Method for constructing hepatitis C virus specific ribozyme M1GS-hcv/C76 and uses thereof
A technology of hepatitis C virus and construction method, applied in the field of biomedicine, can solve the problems of easy recurrence, unsatisfactory curative effect of interferon therapy, and no specific and effective prevention and treatment methods
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[0031] Example 1 Construction of hepatitis C virus-specific ribozyme M1GS-hcv / c76 and determination of cleavage activity
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[0034] 1.1 Enzymes and reagents Restriction endonuclease (KpnI, HindIII, XbaI), T7 RNA polymerase, DNase I (RNase free) were purchased from Fermentas (MBI) company; Ex Taq enzyme, mung bean sprout nuclease, T4 DNA ligase, RNase inhibitors were purchased from TaKaRa Bao Biology (Dalian) Technology Co., Ltd.; α- 32 UTP marked with P was purchased from Beijing Furui Company.
[0035] 1.2 Plasmids and pFL117 plasmids containing M1RNA gene in nucleotide fragments were donated by Professor Liu Fenyong (Fenyong Liu, University of California, Berkeley) from the University of California, Berkeley; pUC19 and pGEM3z plasmids were purchased from TaKaRabao Biological (Dalian) Technology Co., Ltd.; the pGEM-HC plasmid containing the 5'UTR of the HCV genome was constructed by the laboratory of Guangdong Pharmaceutical University o...
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