人巨细胞病毒UL135蛋白结合蛋白的酵母双杂交筛选

Screening of Protein Interacting with HCMV UL135 Protein by Yeast Two Hybrid System from Human Fetus Brain cDNA Library

目的利用酵母双杂交系统从人胎脑cDNA文库中筛选与人巨细胞病毒(HCMV)UL135编码蛋白相互作用的蛋白。方法将成功构建的酵母诱饵表达载体pGBKT7-UL135转化到酵母菌AH109中,再将人胎脑文库DNA转化到含有pGBKT7-UL135的酵母细胞中,筛选与HCMVUL135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60个克隆与HCMVUL135编码蛋白相互作用,其中2个克隆与Thy-1高度同源。结论成功应用酵母双杂交系统筛选出与HCMVUL135编码蛋白相互作用的蛋白,其中Thy-1在HCMV感染致病过程中可能起重要作用。

Objective To screen the proteins which can interact with the human cytomeg a lovirus HCMV UL135 protein from human fetus brain cDNA library with yeast two-hybrid system. Methods The ＂bait plasmid＂（named as pGBKT7-UL135）was constructed. Using HCMV UL135 as a bait,we screened a human fetus brain cDNA library. The proteins interacting with UL135 protein were searched. The positive clones were sequenced and analyzed by bioinformatic methods. Results Ten clones interacting with HCMV UL135 were identified, two of them were Thy-1. Conclusion Some proteins interacting with HCMV UL135 in hm/mn fetus brain eDNA library were suecessfuUy screened. Thy-1 may play an important role in the course of HCMV infection.