Molecular cloning, sequencing and expression of core and NS3 fragments of HCV from patients with HCV infection.
Fragments of core and NS3 of hepatitis C virus-Hunan (HCV-Hun) were cloned by RT-PCR and gene recombinant techniques from blood samples collected in Hunan Province, China. In comparison with sequences of our samples with those of HCV-US and HCV-J, the homologies of nucleotides and amino acids were about 90%, indicating that fragments of core and NS3 of HCV-Hun were in a relative conserved region of HCV. Two fusion proteins containing the peptides coded by HCV core (MBP-HCV core) and HCV. NS3 (MBP-HCV. NS3-Gal) were expressed by Escherichia Coli with recombinant plasmids. The specific HCV antigenicity of the two fusion proteins were identified by western blotting. Therefore, MBP-HCV. core and MBP-HCV.NS3-Gal were found useful for anti-HCV assay.
Tan
D M
DM
Department of Infectious Diseases, First Affiliated Hospital, Hunan Medical University, Changsha.
Hu
G L
GL
Arima
T
T
Zhang
Z
Z
Nanba
T
T
Hatanaka
T
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000235:genetics; D015870:Gene Expression; D016174:Hepacivirus / Q000235:genetics*; D006526:Hepatitis C / Q000382:microbiology*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D014758:Viral Core Proteins / Q000235:genetics*; D017361:Viral Nonstructural Proteins / Q000235:genetics*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D004274:DNA, Recombinant; D004279:DNA, Viral; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C069130:nucleocapsid protein, Hepatitis C virus