Screening of interferon?binding proteins by yeast-two hybrid technique
- VernacularTitle:酵母双杂交技术筛选?干扰素结合蛋白的基因
- Author:
Jian-Hui QU
;
Jun CHENG
;
Ling-Xia ZHANG
;
- Publication Type:Journal Article
- Keywords:
Interferon alpha;
Yeasts;
Two hybrid system techniques;
Cloning,molecular;
Genes
- From:
Chinese Journal of Infectious Diseases
1997;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To screen proteins binding with interferon?(IFN?)from human hepatic cDNA libraty by yeast-two hybrid technique.Methods The IFN?gene was amplified by polymerase chain reaction(PCR)and constructed into pGBKT7 vector as the bait plasmid in yeast-two hybrid system3,pGBKT7-IFN?was then transfected into yeast AH109.The transfected yeast were mated with yeast Y187 containing liver cDNA library plasmid in 2?YPDA medium.Diploid yeast was plated on synthetic dropout nutrient medium(SD/-Trp Leu-His-Ade)and synthetic dropout nutrient medium(SD/-Trp-Leu-His-Ade)containing X-?-gal for selecting.After plasmid extracting and en- zyme cutting analysis,the blue colonies were performed sequence analysis,the results were analyzed by bioinformatics.Results IFN?gene was successfully cloned and expressed in yeast cells.Thirty- four positive colonies were obtained using yeast-two hybrid technique.After sequence analysis,eight clones were found may have a binding effect with IFN protein.Conclusions IFN?genes was success- ful cloned and eight proteins that could bind with IFN?protein were also screened.
