中国兽医科学2011，Vol.41Issue(3)：280-286,7.
布鲁氏菌bp26基因缺失株的构建
Construction of unmarked bp26 gene-deleted strains of Brucella spp.
摘要
Abstract
Three unmarked gene deletion strains of Brucella spp., S19-Δbp26, M5-Δbp26 and S2-Δbp26,were constructed by the allelic exchange introduced by the transformation of the pRE1l2 suicide plasmid containing counter-selection gene sacB.Firstly, the upstream and downstream fragments of bp26 gene were amplified from Brucella spp.genome and then subcloned into suicide plasmid pRE112 to construct the recombinant suicide vector pRE-Δbp26 with 681 bp-deleted bp26 gene fragment.The recombinant suicide vector pREΔbp26 was transformed by electroporation into Brucella spp.and the transformants were selected on TSB-YE plates containing 5 μg/mL chloramphenicol and then 70 g/L sucrose.The successful construction of bp26 gene deletion strains of Brucella spp.were confirmed by the bacterial colony PCR and DNA sequencing.Twenty generations of continuous passage of the S19-Δbp26, M5-Δbp26 and S2-Δbp26 strains showed genetic stability in vitro.The application of pRE112 provided a new suicide plasmid for the construction of unmarked gene deletion strain of Brucella spp.and these strains would be utilized for the study of gene function of Brucella.In addition,these bp26 gene deletion strains could be adapted to develop a live and attenuated Brucella carrier.关键词
布鲁氏菌/bp26基因/自杀质粒/基因缺失株分类
农业科技引用本文复制引用
潘文,王佳莹,赵明秋,珺春梅,易琳,常艳,虞红娇,陈金顶..布鲁氏菌bp26基因缺失株的构建[J].中国兽医科学,2011,41(3):280-286,7.基金项目
广东省动植物防疫检疫研究专项(粤财农[2009]225) （粤财农[2009]225）
国家自然科学基金-广东省自然科学基金联合项目(U0631006) （U0631006）
教育部"长江学者和创新团队发展计划"创新团队项目(IRT0723) （IRT0723）
广东省自然科学基金创新团队项目(5200638) （5200638）
