cagA基因缺失的中国幽门螺杆菌突变菌株的构建及鉴定
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黄志刚, 段广才, 范清堂, 黄学勇
幽门螺杆菌;cagA基因;缺失表达;突变体,黄志刚,段广才,黄学勇,段广才,范清堂,黄志刚,通讯作者:,Constructionand
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黄志刚, 段广才, 黄学勇, 郑州大学公共卫生学院流行病学教研室 河南省郑州市 450052
段广才, 范清堂, 郑州大学河南省分子医学重点实验室 河南省郑州市 450052
黄志刚, 河北工程大学医学院教师, 副教授, 郑州大学公共卫生学院在读博士研究生, 主要从事幽门螺杆菌分子流行病学研究.
通讯作者: 段广才, 450052, 河南省郑州市大学路40号, 郑州大学河南省分子医学重点实验室. gcduan@public.zz.ha.cn
电话: 0371-66912869
收稿日期: 2006-07-29 接受日期: 2006-08-14
Construction and identification of Chinese mutant Helicobacter pylori strain with absent expression of cagA gene
Zhi-Gang Huang, Guang-Cai Duan, Qing-Tang Fan, Xue-Yong Huang
Zhi-Gang Huang, Guang-Cai Duan, Xue-Yong Huang,Department of Epidemiology, College of Public Health, Zhengzhou University, Zhengzhou 450052, Henan Province, China
Guang-Cai Duan, Qing-Tang Fan,Henan Provincial Key Laboratory of Molecular Medicine, Zhengzhou 450052, Henan Province, China
Correspondence to:Guang-Cai Duan, Henan Provincial Key Laboratory of Molecular Medicine, Zhengzhou 450052, Henan Province, China. gcduan@public.zz.ha.cn
Received: 2006-07-29 Accepted: 2006-08-14
Abstract
AIM: To knock out the entire cagA gene of Chinese Helicobacter pylori strain via homologous recombination and construct a cagA-deleted mutant strain of Chinese H pylori.
METHODS: Two DNA fragments locating in the upper and downstream of cagA gene were amplified and a kanamycin resistance gene of pEGFP-N2 between them were engineered into pBluescript SK II(-) plasmid (pBs-cagA-mutant). Electrotransformation of H pylori cells with pBs-cagA-mutant resulted in isolation of kanamycin-resistant H pylori transformants, which was identified by polymerase chain reaction (PCR) and sequencing analysis.
RESULTS: Restriction endonuclease analyses showed that pBs-cagA-mutant vector had been successfully recombined. The cagA-deleted status of Chinese H pylori mutants was confirmed by PCR with primers specific for the genes of cagA and kanamycin resistance. After 25 generations of culture, the constructed Chinese H pylori mutants were confirmed stable ......
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