摘要
目的:构建Cpn0147酵母双杂交诱饵载体,为应用酵母双杂交系统筛选与其相互作用的蛋白奠定基础。方法:应用聚合酶链式反应(polymerase chain reaction,PCR)扩增技术获得Cpn0147基因片段,克隆入pGBKT7载体,鉴定构建是否成功,确定构建成功后将重组载体pGBKT7-Cpn0147转入AH109及Y187酵母中,进行诱饵载体的毒性及自激活活性分析。结果:Cpn0147诱饵载体构建成功,且该诱饵载体无自激活活性,对两种宿主酵母细胞无毒性。结论:诱饵载体pGBKT7-Cpn0147可用于酵母双杂交系统,为进一步筛选与之相互作用蛋白提供了实验基础。
Objective： To construct a bait vector of pGBKT7-Cpn0147 for seeking its ligands with yeast two-hybrid system. Methods： Cpn0147 gene was amplified by polymerase chain reaction （PCR）, and then cloned into pGBKT7 to construct the bait vector. These recombinant plasmids were transformed into yeast cells AH109 and Y187 to test toxic effects and self-activation activity. Results：The pGBKT7-Cpn0147 bait vector was constructed successfully, which neither had self-activation ability nor yeast cell toxicity. Conclusion： The constructed pGBKT7-Cpn0147 could be used as a bait vector in yeast two-hybrid system, and provided an experimental basis for further screening the interaction proteins.
出处
《神经药理学报》
2012年第5期19-23,共5页
Acta Neuropharmacologica
关键词
肺炎衣原体
包涵体膜蛋白
酵母双杂交
诱饵载体
自激活
chlamydia pneumoniae
inclusion membrane protein
yeast two-hybrid system
bait vector
self-activation
