Screening of interferon?binding proteins by yeast-two hybrid technique / 中华传染病杂志
Chinese Journal of Infectious Diseases
; (12)1997.
Article
de Zh
| WPRIM
| ID: wpr-679389
Bibliothèque responsable:
WPRO
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.
Texte intégral:
1
Indice:
WPRIM
Type d'étude:
Diagnostic_studies
/
Screening_studies
langue:
Zh
Texte intégral:
Chinese Journal of Infectious Diseases
Année:
1997
Type:
Article