酵母双杂交筛选与乙型肝炎病毒表面抗原中蛋白相互作用的肝细胞蛋白  

Screening of hepatitis B surface antigen-interacted liver cell proteins by yeast two-hybrid system

作　　者：陈婉南[1] 刘玲玲[1] 吴云丽[1] 焦伯延[1] 林万松[1] 林旭[1] 

机构地区：[1]福建医科大学,教育部消化道急性肺瘤重点实验室,福建省肺瘤微生物学重点实验室,福州350004

出　　处：《中国人兽共患病学报》2010年第10期907-911,共5页Chinese Journal of Zoonoses

摘　　要：目的筛选与乙型肝炎病毒表面抗原中蛋白(Middle hepatitis B surface protein,MHBs protein)相互作用的肝细胞蛋白。方法 PCR扩增MHBs编码基因并将S区起始密码子突变,获得目的基因MHBs-mut,克隆于诱饵载体pGBKT7。在证实MHBs-mut蛋白不具有自激活作用的前提下,以酵母双杂交系统筛查与MHBs-mut蛋白相互作用的肝细胞蛋白,并在酵母细胞内验证蛋白的相互作用。结果构建酵母双杂交诱饵载体,获得重组质粒pGBKT7-MHBs-mut。Western blot显示其在酵母中表达MHBs-mut蛋白。酵母双杂交筛选并验证,得到与MHBs-mut蛋白相互作用的肝细胞蛋白:COP9信号体第五个亚单位(COPS5,又名C-Jun结合蛋白1,c-Jun-activation-domain binding protein 1,JAB1)。结论乙型肝炎病毒表面抗原中蛋白与COPS5在酵母AH109细胞中具有相互作用。To screen the liver cell proteins which interacts with middle hepatitis B surface protein （MHBs protein）, the sequences encoding MHBs was amplified by means of PCR and the start codon of S region ATG was mutated to ACG to generate the target gene MHBs mut, then the MHBs-mut was cloned into the bait vector pGBKT7. After exclusion of self-activation capacities of MHBs-mut protein, a two-hybrid library screening was performed using a pre-transformed human liver cDNA li brary to screen the liver cell proteins interacting with MHBs-mut. Analysis and verification of putative positive clones were also done to further confirm the interactions in AH109 yeast cells. Results indicated that liver cell protein of Homo sapiens COP9 constitutive photomorphogenic homolog subunit 5（COPS5） was found to be able to interact with MHBs-mut in AH109.

关 键 词：乙型肝炎病毒 表面抗原中蛋白 酵母双杂交 

分 类 号：R373.21[医药卫生—病原生物学]