The ORF sequence of HCMV UL23 was cloned into yeast-expressing vector pGBKT7 to construct the vector pGBKTT-UL23. The fusion protein pGBKT7-UL23 expressed in the AH109 yeast strain served as the bait protein. We used a yeast two-hybridization assay to screen the proteins that can interact with HCMV pUL23 from HEK eDNA library. Yeast two-hybridization retransformation experiment was applied to verify the interactions. 79 proteins that interacted with pUL23 were screened out after the construction of pGBKT7-UL23. Among which, eIF3e were verified by the yeast two-hybridization retrans-formation experiment to interact with pUL23. HCMV pUL23 interacting proteins could be screened by u-sing yeast two-hybrid system. These results provide the basis for the funtion of pUL23 in the HCMV lifecycle.%将UL23基因的ORF序列克隆到酵母表达载体pGBKT7构建诱饵质粒pGBKT7-UL23,用酵母双杂交技术筛选人胚肾cDNA文库中与人巨细胞病毒pUL23相互作用的宿主蛋白分子,再通过回复酵母双杂交再次确认两者之间的相互作用.结果表明:酵母双杂交,回复酵母双杂交试验证明宿主蛋白分子elF3e(eukaryotic trans-lation initiation factor 3,subunit E interacting protein)能够与人巨细胞病毒UL23蛋白相互作用.宿主蛋白分子elF3e能够与人巨细胞病毒UL23蛋白相互作用,这为进一步研究pUL23蛋白在HCMV生活周期中的作用机制提供依据.
展开▼
