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Sex identification of pigs using polymerase chain reaction amplification of the amelogenin gene

Published online by Cambridge University Press:  01 November 2008

Shoichiro Sembon ,
Shun-ichi Suzuki ,
Dai-ichiro Fuchimoto ,
Masaki Iwamoto ,
Tatsuo Kawarasaki  and
Akira Onishi
Shoichiro Sembon
Affiliation:
Transgenic Animal Research Center, National Institute of Agrobiological Sciences, Ikenodai 2, Tsukuba, Ibaraki 305–0901, Japan.
Shun-ichi Suzuki
Affiliation:
Transgenic Animal Research Center, National Institute of Agrobiological Sciences, Ikenodai 2, Tsukuba, Ibaraki 305–0901, Japan.
Dai-ichiro Fuchimoto
Affiliation:
Transgenic Animal Research Center, National Institute of Agrobiological Sciences, Ikenodai 2, Tsukuba, Ibaraki 305–0901, Japan.
Masaki Iwamoto
Affiliation:
Prime Tech Ltd, Nakamukaihara 635, Tsuchiura, Ibaraki 300–0841, Japan.
Tatsuo Kawarasaki
Affiliation:
Shizuoka Swine and Poultry Experiment Station, Nishikata 2780, Kikugawa, Shizuoka 439–0037, Japan.
Akira Onishi*
Affiliation:
Transgenic Animal Research Center, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305–0901, Japan. Transgenic Animal Research Center, National Institute of Agrobiological Sciences, Ikenodai 2, Tsukuba, Ibaraki 305–0901, Japan.
*
All correspondence to: Akira Onishi. Transgenic Animal Research Center, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305–0901, Japan. Tel: +81 298 38 8635. Fax: +81 298 38 8635. e-mail: onishi@affrc.go.jp.
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Summary

The amelogenin (AMEL) gene exists on both sex chromosomes of various mammalian species and the length and sequence of the noncoding regions differ between the two chromosome-specific alleles. Because both forms can be amplified using a single primer set, the use of AMEL in polymerase chain reaction (PCR)-based methods has facilitated sex identification in various mammalian species, including cattle, sheep and humans. In this study, we designed PCR primers to yield different-sized products from the AMEL genes on the X (AMELX) and Y (AMELY) chromosomes of pigs. PCR amplification of genomic DNA samples collected from various breeds of pigs (European breeds: Landrace, Large White, Duroc and Berkshire; Chinese breeds: Meishan and Jinhua and their crossbreeds) yielded the expected products. For all breeds, DNA from male pigs produced two bands (520 and 350 bp; AMELX and AMELY, respectively), whereas samples from female pigs generated only the 520 bp product. We then tested the use of PCR of AMEL for sex identification of in vitro-produced (IVP) porcine embryos sampled at 2 or 5 to 6 days after fertilization; germinal vesicle (GV)-stage oocytes and electroactivated embryos were used as controls. More than 88% of the GV-stage oocytes and electroactivated embryos yielded a single 520 bp single band and about 50% of the IVP embryos tested produced both bands. Our findings show that PCR analysis of the AMEL gene is reliable for sex identification of pigs and porcine embryos.

Keywords

AmelogeninEmbryoPCRPorcineSexing
Type
Research Article
Information
Zygote , Volume 16 , Issue 4 , November 2008 , pp. 327 - 332
Copyright
Copyright © Cambridge University Press 2008

