Journal of Integrative Agriculture ›› 2016, Vol. 15 ›› Issue (06): 1330-1337.DOI: 10.1016/S2095-3119(15)61218-7

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

  

  • 收稿日期:2015-03-23 出版日期:2016-06-01 发布日期:2016-06-06

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

GAO Xue1*, DU Xin-hua1, 2*, ZHANG Lu-pei1, CHEN Yan1, GAO Hui-jiang1, XU Shang-zhong1, LI Jun-ya1   

  1. 1 Key Laboratory for Farm Animal Genetic Resources and Utilization, Ministry of Agriculture/Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China
    2 State Key Laboratory of Agro-Biotechnology, China Agricultural University, Beijing 100193, P.R.China
  • Received:2015-03-23 Online:2016-06-01 Published:2016-06-06
  • Contact: XU Shang-zhong, E-mail: xu1950@iascaas.net.cn; Li Jun-ya, E-mail: Jl1@iascaas.net.cn
  • About author:GAO Xue, E-mail: gaoxue76@126.com * These authors contributed equally to this study.
  • Supported by:

    This work was funded by the National Key Project for Production of Transgenic Breeding Plan, China (2013ZX08007-002, 2014ZX08007-002), the Earmarked Fund for Modern Agro-Industry Technology Research System, China (CARS-38), and the Agricultural Science and Technology Innovation Program (ASTIP-IAS03) and the Fundamental Research Budget Increment Project (2013ZL031) of Chinese Academy of Agricultural Sciences.

Abstract:   Omega-3 polyunsaturated fatty acids (ω-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 ω-3 PUFAs, due to the lack of the ω-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-1. We have demonstrated that degree of codon bias of Bfat-1 gene was increased in beef cattle. Moreover, Bfat-1 gene could be transiently expressed in mouse liver and muscle, the ω-6/ω-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 ω-6 PUFAs to ω-3 PUFAs.

Key words: beef cattle , codon bias ,  modified Bfat-1 ,  hydrodynamic tail vein injection ,  mice