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 sequenced. Among them, five colonies were Homo sapiens solute carrier family 25, member 23 (SLC25A23), one was Homo sapiens calreticulin, one was human serum albumin (ALB) gene, one was Homo sapiens metallothionein 2A, two were Homo sapiens betaine-homocysteine methyltransferase, three were Homo sapiens Na(+) and H(+) coupled amino acid transport system N, one was Homo sapiens CD81 antigen (target of anti-proliferative antibody 1) (CD81), three were Homo sapiens diazepam binding inhibitor, two colonies were new genes with unknown function. CONCLUSION:The yeast-two hybrid system is an effective method for identifying hepatocyte proteins interacting with complete S protein of HBV. The complete S protein may bind to different proteins i.e., its multiple functions in vivo.

journal_name

World J Gastroenterol

authors

Bai GQ,Cheng J,Zhang SL,Huang YP,Wang L,Liu Y,Lin SM

doi

10.3748/wjg.v11.i25.3899

keywords:

subject

Has Abstract

pub_date

2005-07-07 00:00:00

pages

3899-904

issue

25

eissn

1007-9327

issn

2219-2840

journal_volume

11

pub_type

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