CN110438102A - 一种甲基化保护宿主菌及其构建方法和应用 - Google Patents
一种甲基化保护宿主菌及其构建方法和应用 Download PDFInfo
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- CN110438102A CN110438102A CN201910828288.4A CN201910828288A CN110438102A CN 110438102 A CN110438102 A CN 110438102A CN 201910828288 A CN201910828288 A CN 201910828288A CN 110438102 A CN110438102 A CN 110438102A
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- ndei
- transmethylase
- restriction enzyme
- ile
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Abstract
本发明提供了一种甲基化保护宿主菌及其构建方法和应用，所述甲基转移酶的氨基酸序列包括如SEQ ID NO:1所示的氨基酸序列，核苷酸序列包括如SEQ ID NO:2所示的核苷酸序列，所述宿主菌转化有甲基转移酶表达载体，实现了对宿主菌基因组DNA的甲基化保护作用，避免了NdeI对宿主DNA的切割作用，可以用于表达包括NdeI在内的多种限制性内切酶。
Description
技术领域
本发明属于生物技术领域，涉及一种宿主菌及其构建方法和应用，尤其涉及一种表达限制性内切酶NdeI的甲基化保护宿主菌及其构建方法和应用。
背景技术
在生物体内有一类酶能够切断外来DNA，限制异源DNA的侵入并使之失去活力，但对自身DNA无损害作用，从而起到保护细胞原有遗传信息的作用。由于这种切割作用是在DNA分子内部进行的，故名限制性内切酶(简称限制酶)。限制性内切酶的发现促进了DNA重组技术的诞生，极大地推动了现代分子生物学和基因工程的发展，是当代基因工程研究不可或缺的基础工具。
细菌体内存在“限制-修饰”现象，内源DNA通过被甲基化酶修饰，避免了限制性内切酶的切割作用，达到了抵御外来噬菌体侵染的目的。在原核表达体系中单一地重组表达限制性内切酶，会导致宿主DNA被切割而死亡。甲基转移酶通过在限制性内切酶的识别序列上进行甲基化修饰，可有效防止限制性内切酶的切割作用。限制-修饰基因紧密连锁，细菌通过调控两者的表达，首先使自身DNA受到甲基化保护，随后再表达限制性内切酶，而入侵的异源DNA由于没有受到保护而被降解(Geoffery G.W.，Nucleic Acids Reseach，2539-2564，1991)。
研究表明，限制-修饰系统在原核细胞中的建立和维持是以甲基转移酶的保护作用为前提的。甲基化修饰是蛋白质修饰的重要形式之一，参与了原核生物和真核生物的多项细胞进程。基于DNA分子的结构与性质，自然界中主要存在三类DNA甲基转移酶，分别是C5胞嘧啶甲基酶、N4胞嘧啶甲基酶和N6腺嘌呤甲基酶。其中，N4胞嘧啶甲基酶和N6腺嘌呤甲基酶属于氨基-甲基转移酶(Malone et al.，J.Mol.Biol.，618-632，1995)。DNA在甲基转移酶的作用下，催化底物S-腺苷甲硫氨酸转移一个甲基至胞嘧啶的C5位置，生成5-甲基胞嘧啶。当DNA被甲基转移酶修饰后，便具有了抵抗限制性内切酶的特性。即通常情况下，如果限制酶的识别序列相同或包含在内，并且甲基化酶修饰位点和方式一致，那么经过修饰的DNA就能够抵抗限制性内切酶的切割作用，从而保护宿主细胞DNA免受破坏。例如AluI的识别序列为5’-AGCT-3’，包含在SacI的识别序列5’-GAGCTC-3’之中，两者的甲基化位点和修饰方式一致(C5胞嘧啶甲基酶)，因此可以通过在宿主菌内表达M.AluI来抵抗限制酶SacI对DNA的酶切。
