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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 no specific and effective prevention and treatment method, easy repetition, unsatisfactory effect of interferon therapy, etc.

Inactive Publication Date: 2010-11-17
GUANGDONG PHARMA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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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 interferon therapy is not satisfactory and it is easy to repeat. Therefore, it is of great significance to explore new prevention and treatment methods

Method used

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  • Method for constructing hepatitis C virus specific ribozyme M1GS-hcv/C76 and uses thereof
  • Method for constructing hepatitis C virus specific ribozyme M1GS-hcv/C76 and uses thereof
  • Method for constructing hepatitis C virus specific ribozyme M1GS-hcv/C76 and uses thereof

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0031] Example 1 Construction of hepatitis C virus-specific ribozyme M1GS-hcv / c76 and determination of cleavage activity

[0032] 1 material

[0033] Material

[0034] 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.

[0035] 1.2 Plasmids and nucleotide fragments The pFL117 plasmid containing the M1 RNA gene was donated by Professor Liu Fenyong (Fenyong Liu, University of California, Berkeley) from the University of California, Berkeley; the pUC19 and pGEM3z plasmids were purchased from TaKaRa Biotech (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...

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Abstract

The invention discloses a structural method of hepatitis C virus-specific ribozymes M1GS-hcv / c76, which is structured by selecting sequence between 77-89nt of genome 5'UTR of hepatitis C virus HCV as target design guide sequence, and covalently linking guide sequence with escherichia coli ribozyme P. The invention selects sequence between 77-89nt of hepatitis C virus as candidate target to successfully design guide sequence to bind with the same. Designed ribozyme by the method displays evident targeting cleavage activity to substrate. The ribozyme provides precise and reliable experimental material for subsequent experiment after its successful structuring, thus lays a foundation for research in novel HCV medicament and 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 / c76 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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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/22C12N15/51C12N15/55C12N15/63A61K38/46A61P31/12
CPCY02A50/30
Inventor 张文军李红枝邓祖军
Owner GUANGDONG PHARMA UNIV
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