丙型肝炎病毒 NS4A蛋白在胰腺细胞 cDNA文库中的结合蛋白基因的筛选

目的:从人胰腺细胞cDNA文库中筛选出与丙型肝炎病毒(HCV)NS4A蛋白存在相关作用并且参与糖、脂类代谢过程的结合蛋白,为研究HCV影响糖、脂类代谢的分子生物学机制提供实验依据.方法:构建pGBKT7-NS4A作为诱饵质粒,转化酵母菌株AH109,用SD/-Trp筛选阳性菌落.配合两种重组酵母菌株AH109与Y187...

Screening of proteins binding to hepatitis C virus NS4A protein from human pancreas cDNA library

Objective：To screen proteins from human pancreas cDNA library, which interact with the protein coded by hepatitis C virus （HCV） NS4A and relate with the metabolism of glucose or lipid. Methods： The recon- structed plasmid pGBKTT-NS4A was transformed into yeast strain AH109 and identified. The transformed AH109 （pGBKT7-NS4A） mated with Y187 （plasmid of pancreatic cDNA library）. The diploid yeast cells were plated on nutrient deficiency medium containing X-a-gal for selecting. The plasmids in diploid yeast cells were extracted and electrotransformed into E. coli DH5a. The plasmids in DHSa were extracted, sequenced and blasted. Results： Five proteins interacting with HCV NS4A were found, including CDK5R1, elastase 3A, pancreatic lipase, KRAS and bile salt-stimulated esterase. Conclusions：The five pancreatic proteins related with metabolisms of glucose and lipid could interact with HCV NS4A, thus disturbing the metabolism of glucose or lipid.