NdeI是一种应用较为广泛的限制性内切酶，可以识别六个碱基的核酸序列(5’-CA^TATG-3’)，不仅可以作为普通的限制性内切酶应用于分子克隆，在生物领域的其他方向也具有重要的应用价值及研究潜力。但是，NdeI限制性内切酶存在表达量低、分离纯化程序繁琐、蛋白得率低等问题，目前市售的NdeI限制酶价格昂贵，限制了其应用与研究。
限制性内切酶具有切割DNA的特性，一旦表达将可能造成宿主细胞的损伤甚至死亡。因此，必须对宿主细胞的DNA进行特异性的甲基化修饰后才能实现重组表达。现有技术方案中，利用NdeI对应的天然特异性甲基转移酶M.NdeI进行宿主DNA的保护，具体步骤包括：提取天然来源菌株Neisseria denitrificans的染色体DNA并制备DNA文库；利用M.NdeI保护DNA不被NdeI切割的特性，从文库中筛选出M.NdeI基因；将带有M.NdeI基因的质粒转化进入表达宿主工程菌株中；获得阳性宿主工程菌后，将NdeI限制酶基因构建于表达载体，转化进入上述菌株，诱导NdeI的重组表达。表达后的纯化步骤涉及亲和层析、离子交换层析、疏水层析、反相层析和凝胶分子排阻层析等纯化过程。然而，该方法的缺点在于：通过繁琐的DNA文库构建和筛选、获得甲基化保护的宿主细胞后，甲基化保护菌株只能特异性保护DNA免于NdeI限制酶的切割，少有能用于其他限制酶的表达，应用范围极为狭窄。
CN 106755002A公开了利用甲基化酶的保护高效重组表达限制性内切酶的方法，所述的甲基化酶是M.CviPI，所述的限制性内切酶C选自：AatI，AscI，BanII，BgII，BmtI，BspQI，BsrDI，BssHII，BtsI，EagI，HindIII，KasI，MluI，NheI，NotI，NruI，PstI，PvuII，SacI，SapI，SbfI，SfiI，SphI。甲基化酶M.CviPI虽然具有广谱保护性，但特异性弱，不能对宿主细胞进行针对特异性限制性内切酶的甲基化修饰，限制性内切酶对宿主细胞DNA具有一定的降解作用，造成表达量低、蛋白得率低等问题。
因此，提供一种兼具特异性与广谱保护性的甲基化保护宿主菌，用于表达限制性内切酶NdeI，在生物技术领域具有重要意义。
发明内容
针对现有技术的不足，本发明提供了一种甲基化保护宿主菌及其构建方法和应用，所述宿主菌转化有具有一定广谱性的甲基转移酶表达载体，可以用于表达包括NdeI在内的多种限制性内切酶。
为达此目的，本发明采用以下技术方案：
第一方面，本发明提供了一种甲基转移酶，所述甲基转移酶的氨基酸序列包括如SEQ ID NO:1所示的氨基酸序列；
所述SEQ ID NO:1的氨基酸序列为：
MWTTSSVAKNENLYNTLDIAINIKFPMAATTVKDISNPDTYKGLYSFHKYWGKKPTESISFFIQNYTSITDIVLDPFLGSGLISRECLSQKRRFIGIDINPFSVEHTKFLLELPEATSFRETFKSVETNIKQKINDTYRTSNGKIASHYLWNGKNLSKIWIKPEVGRSRIEIEPNEFDWENFNSFSQYSVRNIREAIFFTNSRINSNNQMSIYDLFTSRSLHNIDLILDEVKTLPDTLKRAFLLTLTSSSGQMSSMVFAITNRGKAKKQVSNKIEVGSWVIGYWRPHLHFEINVWNCFESRANKLHKTLLNIGRREHPQCESINRLLESTQGALILNEDCLSVMKTIPEKTIKLICTDPPHSDRIPYLELSEMWNSILNEKVCFEKEIIVSNAKERNKKKDEYIKQMKLFINEASRVLTDEGIFLMYFNARDKQSWRFLEVLENSSDLNFVGTFPMEYSANSVVQDNRKGGLKTDYVLVLVKKRFNIKFQHRLDEIPGWSASLPKVGLEP；
