鼠伤寒沙门菌spvB 基因缺陷突变株的构建

生物技术通报 ›› 2012, Vol. 0 ›› Issue (12): 180-183.

鼠伤寒沙门菌spvB 基因缺陷突变株的构建

邱桥成^{1, 2}, 叶颖¹, 储元元¹, 廖莉¹, 吴淑燕¹, 黄瑞¹

1 苏州大学医学部病原生物学系，苏州 215123 ；2 苏州大学附属第一医院江苏省血液研究所，苏州 215006

收稿日期:2012-05-23 修回日期:2013-01-25 出版日期:2012-12-26 发布日期:2013-02-06
作者简介:邱桥成, 男, 硕士, 研究方向:病原分子生物学;E-mail:qiuqiaocheng@163.com
基金资助:
: 国家自然科学基金资助项目（30972768）, 江苏省自然科学基金资助项目（L2134051511）, 江苏省青蓝工程（SR13400211）, 高等学校博士学科点专项科研基金项目（20103201110009）

Construction of the Mutant of the Salmonella typhimurium BacteriaspvB Genetic Defect

Qiu Qiaocheng^1,2, Ye Ying¹, Chu Yuanyuan¹, Liao Li¹, Wu Shuyan¹, Huang Rui¹

1Department of Microbiology of Soochow University School of Medicine， Suzhou 215123 ；2 the First Affiliated Hospital of Soochow University，Jiangsu Institute of Hematology，Suzhou 215006

Received:2012-05-23 Revised:2013-01-25 Published:2012-12-26 Online:2013-02-06

摘要/Abstract

摘要： 高度保守的spvB基因，其编码产物具有ADP 核糖基转移酶活性，是导致受染后细胞发生凋亡的重要毒力因子。为进一步研究该基因致病机制，构建了鼠伤寒沙门菌spvB基因缺陷突变株。根据GenBank 鼠伤寒沙门菌spvB基因序列，用Primer Premier 5.0 设计PCR 特异性引物，获得spvB基因缺陷性核苷酸片段后连接至自杀质粒PGMB151。将构建的自杀载体导入含有spvB基因的鼠伤寒沙门菌标准株SR-11 中进行同源重组，PCR 筛选缺陷突变株。经PCR 和DNA 序列测定，成功获得了spvB基因缺陷的鼠伤寒沙门菌突变株。

关键词: 鼠伤寒沙门菌, spvB, 毒力基因, 同源重组, 基因敲除, 自杀质粒

Abstract: Highly conserved spvB gene is an important virulence factor of infected cells apoptosis, which encoded product with the ADPribosyltransferaseenzymatic activity. To further study the pathogenic mechanism of the spvB gene, this study began to build a mutant of the spvB gene in Salmonella typhimurium. According to the spvB gene sequence in GeneBank, two pair’s primers were designed with Primer Premier 5.0, upper- and down-stream of the spvB gene to amplify two homologous DNA fragments. The homologous recombinant DNA fragment of the defective target gene was cloned into the suicide plasmid PGMB151, which was then transduced into the target cell of S. typhimurium SR-11 for homologous recombination. The recombination was identified by PCR. A deletion mutant of the spvB gene in SR-11 was confirmed by PCR and sequencing analysis.

邱桥成, 叶颖, 储元元, 廖莉, 吴淑燕, 黄瑞. 鼠伤寒沙门菌spvB 基因缺陷突变株的构建[J]. 生物技术通报, 2012, 0(12): 180-183.

Qiu Qiaocheng, Ye Ying, Chu Yuanyuan, Liao Li, Wu Shuyan, Huang Rui. Construction of the Mutant of the Salmonella typhimurium BacteriaspvB Genetic Defect[J]. Biotechnology Bulletin, 2012, 0(12): 180-183.

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