[Construction and expression of a novel HBeAg binding protein 1 of hepatitis B virus in yeast].
OBJECTIVE
To construct the eukaryotic expression vector of HBEBP1 gene and express HBEBP1 recombinant protein in yeast.


METHODS
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.


RESULTS
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).


CONCLUSIONS
The successful expression of HBEBP1 protein in yeast cells lay the foundation for studying biological function of HBEBP1.
Beijing Centre For Physical & Chemical Analysis, Beijing 100094, China.
Yue
Y
Li
Qi
Q
Wang
Jun
J
Cheng
Hui-Juan
HJ
Wu
D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics*; D006513:Hepatitis B e Antigens / Q000378:metabolism*; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D002352:Carrier Proteins; D006513:Hepatitis B e Antigens; D011994:Recombinant Proteins