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Human Bcl-2 and human VEGF165 double-gene co-expression recombinant vector and building method thereof

A technology for recombinant vectors and construction methods, which is applied in botany equipment and methods, biochemical equipment and methods, genetic engineering, etc., can solve unsatisfactory problems, achieve the effects of accelerating recovery, promoting angiogenesis, and broad application prospects

Inactive Publication Date: 2010-02-24
THE SECOND AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV
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Problems solved by technology

Studies have shown that the application of Bcl-2 gene or VEGF gene alone has a significant therapeutic effect on myocardial infarction, but it is still not ideal, and there is room for further improvement

Method used

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  • Human Bcl-2 and human VEGF165 double-gene co-expression recombinant vector and building method thereof
  • Human Bcl-2 and human VEGF165 double-gene co-expression recombinant vector and building method thereof
  • Human Bcl-2 and human VEGF165 double-gene co-expression recombinant vector and building method thereof

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specific Embodiment 1

[0041] Figure 1-11 Constitute the specific embodiment 1 of the present invention.

[0042] Human Bcl-2 and Human VEGF 165 Double-gene co-expression recombinant vector, including human Bcl-2 gene and human VEGF 165 Gene; wherein, the human Bcl-2 gene sequence is described in the sequence table SEQ ID No. 1; human VEGF 165 The gene sequence is as described in SEQ ID No. 2 in the sequence table; the vector is an adenovirus vector, and other vectors such as other adeno-associated viruses, lentiviruses and plasmids can also be used.

[0043] The human Bcl-2 and human VEGF 165 The construction process of the double gene co-expression recombinant adenoviral vector is as follows: figure 1 As shown, its construction method is as follows:

[0044] 1. Main experimental reagents and equipment:

[0045] Biochemical reagents: DNA Marker (Takara, Japan), tryptone, yeast extract, agar powder (Oxoid, Japan), agarose (Biowest, Spain), kanamycin, ampicillin (Amresco, USA) and Trizol reage...

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Abstract

The invention relates to a human Bcl-2 and human VEGF165 double-gene co-expression recombinant vector and a building method thereof, and is characterized in that IRES segment is utilized to connect an hBc1-2 gene and an hVEGF165 gene, and homologous recombination mechanisms in bacterium are utilized to recombine and build the human Bcl-2 and human VEGF165 double-gene co-expression recombinant vector according to an AdEasy system. The invention leads the Bcl-2 gene with anti-apoptosis capacity and the VEGF165 gene which currently promotes angiogenesis cytokine with strongest function to recombine on a same vector to be co-expressed, and solves the problem of low survival rate of transplanted cells under the condition of hypoxia and inflammation, simultaneously plays the function of promoting angiogenesis of VEGF165, plays an important part in the gene therapy of myocardial infarction and the cell transplantation research field, and has wide application prospect.

Description

technical field [0001] The present invention relates to a double-gene co-expression recombinant vector and its construction method, in particular to a kind of human Bcl-2 and human VEGF 165 Double-gene co-expression recombinant vector and its construction method. It belongs to the field of biological gene therapy. Background technique [0002] Myocardium infarction (MI) is a sharp reduction or interruption of coronary artery blood supply on the basis of coronary artery disease, resulting in severe and persistent myocardial ischemia and necrosis of the corresponding myocardium. Cardiomyocytes are terminally differentiated cells, which cannot be repaired by cell regeneration after myocardial injury, but form fibrotic scars. At present, the clinical treatment methods for MI are mainly drug therapy, coronary artery intervention and coronary artery bypass surgery. Although these three methods can recanalize the occluded blood vessels and restore blood perfusion in some patients...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N15/861
Inventor 陈敏生倪晓彬何晓青刘世明罗承锋
Owner THE SECOND AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV
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