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Modified Bfat-1 gene and its biological verification in mice by hydrodynamic tail vein injection

Modified Bfat-1 gene and its biological verification in mice by hydrodynamic tail vein injection
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摘要 Omega-3 polyunsaturated fatty acids (w-3 PUFAs) are essential components required for normal cellular function and have been shown to have important therapeutic and nutritional benefits in humans. But humans or mammals cannot naturally produce w-3 PUFAs, due to the lack of the co-3 fatty acid desaturase gene (fat-1 gene). Previously, fat-1 gene has been cloned from Caenorhabditis elegans and transferred into mice, pigs and sheep, but not yet into beef cattle. We attempt to transfer it into beef cattle. The object of this paper is to edit the fat-1 gene from C. elegans to express more efficiently in beef cattle and verify its biological function in mice model. As a result, the fat-1 gene from C. elegans was modified by synonymous codon usage and named it Bfat-l. We have demonstrated that degree of codon bias of Brat-1 gene was in- creased in beef cattle. Moreover, Bfat-1 gene could be transiently expressed in mouse liver and muscle, the w-6/co-3 PUFAs ratio of 18 and 20 carbon was decreased significantly in liver (P〈0.05), and the ratio of 20 carbon decreased significantly in muscle 24 and 72 h after injection (P〈0.05). This confirms that the Bfat-1 gene modification was successful, and the protein encoded was able to catalyze the conversion of w-6 PUFAs to e0-3 PUFAs. Omega-3 polyunsaturated fatty acids (w-3 PUFAs) are essential components required for normal cellular function and have been shown to have important therapeutic and nutritional benefits in humans. But humans or mammals cannot naturally produce w-3 PUFAs, due to the lack of the co-3 fatty acid desaturase gene (fat-1 gene). Previously, fat-1 gene has been cloned from Caenorhabditis elegans and transferred into mice, pigs and sheep, but not yet into beef cattle. We attempt to transfer it into beef cattle. The object of this paper is to edit the fat-1 gene from C. elegans to express more efficiently in beef cattle and verify its biological function in mice model. As a result, the fat-1 gene from C. elegans was modified by synonymous codon usage and named it Bfat-l. We have demonstrated that degree of codon bias of Brat-1 gene was in- creased in beef cattle. Moreover, Bfat-1 gene could be transiently expressed in mouse liver and muscle, the w-6/co-3 PUFAs ratio of 18 and 20 carbon was decreased significantly in liver (P〈0.05), and the ratio of 20 carbon decreased significantly in muscle 24 and 72 h after injection (P〈0.05). This confirms that the Bfat-1 gene modification was successful, and the protein encoded was able to catalyze the conversion of w-6 PUFAs to e0-3 PUFAs.
出处 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2016年第6期1330-1337,共8页 农业科学学报(英文版)
基金 funded by the National Key Project for Production of Transgenic Breeding Plan, China (2013ZX08007002, 2014ZX08007-002) the Earmarked Fund for ModernAgro-Industry Technology Research System, China (CARS38) the Agricultural Science and Technology Innovation Program (ASTIP-IAS03) the Fundamental Research Budget Increment Project (2013ZL031) of Chinese Academy of Agricultural Sciences
关键词 beef cattle codon bias modified Brat-l hydrodynamic tail vein injection MICE beef cattle, codon bias, modified Brat-l, hydrodynamic tail vein injection, mice
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参考文献31

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