摘要
利用Sos招募系统(SRS),通过PCR扩增羊布鲁菌16M VjbR基因的编码序列,定向克隆到酵母表达载体pSos中,构建诱饵重组质粒pSos-VjbR,经测序正确后其将转入酵母菌cdc25H(α)感受态细胞,检测其表达产物对酵母细胞有无毒性作用及对报告基因有无自激活作用。结果表明,经序列测定证实重组诱饵质粒pSos-VjbR构建成功。重组质粒转化入酵母细胞后,经检测其表达产物对cdc25H(α)酵母细胞无毒性作用,对报告基因亦无自激活作用。可以利用SRS研究与羊布鲁菌16M VjbR蛋白相互作用的蛋白,为进一步研究羊布鲁菌致病机制奠定了基础。
To construct a yeast expression vector of Brucella melitensis 16M VjbR gene using the Sos-recruitment system（SRS）,the coding sequence of Brucella melitensis 16M VjbR gene was amplified by PCR and cloned into the yeast expression plasmid pSos.The recombinant bait plasmid pSos-VjbR was verified by sequencing before transformation into competent yeast cells.The effects of the expression product on the yeast cell growth and activation of the reporter gene were evaluated.The results show the yeast expression vector of Brucella melitensis 16M VjbR gene was constructed sucessfully.The recombinant bait plasmid showed no toxic effect on yeast cdc25H cells without a self-activation of the reporter gene.The SRS can be used to study the proteins interacting with Brucella melitensis 16M VjbR protein and provides a means for obtaining insight into the pathogenic mechanism of Brucella melitensis 16M.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2013年第9期1378-1382,共5页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(30972198/C180503)
国家科技支撑计划资助项目(2010BAD04B03)
