Huang YP, Cheng J, Zhang SL, Wang L, Guo J, Liu Y, Yang Y, Zhang LY, Bai GQ, Gao XS, Ji D, Lin SM, Shao Q. Screening of hepatocyte proteins binding to F protein of hepatitis C virus by yeast two-hybrid system. World J Gastroenterol 2005; 11(36): 5659-5665 [PMID: 16237761 DOI: 10.3748/wjg.v11.i36.5659]
Corresponding Author of This Article
Yan-Ping Huang, Department of Pediatrics, the First Hospital of Xi’an Jiaotong University, Xi’an 710061, Shaanxi Province, China. huangyp423@126.com
Article-Type of This Article
Viral Hepatitis
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World J Gastroenterol. Sep 28, 2005; 11(36): 5659-5665 Published online Sep 28, 2005. doi: 10.3748/wjg.v11.i36.5659
Screening of hepatocyte proteins binding to F protein of hepatitis C virus by yeast two-hybrid system
Yan-Ping Huang, Jun Cheng, Shu-Lin Zhang, Lin Wang, Jiang Guo, Yan Liu, Yuan Yang, Li-Ying Zhang, Gui-Qin Bai, Xue-Song Gao, Dong Ji, Shu-Mei Lin, Qing Shao
Yan-Ping Huang, Shu-Lin Zhang, Yuan Yang, Gui-Qin Bai, Shu-Mei Lin, The First Hospital of Xi’an Jiaotong University, Xi’an 710061, Shaanxi Province, China
Jun Cheng, Lin Wang, Jiang Guo, Yan Liu, Li-Ying Zhang, Xue-Song Gao, Dong Ji, Qing Shao, 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, Nos. C03011402 and C30070690; and the Research and Technique Foundation of PLA during the 9th-Five year plan period, No. 98D063; and the Launching Foundation for Student Studying Abroad of PLA, No. 98H038; and the Youth Research and Technique Foundation of PLA during the 10th-Five Year Plan Period, No. 01Q138; and the Research and Technique Foundation of PLA during the 10th-Five Year Plan Period, No. 01MB135
Correspondence to: Yan-Ping Huang, Department of Pediatrics, the First Hospital of Xi’an Jiaotong University, Xi’an 710061, Shaanxi Province, China. huangyp423@126.com
Telephone: +86-29-85324065
Received: July 17, 2004 Revised: August 23, 2004 Accepted: August 25, 2004 Published online: September 28, 2005
Abstract
AIM: To investigate the biological function of F protein by yeast two-hybrid system.
METHODS: We constructed F protein bait plasmid by cloning the gene of F protein into pGBKT7, then 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 plasmids from positive (blue) colonies, we underwent sequence analysis by bioinformatics.
RESULTS: Thirty-six colonies were selected and sequenced. Among them, 11 colonies were zymogen granule protein, 5 colonies were zinc finger protein, 4 colonies were zinc-α-2-glycoprotein, 1 colony was sialyltransferase, 1 colony was complement control protein factor I, 1 colony was vitronectin, and 2 colonies were new genes with unknown function.
CONCLUSION: The yeast two-hybrid system is an effective method for identifying hepatocyte proteins interacting with F protein of hepatitis C virus. F protein may bind to different proteins.
