Hepatitis C virus core protein: synthesis, affinity purification and immunoreactivity with infected human sera.
The genomic region encoding the core (C) protein (amino acids 1-162) of hepatitis C virus (HCV) was expressed in Escherichia coli as a recombinant (re-) protein with the maltose-binding protein (MBP) using the prokaryotic expression vector pMAL-CR1. The fusion protein (C::MBP) was identified as a approx. 62-kDa polypeptide by immunoblot analysis using antiserum to MBP and HCV-infected human sera. The size of C::MBP corresponded to the calculated combined molecular mass of the approx. 20-kDa HCV C protein and the approx. 42-kDa MBP. The approx. 62-kDa C::MBP was purified using amylose resin as a matrix in affinity chromatography, and showed specific reactivity with HCV-infected human sera. These results suggest that C::MBP may serve as a source of the core antigen for immunological studies on HCV infection.
Khanna
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Virology Laboratory, National Institute of Immunology, New Delhi, India.
Ray
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R
D018528:ATP-Binding Cassette Transporters; D000688:Amylose; D001426:Bacterial Proteins / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D015870:Gene Expression; D016174:Hepacivirus / Q000737:chemistry* / Q000276:immunology; D006508:Hepatitis Antibodies / Q000276:immunology; D006526:Hepatitis C / Q000276:immunology*; D018937:Hepatitis C Antibodies; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D058619:Maltose-Binding Proteins; D008970:Molecular Weight; D009004:Monosaccharide Transport Proteins; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis / Q000737:chemistry; D014758:Viral Core Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D006508:Hepatitis Antibodies; D018937:Hepatitis C Antibodies; D007106:Immune Sera; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; C461599:maltose transport system, E coli; C069130:nucleocapsid protein, Hepatitis C virus; D000688:Amylose