| 同源重组法构建枯草芽孢杆菌224 yugS基因缺失突变株 |
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| 引用本文： | 刘杰,房春红,李琬,矫洪涛,喻江,李景鹏.同源重组法构建枯草芽孢杆菌224 yugS基因缺失突变株[J].生物技术通报,2007,4(4):148-151. |
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| 作者姓名： | 刘杰 房春红 李琬 矫洪涛 喻江 李景鹏 |
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| 作者单位： | 东北农业大学生命科学学院,哈尔滨,150030 |
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| 摘 要： | 从枯草芽孢杆菌224的基因组DNA中分别扩增出溶血样基因yugS的上、下游片段,并利用携带新霉素抗性基因(neor)的重组质粒pMD18-T-neo作为骨架,构建了基于yugS基因位点的基因阻断质粒pMD18-T-neo-yugS,线性化后电转化入枯草杆菌224,从新霉素抗性平板上挑取转化子,通过对基因组的PCR鉴定和核苷酸测序证明,确定转化子yugS156为yugS基因缺失突变株。
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| 关 键 词： | 枯草杆菌224 yugS 基因 同源重组 |
| 修稿时间： | 2007-02-13 |
Construction of Bacillus Subtilis 224yugS Mutant by Homologous Recombination Method |
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| Liu Jie,Fang Chunhong,Li Wan,Jiao Hongtao,Yu Jiang,Li Jingpeng.Construction of Bacillus Subtilis 224yugS Mutant by Homologous Recombination Method[J].Biotechnology Bulletin,2007,4(4):148-151. |
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| Authors: | Liu Jie Fang Chunhong Li Wan Jiao Hongtao Yu Jiang Li Jingpeng |
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| Institution: | college of life science,Northeast Agriculture University,Harbin 150030 |
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| Abstract: | Upstream and downstream fragments of yugS gene which might be related to hemolysis were amplified by PCR using chromosomal DNA of Bacillus Subtilis 224 as a template,gene disruption plasmid pMD18-T-neo-yugS which can be integrated into yugS locus within Bacillus Subtilis 224 chromosome was constructed by using recombination plasmid pMD18-T-neo,which carried the neomycin resistance gene(neor)as a resistant marker,as a skelecton.Then BS224 was electro-transformated with linear pMD18-T-neo-yugS.The neor transformants was verified by genome PCR identification and nucleotide sequence evidence,finally transformant yugS156 was testified as a yugS gene imperfection strain. |
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| Keywords: | Bacillus Subtilis 224 yugS gene Homologous recombination |
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