以克隆载体为自杀载体快速构建布鲁氏菌的无痕缺失突变株

Construction of Brucella Unmarked Deletion Mutant by Using Conventional Cloning Vector as Suicide Plasmid

摘要:

构建突变株是病原微生物致病机理研究的重要手段。以往研究中布鲁氏菌的无痕缺失突变株都采用传统的自杀载体来构建，效率低下。首先对布鲁氏菌的电击转化条件进行了优化，然后选择含有反向筛选基因sacB的pEX18Gm质粒作为自杀载体，构建了缺失Ⅳ型启动子区的布鲁氏菌无痕缺失突变株。这不仅为构建布鲁氏菌的突变株提供了一个快速有效的技术平台，也为深入研究Ⅳ型分泌系统的功能奠定了基础。

Abstract:

Construction of mutant strain is an essential method in pathogenesis researches. The conventional method for Brucella unmarked deletion mutant construction is based on suicide plasmid, but the efficiency is very low. In the present study, we first optimized the electroporation parameters, and then, the cloning plasmid pEX18Gm containing sacB was successfully used to construct unmarked deletion mutant of the type IV secretion system. This indicated that by using conventional cloning plasmid as suicide plasmid in Brucella, unmarked deletion mutants can be constructed with high efficiency.