所述甲基转移酶的核苷酸序列包括如SEQ ID NO:2所示的核苷酸序列；
所述SEQ ID NO:2的核苷酸序列为：
atgtggaccaccagtagcgttgcgaaaaacgaaaacctgtacaacaccctggatatcgcgatcaatatcaagttcccgatggcggcaaccaccgttaaagacatcagcaacccggatacctacaaaggcctgtacagcttccacaagtactggggcaaaaaaccgaccgaaagcattagcttcttcatccagaactacaccagcatcaccgacattgttctggatccgtttctgggtagcggtctgatttctcgcgagtgtctgagtcaaaagcgccgtttcatcggcatcgatatcaacccgttcagcgtcgaacacaccaaatttctgctggaactgccggaagcaacctcttttcgcgaaaccttcaagagcgtcgagaccaacatcaagcagaagatcaacgacacctaccgtaccagcaacggcaaaattgcgtcccattacctgtggaacggcaaaaacctgagcaagatctggatcaaaccggaagttggtcgtagccgcattgaaattgaaccgaacgagttcgactgggagaacttcaacagcttcagccagtacagcgtccgtaacatccgcgaagcgatctttttcaccaacagccgcatcaactccaacaaccagatgagcatctacgacctgtttaccagccgcagtctgcataacatcgacctgatcctggacgaagttaaaaccctgccggataccctgaaacgcgcatttctgctgaccctgacctcttcttctggtcaaatgagcagcatggtctttgcgatcaccaaccgcggtaaagcgaaaaaacaggtcagcaacaaaatcgaagtcggcagctgggttatcggttattggcgtccgcatctgcatttcgagatcaacgtctggaactgcttcgaaagccgcgcaaacaaactgcataaaaccctgctgaacattggtcgtcgcgaacatccgcagtgcgaaagcattaaccgtctgctggaaagcacccaaggcgcactgattctgaacgaagattgcctgagcgtcatgaaaaccatcccggaaaagaccatcaaactgatttgcaccgatccgccgcatagcgatcgtattccgtacctggaactgtccgaaatgtggaacagcatcctgaacgagaaggtctgcttcgagaaagaaattatcgtcagcaacgcgaaagagcgcaacaagaagaaggacgagtacatcaagcagatgaagctgttcattaacgaagcgagtcgcgttctgaccgatgaaggcatcttcctgatgtacttcaacgcgcgcgataaacagagttggcgtttcctggaagtcctggaaaacagcagcgatctgaacttcgttggcacctttccgatggaatatagcgcgaacagcgtcgttcaggataaccgtaaaggcggtctgaaaaccgactacgttctggttctggtcaagaagcgcttcaacatcaagttccagcaccgtctggacgaaattccgggttggtctgcaagtctgccgaaagttggtctggaaccg.
