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Polyunsaturated fatty acids (PUFAs) is an important component of the cell membrane, plays an important function in the regulation of the metabolism of the body's hormones, and a variety of activity, cause the body once the lack of physiological disorder caused by a variety of diseases, including most biological function is the ω-3PUFAs and ω-6 PUFAs of. The study found that the ω-6 PUFAs little significance to the human body, its excessive intake it will harm human health. Ω-3 PUFAs on the human body has a positive significance, has been confirmed in the prevention and treatment of cardiovascular disease, arthritis, cancer and other diseases play an important role. Mammals and humans can not be in the in vivo synthesis of ω-3 PUFAs and ω-6PUFAs from food intake to meet their needs. Nature omega-6 PUFAs in content-rich ω-3 PUFAs content is extremely low, leading to excessive intake of ω-6 PUFAs and ω-3 PUFAs intake is a serious shortage of body so that the body ω-6/ω-3 ratio of up to 16 or more. According to the research, ω-6 and ω-3 PUFAs proportion should be in a suitable body usually in the state of inadequate supply of ω-3 PUFAs. Mammals maintain a certain amount of ω-3 PUFAs can play to prevent cardiovascular disease, neurodegenerative diseases and even cancer role, ω-6 with ω-3PUFA balance the body's normal growth and a stable environment within an important role, how to increase the intake of ω-3 PUFAs body became rational use of key fatty acids. ω-3 fatty acid desaturase gene is critical gene synthesized by the ω-3 PUFAs, which uses the ω-6 PUFAs as substrates, generated at the third carbon of the methyl end of the fatty acid carbon chain catalytic one unsaturated bond, and thus the synthesis of the corresponding ω-3 PUFAs. Fat-1 gene exist widely in fungi, plants, and lower animals, it can produce a ω-3 fatty acid desaturase from the ω-6 polyunsaturated fatty acid is converted to ω-3 form and function. Researchers have genetically modified animals, such as: mouse, pig, expressed Fat-1 gene, these studies provide efficient and accurate medical nutrition animal models for exploring the ω-3 PUFAs function. In this study, using technical means such as molecular biology research nematodes omega-3 fatty acid desaturase gene. According to that reported in GenBank nematode Caenorhabditis Briggsae ω-3 fatty acid dehydrogenase gene sequences sFat-1, to optimize its codon introduced upstream of the gene sFat-1 enhancer SP163 sequence, named as SP163-sFat-1 . Synthetic SP163-sFat-1, and then proceed to the cloning, sequencing, results in the correct sequence. Using molecular biology techniques, SP163-sFat-1 gene was constructed in the eukaryotic expression vector pcDNA3.1 downstream of the CMV promoter, the sFat-1 gene eukaryotic expression vector pCDNA3.1 () the-sFat-SP163. Liposome-mediated method, pCDNA3.1 ()-sFat-SP163 transfected CHO cells, while pCDNA3.1 () as a control. G418 transfected CHO cells were used for resistance screening, stable hybrid cell lines, the positive rate of 90%. PCDNA3.1 ()-sFat-SP163 transfected cell lines named sFat-1-CHO, pcDNA3.1 () named pcDNA3.1-CHO transfected cells. Detection by fluorescence quantitative RT-PCR method and Western-blot method, sFat-1-CHO pCDNA3.1-CHO, results of target gene transcription, the pCDNA3.1-CHO high 10 to 5; simultaneously purpose gene expression, sFat-1-CHO successfully expressed sFat-1 protein, using gas chromatography, for fatty acid analysis on the sFat-1-CHO pCDNA3.1-CHO results sFat-1-CHO in the ω-3 PUFAs The content ratio pCDNA3.1-CHO high, and of ω-6/ω-3 the proportion decline that sFat-1-CHO recombinant protein with biological activity. The study further the use of transgenic technology to promote animal test data from the synthesis of polyunsaturated fatty acids in health food.
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