Affinity between hepatitis B virus e antigen and protein AK026018 / 中华传染病杂志

ABSTRACT

Objective To investigate the intracellular and extracellular affinity between hepatitis B virus e antigen (HBeAg) and protein AK026018. Methods The gene AK026018 was amplified by reverse transcription polymerase chain reaction (RT-PCR) technique from HepG2 cell. The expressive vector of pGADT7-AK was constructed by routine molecular biological methods. The auxotroph yeast cells were cotransfected with pGADT7-AK and pGBKT7-eAg and plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-?-gal. To further prove the affinity between HBeAg and protein AK026018, translation was performed by using reticulocyte lysate, and immunoprecipitation was showed in vitro. Results The expressive vector was constructed and confirmed by DNA sequencing analysis and restriction enzyme digestion.The positive clones could grow on SD/-Trp-Leu-His-Ade/x-?-gal medium and were blue. The radioautographic band were found in immunoprecipitation analysis. Conclusions HBeAg could bind protein AK026018 in vivo and in vitro.