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References

Bondioli, K.R. (1992). Embryo sexing: a review of current techniques and their potential for commercial application in livestock production. J. Anim. Sci. 70, 19–26.CrossRefGoogle Scholar
Chapman, V.M., Keitz, B.T., Disteche, C.M., Lau, E.C. & Snead, M.L. (1991). Linkage of amelogenin (AMEL) to the distal portion of the mouse X chromosome. Genomics 10, 23–8.CrossRefGoogle Scholar
Chen, C.M., Hu, C.L., Wang, C.H., Hung, C.M., Wu, H.K., Choo, K.B. & Cheng, W.T. (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, 209–14.3.0.CO;2-6>CrossRefGoogle ScholarPubMed
Ennis, S. & Gallagher, T.F. (1994). A PCR-based sex determination assay in cattle based on the bovine amelogenin locus. Anim. Genet. 25, 425–7.CrossRefGoogle ScholarPubMed
Fajfar-Whetstone, C.J., Rayburn, A.L., Schook, L.B. & Weeler, M.B. (1993). Sex determination of porcine preimplantation embryos via Y-chromosome specific DNA sequences. Anim. Biotechnol. 4, 183–93.CrossRefGoogle Scholar
Gibson, C.W., Golub, E.E., Abrams, W.R., Shen, G., Ding, W. & Rosenbloom, J. (1992). Bovine amelogenin message heterogeneity: alternative splicing and Y-chromosomal gene transcription. Biochemistry 31, 8384–8.CrossRefGoogle ScholarPubMed
Kohn, M., Knauer, F., Stoffella, A., Schroder, W. & Paabo, S. (1995). Conservation genetics of the European brown bear—a study using excremental PCR of nuclear and mitochondrial sequences. Mol. Ecol. 4, 95–103.CrossRefGoogle ScholarPubMed
Lyngstadaas, S.P., Risnes, S., Nordbo, H. & Flones, A.G. (1990). Amelogenin gene similarity in vertebrates: DNA sequences encoding amelogenin seem to be conserved during evolution. J. Comp. Physiol. B 160, 469–72.CrossRefGoogle ScholarPubMed
Peters, R.M. & Wells, K.D. (1993). Culture of pig embryos. J. Reprod. Fertil. 48 (Suppl), 61–73.Google Scholar
Pfeiffer, I. & Brenig, B. (2005). X- and Y-chromosome-specific variants of the amelogenin gene allow sex determination in sheep (Ovis aries) and European red deer (Cervus elaphus). BMC Genet. 6, 16.CrossRefGoogle Scholar
Pomp, D., Good, B.A., Geisert, R.D., Corbin, C.J. & Conley, A.J. (1995). Sex identification in mammals with polymerase chain reaction and its use to examine sex effects on diameter of day-10 or -11 pig embryos. J. Anim. Sci. 73, 1408–15.CrossRefGoogle ScholarPubMed
Salido, E.C., Yen, P.H., Koprivnikar, K., Yu, L.C. & Shapiro, L.J. (1992). The human enamel protein gene amelogenin is expressed from both the X and Y chromosomes. Am. J. Hum. Genet. 50, 303–16.Google ScholarPubMed
Sullivan, K.M, Mannucci, A., Kimpton, C.P. & Gill, P. (1993). A rapid and quantitative DNA sex test: fluorescence-based PCR analysis of X-Y homologous gene amelogenin. Biotechniques 15, 636–41.Google ScholarPubMed
Taberlet, P., Mattock, H., Dubois-Paganon, C. & Bouvet, J. (1993). Sexing free-ranging brown bears Ursus arctos using hairs found in the field. Mol. Ecol. 2, 399–403.CrossRefGoogle ScholarPubMed
Wasser, S.K., Houston, C.S., Koehler, G.M., Cadd, G.G. & Fain, S.R. (1997). Techniques for application of faecal DNA methods to field studies of Ursids. Mol. Ecol. 6, 1091–7.CrossRefGoogle ScholarPubMed
Watson, J.M., Spencer, J.A., Graves, J.A., Snead, M.L. & Lau, E.C. (1992). Autosomal localization of the amelogenin gene in monotremes and marsupials: implications for mammalian sex chromosome evolution. Genomics 14, 785–9.CrossRefGoogle ScholarPubMed
Yamamoto, K., Tsubota, T., Komatsu, T., Katayama, A., Murase, T., Kita, I. & Kudo, T. (2002). Sex identification of Japanese black bear, Ursus thibetanus japonicus, by PCR based on amelogenin gene. J. Vet. Med. Sci. 64, 505–8.CrossRefGoogle ScholarPubMed
Yoshioka, K., Suzuki, C., Tanaka, A., Anas, I.M. & Iwamura, S. (2002). Birth of piglets derived from porcine zygotes cultured in a chemically defined medium. Biol. Reprod. 66, 112–9.CrossRefGoogle Scholar