SCREENING AND CLONING OF THE GENES OF PROTEIN INTERACTING WITH THE NONSTRUCTURAL PROTEIN 5B OF HEPATITIS C VIRUS
- VernacularTitle:丙型肝炎病毒非结构蛋白5B结合蛋白的酵母双杂交筛选研究
- Author:
Jian ZHANG
;
Jun CHENG
;
Lin WANG
- Publication Type:Journal Article
- Keywords:
hepatitis C virus;
viral nonstructural proteins;
yeast two hybrid
- From:
Medical Journal of Chinese People's Liberation Army
2001;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
Protein-protein binding is the basis of virus and host cell interactions. With the application of technology of studying protein interactions, more knowledge of replication and pathogenesis of hepatitis C virus (HCV) was acquired. Non-structure protein 5B(NS5B) of HCV is a kind of viral protein, which plays an important role in replication of HCV. However, the effect of NS5B is not clear. To investigate the biological function of NS5B, we performed yeast two hybrid to look for proteins in hepatocytes interacting with NS5B. We constructed NS5B bait plasmid by cloning the gene of NS5B into pGBKT7, then transformed it into yeast AH109(a type). The transformed yeast was mated with yeast Y187(? type)containing liver cDNA library plasmid in 2?YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-?-gal for screening. Thirty-three colonies were selected and sequenced. Among them, two colonies were new genes with unknown function. The preliminary successful cloning of gene of protein interacting with NS5B paved the way for the study of the physiological function of NS5B and its associated protein.
