Abstract
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.
MeSH terms
-
ATP-Binding Cassette Transporters*
-
Amylose
-
Bacterial Proteins / genetics
-
Carrier Proteins / genetics
-
Chromatography, Affinity
-
Escherichia coli / genetics
-
Escherichia coli Proteins*
-
Gene Expression
-
Hepacivirus / chemistry*
-
Hepacivirus / immunology
-
Hepatitis Antibodies / immunology
-
Hepatitis C / immunology*
-
Hepatitis C Antibodies
-
Humans
-
Immune Sera / immunology
-
Maltose-Binding Proteins
-
Molecular Weight
-
Monosaccharide Transport Proteins*
-
Recombinant Fusion Proteins / analysis
-
Recombinant Fusion Proteins / biosynthesis
-
Recombinant Fusion Proteins / chemistry
-
Viral Core Proteins* / biosynthesis
-
Viral Core Proteins* / genetics
-
Viral Core Proteins* / immunology
-
Viral Core Proteins* / isolation & purification
Substances
-
ATP-Binding Cassette Transporters
-
Bacterial Proteins
-
Carrier Proteins
-
Escherichia coli Proteins
-
Hepatitis Antibodies
-
Hepatitis C Antibodies
-
Immune Sera
-
Maltose-Binding Proteins
-
Monosaccharide Transport Proteins
-
Recombinant Fusion Proteins
-
Viral Core Proteins
-
maltose transport system, E coli
-
nucleocapsid protein, Hepatitis C virus
-
Amylose