本发明中，甲基转移酶M.Mae2481ORF83P针对NdeI和FauNDI限制性内切酶均具有宿主保护作用。甲基转移酶M.Mae2481ORF83P的识别序列为5’-CATATG-3’，对第4位碱基A具有甲基化修饰作用，NdeI和FauNDI对DNA的特异性识别序列为5’-CATATG-3’，切割位点在AT碱基间。本发明利用M.Mae2481ORF83P对5’-CATATG-3中碱基A的特异性甲基化修饰，实现了对宿主菌基因组DNA的甲基化保护作用，避免了NdeI和FauNDI对宿主DNA的切割作用。
第二方面，本发明提供了一种甲基转移酶表达载体，包括如第一方面所述的甲基转移酶的核苷酸序列。
第三方面，本发明提供了一种甲基化保护宿主菌，所述甲基化保护宿主菌包括如第二方面所述的甲基转移酶表达载体。
本发明中，转化有M.Mae2481ORF83P甲基转移酶的宿主菌的DNA被甲基化修饰，抵抗了限制性内切酶NdeI对宿主菌DNA的切割作用，实现了NdeI限制性内切酶在宿主菌中的高效表达，同时可应用于其他具有类似识别位点的FauNDI限制性内切酶的重组表达。
第四方面，本发明提供了一种甲基化保护宿主菌的构建方法，所述方法包括：
构建如第二方面所述的甲基转移酶表达载体，转化到宿主菌，抗性筛选后得到所述甲基化保护宿主菌。
优选地，所述构建方法包括以下步骤：
(1)采用如SEQ ID NO:3-4所示的引物对，从菌株Microcystis aeruginosaNIES-2481的基因组中获得甲基转移酶M.Mae2481ORF83P基因；
(2)将M.Mae2481ORF83P基因插入pACYC184的NdeI和XhoI酶切位点，转化入大肠杆菌克隆菌株，氯霉素抗性筛选后提取质粒，得到甲基转移酶表达载体；
(3)将甲基转移酶表达载体转化入大肠杆菌表达菌株，氯霉素抗性筛选后得到所述甲基化保护宿主菌；
所述SEQ ID NO:3的核酸序列为：
5’-CTTTAAGAAGGAGATATACCAATGTGGACCACCAGTAG-3’；
所述SEQ ID NO:4的核酸序列为：
5’-CCGCATTAAAGCTTCTTACATATGTTACGGTTCCAGAC-3’.
本发明中，采用如SEQ ID NO:3-4所示的引物对特异性扩增M.Mae2481ORF83P，同时引入了NdeI的酶切位点CATATG，方便后续进行筛选实验，验证甲基转移酶表达载体成功导入宿主菌中。
根据本发明，过高的甲基化酶表达和修饰水平对宿主同样具有致死效果，本发明将M.Mae2481ORF83P基因序列插入低拷贝表达载体pACYC184质粒中，实现了甲基化酶在宿主菌中的正常表达。
第五方面，本发明提供了一种NdeI限制性内切酶的制备方法，所述方法包括：
将包含NdeI核苷酸序列的重组质粒转化入如第三方面所述的甲基化保护宿主菌，抗性筛选后得到重组菌株；诱导培养，得到所述NdeI限制性内切酶。
优选地，所述制备方法包括以下步骤：
(1’)采用如SEQ ID NO:5-6所示的引物对，从菌株Neisseria denitrificans的基因组中获得限制性内切酶NdeI基因；
(2’)将NdeI基因插入pBAD的NdeI和HindIII酶切位点，转化入大肠杆菌克隆菌株，氨苄青霉素抗性筛选后提取质粒，得到包含NdeI核苷酸序列的重组质粒；
(3’)将包含NdeI核苷酸序列的重组质粒转化入如第三方面所述的甲基化保护宿主菌，氨苄青霉素和氯霉素抗性筛选后得到重组菌株；
(4’)对所述重组菌株进行阿拉伯糖诱导培养，得到所述NdeI限制性内切酶。
所述SEQ ID NO:5的核酸序列为：
5’-TAACAGGAGGAATTAACCATGGGCAAAAACCTGAGTTTTAGCCAGC-3’；
所述SEQ ID NO:6的核酸序列为：
5’-CAAAACAGCCAAGCTTCGAATTCTTCAGGTCTGCTTAATAAACAGAATATC-3’.
