Method for constructing interference plasmid of human fibrinogen hf12 shRNA, and pharmic usage

A technology of cellulose intermediary and construction method, applied in the field of bioengineering, can solve the problem of no definite treatment method for severe hepatitis

Inactive Publication Date: 2007-05-09
TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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Problems solved by technology

So far, there is no confirmed effective treatment for severe hepatitis in the world, and exploring the pathogenesis

Method used

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  • Method for constructing interference plasmid of human fibrinogen hf12 shRNA, and pharmic usage
  • Method for constructing interference plasmid of human fibrinogen hf12 shRNA, and pharmic usage
  • Method for constructing interference plasmid of human fibrinogen hf12 shRNA, and pharmic usage

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Embodiment Construction

[0042] Below in conjunction with specific embodiment the present invention is described in further detail:

[0043] The present invention proposes the method for constructing hfgl2 shRNA interference plasmid and confirms its interference effect at cellular level and comprises the following steps:

[0044] 1. The construction part of the carrier

[0045]Method for constructing hfgl2 shRNA interference plasmid:

[0046] 1. Screen the target sequence according to the hfgl2 prothrombinase cDNA sequence and design principles, design the hfgl2 shRNA template according to the target sequence, then design the template in the downstream primers, amplify the U6 promoter by PCR, and introduce restriction sites at both ends BamH I and Hind III (see Figure 1 for the two target sequences of hfgl2 and the upstream primers of the U6 promoter).

[0047] 2. Insert the U6 promoter with the hfgl2 shRNA template downstream into the pMSCVneo vector (Figure 2) corresponding restriction sites BamH ...

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Abstract

This invention discloses a method for constructing shRNA interference plasmid targeting human fibrinogen-like protein 2 (hfgl2) and its medical application. The method comprises: screening the target sequence according to hfgl2 cDNA sequence, designing a template of hfgl2 cDNA according to the target sequence, designing a reverse primer containing the template, performing PCR to amplift U6 promoter, inserting BamH I and Hind III sites at both ends respectively, cutting pMSCVneo plasmid with BamH I and Hind III, and inserting U6 promoter sequence with shRNA template sequence at the downstream into pMSCVneo. It is proved by cell level experiments that hfgl2 shRNA interference plasmid can effectively inhibit the expression of hfgl2 gene.

Description

technical field [0001] The invention relates to the field of bioengineering, specifically to construct a new biological agent: hfgl2 shRNA interference plasmid, and its pharmaceutical application for treating virus-induced severe hepatitis and other diseases whose main pathophysiological feature is microcirculation disturbance. Background technique [0002] As we all know, viral hepatitis has become an important disease that seriously affects people's health and living standards and restricts economic development due to its high incidence and great harm. So far, there is no confirmed effective treatment for severe hepatitis in the world. Exploring the pathogenesis and prevention methods of severe hepatitis is an important topic in the field of applied basic and clinical research. [0003] Infecting different strains of inbred mice with mouse hepatitis virus-3 (MHV-3) has successfully established various clinical types of hepatitis similar to humans, includ...

Claims

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Application Information

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IPC IPC(8): C12N15/66C12N15/57A61K48/00C12N1/21A61P1/16C12R1/19
Inventor 宁琴罗小平李黎习东朱传龙严伟明
Owner TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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