Huang YP, Zhang SL, Cheng J, Wang L, Guo J, Liu Y, Yang Y, Zhang LY, Bai GQ, Gao XS, Ji D, Lin SM, Zhong YW, Shao Q. Screening of genes of proteins interacting with p7 protein of hepatitis C virus from human liver cDNA library by yeast two-hybrid system. World J Gastroenterol 2005; 11(30): 4709-4714 [PMID: 16094715 DOI: 10.3748/wjg.v11.i30.4709]
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. Aug 14, 2005; 11(30): 4709-4714 Published online Aug 14, 2005. doi: 10.3748/wjg.v11.i30.4709
Screening of genes of proteins interacting with p7 protein of hepatitis C virus from human liver cDNA library by yeast two-hybrid system
Yan-Ping Huang, Shu-Lin Zhang, Jun Cheng, Lin Wang, Jiang Guo, Yan Liu, Yuan Yang, Li-Ying Zhang, Gui-Qin Bai, Xue-Song Gao, Dong Ji, Shu-Mei Lin, Yan-Wei Zhong, 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, Yan-Wei Zhong, 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 Scientific Foundation, No. C03011402, No. C30070690; the Research and Technique Foundation of PLA during the 9th-five year plan period, No. 98D063; 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: October 19, 2004 Revised: December 23, 2004 Accepted: December 26, 2004 Published online: August 14, 2005
Abstract
AIM: To investigate the biological function of p7 protein and to look for proteins interacting with p7 protein in hepatocytes.
METHODS: We constructed p7 protein bait plasmid by cloning the gene of p7 protein into pGBKT7, then transformed it into yeast AH109 (a type). The transformed yeast was mated with yeast Y187 (α type) containing liver cDNA library plasmid, pACT2 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 blue colonies, we performed sequence analysis by bioinformatics.
RESULTS: Fifty colonies were selected and sequenced. Among them, one colony was Homo sapiens signal sequence receptor, seven colonies were Homo sapiens H19, seven colonies were immunoglobulin superfamily containing leucine-rich repeat, three colonies were spermatid peri-nuclear RNA binding proteins, two colonies were membrane-spanning 4-domains, 24 colonies were cancer-associated antigens, four colonies were nucleoporin 214 ku and two colonies were CLL-associated antigens.
CONCLUSION: The successful cloning of gene of protein interacting with p7 protein paves a way for the study of the physiological function of p7 protein and its associated protein.
