Screening of hepatocyte proteins binding to complete S protein of hepatitis B virus by yeast-two hybrid system.

ABSTRACT:

Aim

To investigate the biological function of complete S protein and to look for proteins interacting with complete S protein in hepatocytes.

Methods

We constructed bait plasmid expressing complete S protein of HBV by cloning the gene of complete S protein into pGBKT7, then the recombinant plasmid DNA was transformed into yeast AH109 (a type). The transformed yeast AH109 was mated with yeast Y187 (alpha type) containing liver cDNA library plasmid in 2XYPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X-alpha-gal for selection and screening. After extracting and sequencing of plasmids from positive (blue) colonies, we underwent sequence analysis by bioinformatics.

Results

Nineteen colonies were selected and sequ

SUBMITTER: Bai GQ 

PROVIDER: S-EPMC4504893 | biostudies-literature | 2005 Jul

REPOSITORIES: biostudies-literature