Phua Alice Choon Yen, Abdullah Ramli Bin, Mohamed Zulqarnain
Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.
J Reprod Dev. 2003 Aug;49(4):307-11. doi: 10.1262/jrd.49.307.
Sex determination of livestock is performed to achieve the objectives of livestock breeding programmes. Techniques for sex determination have evolved from karyotyping to detecting Y-specific antigens and recently to the polymerase chain reaction (PCR), which appears to be the most sensitive, accurate, rapid and reliable method to date. In this study, a PCR-based sex determination method for potential application in goat breeding programmes was developed. Primers were designed to amplify a portion of the X amelogenin gene (Aml-X) on the X chromosome to give a 300 bp product and Sry gene on the Y chromosome to give a 116 bp product. PCR optimization was performed using DNA template extracted from a whole blood sample of Jermasia goats (German Fawn x Katjang) of both sexes. It was possible to identify the sex chromosomes by amplifying both male- and female-specific genes simultaneously in a duplex reaction with males yielding two bands and females yielding one band. The Aml-X primer set, which served as an internal control primer, did not interfere with amplification of the Y-specific sequence even when a low amount of DNA (1 ng) was used. The duplex reaction subjected to a blind test showed 100% (14/14) concordance, proving its accuracy and reliability. The primer sets used were found to be highly specific and were suitable for gender selection of goats.
进行家畜性别鉴定是为了实现家畜育种计划的目标。性别鉴定技术已从染色体组型分析发展到检测Y特异性抗原，最近又发展到聚合酶链反应（PCR），这似乎是迄今为止最灵敏、准确、快速且可靠的方法。在本研究中，开发了一种基于PCR的性别鉴定方法，用于山羊育种计划的潜在应用。设计引物以扩增X染色体上的X牙釉蛋白基因（Aml-X）的一部分，得到300 bp的产物，以及Y染色体上的Sry基因，得到l l6 bp的产物。使用从两性的杰尔马西亚山羊（德国小鹿×卡jang）全血样本中提取的DNA模板进行PCR优化。通过在双重反应中同时扩增雄性和雌性特异性基因来鉴定性染色体是可行的，雄性产生两条带，雌性产生一条带。作为内部对照引物的Aml-X引物组即使在使用少量DNA（1 ng）时也不会干扰Y特异性序列的扩增。经过盲测的双重反应显示一致性为100%（14/14），证明了其准确性和可靠性。所使用的引物组具有高度特异性，适用于山羊的性别选择。
