glu Glu Tyr Phe Glu
1280 1285 1290
Phe Val Gln Asn Arg Asn Asn
1295 1300
<210> 3
<211> 297
<212> DNA
<213> 人工合成
<400> 3
ggatcatgaa ccaacggcct ggctgtattt ggtggttgtg tagggagatg gggagaagaa 60
aagcccgatt ctcttcgctg tgatgggctg gatgcatgcg ggggagcggg aggcccaagt 120
acgtgcacgg tgagcggccc acagggcgag tgtgagcgcg agaggcggga ggaacagttt 180
agtaccacat tgcccagcta actcgaacgc gaccaactta taaacccgcg cgctgtcgct 240
tgtgtgtaat ttctactgtt gtagatggga atgctggaac tgcaatgcgt ttttttt 297
<210> 4
<211> 22
<212> DNA
<213> 人工合成
<400> 4
gaacttgccc cgtcttttga at 22
<210> 5
<211> 20
<212> DNA
<213> 人工合成
<400> 5
gcaagtgaat ggtggtgtct 20
<210> 6
<211> 20
<212> DNA
<213> 人工合成
<400> 6
ccttgacaaa gaagcgtgcc 20
<210> 7
<211> 20
<212> DNA
<213> 人工合成
<400> 7
ccagtttgga ttatctgggc 20
<210> 8
<211> 45
<212> DNA
<213> 人工合成
<400> 8
agcgctacgc ctgaacttaa gatgtccatc taccaggagt tcgtg 45
<210> 9
<211> 44
<212> DNA
<213> 人工合成
<400> 9
gctgccgccg gagtcacgcg tgttgttcct gttctgcaca aact 44
<210> 10
<211> 22
<212> DNA
<213> 人工合成
<400> 10
accacagctg accttcttca cc 22
<210> 11
<211> 41
<212> DNA
<213> 人工合成
<400> 11
acgacggcca gtgccaagct tggatcatga accaacggcc t 41
<210> 12
<211> 45
<212> DNA
<213> 人工合成
<400> 12
gcactgcagg catgcaagct taaaaaaaag caacatatcc aagat 45
<210> 13
<211> 28
<212> DNA
<213> 人工合成
<400> 13
ctatgtgcat ctacaacagt agaaatta 28
<210> 14
<211> 20
<212> DNA
<213> 人工合成
<400> 14
gaaactccat cacattatgc 20
<210> 15
<211> 20
<212> DNA
<213> 人工合成
<400> 15
agcagaagga ttgtcccctc 20
<210> 16
<211> 22
<212> DNA
<213> 人工合成
<400> 16
ccttgccgtc agcttattag at 22
<210> 17
<211> 20
<212> DNA
<213> 人工合成
<400> 17
tcctgttatc agcatgaact 20
<210> 18
<211> 19
<212> DNA
<213> 人工合成
<400> 18
ggcgaaggtg acgaagagt 19
<210> 19
<211> 21
<212> DNA
<213> 人工合成
<400> 19
tacaactcta gcacacgcct c 21
<210> 20
<211> 27
<212> DNA
<213> 人工合成
<400> 20
ttagcacata gatcttggat atgttgt 27
<210> 21
<211> 27
<212> DNA
<213> 人工合成
<400> 21
ttagcacata tatcttggag acgttac 27
<210> 22
<211> 27
<212> DNA
<213> 人工合成
<400> 22
tttacacata tatcttgggt ttgctgc 27
Claims (10)
1.一种单碱基编辑载体，其包含dFnCpf1序列和crRNA序列。
2.根据权利要求1所述的单碱基编辑载体，其为dFnCpf1-CBE-BT2，载体结构如图2B所示。
3.包含权利要求1或2所述的单碱基编辑载体的宿主细胞。
4.根据权利要求3所述的宿主细胞，其为玉米原生质体。
5.一种单碱基编辑载体的构建方法，其包括如下步骤：
(1)构建包含dFnCpf1序列的载体；
(2)合成包含crRNA序列的表达框；
(3)构建包含dFnCpf1序列和crRNA序列的单碱基编辑载体。
6.根据权利要求5所述的单碱基编辑载体的构建方法，其包括如下步骤：
