purD和purF基因的突变进一步减弱了布鲁氏菌流产菌株RB51。
Mutation of purD and purF genes further attenuates Brucella abortus strain RB51.
作者信息
Truong Quang Lam, Cho Youngjae, Barate Abhijit Kashinath, Kim Suk, Watarai Masahisa, Hahn Tae-Wook
机构信息
College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University, Chuncheon 200-701, South Korea.
College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, South Korea.
出版信息
Microb Pathog. 2015 Feb;79:1-7. doi: 10.1016/j.micpath.2014.12.003. Epub 2014 Dec 27.
In the present study, transposon mutagenesis was used to further attenuate Brucella abortus RB51 vaccine strain. Two purD and purF mutants were constructed, characterized and evaluated for attenuation via intracellular survival in murine macrophage-like RAW264.7 and HeLa cells, and by clearance in BALB/c mice. The purD and purF mutants showed significantly decreased intracellular survival, and complementation of these mutants with intact copies of purD or purF genes of RB51 strain was able to restore these defects. In addition, the pur mutants presented significantly lowered persistence in mice. Immunization with purD and purF mutants protected mice against a challenge with the virulent B. abortus strain 544 at a level similar to that of the parent RB51. These data suggest that genes encoding the early stages of purine biosynthesis (purD and purF) are required for intracellular survival and virulence of B. abortus.
在本研究中，转座子诱变被用于进一步减毒流产布鲁氏菌RB51疫苗株。构建了两个purD和purF突变体，通过在小鼠巨噬细胞样RAW264.7和HeLa细胞中的细胞内存活情况以及在BALB/c小鼠中的清除情况对其进行表征和减毒评估。purD和purF突变体的细胞内存活能力显著降低，用RB51菌株完整的purD或purF基因拷贝对这些突变体进行互补能够恢复这些缺陷。此外,pur突变体在小鼠体内的持续性显著降低。用purD和purF突变体免疫可保护小鼠抵抗强毒流产布鲁氏菌544菌株的攻击，保护水平与亲本RB51相似。这些数据表明，编码嘌呤生物合成早期阶段的基因（purD和purF）是流产布鲁氏菌细胞内存活和毒力所必需的。
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