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Use of the Non-electrophoretic Method to Detect Testis Specific Protein Gene for Sexing in Preimplantation Bovine Embryos
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 Title & Authors
Use of the Non-electrophoretic Method to Detect Testis Specific Protein Gene for Sexing in Preimplantation Bovine Embryos
Huang, Jinming; You, Wei; Wu, Naike; Tan, Xiuwen;
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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.
Bovine Embryo;Sex Determination;TSPY Gene;PCR;
 Cited by
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Asian-Australasian Journal of Animal Sciences, 2007. vol.20. 11, pp.1689-1693 crossref(new window)
Aasen, E. and J. F. Medrano. 1990. Amplification of the ZFY and ZFX genes for sex identification in humans, cattle, sheep and goats. Biotechnol. 8(12):1279-1281. crossref(new window)

Affara, N., C. Bishop and W. Brown. 1996. Report of the second international workshop on Y chromosome mapping. Cytogenet Cell Genet.73:33-76. crossref(new window)

Bradbury, M. W., L. M. Isola and J. W. Gordon. 1990. Enzymatic amplification of a Y chromosome repeat in a single blastomere allows identification of the sex of preimplantation mouse embryos. Proc. Natl. Acad. Sci. 87:4053-4057.

Bredbacka, P., A. Kankaanpaa and J. Peippo. 1995. PCR-sexing of bovine embryos: A simplified protocol. Theriogenol. 44(2):167-176. crossref(new window)

Bredbacka, P. 1998. Recent developments in embryo sexing and its field application. Reprod. Nutr. Dev. 38(6):605-613. crossref(new window)

Chen,C. M., C. L. Hu, C. H. Wang, C. M. Hung, H. K. Wu, K. B. Choo and W. T. Cheng. 1999. Gender determination in single bovine blastomeres by polymerase chain reaction amplification of sex-specific polymorphic fragments in the amelogenin gene. Mol. Reprod. Dev. 54(3):209-214. crossref(new window)

Chen, C. Y., L. S. Huang, J. B. Chen, N. S. Ding, L. H. Zhou and H. X. Xiao. 2003. Establishing and optimizing the system for sex determination of bovine preimplantation embryos. J. Agric. Biotech. 11(4):399-402.

Chrenek, P. and J. Bulla. 2002. Simultaneous analysis of sex determination and K-casein genotypes from bovine preimplantation embryos. Czech J. Anim. Sci. 47(8):1-5.

Ennis, S. and T. F. Gallagher. 1994. A PCR-based sex-determination assay in cattle based on the bovine amelogenin locus. Anim. Genet. 25:425-427. crossref(new window)

Gong, R. C., Y. O. Xu and R. Tang. 1997. ZFY gene sequences in bovine by PCR.J of Southwest University for Nationalities (Natural Science Edition). 17(3):234-237.

Hasler, J. F., E. Cardey, J. E. Stokes and P. Bredbacka. 2002. Nonelectrophoretic PCR-sexing of bovine embryos in a commercial environment. Theriogenol. 58(8):1457-1469. crossref(new window)

Higuchi, R., G. Dollinger, P. S. Walsh and R. Griffith. 1992. Simultaneous amplification and detection of specific DNA sequences. Biotechnol. 10(4):413-417. crossref(new window)

Hirayama, H., S. Kageyama, S. Moriyasu, K. Sawai, S. Onoe, Y. Takahashi, S. Katagiri, K. Toen, K. Watanabe, T. Notomi, H. Yamashina, S .Matsuzaki and A. Minamihashi. 2004. Rapid sexing of bovine preimplantation embryos using loopmediated isothermal amplification. Theriogenol. 62(5):887-896. crossref(new window)

Hochman, D., Y. Zaron, I. Dekel, E. Feldmesser, J. F. Medrano, M. Shani and M. Ron. 1996. Multiple genotype analysis and sexing of IVF bovine embryos. Theriogenol. 46:1063-1075. crossref(new window)

Huang, J. M., J. Xiong, Y. B. Liu and W. You. 2005. Sex determination effect of bovine preimplantation embryo by LAMP method. Ecol. Dome. Anim. 26:20-22.

