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首页> 外文期刊>Infection and immunity >Deletion of the BCSP31 gene of Brucella abortus by replacement.
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Deletion of the BCSP31 gene of Brucella abortus by replacement.

机译：通过置换缺失流产布鲁氏菌BCSP31基因。

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The 31-kDa salt-extractable immunogenic protein, BCSP31, was deleted from several Brucella abortus strains by replacement with a marker gene encoding resistance to the antibiotics kanamycin and neomycin. The BCSP31 gene replacement plasmids, constructed with ColE1-derived vectors, were introduced by electroporation into B. abortus strain 19 (S19), into a rough variant of B. abortus S19, and into B. abortus S2308, and antibiotic-resistant transformants were isolated. B. abortus S19 is an attenuated strain used as a vaccine for prevention of bovine brucellosis in the United States, and B. abortus S2308 is a commonly used challenge strain. The antibiotic-resistant isolates were all obtained by recombination; none were spontaneous mutants. Loss of the gene encoding BCSP31 and presence of the marker gene were confirmed by Southern analysis. Vector sequences were either absent or linked to the genome, indicating that ColE1-derived plasmids are not maintained in B. abortus. Survival of B. abortus mutant strains in the macrophagelike cell line J774 and in HeLa cells was examined and shown to be indistinguishable from that of the parental strain.
机译：通过用编码对卡那霉素和新霉素具有抗性的标记基因替代，从数种布鲁氏流产菌株中删除了31 kDa的盐可提取免疫原性蛋白BCSP31。将通过ColE1衍生载体构建的BCSP31基因置换质粒通过电穿孔导入流产双歧杆菌菌株19（S19），流产双歧杆菌S19的粗糙变体和流产双歧杆菌S2308中，并将抗生素抗性转化子导入隔离的。流产双歧杆菌S19是减毒株，在美国用作预防牛布鲁氏菌病的疫苗，而流产双歧杆菌S2308是常用的挑战株。耐药菌株均通过重组获得;没有一个是自发突变体。通过Southern分析证实了编码BCSP31的基因的丢失和标记基因的存在。载体序列不存在或与基因组连接，表明在流产芽孢杆菌中未保留源自ColE1的质粒。检查了巨噬细胞样细胞系J774和HeLa细胞中流产芽孢杆菌突变株的存活情况，显示与亲本株没有区别。

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