Asian-Australasian Journal of Animal Sciences

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
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Sexing of Sheep Embryos Produced In vitro by Polymerase Chain Reaction and Sex-specific Polymorphism

  • Saravanan, T. (Molecular Virology Lab, Indian Veterinary Research Institute) ;
  • Nainar, A. Mahalinga (Transgenic lab, Department of Animal Biotechnology, Madras Veterinary College) ;
  • Kumanan, K. (Transgenic lab, Department of Animal Biotechnology, Madras Veterinary College) ;
  • Kumaresan, A. (Division of Animal Reproduction, Indian Veterinary Research Institute)
  • Received : 2002.05.27
  • Accepted : 2002.11.04
  • Published : 2003.05.01
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Abstract

The accuracy of Polymerase chain reaction (PCR) assay in sexing of sheep embryos was assessed in this study. A total of 174 ovine embryos produced in vitro at different stages of development (2, 4-8 cell stages, morula and blastocyst) were sexed. The universal primers (P1-5EZ and P2-3EZ) used in this assay amplified ZFY/ZFX-specific sequences and yielded a 445 bp fragment in both sexes. Restriction enzyme analysis of ZFY/ZFX-amplified fragments with Sac I exhibited polymorphism between sexes, three and two fragments in males and in females, respectively. For verification of accuracy, blood samples of known sex were utilized as positive controls in each test. The mean percentages of sex identification by this method at 2 cell, 4-8 cell, morula and blastocyst were $73.00{\pm}5.72$, $89.77{\pm}3.79$, $3.33{\pm}8.08$ and $79.6{\pm}9.09$, espectively with the over all male to female ratio of 1:0.87. It is concluded that the ZFY/ZFX based method is highly reliable for the sexing of sheep embryos.

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References

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