Asian-Australasian Journal of Animal Sciences

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
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Production of Transgenic Chimeric Chickens Using Blastodermal Cells

  • Yan, Haifeng (Lab for Molecular Biology Applied to Animal Production, Hunan Institute of Animal and Veterinary Science) ;
  • Lee, Chaeyoung (Ilsong Institute of Life Science, Hallym University) ;
  • Xiao, Bingnan (Lab for Molecular Biology Applied to Animal Production, Hunan Institute of Animal and Veterinary Science) ;
  • Trefil, Pavel (Research Institute of Biopharmacy and Veterinary Drugs) ;
  • Liu, Shixun (Lab for Molecular Biology Applied to Animal Production, Hunan Institute of Animal and Veterinary Science) ;
  • Kim, Younyoung (Ilsong Institute of Life Science, Hallym University) ;
  • Wu, Xiaolin (Lab for Molecular Biology Applied to Animal Production, Hunan Institute of Animal and Veterinary Science)
  • Received : 2004.04.09
  • Accepted : 2004.09.06
  • Published : 2005.02.01
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Abstract

A practical approach was proposed to produce transgenic chimeric chickens using blastodermal cells (BCs). The chicken BCs were mechanically dissociated and transferred into the recipient eggs that had been exposed to 500 rads irradiation of$^{60}Co$ and windowed on the equatorial plane. Chimeric chickens were generated using two models: the crosses (MXL) from Black Minors (ii,EE,b/b) ♂${\times}$Barred Leghorns (ii,ee,B/-) ♀ as donors and White Leghorns (WL, II) as acceptors (Model 1), or the Black Heifengs (BH, ii,EE,bb) as donors and Hua-xing white (HW, II) as recipients (Model 2). The treated eggs were incubated in their original shells in normal conditions until hatching. Green fluorescent protein (GFP) gene was transferred into the BCs derived from MXL and BH via lipofectamine and the pEGFP-C1, and transfection efficiency into the BCs was examined under a fluorescent microscope. Potential transgenic chimeras were selected based on the proposed methods in this study. Using the fresh BCs, the best rate of phenotypic chimeras was 6.7% and 26.0% in model-1 groups, and model-2 groups, respectively. We also described the optimized conditions for transfection. Although 30% of the BCs transfected in vitro emitted green light under an inverted fluorescent microscope, no embryos injected with the transfected BCs expressed foreign GFP gene at 3-4 days.

Keywords

References

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