Construction and expression of a novel HBeAg binding protein 1 of hepatitis B virus in yeast / 中华实验和临床病毒学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To construct the eukaryotic expression vector of HBEBP1 gene and express HBEBP1 recombinant protein in yeast.</p><p><b>METHODS</b>PCR was performed to amplify the gene of HBEBP1 from the cDNA template origining from HepG2, and the gene was cloned into pGEM-T vector. After sequencing, the correct DNA fragment was cut from pGEM-T-HBEBP1 and inserted into yeast expression plasmid pGBKT7. The reconstructed plasmid pGBKT7-HBEBP1 was transformed into yeast cell AH109 and screened on the synthetic dropout nutrient medium (SD/-Trp/Kana). The yeast protein was isolated and analyzed with SDS-PAGE and Western Blot.</p><p><b>RESULTS</b>The eukaryotic expressive vector was constructed successfully. The results of Western Blot showed HBEBP1 protein was existed within yeast cells and the molecular weight of it was about 33 x 10(3).</p><p><b>CONCLUSIONS</b>The successful expression of HBEBP1 protein in yeast cells lay the foundation for studying biological function of HBEBP1.</p>