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Method for preparing transgenic mouse with central nervous system specific expression Cre recombinase

A transgenic mouse and central nervous system technology, applied in botany equipment and methods, biochemical equipment and methods, and other methods of inserting foreign genetic materials, can solve problems such as inability to obtain adult animals, death, mouse embryo deformities, etc.

Inactive Publication Date: 2008-05-14
INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE
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AI Technical Summary

Problems solved by technology

Traditional gene knockout and tissue-specific knockout mediated by Cre / LoxP or Red recombination system have played an important role in this area as an effective research tool, but deletion of important genes related to embryonic development often results in small Mouse embryos are malformed or die early in the embryo so that adult animals cannot be obtained

Method used

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  • Method for preparing transgenic mouse with central nervous system specific expression Cre recombinase
  • Method for preparing transgenic mouse with central nervous system specific expression Cre recombinase
  • Method for preparing transgenic mouse with central nervous system specific expression Cre recombinase

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Experimental program
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Effect test

Embodiment Construction

[0019] Materials and methods

[0020] 1 Plasmids and strains

[0021] The pancreas-specific Cre recombinase expression plasmid pRCH was donated by Dr. Zhou Jiang of our institute, and the pBC1 plasmid, pGEM-5Z plasmid, and DH10β strain were preserved by our laboratory.

[0022] 2 Tool Enzymes and Reagents

[0023] The restriction endonuclease, Klenow fragment enzyme, T4 DNA polymerase and dNTP used were purchased from TaKaRa Company. T4 DNA ligase and Taq DNA polymerase were purchased from Huamei Company, and proteinase K was purchased from Merck Company. M2, M16 and paraffin oil were purchased from Sigma Company. X-gal, Wizard DNA Clean-up System, and Prime-A-Gene Labeling System were purchased from Promega. QIAprep Spin Miniprep Kit was purchased from QIAGEN.

[0024] Construction of 3pGFAP-Cre transgenic vector

[0025] The genomic library of 129sv mice was screened by PCR method, and the 1.8Kb regulatory sequence of the 5' flanking region of 129sv mouse glial cell ac...

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Abstract

A process for preparing the transgenic mouse able to specifically express Cre recombinant enzyme in cental nerve system includes such steps as using the terminals' regulatory sequence of 1.8 kb GFAP gene to configure the transgenic carrier pGFAP-Cre-hGH containing two beta-glubolin insulators, GFAP5' terminal regulatory region, Cre recombinant enzyme gene and human growth hormone gene, injecting the 7.6 kbtransgenic fragment beta globin insulator-pGFAP-Cre-hGH into mouse's fertilizer eggs, transplating them in the deference duct of male mouse, and developing to obtain the target mouses.

Description

technical field [0001] The present invention relates to the 1.8kb 5' end regulatory region of glial cell acidic protein (glial fibrillary acidic protein, GFAP) gene cloned from 129sv mouse genome library by PCR screening method and using the regulatory sequence to prepare central A method for preparing transgenic mice specifically expressing Cre recombinase in the nervous system. Background technique [0002] The central nervous system, including the brain and spinal cord, is the high-level center of various functional activities of human beings and the material basis of human thought and consciousness activities. In 1989, the United States took the lead in launching a nationwide brain science plan focused on protecting the brain and preventing brain diseases, and named the last decade of this century the "Decade of the Brain". This move was immediately responded by the International Brain Research Organization and many international academic organizations. Therefore, the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63C12N15/12C12N15/52C12N15/89A01K67/027
Inventor 绳纪坡邓继先杨晓
Owner INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE
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