Jakubiczka, S., F. Schnieders and J. Schmidtke. 1993. A bovine homologue of the human TSPY gene. Genomics. 17:732-735. crossref(new window)

Kageyama, S., I. Yoshida, K. Kawakura and K. Chikuni. 2004. A novel repeated sequence located on the bovine Y chromosome: its application to rapid and precise embryo sexing by PCR. J. Vet. Med. Sci. 66(5):509-514. crossref(new window)

Levinson, G., R. A. Fields, G. L. Harton, F. T. Parmer, A. Maddalena, E. F. Fugger and J. D. Schulman. 1992. Reliable gender screening for human preimplantation embryos using multiple DNA target sequences. Human Reprod. 7:1304-1313.

Lindner, G. M. and R. W. Wright. 1983. Bovine embryo morphology and evaluation. Theriogenol. 20:407-416. crossref(new window)

Liu, Y. B. and J. M. Huang. 2004. Experiment of superovulation of Holstein heifer. J. China Herbivores. 24(01):14-16.

Machaty, Z., A. Paldi, T. Csaki, Z. Varga, I. Kiss, Z. Barandi and G. Vajta. 1992. Biopsy and sex determination by PCR of IVF bovine embryos. J. Reprod. Fertil. 98(2):467-470.

Manz, E., T. Vogel and P. Glatzel. 1998. Identification of an equine Y chromosome specific gene locus (eTSPY) with potential in preimplantation sex diagnosis.Theriogenol. 49:364. crossref(new window)

Park, J. H., J. H. Lee, K. M. Choi, S. Y. Joung, J. Y. Kim, G. M. Chung, D. I. Jin and K. S. Im. 2001. Rapid sexing of preimplantation bovine embryo using consecutive and multiplex polymerase chain reaction (PCR) with biopsied single blastomere. Theriogenol. 55(9):1843-1853. crossref(new window)

Peura, T., J. M. Hyttinen, M. Turunen and J. Janne. 1991. A reliable sex determination assay for bovine preimplantation embryos using the polymerase chain reaction. Theriogenol. 35(3):547-555. crossref(new window)

Pierce, K. E., J. E. Rice and J. A. Sanchez. 2000. Real-time PCR using molecular beacons for accurate detection of the Y chromosome in single human blastomere. Mol. Hum. Reprod. 6:1155-1164. crossref(new window)

Plucienniczak, A. S. J. and J. Jaworski. 1982. Nucleotide sequence of bovine 1.715 satellite DNA and its relation to other bovine satellite sequences. J. Mol. Biol. 158:293-304. crossref(new window)

Schempp, W., A. Binkele, J. Arnemann, B. Glaser, K. Ma, K. Taylor, R. Toder, J. Wolfe and A. C. Chandley. 1995. Comparative mapping of YRRM- and TSPY-related cosmids in man and hominoid apes. Chromosome Res. 3:227-234. crossref(new window)

Shea, B. F. 1999. Determining the sex of bovine embryos using polymerase chain reaction results: A six-year study. Theriogenol. 51:841-854. crossref(new window)

Thibier, M. and M. Nibart. 1995. The sexing of bovine embryos in the field. Theriogenol. 43:71-80. crossref(new window)

Utsumi, K., H. Takeda and M. Yamada. 1994. Sexing of bovine embryos by PCR using Sry primer. Theriogenol. 41:323(abstract). crossref(new window)

Virta, J., J. Markola and J. Peippo. 2002. Sex determination of bovine embryo blastomeres by fluorogenic probes. Theriogenol. 57:2229-2236. crossref(new window)

Vogel, T., F. Dechend and E. Manz. 1997. Organization and expression of bovine TSPY. Mamm. Genome. 8:491-496. crossref(new window)

Zeng, Y. T., M. L. Zhang and M. Y. Chen. 1992. Using the polymerase chain reaction (PCR) amplify the cow Y chromosome the sex determination area (SRY) sequence to sex cow embryo. Chinese Science Bulletin 37(5):97-98.