摘要
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)UL131A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pGB-KT7-UL131A转化到酵母菌AH109中,再将人胎脑文库DNA转化到含有pGBKT7-UL131A的酵母细胞中,筛选与HCMV UL131A编码蛋白相互作用的细胞蛋白,并对筛选得到的阳性克隆进行测序和生物信息学分析.结果:成功构建酵母诱饵表达载体pGBKT7-UL131A,并将其与人胎脑cDNA文库共转化到酵母细胞AH109中,最终确认有23种人胎脑文库蛋白与HCMV UL131A编码蛋白相互作用,其中一种与Thy-1蛋白高度同源,其同源性高达99%.结论:成功应用酵母双杂交系统筛选出与HCMV UL131A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.
AIM： To screen proteins that interact with the human cytomegalovirus （HCMV） UL131A protein from a human fetal brain cDNA library using yeast two-hybrid system.METHODS： The ＂bait plasmid＂ （named pGBKT7-UL131A） was constructed and used as a bait to screen a human fetal brain cDNA library to find proteins interacting with the UL131A protein.The positive clones were sequenced and analyzed using bioinformatic methods.RESULTS： The ＂bait plasmid＂ was constructed successfully and co-transformed together with a human fetus brain cDNA library into yeast cells.At last twenty-three proteins interacting with the HCMV UL131A were identified,and one of them shares 99% homology with the Thy-1 gene.CONCLUSION： Some proteins interacting with HCMV UL131A have been successfully screened from a human fetal brain cDNA library.
出处
《世界华人消化杂志》
CAS
北大核心
2010年第24期2584-2588,共5页
World Chinese Journal of Digestology
基金
国家自然科学基金资助项目
No.30770109~~
