人巨细胞病毒UL145相互作用蛋白的筛选和功能分析

摘要

目的 应用酵母双杂交系统筛选与人巨细胞病毒(HCMV)UL145编码蛋白相互作用的来源于人胎脑cDNA文库的组织蛋白.方法 采用聚合酶链式反应扩增HCMV UL145片段,酶切及纯化扩增的目的片段及酵母表达载体pGBKT7,连接HCMVUL145片段及载体pGBKT7,将连接后的重组诱饵表达载体pGBKT7-UL145转化到酵母菌AH109中,再将人胎脑文库DNA也转化到酵母AH109中,筛选与HCMV UL145编码蛋白相互作用的蛋白,并对阳性克隆进行测序和生物信息学分析.结果 最终确认多个克隆与HCMV UL145编码蛋白相互作用,BLAST结果显示3个与FOXG1高度同源.结论 利用酵母双杂交系统成功筛选出与HCMV UL145编码蛋白相互作用的人胎脑cDNA文库蛋白FOXG1,推测该目的蛋白在HCMV感染所致神经系统损伤过程中起重要作用.%Objective To screen a human fetal brain cDNA library for proteins that can interact with HCMV UL145 using a yeast two-hybrid system.Methods A bait plasmid (pGBKT7-UL145) was constructed.Using HCMV UL145 as bait,a human fetal brain cDNA library was screened and proteins interacting with UL145 were identified using bioinformatic methods to sequence and analyze the positive clones.Results Three clones interacting with HCMV UL145 were found,and identified as FOXG1.Conclusion Several proteins interacting with HCMV UL145 in the human fetal brain cDNA library were identified as FOXG1,indicating that this protein may play an important role in the course of HCMV infection.