(1)以PB-nCas9-PBE载体为骨架，切除nCas9序列，以pUC57-dFnCpf1载体为模板，扩增dFnCpf1序列，将dFnCpf1序列连入线性化的PB-nCas9-PBE载体，构建dFnCpf1-PBE载体，并切除OsU3-sgRNA-scaffold表达框；
(2)选择OsU6启动子，mature-crRNA序列，合成OsU6-crRNA-polyT表达框，并放入pUC57-simple载体；
(3)设计包含玉米bt2基因靶序列及同源臂的引物，以OsU6-crRNA-polyT载体为模板进行扩增，连入经酶切线性化的dFnCpf1-PBE载体，构建dFnCpf1-CBE-BT2载体。
7.权利要求1或2所述的单碱基编辑载体在对靶序列上特定碱基进行定向替换中的应用，其能够识别TTN作为原型间隔序列毗邻基序(PAM)，并对靶位点进行单碱基编辑。
8.根据权利要求7所述的应用，其中所述单碱基编辑发生在靶位点5’端8-12bp的编辑框内，单碱基编辑类型是将鸟嘌呤(G)转换为胸腺嘧啶(T)，或者将胞嘧啶(C)转换为腺嘌呤(A)。
9.根据权利要求8所述的应用，其中所述的单碱基编辑为将靶位点5’端第11位碱基的胞嘧啶(C)转换为腺嘌呤(A)。
10.采用权利要求1或2所述的单碱基编辑载体转化的植物体，优选玉米，更加优选玉米原生质体。
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114836459A (zh) * | 2022-03-17 | 2022-08-02 | 江南大学 | 一种胞嘧啶碱基编辑系统及其应用 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108795972A (zh) * | 2017-05-05 | 2018-11-13 | 中国科学院遗传与发育生物学研究所 | 不使用转基因标记序列分离细胞的方法 |
| CN109957569A (zh) * | 2017-12-22 | 2019-07-02 | 中国科学院遗传与发育生物学研究所 | 基于cpf1蛋白的碱基编辑系统和方法 |
| CN111788232A (zh) * | 2018-02-23 | 2020-10-16 | 上海科技大学 | 用于碱基编辑的融合蛋白 |
-
2021
- 2021-11-12 CN CN202111339747.6A patent/CN114045302A/zh active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108795972A (zh) * | 2017-05-05 | 2018-11-13 | 中国科学院遗传与发育生物学研究所 | 不使用转基因标记序列分离细胞的方法 |
| CN109957569A (zh) * | 2017-12-22 | 2019-07-02 | 中国科学院遗传与发育生物学研究所 | 基于cpf1蛋白的碱基编辑系统和方法 |
| CN111788232A (zh) * | 2018-02-23 | 2020-10-16 | 上海科技大学 | 用于碱基编辑的融合蛋白 |
| US20210163913A1 (en) * | 2018-02-23 | 2021-06-03 | Shanghaitech University | Fusion proteins for base editing |
Non-Patent Citations (3)
| Title |
|---|
| CHEN Z等: "Engineered DNase-inactive Cpf1 variants to improve targeting scope for base editing in E.coli", 《SYNTH SYST BIOTECHNOL》, vol. 6, no. 4, pages 326 - 334, XP093051336, DOI: 10.1016/j.synbio.2021.09.002 * |
| ZONG, Y.等: "Precise base editing in rice, wheat and maize with a Cas9-cytidine deaminase fusion", 《NAT BIOTECHNOL》, vol. 35, pages 438 - 440, XP055850721, DOI: 10.1038/nbt.3811 * |
| 李树磊: "FnCpf1介导的基因组定点编辑技术研究", 《中国优秀硕士学位论文全文数据库基础科学辑》, no. 2021, pages 006 - 32 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114836459A (zh) * | 2022-03-17 | 2022-08-02 | 江南大学 | 一种胞嘧啶碱基编辑系统及其应用 |
| CN114836459B (zh) * | 2022-03-17 | 2024-01-26 | 江南大学 | 一种胞嘧啶碱基编辑系统及其应用 |
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