论文部分内容阅读
目的应用酵母双杂交技术,在人胎脑文库中筛选与人类巨细胞病毒(human cytomegalovirus,HCMV)UL146基因编码蛋白相互作用蛋白,以探讨HCMV-UL146的生物功能。方法应用酵母双杂交系统,构建pGBKT7-UL146诱饵质粒,转化酵母AH109,与含人胎脑cDNA文库质粒的酵母相互作用,于涂有X-α-gal营养缺陷型培养基上筛选生长。挑选蓝色克隆,提取此酵母克隆的质粒转化大肠杆菌,提取质粒DNA进行测序分析。结果 pGBKT7-UL146构建成功。经严格筛选,共有200余个阳性克隆,分离测序80个克隆,成功筛选出30个与HCMV-UL146特异性相互作用的蛋白。结论 HCMV-UL146与多种蛋白有相互作用,提示其生物学功能复杂。此结果为进一步研究HCMV-UL146基因功能提供依据。
Objective To investigate the biological function of HCMV-UL146 by screening human fetal brain library with yeast-two-hybrid technique and interacting protein encoded by human cytomegalovirus (HCMV) UL146 gene. Methods The bait plasmid pGBKT7-UL146 was constructed by yeast two-hybrid system and transformed into yeast AH109. The bait plasmid was transformed into yeast containing human fetal brain cDNA library and screened on X-α-gal auxotrophic medium. The blue clones were picked, the plasmid of this yeast clone was extracted and transformed into E. coli, and the plasmid DNA was extracted for sequencing analysis. Results pGBKT7-UL146 was successfully constructed. After rigorous screening, a total of more than 200 positive clones, 80 clones were isolated and sequenced, and 30 proteins that specifically interacted with HCMV-UL146 were successfully screened. Conclusion HCMV-UL146 interacts with many proteins, suggesting that the biological function of HCMV-UL146 is complex. The results provide the basis for further study on the function of HCMV-UL146 gene.