中文摘要：

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。

英文摘要：

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.