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Rastgoo N, Karami A, Sabahi F. Cloning, Sequencing and Expression of Core Gene of Hepatitis C Virus from an Iranian Patient with Chronic Hepatitis. mjms. 2010; 9 :9-18
URL: http://mjms.modares.ac.ir/article-30-3659-en.html
1- M.Sc. in Medical Biotechnology, Tarbiat Modares University, Tehran, Iran.
2- Ph.D. in Biotechnology, Associate Professor, Bagyatallah University, Tehran, Iran.
3- Ph.D. in Virology, Associate Professor, Tarbiat Modares University, Tehran, Iran
Abstract:   (4066 Views)
Introduction: Hepatitis C virus (HCV) is the major cause of acute and chronic hepatitis, cirrhosis, and hepatocellular carcinoma worldwide. Because of increasing number of infected patients throughout the world, various studies are concentrated on development of high quality diagnostic tests and genotyping in various regions of the world. Methods: In this study, we used RT-PCR to isolate and amplify the core gene sequence of the hepatitis C virus that is very conserved among various genotypes, from an isolate derived from an Iranian patient with chronic hepatitis. Then it was cloned in pUC18 vector and sequenced with universal primers. In order to produce recombinant core protein, the core gene was cloned in PET28A expression vector and the recombinant plasmid was transformed into BL21 (DE3) bacteria. Results and Discussion: The results of sequencing showed that the core gene of the hepatitis C virus from an isolate derived from an Iranian patient with chronic hepatitis has a higher similarity with 1a (96%) and 1b (95%) strains of the virus. This result is similar to those obtained by previous studies. The presence of a 23.5 KD band in SDS-PAGE and Western blot using monoclonal antibody, proved the expression of Core protein in PET28A vector. Mass production of this protein could lead to its use in detection of anti-HCV antibodies in infected patients by immunoassays.
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Received: 2005/04/2 | Accepted: 2006/04/2

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