【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。更多还原

【Abstract】 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.更多还原