CHARACTERIZATION,CLONING AND EXPRESSION OF NS3 PROTEIN GENE OF HEPATITIS C GENOTYPE 4A
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Date
2009
Journal Title
Journal ISSN
Volume Title
Type
Article
Publisher
Egyptian Society of Parasitology
Series Info
Journal of the Egyptian Society of Parasitology;39 (3), 2009: 865-880
Doi
Scientific Journal Rankings
Abstract
Clone and express NS3 gene of the Egyptian strain ED43 of HCV genotype
4a in E. coli was studied. Gene and protein sequences of NS3 gene of the ED43
strain were first analyzed using PC/GENE program. DNA homology was 89%
the homologies and that of the protein was 78.8% indicating that NS3 gene of
the genotype 4a is different from those isolated from other strains.
DNA of NS3 region of genotype 4a was amplified from HCV_ED43/PUC19
plasmid. The PCR product was cloned and expressed in E. coli M15 using
pQE-30 vector. Fusion protein containing the peptides coded by HCV NS3
(NS3_4a) was expressed by Escherichia coli. The specific HCV antigenicity
of the NS3_4a fusion protein was identified by western blotting.
Description
MSA Google Scholar
Keywords
HCV,, Genotype 4a, Egyptian strain ED43, NS3_4a, Cloning, Expression
Citation
Alfonso, V.; Flichman, D.; Sookoian, S.; Mbayed, V. A.; Campos, R. H.; 2001: Phylogenetic characterization of genotype 4 hepatitis C virus isolates from Argentina. J. Clin. Microbiol. 39, 5:1989-92. Altschul, S.F.; Madden, T.L.; Schäffer, A.A.; Zhang, J.; Zhang, Z.; Miller, W.; Lipman, D.J.; 1997: Gapped BLAST nad PSIBLAST: a new generation of protein database search programs. Nucl. Acids Res., 25:3389-402. Bartenschlager, R.; 1997: Molecular targets in inhibition of hepatitis C virus replication. Antivir. Chem. Chemother. 8:281-301. Bukh, J.; Miller, R.H.; Purcell, R.H.; 1995: Defining the genotypes of hepatitis C virus and their epidemiology. INTER-action 3:10-15. Cheng, J.; Zhong, Y.; Liu, Y.; Dong, J., Yang, J.; Yang, S.; 2002: Inducible expression of nonstructural protein 3 of hepatitis C virus in E. coli. Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi, 16, 1:85-7. De Francesco, R.; Steinkühler, C.; 2000: Structure and function of the hepatitis C virus NS3-NS4A serine proteinase. Curr. Top. Microbiol. Immunol. 242:149-69. Du, G.X.; Hou, L.H.; Guan, R.B.; Tong, Y.G.; Wang, H.T.; 2002: Establishment of a simple assay in vitro for hepatitis C virus NS3 serine protease based on recombinant substrate and single-chain protease. Wld. J Gastroenterol. 8, 6:1088-93. Dusheiko, G.; Weiss, H.; Brown, D.; McOmish, F.; Yap, P.L.; et al.; 1994: Hepatitis C virus genotypes: an investigation of typespecific differences in geographic origin and disease. Hepatol., 19:13- 8. Frick, D.N.; 2007: The hepatitis C virus NS3 protein: a model RNA helicase and potential drug target: Review. Mol. Biol., 9, 1:1-20. Hong, Z.; Standring, D.N.; Baroudy, B.; Lau, J.Y.; 2000: Development of novel anti-HCV therapies: HCV protease, helicase, and polymerase as thera-peutic targets. Acta Gastroenterol. Belgica, 63:210-2. Jiao, X.; Wang, R.Y.; Feng, Z.; Hu, G.; Alter, H.J.; Shih, J.; 2004: DNA immunization encoding the secreted nonstructural protein 3 (NS3) of hepatitis C virus and enhancing the Th1 type immune response. J. Viral. Hepat. 