Method for constructing human neurenergen-3 receptor gene recombination adenovirus

A technology of recombinant adenovirus and receptor gene, which is applied in the fields of genetic engineering, plant genetic improvement, botany equipment and methods, etc., can solve the problem of weak effect of neurotrophin-3, so as to promote social and economic development and relieve social and family burden. Burden, the effect of improving the quality of life

Inactive Publication Date: 2005-10-19
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
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Problems solved by technology

Brain-derived neurotrophic factor, neurotrophin-4 / 5, and neurotrophin-3 all bind and activate TrkB, but neurotrophin-3 is less potent

Method used

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Examples

Experimental program
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Embodiment Construction

[0017] Below by specific embodiment main instrument used in the present invention and reagent are described in detail:

[0018] 1. Main instruments

[0019] PCR instrument (Gene Amp pcr system 9700), low-temperature centrifuge (Beckman), ultra-high-speed centrifuge (Beckman, USA), CO 2 Incubator (NAPCO), PRISM377 DNA sequencer (ABI, USA), high-speed desktop centrifuge 5415D (eppendorf, Germany) and BeckmanDV640 UV spectrophotometer (Beckman, USA).

[0020] 2. Vector and strain

[0021] Adeno-X TM The Expressien System kit is a product of Clontech, and the HEK293 cell line and Escherichia coli DH5α are gifts from Professor Huang Wenlin's laboratory of the Cancer Prevention and Control Center.

[0022] 3. Main reagents for gene recombinant adenovirus expression vector

[0023] RT-PCR kits and Trizol are products of Promege; restriction enzymes XbaI, KpnI, DNA MarkerDL2000, and DNA Ligation Kit Ver.2 are products of Dalian Takara Company; DNA Marker 1 KB Ladder is a product ...

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PUM

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Abstract

The present invention relates to the construction method and application of one kind of human neurenergen-3 receptor gene (human TrkC gene) recombined adenovirus expressing vector. The basic scheme of the present invention includes: designing and synthesizing primer, RT-PCR process, construction and identification of cloned shuttle vector pShuttle-TrkC, construction and identification of recombined adenovirus vector pAdeno-TrkC, 293 cell packaging, identification of recombined adenovirus, bioactivity detection of human neurenergen-3 receptor gene recombined adenovirus expressing vector and other steps. The present invention is superior in that one kind gene recombined adenovirus expressing vector is selected to promote the survival of damaged central neure and the regeneration of axone, to promote nerve stem cell to differentiate into more neures to replace damaged or dead neures.

Description

technical field [0001] The invention relates to a method for constructing a recombinant adenovirus expression vector, in particular to a method for constructing an adenovirus expression vector recombined with human neurotrophin-3 receptor gene (human TrkC gene) and its application. Background technique [0002] Neurotrophic factors are a class of chemical factors that can support the survival of neurons in the body, promote their growth, differentiation, and maintain their functions. Before neurotrophic factors exert cytological effects on their target cells, they must first bind to receptors on the target cells. Currently known neurotrophic factors have two receptors: tyrosine protein kinase receptor (high affinity receptor) and p75 receptor (low affinity receptor). Tyrosine protein kinase receptor is a receptor encoded by the proto-oncogene Trk, also known as Trk receptor. Trk receptors can be divided into three types: TrkA, TrkB and TrkC. [0003] Neurotrophic factors ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/861C12Q1/68
Inventor 曾园山王俊梅
Owner SUN YAT SEN UNIV
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