流产布鲁氏菌2308株磷酸甘露糖变位酶pmm基因缺失株构建及鉴定  

Construction and Indentification of Phosphomannomutase Gene (pmm) Deletion Mutants of Brucella abortus Strain 2308

作　　者：李亚[1] 张辉[1] 郭飞[1] 陈创夫[1] 张科[1] 桂丹[1] 温书香 纪太旺 张欢[1] 

机构地区：[1]石河子大学动物科技学院,新疆石河子832000

出　　处：《西北农业学报》2014年第6期29-32,共4页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金(31260596;31360610);兵团国际科技合作项目(2013BC005)

摘　　要：旨在构建流产布鲁氏菌2308株pmm基因缺失株。PCR扩增流产布鲁氏菌2308株pmm基因的上、下游同源臂片段,并与枯草芽孢杆菌上SacB基因片段融合。连接至自杀载体PGEM-7Zf+,构建自杀载体PGEM-7Zf+Δpmm-SacB。电转化至流产布鲁氏菌2308株感受态细胞内,通过氨苄抗性和蔗糖敏感性筛选,获得基因缺失株2308Δpmm,对其进行PCR鉴定和稳定性检测。成功构建可稳定遗传的2308Δpmm基因缺失株,并在15代内未发生回复。在此基础上可以对2308Δpmm进行深入研究。To construct Brucella abortus strain 2308Δpmm mutants,the upstream and downstream homology arm fragments of pmmgene from B.abortus strain 2308were cloned by PCR,and fusion with the fragments of sacBfromBacillus subtilis.The fusion product was connected with the suicide vector PGEM-7Zf＋.The recombination plasmid PGEM-7Zf＋Δpmm-sacB was constructed,and electroporated into B.abortus strain 2308competent cells.We screened Brucellagene deletion mutant by ampicillin resistance screened and sugar continuous bacteria culture.The 2308Δpmmwas identified by PCR,and its stability was detected by continuous bacteria culture.We obtained a stable genetic2308Δpmmsuccessfully.The reverse mutation did not occur within 15generations.We can make further research based on the mutant strain 2308Δpmm.

关 键 词：流产布鲁氏菌2308株 pmm基因 基因缺失株 

分 类 号：S852.61[农业科学—基础兽医学]