title: Sex identification of pigs using polymerase chain reaction amplification of the amelogenin gene.
abstract: 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.
authors.affiliation: Transgenic Animal Research Center, National Institute of Agrobiological Sciences, Ikenodai 2, Tsukuba, Ibaraki 305-0901, Japan.
authors.forename: Shoichiro
authors.initials: S
authors.lastname: Sembon
authors.forename: Shun-ichi
authors.initials: S
authors.lastname: Suzuki
authors.forename: Dai-ichiro
authors.initials: D
authors.lastname: Fuchimoto
authors.forename: Masaki
authors.initials: M
authors.lastname: Iwamoto
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Kawarasaki
authors.forename: Akira
authors.initials: A
authors.lastname: Onishi
mesh_terms: D053523:Amelogenin / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D017931:DNA Primers; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology; D005260:Female; D005306:Fertilization; D005784:Gene Amplification; D008297:Male; D008969:Molecular Sequence Data; D009865:Oocytes / Q000166:cytology / Q000502:physiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012732:Sex Determination Analysis; D013045:Species Specificity; D013552:Swine; D014960:X Chromosome; D014998:Y Chromosome
chemical_list: D053523:Amelogenin; D017931:DNA Primers; D004247:DNA