本发明中，利用阿拉伯糖操纵子表达系统严苛控制了限制性内切酶NdeI的本底表达水平。
第六方面，本发明提供了一种NdeI限制性内切酶，采用如第五方面所述的方法制备得到。
优选地，所述NdeI限制性内切酶的氨基酸序列如SEQ ID NO:7所示；
所述SEQ ID NO:7的氨基酸序列为：
GKSLGFQQLDLFERGDDEPIIFQHQYLIQYIKLQSMSQKFLMFFEERFYFLDLSKIKSKFYEQYAEFNLDLTLFSLNSELFKKYRLFQTDQIIRFDYIVDRIQIRYEWGGSEGNSIEKNIVDNYVKKIQSFDIINKKIFTFIFSSIQGYTINSWYNHWTSIIIFDIFKDHANVIPTIGIIKKIDFFINFIPFDIKVTYFPFQFIAFKIKQKGFGNFITQIKQICQKINIIIPNDMSDKNIKIHIYTKVSFCHHKFAKFIINFINKIKKQIIQFAFQNSDFIKVWIYFNQGFAQFDASNQFFIIITDFTNINDSWKIKQNIKFIQFKIHSHIDSIKIDINKINTKFYWKKTNFHFNCKSDIIFIKQT.
第七方面，本发明提供了一种如第一方面所述的甲基转移酶、如第二方面所述的甲基转移酶表达载体、如第三方面所述的甲基化保护宿主菌或如第六方面所述的NdeI限制性内切酶在制备分子检测试剂盒中的应用。
与现有技术相比，本发明具有如下有益效果：
(1)本发明通过向宿主菌中转化入M.Mae2481ORF83P甲基转移酶表达载体，构建得到甲基化保护菌株，可稳定高效表达限制性内切酶NdeI，抵抗了限制性内切酶NdeI对宿主基因组DNA的酶切作用；
(2)本发明利用阿拉伯糖操纵子表达系统，严苛控制了限制性内切酶NdeI的本底表达水平，对宿主菌无损害作用；
(3)本发明的甲基化保护宿主菌不仅可以用于表达NdeI限制性内切酶，而且可以应用于其他具有类似识别位点的限制酶的重组表达，比如FauNDI。
附图说明
图1为pACYC184-M.Mae2481ORF83P载体图谱；
图2为pBAD-R.NdeI载体图谱；
图3为限制性内切酶NdeI甲基化保护菌株的筛选电泳图，其中，1-NdeI酶切线性化质粒λ，2-对照质粒pACYC184-M.Mae2481ORF83P，3-NdeI酶切质粒pACYC184-M.Mae2481ORF83P；
图4为将NdeI原液进行不同梯度稀释后的特异性酶切活力检测电泳图，其中，1-4倍稀释，2-8倍稀释，3-16倍稀释，4-32倍稀释，5-64倍稀释，6-128倍稀释。
具体实施方式
为进一步阐述本发明所采取的技术手段及其效果，以下结合实施例和附图对本发明作进一步地说明。可以理解的是，此处所描述的具体实施方式仅仅用于解释本发明，而非对本发明的限定。
实施例中未注明具体技术或条件者，按照本领域内的文献所描述的技术或条件，或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者，均为可通过正规渠道商购获得的常规产品。
1、菌株与质粒
Microcystis aeruginosaNIES-2481菌株来自美国菌种保藏中心(ATCC)；
Neisseria denitrificans菌株来自美国菌种保藏中心(ATCC)；
Trans1 T1购自北京全式金生物技术有限公司；
ER2566购自赛默飞世尔科技有限公司(Thermo Fisher Scientific)；
pBAD、pACYC184、pUC19购自淼灵生物质粒平台；
酶活检测底物λDNA购自赛默飞世尔科技有限公司(Thermo Fisher Scientific)；
2、仪器与试剂
JN-02C低温超高压连续流细胞破碎仪购自广州聚能纳米生物科技股份有限公司；
蛋白分子量标准(PAGE-MASTER Protein Standard Plus)购自金斯瑞生物科技有限公司；
限制性内切酶FlashCutTM NdeI，FlashCutTM HindIII，FlashCutTM XhoI购自莫纳生物科技有限公司；
NdeI购自New England Biolabs(NEB)；
MonHI-FI DNA Polymerase购自莫纳生物科技有限公司；
