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Recombinant virus with coexpression of anthropogenic glutamate decarboxylase and cell factor

A technology of glutamate desustriase and cytokines, which is applied in the direction of virus/bacteriophage, recombinant DNA technology, gene therapy, etc., can solve the problems that cannot change the clinical symptoms of Parkinson's pathological process

Inactive Publication Date: 2009-11-25
王尚武 +2
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Problems solved by technology

However, it is clear that this treatment is a cure for improving the symptoms of Parkinson's disease, and cannot change the pathological process of Parkinson's disease and other clinical symptoms resulting from it.

Method used

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  • Recombinant virus with coexpression of anthropogenic glutamate decarboxylase and cell factor
  • Recombinant virus with coexpression of anthropogenic glutamate decarboxylase and cell factor
  • Recombinant virus with coexpression of anthropogenic glutamate decarboxylase and cell factor

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Embodiment Construction

[0037] specific implementation

[0038] In order to facilitate the description of the specific implementation process of constructing a recombinant virus that co-expresses human GAD and human GDNF expression structures, the present invention uses an adeno-associated virus (AAV, the same below) expression system, more specifically a helper-free AAV expression system, AAV recombinant virus co-expressing GAD and GDNF expression constructs was constructed.

[0039] The specific implementation process of constructing the AAV recombinant virus co-expressing GAD and GDNF expression structure is as follows:

[0040] 1. Cloning of human glutamic acid decarboxylase GAD65 gene:

[0041] 1). Considering the large size of the GAD65 gene, it was decided to divide the gene into two fragments for cloning, and then use appropriate endonuclease sites to complete the splicing of the full-length GAD65 gene.

[0042] Firstly, total RNA (total RNA) of brain tissue was extracted, and cDNA of brain...

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Abstract

The invention relates to a recombinant virus containing a gene expression box expressing an anthropogenic GAD gene and a cell factor, in particular to a method for constructing a recombinant adeno-associated virus with the coexpression of an anthropogenic GAD 65 gene and a GDNF gene, an application and a meaning thereof. Anthropogenic GAD 65 gene and the GDNF gene protein products generated by the expression of the recombinant virus have favorable synergistic effect on the treatment of diseases relative to Parkinson's disease and the central nervous system and have important values for treatment of relative diseases.

Description

technical field [0001] The invention relates to the construction and preparation method of a recombinant virus co-expressing glutamic acid decarboxylase and cytokines, especially the recombinant adeno-associated virus, and its technical field is related to life science and biomedicine technology. Background technique [0002] Studies have proved that the neurotransmitter of most neurons in the striatum and striatum projection area of ​​the mammalian brain is γ-aminobutyric acid (GABA, the same below), and GABA is the main inhibitory neurotransmitter of the mammalian brain tissue. And glutamic acid decarboxylase (GAD, the same below) is the rate-limiting enzyme of synthetic GABA, and GAD has two kinds of isozymes of GAD 65 and GAD 67 ( Lindefors N . et al: Prog. Neuropsycho. Pharmacol. Biol Psychiatry. 1993;17(6):887-903). Some researchers applied the adeno-associated virus (AAV, the same below) expressing the GAD65 gene to conduct therapeutic research on animal models o...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/60C12N15/12C12N15/62C12N15/10A61K38/17A61K48/00A61P25/00A61P25/16
Inventor 王尚武朱雅南徐评议
Owner 王尚武
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