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Iranian Journal of Biotechnology، جلد ۱۳، شماره ۴، صفحات ۳۲-۳۷
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عنوان انگلیسی Analysis of Immumoreactivity of Heterologously Expressed Non-structural Protein 4B (NS4B) from Hepatitis C Virus (HCV) Genotype 1a
چکیده انگلیسی مقاله Background: Detection of hepatitis C virus specific antibodies is the initial step in chronic HCV diagnosis. HCV NS4B is among the most immunogenic HCV antigens and has been widely used in commercial Enzyme Immunoassays (EIA). Additionally, NS4B, a key protein in the virus replication, can be an alternative target for antiviral therapy. Objectives: Development of a new method for high-level expression and purification of NS4B coding region was the aim of the report. Materials and Methods: Viral RNA was purified from the serum of an HCV positive patient and NS4B coding region was amplified using nested RT-PCR. PCR products were cloned into pET102/D-TOPO expression vector and transformed into E. coli BL21. Induction was performed by adding 1mM isopropyl-b-D-thiogalactopyranoside (IPTG) to the culture medium. Immunoreactivity of the purified recombinant proteins was evaluated by immunoblotting and indirect enzyme-linked immunosorbent assay (ELISA). Results: The recombinant NS4B protein was expressed and its immunoreactivity was confirmed by ELISA and western blotting. Conclusions: The directional TOPO cloning provides an efficient and easy platform for heterologous expression of immunoreactiveHCV NS4B.
کلیدواژه‌های انگلیسی مقاله Hepatitis C Virus,NS4B,recombinant antigen,Serodiagnosis
نویسندگان مقاله امیر savardashtaki |
department of medical biotechnology, school of allied medical sciences, iran university of medical sciences, tehran, iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی ایران (Iran university of medical sciences)

زهره شریفی |
department of virology, iranian blood transfusion organization, tehran, iran.
سازمان اصلی تایید شده: سازمان انتقال خون ایران (Blood transfusion research center)

سپیده حمزه لو |
department of medical biotechnology, school of allied medical sciences, iran university of medical sciences, tehran, iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی ایران (Iran university of medical sciences)

محمد مهدی فرج اللهی | mohammad m
department of medical biotechnology, school of allied medical sciences, iran university of medical sciences, tehran, iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی ایران (Iran university of medical sciences)

نشانی اینترنتی http://www.ijbiotech.com/article_11960_bf182aa02e113ccce19a3c295c66ab41.pdf
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