MonCloneTM Fast AP购自莫纳生物科技有限公司；
MonClone Hi-Fusion Cloning Mix购自莫纳生物科技有限公司；
10×FlashOneTM Buffer购自莫纳生物科技有限公司。
实施例1甲基转移酶M.Mae2481ORF83P宿主菌的构建
(1)获取甲基转移酶M.Mae2481ORF83P基因
采用PCR扩增的方法，从菌株Microcystis aeruginosaNIES-2481的基因组中使用甲基转移酶M.Mae2481ORF83P基因的特异性引物，获得甲基转移酶M.Mae2481ORF83P基因，具体步骤为：
去离子水重悬菌株Microcystis aeruginosaNIES-2481的平板培养菌落，95℃温育10min，作为PCR模板，SEQ ID NO:3-4的核酸序列作为PCR引物对，使用MonHI-FI DNAPolymerase，按照标准流程进行PCR反应，获得了与理论值大小一致的M.Mae2481ORF83P基因片段。
(2)构建甲基转移酶表达载体
pACYC184质粒采用FlashCutTM NdeI和FlashCutTM XhoI双酶切后，以MonCloneTMFast AP进行去磷酸化处理，酶切产物进行琼脂糖凝胶电泳，割胶回收线性化的pACYC184载体片段，溶于TE Buffer中；
采用MonClone Hi-Fusion Cloning Mix，将步骤(1)获得的M.Mae2481ORF83P基因片段与线性化的pACYC184载体无缝克隆连接，在50℃下反应15min，使用化学转化法转化进入Trans1 T1感受态细胞，氯霉素筛选后获得重组表达质粒pACYC184-M.Mae2481ORF83P，如图1所示为pACYC184-M.Mae2481ORF83P的载体图谱，其序列经过测序确认；
(3)甲基化保护宿主菌的筛选
将步骤(2)得到的测序正确的pACYC184-M.Mae2481ORF83P通过化学转化法转化进入大肠杆菌ER2566，在氯霉素筛选条件下进行细胞培养，挑选单克隆，在含有35μg/mL氯霉素的琼脂平板上划线，编号后于37℃、200rpm培养16h，收取菌液提取质粒；
在10μL反应体系中，取0.2μL限制性内切酶NdeI，在缓冲条件下，与100ng pACYC184-M.Mae2481ORF83P在37℃温育1h，使用0.8％琼脂糖凝胶电泳检测底物被NdeI消化的情况。
结果如图3所示，采用NdeI酶切的重组质粒pACYC184-M.Mae2481ORF83P与未经NdeI酶切的对照质粒的条带完全一致，表明pACYC184-M.Mae2481ORF83P在宿主菌体内受到M.Mae2481ORF83P甲基转移酶的完全保护，默认宿主菌基因组DNA同样可受到M.Mae2481ORF83P甲基转移酶的保护。经再次验证保护性后，将平板上相应编号的菌株使用Inoue处理，制备成感受态细胞[pACYC184-M.Mae2481ORF83P,ER2566]。
实施例2限制性内切酶NdeI重组表达菌株的构建
(1)获取限制性内切酶NdeI基因
采用PCR扩增的方法，从菌株Neisseria denitrificans的基因组中使用限制性内切酶NdeI基因的特异性引物，获得限制性内切酶NdeI基因，具体步骤为：
去离子水重悬菌株Neisseria denitrificans的平板培养菌落，95℃温育10min，作为PCR模板，SEQ ID NO:5-6的核酸序列作为PCR引物对，使用MonHI-FI DNA Polymerase，按照标准流程进行PCR反应，获得了与理论值大小一致的R.NdeI基因片段。
(2)构建限制性内切酶NdeI表达载体
pBAD质粒采用FlashCutTM NdeI和FlashCutTM HindIII双酶切后，以MonCloneFast AP进行去磷酸化处理，酶切产物进行琼脂糖凝胶电泳，割胶回收线性化的pBAD载体片段，溶于TE Buffer中；
采用MonClone Hi-Fusion Cloning Mix，将步骤(1)获得的R.NdeI基因片段与线性化的pBAD载体片段无缝克隆连接，在50℃下反应15min，使用化学转化法转化进入Trans1T1感受态细胞，氨苄青霉素筛选后获得重组表达质粒pBAD-R.NdeI，如图2所示为pBAD-R.NdeI的载体图谱，其序列经过测序确认；