11, 1:18-26. Johnson, D.R.; Bhatnagar, R.S; Knoll, L.J.; Gordon, J.I.; 1994: Genetic and biochemical studies of protein N-myristoylation. Ann. Rev. Biochem., 63: 869-914. Kwong, A.D.; Kim, J.L.; Rao, G.; Lipovsek, D.; Raybuck, S.; 1998: He-patitis C virus NS3/4A protease. Antivir. Res. 40:1-18. Lavanchy, D.; McMahon, B.; 2000: Worldwide Prevalence and Prevention of HC. In: Hepatitis C. By Liang TJ, Hoofnagle JH, San Diego, Academic Press. Lodrini, S.; Bagaglio, S.; Canducci, F.; De Mitri, M.; Andreone, P.; et al.; 2003: Sequence analysis of NS3 protease gene in clinical strains of hepatitis C virus. J. Biol. Regul. Homeost. Agents 17, 2:198-204. Misialek, S.; Rajagopalan, R.; Stevens, S.K.; Beigelman, L.; Seiwert, S.D.; Kossen, K.; 2009: Optimization of the multiple enzymatic activities of the hepatitis C virus NS3 protein. Anal. Biochem., 394,1:138-40. Penin, F.; Dubuisson, J.; Rey, F. A.; Moradpour, D.; Pawlotsky, J. M.; 2004: Structural biology of hepatitis C virus. Hepatol., 39, 1:5-19. Pless, D.D.; Lennarz, W. J.; 1977: Enzymatic conversion of proteins to glycoproteins. Proc. Natl. Acad. Sci. USA., 74, 1:134-8. Poliakov A.; Hubatsch I.; Shuman C.F.; Stenberg G.; Danielson U. H.; 2002: Expression and purification of recombinant full-length NS3 protease-helicase from a new variant of Hepatitis C virus. Protein Exp. Purif., 25, 3: 363-71. Ray, S.C.; Arthur, R.R.; Carella, A.; Bukh, J.; Thomas, D.L.; 2000: Genetic epidemiology of hepatitis C virus throughout Egypt. J. Infect. Dis., 182: 698-707. Sambrook, J.; Fritsch, E.F.; Maniatis, T.; 1989: Molecular Cloning: Cold Spring Harbor Laboratory Press. Sillanpää, M. ; Melén, K.; Porkka, P.; Fagerlund, R.; Nevalainen, K.; et al.; 2009 : HCV core, NS3, NS4B and NS5A are the major immunegenic proteins in humoral immunity in chronic HCV infection. Virol. J., 24, 6:86. Stuyver, L.; Van Arnhem, W.; Wyseur, A.; Hernandez, F.; Delaporte, E.; Maertens, G.; 1994: Classification of hepatitis C viruses based on phylo-genetic analysis of the envelope 1 and nonstructural 5B regions and identi-fication of five additional subtypes. Proc. Natl. Acad. Sci. USA, 91, 10:104-13. Tai, C.L.; Chen, D.S.; Hwang, L.H.; 1996: The helicase activity associated with hepatitis C virus nonstructural protein 3 (NS3). J. Virol., 70:8477-84. Tanaka, T.; Tsukiyama, K.; Yamaguchi, K.; Yagi, S.; Tanaka, S.; et al.; 1994: Significance of specific antibody assay for genotyping of hepatitis C virus. Hepatol., 19, 6:1347-53. Tellinghuisen, T.L.; Rice, C.M.; 2002: Interaction between hepatitis C virus proteins and host cell factors. Curr Opin Microbiol., 5:419- 27. Thibeault, D.; Bousquet, C.; Gingras, R.; Lagace, L.; Maurice, R.; et al.; 2004: Sensitivity of NS3 serine proteases from hepatitis C virus genotypes 2 and 3 to the inhibitor BILN 2061. J. Virol., 78, 14:7352- 9. Vishnuvardhan, D.; Kakiuchi, N.; Urvil, P.T.; Shimotohno, K.; Kumar, P.; Nishikawa, S.; 1997: Expression of highly active recombinant NS3 protease domain of hepatitis C virus in E. coli. FEBS Letters, 402, 2-3:209-12.