Method for constructing hepatitis C virus specific ribozyme M1GS-hcv/C20 and uses thereof

A technology for hepatitis C virus and construction method, applied in the field of biomedicine, can solve the problems of easy repetition, no specific and effective prevention and treatment method, and unsatisfactory effect of interferon therapy.

Inactive Publication Date: 2008-08-27
GUANGDONG PHARMA UNIV
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  • Abstract
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Problems solved by technology

At present, there is no specific and effective prevention and treatment method for this virus. The curative effect of conventional inte

Method used

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  • Method for constructing hepatitis C virus specific ribozyme M1GS-hcv/C20 and uses thereof
  • Method for constructing hepatitis C virus specific ribozyme M1GS-hcv/C20 and uses thereof
  • Method for constructing hepatitis C virus specific ribozyme M1GS-hcv/C20 and uses thereof

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Embodiment 1

[0031] The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments. Example 1 Construction of hepatitis C virus-specific ribozyme M1GS-hcv / c20 and determination of cleavage activity

[0032] 1 material

[0033] 1.1 Enzymes and reagents Restriction endonuclease (Kpn I, Hind III, Xba I), T7 RNA polymerase, DNase I (RNase free) were purchased from Fermentas (MBI); Ex Taq enzyme, mung bean sprout nuclease, T4DNA Ligase and RNase inhibitors were purchased from TaKaRa Bao Biological (Dalian) Technology Co., Ltd.; α- 32 UTP marked with P was purchased from Beijing Furui Company.

[0034] 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 Biology (Dalian) Technology Co., Ltd.; the pGEM-HC plasmi...

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Abstract

The invention discloses a method for constructing hepatic c virus specific ribozyme MIGS-hcv/C20, which comprises: selecting a sequence between 21nt and 36nt of hepatitis c virus HCV genomere 5' UTR as a target site to design a guide sequence and obtaining through associating and constructing the guide sequence and escherichia coli ribozyme P covalently. The guide sequence is designed to be effectively combined with the hepatic c virus specific ribozyme successfully through selecting a single-stranded region sequence between the 21nt-36nt of the hepatitis c virus as a candidate target site, the ribozyme which is designed accordingly displays clear targeting cutting activity for a substrate, successful construction of the ribozyme provides experimental materials which are accurate and can be depended for subsequent experiments, and thereby the method lays a foundation for researching new anti-HCV medicine and an antisense gene therapy.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a hepatitis C virus-specific ribozyme M1GS-hcv / c20 and its construction method and application. Background technique [0002] At present, the antisense technology based on ribozyme P (RNase P) has received extensive attention in the field of antiviral research. A variety of viruses have been studied, and it has been found that it can efficiently and specifically inhibit the expression of specific virus genes, which shows the great potential of this technology in the field of anti-virus. Ribozyme P is a natural macromolecular ribozyme widely present in various biological cells (including eukaryotic cells, prokaryotic cells and archaea). Its main biological function is to be responsible for the specific excision of the 5' leader sequence of the precursor tRNA (ptRNA) to promote the maturation of tRNA. For any target mRNA whose sequence is known, theoretically, a c...

Claims

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Application Information

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IPC IPC(8): C12N9/22C12N15/51C12N15/55C12N15/63A61K38/46A61P31/12
CPCY02A50/30
Inventor 李红枝张文军陈伟强
Owner GUANGDONG PHARMA UNIV
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