(3)限制性内切酶NdeI重组表达菌株的筛选
将步骤(2)得到的测序正确的pBAD-R.NdeI转化进入感受态细胞[pACYC184-M.Mae2481ORF83P，ER2566]，在氨苄青霉素和氯霉素筛选条件下进行细胞培养，获得限制性内切酶NdeI的重组表达菌株。
(4)限制性内切酶NdeI的重组表达
将限制性内切酶NdeI的重组表达菌株接种到LB培养基中，在37℃摇床中以200rpm培养至OD＝0.8，加入终浓度为0.2％的阿拉伯糖诱导剂，并在16℃摇床中以200rpm培养16h，得到的细菌蛋白粗提液中具有限制性内切酶NdeI的活性。
实施例3限制性内切酶NdeI的活力检测
限制性内切酶NdeI的酶活定义为：37℃下，在20μL反应体系中，1μL限制性内切酶NdeI能够在15min内完全消化1μgλDNA。
本实施例对经纯化的限制性内切酶NdeI进行梯度稀释，在1×FlashOneTM Buffer(莫纳生物科技有限公司)缓冲条件下、20μL反应体系中，与底物λDNA在37℃下温育60min，然后以1％琼脂糖凝胶电泳检测底物的酶切情况。结果如图4所示，纯化获得的限制酶NdeI具有良好的酶切活力，比活力约为900,000U/mg诱导菌液。
综上所述，本发明通过向宿主菌中转化入M.Mae2481ORF83P甲基转移酶表达载体，构建得到甲基化保护菌株，可稳定高效表达限制性内切酶NdeI，抵抗了限制性内切酶NdeI对宿主基因组DNA的酶切作用；本发明的甲基化保护宿主菌不仅可以用于表达NdeI限制性内切酶，而且可以应用于其他具有类似识别位点的限制酶的重组表达；本发明利用阿拉伯糖操纵子表达系统，严苛控制了限制性内切酶NdeI的本底表达水平，对宿主菌无损害作用。
申请人声明，本发明通过上述实施例来说明本发明的详细方法，但本发明并不局限于上述详细方法，即不意味着本发明必须依赖上述详细方法才能实施。所属技术领域的技术人员应该明了，对本发明的任何改进，对本发明产品各原料的等效替换及辅助成分的添加、具体方式的选择等，均落在本发明的保护范围和公开范围之内。
SEQUENCE LISTING
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Claims (10)
1.一种甲基转移酶，其特征在于，所述甲基转移酶的氨基酸序列包括如SEQ ID NO:1所示的氨基酸序列；
所述甲基转移酶的核苷酸序列包括如SEQ ID NO:2所示的核苷酸序列。
2.一种甲基转移酶表达载体，其特征在于，包括如权利要求1所述的甲基转移酶的核苷酸序列。
3.一种甲基化保护宿主菌，其特征在于，所述甲基化保护宿主菌包括如权利要求2所述的甲基转移酶表达载体。
4.一种甲基化保护宿主菌的构建方法，其特征在于，所述方法包括：
构建如权利要求2所述的甲基转移酶表达载体，转化到宿主菌，抗性筛选后得到所述甲基化保护宿主菌。
5.根据权利要求4所述的方法，其特征在于，所述构建方法包括以下步骤：
(1)采用如SEQ ID NO:3-4所示的引物对，从菌株Microcystis aeruginosaNIES-2481的基因组中获得甲基转移酶M.Mae2481ORF83P基因；
(2)将M.Mae2481ORF83P基因插入pACYC184的NdeI和XhoI酶切位点，转化入大肠杆菌克隆菌株，氯霉素抗性筛选后提取质粒，得到甲基转移酶表达载体；
(3)将甲基转移酶表达载体转化入大肠杆菌表达菌株，氯霉素抗性筛选后得到所述甲基化保护宿主菌。
6.一种NdeI限制性内切酶的制备方法，其特征在于，所述方法包括：
将包含NdeI核苷酸序列的重组质粒转化入如权利要求3所述的甲基化保护宿主菌，抗性筛选后得到重组菌株；诱导培养，得到所述NdeI限制性内切酶。
7.根据权利要求6所述的方法，其特征在于，所述制备方法包括以下步骤：
(1’)采用如SEQ ID NO:5-6所示的引物对，从菌株Neisseria denitrificans的基因组中获得限制性内切酶NdeI基因；
(2’)将NdeI基因插入pBAD的NdeI和HindIII酶切位点，转化入大肠杆菌克隆菌株，氨苄青霉素抗性筛选后提取质粒，得到包含NdeI核苷酸序列的重组质粒；
