Pomp D, Good B A, Geisert R D, Corbin C J, Conley A J
Department of Animal Science, Oklahoma State University, Stillwater 74078, USA.
J Anim Sci. 1995 May;73(5):1408-15. doi: 10.2527/1995.7351408x.
The objectives of this study were to develop a rapid method for sex determination for several mammalian species using polymerase chain reaction (PCR) and to use this method to determine whether there is a significant developmental difference in spherical diameter between male and female d-10 or -11 porcine embryos. The PCR system was developed and verified using genomic DNA from pigs of known sex, then it was tested with genomic DNA from several other mammalian species. Sex is determined by amplification of two genes in a single reaction. The presence or absence of a region of the Sry (sex-determining region Y) gene determines sex, and amplification of the Zfy (male) or Zfx (female) genes acts as a positive control for PCR. Sex determination was successful for all animals tested, including pigs, cattle, sheep, goats, llamas, horses, humans, baboons, dogs, cats, rats, and mice. A total of 209 embryos were collected from 21 crossbred gilts on d 10 or 11 of gestation, and their diameters were measured. No significant difference in embryo diameter was detected between male and female embryos, indicating that sexual dimorphism in embryonic growth in pigs does not occur before the period of rapid embryo elongation. The present sexing technique using PCR is rapid (approximately 6 h from receipt of embryos to results), and it may be useful for examining the effects of sex on any trait of interest in early porcine embryos and embryos from several other mammals.
本研究的目的是开发一种利用聚合酶链反应(PCR)对几种哺乳动物进行性别鉴定的快速方法,并使用该方法确定10或11日龄猪胚胎的雄性和雌性之间在球形直径上是否存在显著的发育差异。PCR系统是利用已知性别的猪的基因组DNA开发并验证的,然后用其他几种哺乳动物的基因组DNA进行测试。通过在单一反应中扩增两个基因来确定性别。Sry(Y染色体性别决定区)基因一个区域的存在或缺失决定性别,Zfy(雄性)或Zfx(雌性)基因的扩增作为PCR的阳性对照。对所有测试动物,包括猪、牛、羊、山羊、美洲驼、马、人、狒狒、狗、猫、大鼠和小鼠,性别鉴定均成功。在妊娠第10或11天从21头杂种母猪收集了总共209个胚胎,并测量了它们的直径。未检测到雄性和雌性胚胎在直径上有显著差异,这表明猪胚胎生长中的性二态性在胚胎快速伸长之前不会出现。目前使用PCR的性别鉴定技术速度很快(从收到胚胎到得出结果约6小时),它可能有助于研究性别对早期猪胚胎以及其他几种哺乳动物胚胎中任何感兴趣性状的影响。