In vitro activity measuring method for hepatitis C virus RNA depending RNA polymerase and application thereof
A hepatitis C virus, activity measurement technology, applied in the field of genetic engineering technology, can solve the problems of impracticality and complex activity measurement methods, and achieve the effect of simple method, enrichment and amplification of detection signals
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[0049] Detailed description of the embodiment of the present invention:
[0050] Cloning, expression and identification of hepatitis C virus C-terminal truncated NS5B protein with 21 amino acids
[0051] PCR technology was used to amplify HCV NS5B-C21, the gene fragment of NS5B protein truncated by 21 amino acids at the C-terminus, the gene fragment was cloned into the BamHI / Xho I site in plasmid pET28a(+), and the recombinant plasmid was transformed into Escherichia coli BL21( DE3), screened and obtained engineering bacteria expressing NS5B highly, and the expressed NS5B protein accounted for about 20% of the total bacterial protein. IPTG induces expression, and the target protein expression product is purified by affinity chromatography and identified by SDS-PAGE electrophoresis.
[0052] Materials and methods
[0053] 1. Plasmids and bacterial strains: Escherichia coli BL21 (DE3), HCV full-length cDNA plasmids, and prokaryotic expression vector pET-28a (+) are all preserv...
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