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Hyperlipemia transgenic miniature pig and preparation method thereof

A technology of hyperlipidemia and transgenic pigs, applied in the field of biomedicine, can solve unseen problems

Inactive Publication Date: 2011-06-08
JILIN UNIV
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Problems solved by technology

But there is no report of transgenic pig expressing human ApoCIII gene

Method used

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  • Hyperlipemia transgenic miniature pig and preparation method thereof
  • Hyperlipemia transgenic miniature pig and preparation method thereof
  • Hyperlipemia transgenic miniature pig and preparation method thereof

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Experimental program
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Embodiment 1

[0047] 1. Construction of pcDNA-CIII vector

[0048] Connect human apolipoprotein CIII to pcDNA3.1(+) and send it to Beijing Tiangen Company for sequencing. After identification by enzyme digestion and correct sequencing, it is named pcDNA-CIII. Extracted, linearized with NotI single enzyme digestion, and used for cell transfection. Schematic such as figure 1 .

[0049] 2. Cell transfection and screening

[0050] The porcine embryonic fibroblasts were transfected with DNA mediated by Roche's FU GENE HD transfection reagent, and the operation steps were carried out according to the instructions of FU GENE HD. The day before the transfection, the primary cultured embryonic fibroblasts were resuscitated into a 60mm plate with antibiotic-free culture medium, and the cells could be transfected when the cells reached 80-90% confluence. 24 hours after transfection, the cells were passaged by trypsinization 1:18. 48 hours after transfection, G418 selection medium (400 μg / ml conce...

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Abstract

The invention provides a transgenic miniature pig for preparing a human hyperlipemia disease animal model and a preparation method thereof. A human apolipoprotein CIII expression vector pcDNA-CIII is constructed and expressed by using a gene engineering method; a miniature pig fibroblast capable of expressing the human apolipoprotein CIII is screened out by using cell transfection and cell screening technologies; and the miniature pig is cloned by using a somatic cell nuclear transfer technology. The cloned miniature pig can be used for research into pathogenesis of human hyperlipemia related diseases, and screening and preclinical pretesting of medicine intervention and treatment of related diseases.

Description

Technical field: [0001] The invention provides a hyperlipidemia transgenic mini-pig, and also discloses its preparation method, which is used to study the pathogenesis of human hyperlipidemia-related diseases and the screening and preclinical pre-test of drug intervention and treatment of related diseases. The animal model is provided for establishing human disease hyperlipidemia, and belongs to the field of biomedical technology. Background technique: [0002] The main biochemical indicators and metabolic characteristics of pigs are more similar to those of humans, and their lipoprotein structure and composition, lipid metabolism mechanism are also similar to humans, and pigs can also develop spontaneous atherosclerosis with age. Therefore, the research on genetically modified pigs has won the favor of many scientists and investors, and the successful cases have also proved that it has unique advantages and practical value in life science research. Miniature pigs are small...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/85A01K67/027
Inventor 邓旭明魏静元欧阳红生刘国庆
Owner JILIN UNIV
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