| 利用Red重组系统敲除大肠杆菌菌株ClpP基因方法的研究 |
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| 引用本文： | 司微,刘慧芳,王春来,彭玮,赫明雷,杜艳芬,赵海玲,王聃,杨金国,林靖凯,倪洪波,刘思国.利用Red重组系统敲除大肠杆菌菌株ClpP基因方法的研究[J].黑龙江畜牧兽医,2011(15). |
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| 作者姓名： | 司微 刘慧芳 王春来 彭玮 赫明雷 杜艳芬 赵海玲 王聃 杨金国 林靖凯 倪洪波 刘思国 |
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| 作者单位： | 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室动物细菌病研究室; |
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| 基金项目： | 中国农业科学院哈尔滨兽医研究所中央级公益性科研院所基本科研业务费专项(2008-14) |
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| 摘 要： | 为了研究ClpP基因的功能,试验利用Red系统的重组功能,通过PCR扩增出两端与大肠杆菌菌株ClpP基因上、下游序列同源,中间为氯霉素抗性基因cat的DNA片段,电击转化含质粒pKD46的大肠杆菌菌株DH091,筛选替换ClpP基因的阳性转化子,再导入质粒pCP20去除氯霉素抗性基因。结果表明:成功敲除大肠杆菌菌株ClpP基因。
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| 关 键 词： | Red重组系统 ClpP基因 基因敲除 |
The research on the deletion of ClpP gene in chromosome of E. coli by Red recombination system |
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| SI Wei,LIU Hui-fang,WANG Chun-lai,PENG Wei,HE Ming-lei,DU Yan-fen,ZHAO Hai-ling,WANG Dan,YANG Jin-guo,LIN Jing-kai,NI Hong-bo,LIU Si-guo,Harbin ,China.The research on the deletion of ClpP gene in chromosome of E. coli by Red recombination system[J].Heilongjiang Animal Science And veterinary Medicine,2011(15). |
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| Authors: | SI Wei LIU Hui-fang WANG Chun-lai PENG Wei HE Ming-lei DU Yan-fen ZHAO Hai-ling WANG Dan YANG Jin-guo LIN Jing-kai NI Hong-bo LIU Si-guo Harbin China |
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| Institution: | SI Wei,LIU Hui-fang,WANG Chun-lai,PENG Wei,HE Ming-lei,DU Yan-fen,ZHAO Hai-ling,WANG Dan,YANG Jin-guo,LIN Jing-kai,NI Hong-bo,LIU Si-guo(Division of Bacterial Diseases,National Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences(CAAS),Harbin 150001,China) |
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| Abstract: | With the help of Red recombination system,the chloramphenicol resistance gene flanked by homologues of the ClpP gene was amplified by PCR.The PCR products were electro-transformed into E.coli DH091 strain with pKD46.After ClpP gene was replaced by the chloramphenicol gene,the resistance gene was then eliminated by using pCP20,then the ClpP gene was completely deleted by this procedure.This research can provide a basis on the function of the ClpP gene. |
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| Keywords: | Red recombination system ClpP gene gene deletion |
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