ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2009, Vol. 40 ›› Issue (5): 633-638.doi:

• 遗传繁育 • Previous Articles     Next Articles

Production of ω-3 Fatty Acid Desaturase Gene (sFat-1) Transgenic Embryos by Somatic Cell Nuclear Transfer in Pig

FENG Chong1, 2, ZHOU Yan-rong3, LONG Chuan2, LIU Xiao2, CHEN Hong-xing3*, PAN Deng-ke2*, YANG Bo-hui4   

  1. 1. College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070,China; 2. The Key Laboratory of Farm Animal Genetic Resources and Utilization of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193,China; 3. Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071,China; 4. Institute of Animal Science and Veterinary Pharmaceutics,Chinese Academy of Agricultural Sciences, Lanzhou 730050,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-05-25 Published:2009-05-25

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Abstract: In this study, fetal fibroblast cells of Large White pig were transfected with sFat-1 gene from round-worm C. Briggsae by lipofectamine mediated transfection. After selected by 600 μg·mL-1 G418 for 10 days and analyzed by PCR, RT-PCR, 11 positive transgenic cell clones were collected. Reconstructed embryos were combined by porcine oocytes which were matured for 42 h and transgenic cells. After culture, the cleavage percentage of embryos (76.6%±4.1% vs. 81.6%±3.1%) and blastula (10%±1.97% vs. 9.7%±1.4%) have no significant difference between the transgenic cloning embryos and nontransgenic cloning embryos (P>0.05). In assistant activation, CHX was used and make a higher (20.6%±0.89% vs. 10%±1.97%, P<0.05) percentage of blastula than the electrical activation, but the cleavage percentage was insignificant between them (72.4%±4.96% vs. 76.6%±4.1%, P>0.05). As a result, fetal fibroblast sFat-1 transgenic cell line of Large White pig can be obtained by lipofectamine mediate transfection, and the cell line used as nuclear donors make no significant difference in the development of the transgenic cloning embryos and non-transgenic cloning embryos; Then using CHX in assistant activation can significantly raise the percentage of blastula.