Viral Hepatitis
Copyright ©The Author(s) 2005. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jul 7, 2005; 11(25): 3899-3904
Published online Jul 7, 2005. doi: 10.3748/wjg.v11.i25.3899
Screening of hepatocyte proteins binding to complete S protein of hepatitis B virus by yeast-two hybrid system
Gui-Qin Bai, Jun Cheng, Shu-Lin Zhang, Yan-Ping Huang, Lin Wang, Yan Liu, Shu-Mei Lin
Gui-Qin Bai, Shu-Lin Zhang, Yan-Ping Huang, Shu-Mei Lin, First Hospital of Xi’an Jiaotong University, Xi’an 710004, Shaanxi Province, China
Jun Cheng, Lin Wang, Yan Liu, Institute of Infectious Diseases, Ditan Hospital, Anwai Street, Beijing 100011, China
Author contributions: All authors contributed equally to the work.
Supported by the National Natural Science Foundation of China, No. C03011402, No. C30070690; the Science and Technique Foundation of PLA during the 9th Five-year plan period, No. 98D063; the Launching Foundation for Students Studying Abroad of PLA, No. 98H038; the Youth Science and Technique Foundation of PLA during the 10th Five-year plan period, No. 01Q138; and the Science & Technique Foundation of PLA during the 10th Five-year plan period, No. 01MB135
Correspondence to: Dr. Gui-Qin Bai, Department of Obstetrics and Gynecology of First Hospital, Xi’an Jiaotong University, Jiankang Road 1, Xi’an 710061, Shaanxi Province, China
Telephone: +86-29-85213194 Fax: +86-29-85252812
Received: November 8, 2004
Revised: December 23, 2004
Accepted: December 26, 2004
Published online: July 7, 2005
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 (α type) containing liver cDNA library plasmid in 2 ×YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X-α-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.

Keywords: Complete S protein; Yeast-two hybrid system; Hepatitis B virus