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Use of the Non-electrophoretic Method to Detect Testis Specific Protein Gene for Sexing in Preimplantation Bovine Embryos

  • Huang, Jinming (Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences) ;
  • You, Wei (Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences) ;
  • Wu, Naike (Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences) ;
  • Tan, Xiuwen (Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences)
  • Received : 2006.12.13
  • Accepted : 2007.02.26
  • Published : 2007.06.01

Abstract

Testis-specific protein (TSPY) is a Y-specific gene, with up to 200 copy numbers in bulls. In order to make bovine embryo sexing under farm condition more feasible, the possibility of using a non-electrophoretic method to detect the TSPY gene for sexing bovine early embryos was examined. Primers were designed to amplify a portion of the TSPY gene and a common gene as an internal control primer. PCR optimization was carried out using a DNA template from bovine whole blood. Furthermore, embryo samples were diagnosed by this method and the sexing results were contrasted with those of the Loop-Mediated Isothermal Amplification (LAMP) method. The results showed that TSPY was as reliable a sexing method as LAMP. Forty-three morula and blastocyst embryos collected from superovulated donor dairy cattle were sexed by this method, and twenty-one embryos judged to be female embryos were transferred non-surgically to recipients 6 to 8 days after natural estrus. Out of 21 recipients, 9 were pregnant (42.86%) and all delivered female calves. The results showed that the sex predicted by this protocol was 100% accurate. In conclusion, the TSPY gene was a good male specific marker and indicated that a non-electrophoretic method was feasible and accurate to detect the TSPY gene for sexing preimplantation bovine embryos.

Keywords

References

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