[Study on binding of HBeAg to CD81].
OBJECTIVE
To investigate the interaction between HBeAg and CD81.


METHODS
The CD81 gene was amplified by RT-PCR from HepG2 cells. The recombinant expression vector pGADT7-CD81 was constructed by routine molecular biological method. The auxotroph yeast cells were cotransfected with pGADT7-CD81 and pGBKT7-eAg and plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal. The binding of HBeAg to CD81 was also detected in vitro translation and coimmunoprecipitation test.


RESULTS
The recombinant expression vector was constructed and confirmed by restriction enzyme (EcoR I and BamH I) digestion and DNA sequencing analysis. The positive yeast clones could grow and from blue colonies on SD/-Trp-Leu-His-Ade/x-alpha-gal medium. The result of immunocoprecipitation showed that HBeAg could bind to CD81.


CONCLUSIONS
HBeAg can bind to CD81, suggesting that CD81 plays an important role in the pathogenesis of HBV.
Li
Bo-an
BA
Institute of Infectious Diseases, 302th Hospital of PLA, Beijing 100039, China. lba@263.net
Liu
Yan
Y
Li
Jing
J
Shu
Cui-li
CL
He
Wei-ping
WP
Hou
Jun
J
Cheng
Yun
Y
D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D005822:Genetic Vectors; D006513:Hepatitis B e Antigens / Q000235:genetics / Q000378:metabolism*; D018197:Hepatoblastoma / Q000473:pathology; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology; D011995:Recombination, Genetic; D060225:Tetraspanin 28; D014162:Transfection; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics / Q000378:metabolism*
D015703:Antigens, CD; C554798:CD81 protein, human; D006513:Hepatitis B e Antigens; D060225:Tetraspanin 28