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Establishment of Mongolian Sheep Fetal Fibroblast Cell Line Transfected Fat-1 Gene

Author: XueHongYan
Tutor: ZhouHuanMin
School: Inner Mongolia Agricultural University
Course: Animal Genetic Breeding and Reproduction
Keywords: ω-3 polyunsaturated fatty acids Fat-1 pTEK carrier Expression
CLC: S826
Type: Master's thesis
Year: 2011
Downloads: 44
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Abstract


ω-3 multi unsaturated fatty acids, it is essential to maintain normal growth and development of the organism , however of ω-3PUFAs the body content of a serious shortage of , ω - 6PUFAs the content is very high . As the ω-6 and ω-3 fatty acids in the metabolism and function of different, and they have many opposing physiological functions , and thus the balance of the two organism's own stability and normal development is very important. In this study, starting from the livestock breeding angle control ω-6 polyunsaturated fatty acids into the ω-3 polyunsaturated fatty acids , gene Fat - 1 Import sheep somatic cells using somatic cell cloning technology to create a new type of sheep of ω-3PUFAs - rich , nurturing switch to new varieties of Fat - 1 gene high-quality sheep . Experiments established male Mongolian sheep fetal fibroblast cell line , Caenorhabditis elegans total RNA as template synthesis of the first strand cDNA reference design good primers cFat-1 gene , purified connection built to pTEK carrier pTFK-c reorganization the plasmid, transformed DH - 5α competent cells after extraction and sequencing of the recombinant plasmid . And then check the correct cFat-1 gene codon optimization oFat-1, oFat-1 is connected to the carrier pTEK recombinant plasmid construct pTEK-o method of liposome-mediated transfection Mongolian sheep fetal fibroblasts Mongolian sheep fetal fibroblasts obtained by antibiotic selection of stable integration of Fat-1 gene . PCR, RT-PCR identification results prove Fat-1 gene has been integrated into the cellular genome . As the initial results , the the transgenic positive cells obtained in this experiment can be used as a donor somatic cell nuclear transplantation , the integration of Fat-1 gene cloned embryos , has laid a solid foundation for exploring by means of genetically modified to improve lamb quality .

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Livestock > Sheep
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