ATPase抑制因子1是与HCMV pUL23蛋白相作用的宿主蛋白分子-酵母双杂交技术筛选与鉴定  

ATPase inhibitory factor 1-a host cell protein molecule interacting with human cytomegalovirus pUL23 protein

作　　者：曾涛[1] 李实骞[1] 姚伙旺[1] 邹奕[1] 周天鸿[1] 李弘剑[1] 

机构地区：[1]暨南大学生命科学技术学院,生物工程学系,广东广州510632

出　　处：《中国病理生理杂志》2009年第7期1360-1364,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金项目(No.30770106);广东省自然科学基金项目(No.06025162)资助项目;教育部科学技术研究重点项目(No.207114)

摘　　要：目的:利用酵母双杂交技术筛选与人巨细胞病毒相互作用的宿主蛋白分子,为探讨人巨细胞病毒pUL23蛋白在HCMV生活周期中的作用机制提供依据。方法:利用GAL4酵母双杂交系统筛选人胚肾cDNA文库,以获得与人巨细胞病毒pUL23蛋白相互作用的宿主蛋白分子,再通过回交试验和体外GST-pulldown试验验证两者之间的相互作用。结果:酵母双杂交筛选得到宿主蛋白分子ATPase inhibitory factor1(ATIF1),回交试验和体外GST-pulldown试验再次确认ATIF1能够与人巨细胞病毒pUL23蛋白相互作用。结论:pUL23确实能够与ATIF1相互作用,它们之间的相互作用可能为研究pUL23在病毒生活周期发挥的功能提供依据。AIM： pUL23, the product of human cytomegalovirus （HCMV） gene UL23 was identified as one of tegument proteins. The aim of this research is to investigate the function of pUL23 during HCMV life cycle. METHODS： GAIA yeast two - hybrid assay was performed to screen the human fetal kidney cDNA library to obtain host cell protein molecules which interact with pUL23 of HCMV. Then the GST - pulldown experiment was applied to confirm the protein interactions identified by yeast two - hybrid. RESULTS ： ATPase inhibitory factor 1 （ ATIF! ） was selected from host cells using yeast two - hybrid assay. GST - pulldown experiments in vitro further confirmed the interaction between ATIF1 and pUL23. CONCLUSION： pUL23 of HCMV can interact with ATIF1 in host cell, which may provide the evidence for understanding the function of pUL23 in the life cycle of HCMV.

关 键 词：巨细胞病毒 蛋白质pUL23 酵母双杂交实验 ATP酶抑制因子1 

分 类 号：Q71[生物学—分子生物学]