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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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