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Construction method and pharmaceutical application of hFas microRNA adenovirus expression plasmid relevant with severe hepatitis

A technology for expressing plasmids and severe hepatitis, applied in DNA/RNA fragments, recombinant DNA technology, genetic engineering, etc., can solve the problems of lack of specific and effective treatment methods, liver cell death, and dangerous disease

Inactive Publication Date: 2012-07-04
TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] Severe hepatitis can be caused by a variety of etiologies clinically, and its onset is dangerous. Patients may experience large areas of liver cell death in a short period of time. Currently, there is no specific and effective treatment

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  • Construction method and pharmaceutical application of hFas microRNA adenovirus expression plasmid relevant with severe hepatitis
  • Construction method and pharmaceutical application of hFas microRNA adenovirus expression plasmid relevant with severe hepatitis
  • Construction method and pharmaceutical application of hFas microRNA adenovirus expression plasmid relevant with severe hepatitis

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Embodiment Construction

[0070] The present invention will be further described in detail below in conjunction with specific embodiments.

[0071] The present invention constructs the miRNA expression plasmid of hfg12 prothrombinase, hFas and hTNFR1, and confirms its interference effect at the cellular level, including the following steps (taking the construction of hFas-microRNA expression plasmid as an example):

[0072] 1. Construction of pcDNA expression vector

[0073] Construction of hFas-miRNA expression plasmid: According to the hFas cDNA sequence on PubMed, use the miRNA design software of Invitrogen Company to design and synthesize 3 pairs of complementary oligonucleotide single strands. According to the molecular cloning procedure, insert ds Oligo into the microRNA expression vector pcDNA TM 6.2-GW / EmGFP-miR, construct three hFas-miRNA plasmids: p-hFasmi-RNA1, p-hFasmiRNA2 and p-hFasmiRNA3. The miRNA expression plasmids of the unmutated hFas gene were screened by sequencing.

[0074] 2. ...

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Abstract

The invention discloses a construction method and pharmaceutical application of hFas microRNA adenovirus expression plasmid relevant with severe hepatitis and relates to a method for constructing a group of gene biological preparations, i.e. the microRNA adenovirus expression plasmid of hFas gene relevant with severe hepatitis can be constructed. The single-stranded oligonucleotide sequence of the generated hFas microRNA is 5'-TGCTGTTAAGTTGGAGATTCATGAGTTTTGGCCACTGACTGACTCTCATGACTCCAACCTTA-3'. By using the invention, the effectiveness and the specificity of the microRNA interfering plasmid canbe detected in the cell level and the hFas microRNA adenovirus expression plasmid can be established and applied to treating the severe hepatitis.

Description

[0001] The present invention is a divisional application for the construction method of severe hepatitis-related genes hfg12, hFas and hTNFR1 microRNA adenovirus expression plasmids and their pharmaceutical application. The original application number is 200810133445.1, and the application date is July 18, 2008. technical field [0002] The present invention relates to the construction of a group of gene biological preparations, that is, the construction of human fg12 (Human Fibrinogen-like protein 2, hfg12) prothrombinase, Fas and tumor necrosis factor receptor I (tumor necrosis factor, TNFR1) genes associated with severe hepatitis The microRNA adenovirus expression plasmid is combined with three plasmids for treatment of severe hepatitis, and its pharmaceutical use is verified. Background technique [0003] Due to the high incidence and great harm of viral hepatitis, it has become an important disease that seriously affects people's health and living standards and restricts...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/861C12N15/63C12N15/113
Inventor 宁琴罗小平习东高随朱传龙郭健文严伟明王鸣
Owner TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH