Original Articles
Copyright ©The Author(s) 2000. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Apr 15, 2000; 6(2): 220-222
Published online Apr 15, 2000. doi: 10.3748/wjg.v6.i2.220
Cloning and expression of core gene cDNA of Chinese hepatitis C virus in cosmid pTM3
Rong Long Jiang, Qiao Sheng Lu, Kang Xian Luo
Rong Long Jiang, Qiao Sheng Lu, Kang Xian Luo Department of Infectious Diseases, Nanfang Hospital, Guangzhou 510515, Guangdong Province, China;
Rong Long Jiang, graduated from the First Military Medical University in 1987, now a lecturer of medicine, majoring hepatitis B pathogenesis, having 10 papers published.
Author contributions: All authors contributed equally to the work.
Correspondence to: Dr. Rong Long Jiang, Department of Infectious Diseases, Nanfang Hospital, Guangzhou 510515, Guangdong Province, China. Jiangl@fimmu.edu.cn
Telephone: 0086-20-85147289, Fax. 0086-20-87636914
Received: May 19, 1999
Revised: December 6, 1999
Accepted: December 24, 1999
Published online: April 15, 2000
Abstract

AIM: To clone core gene cDNA of Chinese hepatitis C virus (HCV) into eukaryotic expression vector cosmid pTM3 and to express HCV core antigen in HepG2 cells.

METHODS: Core gene cDNA of HCV was introduced into eukaryotice xpression vector cosmid pTM3. Using vaccinia virus/bacterio phage T7 hybrid expression system, HepG2 cells were transfected with the recombinant plasmid pTM3-Q534 by lipofectin.

RESULTS: From the transfected bacteria Top10F′, 2 pTM3-Q534 clones containing the recombinant plasmid were identified from randomly selected 10 ampicillin-resistant colonies. By reverse transcription PCR and indirect immunofluorescence technique, HCV RNA and core protein was identified in HepG2 cells transfected with the recombinant plasmid.

CONCLUSION: The construction of a recombinant plasmid and the expression of core gene cDNA of HCV in HepG2 was successful.

Keywords: hepatitis C virus; gene; viral; cDNA; cosmid vector; gene expression