Construction and expression of a novel HBeAg binding protein 1 of hepatitis B virus in yeast.
- VernacularTitle:乙型肝炎病毒e抗原结合蛋白1“饵”载体的构建及在酵母中的表达
- Author:
Yue LI
1
;
Qi WANG
;
Jun CHENG
;
Hui-Juan WU
Author Information
- Publication Type:Journal Article
- MeSH: Blotting, Western; Carrier Proteins; genetics; Hepatitis B e Antigens; metabolism; Plasmids; Recombinant Proteins; biosynthesis; Saccharomyces cerevisiae; genetics
- From: Chinese Journal of Experimental and Clinical Virology 2011;25(6):489-491
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct the eukaryotic expression vector of HBEBP1 gene and express HBEBP1 recombinant protein in yeast.
METHODSPCR 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.
RESULTSThe 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).
CONCLUSIONSThe successful expression of HBEBP1 protein in yeast cells lay the foundation for studying biological function of HBEBP1.
