Sex Determination of Cattle Meat by Polymerase Chain Reaction Amplification of the DEAD Box Protein (DDX3X/DDX3Y) Gene

  • Gokulakrishnan, P. (Division of Livestock Products Technology, Indian Veterinary Research Institute) ;
  • Kumar, R.R. (Division of Livestock Products Technology, Indian Veterinary Research Institute) ;
  • Sharma, B.D. (Division of Livestock Products Technology, Indian Veterinary Research Institute) ;
  • Mendiratta, S.K. (Division of Livestock Products Technology, Indian Veterinary Research Institute) ;
  • Sharma, D. (Genome Mapping Laboratory, Central Avian Research Institute)
  • Received : 2012.01.01
  • Accepted : 2012.03.17
  • Published : 2012.05.01


Determination of sex origin of cattle meat by fast and reliable molecular methods is an important measure to ensure correct allocation of export refunds particularly in European countries and also female cattle (cow) slaughter is legally banned in India because of religious beliefs. Based on the DEAD box protein gene located on the X and Y chromosomes, 2 pair of primers were designed and the system of PCR was optimized. Upon PCR amplification, male tissue showed 2 bands, while female tissue resulted in only one band. The accuracy and specificity of the primers was assessed using DNA template extracted from cattle meat of known sex. The protocol was subjected to a blind test and showed 100% concordance, proving its accuracy and reliability.


  1. Aasen, E. and J. F. Medrano. 1990. Amplification of the ZFY and ZFX genes for sex identification in humans, cattle, sheep and goats. Nat. Biotechnol. 8:1279-1281.
  2. Abdelhaleem, M. 2005. RNA helicases: Regulators of differentiation. Clin. Biochem. 38:499-503.
  3. Appa Rao, K. B. C., V. Kesava Rao, B. N. Kowale and S. M. Totey. 1995. Sex specific identification of raw meat from cattle, buffalo, sheep and goat. Meat Sci. 39:123-126.
  4. Ballin, N. Z. and K. G. Madsen. 2007. Sex determination in beef by melting curve analysis of PCR amplicons from the amelogenin locus. Meat Sci. 77:384-388.
  5. Benz, J., H. Trachsel and U. Baumann. 1999. Crystal structure of the ATPase domain of translation initiation factor 4a from Saccharomyces cerevisiae - the prototype of the DEAD box protein family. Structure 7:671-679.
  6. 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. 54:209-214.<209::AID-MRD1>3.0.CO;2-6
  7. Draisci, R., L. Palleschi, E. Ferretti, L. Lucentini and P. Cammarata. 2000. Quantitation of anabolic hormones and their metabolites in serum and urine by liquid chromatography-tandem mass spectrometry. J. Chromatogr. A. 870:511-522.
  8. 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.
  9. Fontanesi, L., E. Scotti and V. Russo. 2008. Differences of the porcine amelogenin X and Y chromosome genes (AMELX and AMELY) and their application for sex determination in pigs. Mol. Reprod. Dev. 75:1662-1668.
  10. Fu, Q., M. Zhang, W. S. Qin, Y. Q. Lu, H. Y. Zheng, B. Meng, S. S. Lu and K. H. Lu. 2007. Cloning the swamp buffalo SRY gene for embryo sexing with multiplex nested PCR. Theriogenology 68:1211-1218.
  11. Hartwig, M., S. Hartmann and H. Steinhart. 1997. Physiological quantities of naturally occurring steroid hormones (androgens and progestogens), precursors and metabolites in beef of differing sexual origin. Eur. Food Res. Technol. 205:5-10.
  12. Kirkpatrick, B. W. and R. L. Monson. 1993. Sensitive sex determination assay applicable to bovine embryos derived from IVM and IVF. J. Reprod. Fertil. 98:335-340.
  13. Liu, W. S., A. Wang, Y. Yang, T. C. Chang, E. Landrito and H. Yasue. 2009. Molecular characterization of the DDX3Y gene and its homologs in cattle. Cytogenet. Genome Res. 126:318-328.
  14. Lockley, A. K., J. S. Bruce, S. J. Franklin and R. G. Bardsley. 1997. Simultaneous detection of a sex specific sequence and the Ryr1 point mutation in porcine genomic DNA. Meat Sci. 45:485-490.
  15. Lu, W., N. Rawlings, J. Zhao and H. Wang. 2007. Amplification and application of the HMG box of bovine SRY gene for sex determination. Anim. Reprod. Sci. 100:186-191.
  16. Matthews, M. E. and K. C. Reed. 1992. Sequences from a family of bovine Y chromosomal repeats. Genomics 13:1267-1273.
  17. Miller, J. R. and M. Koopman. 1990. Isolation and characterization of two male specific DNA fragments from the bovine gene. Anim. Genet. 21:77-82.
  18. Parati, K., G. Bongioni, R. Aleandri and A. Galli. 2006. Sex ratio determination in bovine semen: A new approach by quantitative real time PCR. Theriogenology 66:2202-2209.
  19. Rosner, A. and B. Rinkevich. 2007. The DDX3 subfamily of the DEAD box helicases: divergent roles as unveiled by studying different organisms and in vitro assays. Curr. Med. Chem. 14:2517-2525.
  20. Shi, L., W. Yue, Y. Ren, F. Lei and J. Zhao. 2008. Sex determination in goat by amplification of the HMG box using duplex PCR. Anim. Reprod. Sci. 105:398-403.
  21. Simontacchi, C., L. Marinelli, G. Gabai, G. Bono and R. Angeletti. 1999. Accuracy in naturally occurring anabolic steroid assays in cattle and first approach to quality control in Italy. Analyst 124:307-312.
  22. Tagliavini, J., A. Bolchi, P. G. Bracchi and S. Ottonello. 1993. Sex determination on samples of bovine meat by polymerase chain reaction. J. Food Sci. 58:237-238.
  23. Zeleny, R. and H. Schimmel. 2002. Sexing of beef- A survey of possible methods. Meat Sci. 60:69-75.
  24. Zeleny, R., A. Bernreuther, H. Schimmel and J. Pauwels. 2002. Evaluation of PCR based beef sexing methods. J. Agric. Food Chem. 50:4169-4175.
  25. Zinovieva, N., G. Palma, M. Muller and G. Brem. 1995. A rapid sex determination test for bovine blastomeres using allele-specific PCR primers and capillary PCR. Theriogenology 43:365.

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