(3’)将包含NdeI核苷酸序列的重组质粒转化入如权利要求3所述的甲基化保护宿主菌，氨苄青霉素和氯霉素抗性筛选后得到重组菌株；
(4’)对所述重组菌株进行阿拉伯糖诱导培养，得到所述NdeI限制性内切酶。
8.一种NdeI限制性内切酶，其特征在于，采用如权利要求6或7所述的方法制备得到。
9.根据权利要求8所述的NdeI限制性内切酶，其特征在于，所述NdeI限制性内切酶的氨基酸序列如SEQ ID NO:7所示。
10.一种如权利要求1所述的甲基转移酶、如权利要求2所述的甲基转移酶表达载体、如权利要求3所述的甲基化保护宿主菌或如权利要求8或9所述的NdeI限制性内切酶在制备分子检测试剂盒中的应用。
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5139942A (en) * | 1988-05-19 | 1992-08-18 | New England Biolabs, Inc. | Method for producing the nde i restriction endonuclease and methylase |
| US6869786B1 (en) * | 2003-01-08 | 2005-03-22 | New England Biolabs, Inc. | Method for cloning and expression of BsrGI restriction endonuclease and BsrGI methyltransferase in E. coli |
| CN106755002A (zh) * | 2016-11-25 | 2017-05-31 | 江苏愚公生命科技有限公司 | 利用甲基化酶的保护高效重组表达限制性内切酶的方法 |
| CN107557372A (zh) * | 2017-08-30 | 2018-01-09 | 江苏愚公生命科技有限公司 | 用于表达限制性内切酶SacI的甲基化保护菌株的筛选方法 |
-
2019
- 2019-09-03 CN CN201910828288.4A patent/CN110438102A/zh active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5139942A (en) * | 1988-05-19 | 1992-08-18 | New England Biolabs, Inc. | Method for producing the nde i restriction endonuclease and methylase |
| US6869786B1 (en) * | 2003-01-08 | 2005-03-22 | New England Biolabs, Inc. | Method for cloning and expression of BsrGI restriction endonuclease and BsrGI methyltransferase in E. coli |
| CN106755002A (zh) * | 2016-11-25 | 2017-05-31 | 江苏愚公生命科技有限公司 | 利用甲基化酶的保护高效重组表达限制性内切酶的方法 |
| CN107557372A (zh) * | 2017-08-30 | 2018-01-09 | 江苏愚公生命科技有限公司 | 用于表达限制性内切酶SacI的甲基化保护菌株的筛选方法 |
Non-Patent Citations (2)
| Title |
|---|
| H. YAMAGUCHI: "M.Mae2481ORF83P，REBASE Enz Num 154475", 《REBASE》 * |
| ROBERT J. WATSON等: "NdeI: a restriction endonuclease from Neisseria denitrificans which cleaves DNA at 5-CATATG-3 sequences", 《FEBS LETTERS》 * |
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