SCREENING AND IDENTIFICATION OF A NOVEL HEPATITIS B VIRUS E ANTIGEN BINDING PROTEIN E-36 FROM HEPATOCYTE cDNA LIBRARY
- VernacularTitle:肝细胞内与HBeAg结合的新蛋白编码基因E-36的筛选与克隆
- Author:
Yinying LU
;
Yaodong LIANG
;
Jun CHENG
- Publication Type:Journal Article
- Keywords:
hepatitis B e antigens;
genes;
two-hybrid system techniques
- From:
Medical Journal of Chinese People's Liberation Army
2001;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the biological functions of hepatitis B virus e antigen (HBeAg) protein. Methods Yeast-two hybrid technique was performed to look for proteins interacting with HBeAg from hepatocytes. HBeAg bait plasmid was constructed by cloning the HBeAg gene into carrier plasmid pGBKT7, then the yeast AH109 (a type) was transformed. The transformed yeast cells was amplified and mated with yeast cells Y187 (?type) containing liver cDNA library plasmid pCAT2 in 2?YPDA medium. Diploid yeast cells were plated on synthetic dropout nutrient medium and were selected for two times. Results Plasmids of positive colonies were extracted and analyzed by DNA sequencing and BLAST in GenBank. Conclusions After the full sequence of new gene E-36 was amplified from the mRNA of HepG2 cells by reverse transcription polymerase chain reaction (RT-PCR) and cloned into pGADT7 vector, the recombinant plasmid was translated by reticulocyte lysate and analyzed by immuno-coprecipitation technique in vitro together with HBeAg. The result provided some new clues for the study of the biological functions of HBeAg.
