Cloning and Expression of NS5 Region of Korean Type Hepatitis C Virus
Journal of the Korean Society of Virology
;
: 115-128, 1997.
Article
in Korean
| WPRIM
| ID: wpr-36402
ABSTRACT
Three cDNA fragments located within NS5 region of HCV were synthesized by RT using viral RNA extracted from blood sample of Korean patient as a template. The cDNAs were amplified by PCR, cloned into the T-vector, and the nucleotide sequences were determined. Comparative a analysis of the nucleotide and amino acid sequence of NS5 cDNAs showed that it is closely related with HCV type 1b. The cloned NS5 cDNA showed 91-94% homology at the nucleotide sequence level and 96-98% homology at the amino acid sequence level with several strains of the HCV type 1b. The NS5 cDNAs were subcloned into E. coli expression vectors to construct pRSETA5-1, pTHAN5-1, pRSETC5-2, pRSETBB1, pRESTCB1 and PRSETB-H3. Expression of the NS5 proteins was achieved by inducing the promoter with isopropyl-thio-P-D-galactoside (IPTG) and confirmed by SDS-polyacrylamide gel electrophoresis. The NS5 proteins were immunoreactive against sera from Korean hepatitis C patients in Western blot analysis. Among the recombinant NS5 proteins, pRSETA5-1 plasmid derived protein, coded from aa2022 to aa2521 of HCV polyprotein, showed the strongest immunoreactivity against sera from Korean hepatitis C patients in immunoblot analysis. These results suggest that NS5 proteins would be useful as an antigen for detection of antibody against HCV in the blood samples.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Plasmids
/
RNA, Viral
/
Base Sequence
/
Blotting, Western
/
Polymerase Chain Reaction
/
Amino Acid Sequence
/
Clone Cells
/
Hepatitis C
/
DNA, Complementary
/
Hepacivirus
Limits:
Humans
Language:
Korean
Journal:
Journal of the Korean Society of Virology
Year:
1997
Type:
Article
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