Construction of mutant and overexpression strains with small noncoding RNA BSR1526 and study of its virulence in Brucella
XU Xiao-yang1, OU Hong-ping2, PENG Guang-neng1, LEI Shuang-shuang1, TIAN Yi-nan1, CAO Xue-feng1, WANG Yu-fei3, YAN Guang-wen4, ZHONG Zhi-jun1
1.College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China; 2.Chengdu Vocational College of Agricultural Science and Technology, Chengdu 611130, China; 3.Department of Laboratory Medicine, the General Hospital of Chinese People?s Armed Police Forces, Beijing 100039, China; 4.College of Veterinary Medicine,Najing Agricaltural University,Najing 210095,China
Abstract：To evaluate the roles of small noncoding RNA BSR1526 in Brucella intracellular survival, the mutant and overexpression strains of BSR1526 were constructed and their survival capabilities under stresses that simulate intracellular environment and that of in mice were analyzed. Firstly, BSR1526 was verified by Northern blot and its transcription under stress conditions was verified by RT-PCR. The mutant strains of BSR1526 were constructed by fusion PCR. BSR1526 was inserted into the plasmid pBBR1-MCS4, which transformed into Brucella melitensis 16M to generate the overexpression strain16M-BSR1526. At last, the survival capabilities of 16M-△BSR1526 and 16M-BSR1526 in different stimuli resembling that Brucella probably encounters during infection and mice were analyzed. Our results showed that BSR1526 was transcribed in Brucella,and with different transcription levels under the stress conditions. Deletion or overexpression of BSR1526 affected the growth of Brucella in vitro. The 16M-△BSR1526 showed reduced survival under heat shock, oxidative stress and acidic conditions, and significant reduction in mice, indicating that BSR1526 influenced the virulence of Brucella. In summary, small noncoding RNA BSR1526 affects intracellular survival ability of Brucella melitensis 16M. The ability adapting to hostile environments and virulence in mice with mutant BSR1526 strain was decreased.
徐晓阳, 欧红萍, 彭广能, 雷霜霜, 田一男, 曹雪峰, 王玉飞, 严光文, 钟志军. 布鲁氏菌非编码小RNA BSR1526突变株与过表达株的构建及毒力研究[J]. 中国人兽共患病学报, 2016, 32(12): 1083-1090.
XU Xiao-yang, OU Hong-ping, PENG Guang-neng, LEI Shuang-shuang, TIAN Yi-nan, CAO Xue-feng, WANG Yu-fei, YAN Guang-wen, ZHONG Zhi-jun. Construction of mutant and overexpression strains with small noncoding RNA BSR1526 and study of its virulence in Brucella. Chinese Journal of Zoonoses, 2016, 32(12): 1083-1090.
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