摘要
目的利用酵母双杂交技术筛选与人巨细胞病毒(HCMV)UL130表达产物有相互作用的细胞蛋白及其编码基因,了解其在宿主基因组中的地位功能。方法将成功构建的酵母诱饵表达载体pGBKT7-UL130转化到酵母菌AH109中,再将人胎脑文库DNA转化到含有pGBKT7-UL130的酵母细胞中,筛选与HCMVUL130编码蛋白相互作用的细胞蛋白,并对阳性克隆进行测序和生物信息学分析。结果最终确认9个克隆与HCMVUL130编码蛋白相互作用,其中2个克隆与突触小体相关蛋白(SNAP)高度同源。结论成功应用酵母双杂交系统筛选出与HCMVUL130编码蛋白相互作用的蛋白,其中SNAP在HCMV感染致病过程中可能起重要作用。
Objective To screen the human proteins interacting with human cytomegalovirus（HCMV）UL130 from human fetus brain cDNA library by GAL4 two-hybrid system 3 technique and analyze the corresponding coding sequences.Methods The ＂bait plasmid＂（named as pGBKT7-UL130）was constructed.By using HCMV UL130 as the bait,a human fetus brain cDNA library was screened and the proteins interacting with UL130 protein were searched.The positive clones were sequenced and analyzed by bioinformatic methods.Results Nine clones interacting with HCMV UL130 were identified,two of them were synaptosome-associated protein（SNAP）.Conclusion Some proteins interacting with HCMV UL130 in human fetus brain cDNA library were successfully screened.SNAP might play an important role in HCMV infection pathogenesis.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2010年第6期413-416,共4页
Journal of China Medical University
基金
国家自然科学基金资助项目(30770109)
辽宁省博士科研启动基金资助项目(20061039)
