布鲁菌S19疫苗株znuA单基因及bp26/znuA双基因缺失株的构建及鉴定  

Construction and identification of the S19-ΔznuA and the S19-ΔznuA-Δbp26 mutant strains of Brucella abortus

作　　者：马思思[1] 王佳莹[1] 刘翠翠[1] 丁红星[1] 吴云燕[1] 赵明秋[1] 陈金顶[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642

出　　处：《中国兽医学报》2015年第3期418-424,共7页Chinese Journal of Veterinary Science

基　　金：广东省教育部产学研结合资助项目(2011B090400259);公益性行业(农业)科研专项经费资助项目(201203056)

摘　　要：通过等位基因交换,分别敲除牛流产型布鲁菌减毒活疫苗S19株和缺失株S19-Δbp26的znuA基因,构建了布鲁菌znuA基因单缺失株S19-ΔznuA和bp26、znuA基因双缺失株S19-Δbp26-ΔznuA,对获得的2种缺失株进行形态学、生长特性、稳定性和基因测序验证。结果表明,S19株和S19-Δbp26株的znuA基因均成功缺失,生长特性表示2种缺失株与亲本株生长基本无差异;体外连续传至20代,菌落PCR鉴定及基因测序结果显示2种缺失株均遗传稳定。牛流产型布鲁菌单缺失株S19-ΔznuA和双缺失株S19-ΔznuA-Δbp26的成功构建为布鲁菌新型疫苗的研发、布鲁菌感染动物过程中ZnuA蛋白的作用机理及其与Bp26蛋白之间关系的研究奠定了基础。We constructed the suicide vector pRE112-ΔznuA through the genetic engineering method,and a znuA knockout mutation S19-ΔznuA and a double mutant S19-Δbp26-ΔznuA strains were obtained fromBrucella abortus S19 by the allelic exchange.PCR and DNA sequencing detection of the two mutant strains,showed that the znuA was knockout in both S19 strain and theΔbp26 B.abortus S19 mutant.The bacterial forms and growth characteristecs of the two mutant strains were the same as their parent strain S19,and the genetic stability of the two mutant strains were demonstrated in twenty generation by continuous passage in vitro.Thus,acquired two znuA mutation could be provided a chance to study the function of znuA gene and the relationship between znuA and bp26.

关 键 词：布鲁菌 znuA基因 基因缺失株 

分 类 号：S852.614[农业科学—基础兽医学]