Screening of the genes of hepatitis B virus e antigen interacting proteins.
- Author:
Bo-an LI
1
;
Cui-li SHU
;
Jun HOU
;
Yang QI
;
Jing LI
;
Wei-ping HE
;
Yun CHENG
Author Information
- Publication Type:Journal Article
- MeSH: Gene Library; Hepatitis B e Antigens; genetics; metabolism; Humans; Liver; metabolism; Plasmids; genetics; Protein Binding; Transformation, Genetic; Two-Hybrid System Techniques
- From: Chinese Journal of Experimental and Clinical Virology 2005;19(3):271-274
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo screen and clone the genes in hepatocytes which encode protein that can interact with hepatitis B e antigen(HBeAg) by yeast-two hybridization.
METHODSRecombined HBeAg bait plasmid (pGBKT7-eAg) was transformed into yeast AH l09, followed by mating with yeast Yl87 containing liver cDNA library plasmid in 2 x YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-Ade-His) which contains X-a-gal for selecting positive blue clones. Then positive clones were selected and plasmids were prepared and sequenced. Finally, bioinformatics analysis was performed.
RESULTSTotally 245 positive colonies were selected and 101 colonies were sequenced. Through sequences alignment, 6 novel genes and 35 recorded genes were screened.
CONCLUSIONGenes of HBeAg interacting proteins have been cloned successfully, which brings some new clues for further studies on the biological functions of HBeAg and the related proteins.
