Shi Lei, Yue Wenbin, Ren Youshe, Lei Fulin, Zhao Junxing
College of Animal Science and Technology, Shanxi Agricultural University, Taigu, PR China.
Anim Reprod Sci. 2008 May;105(3-4):398-403. doi: 10.1016/j.anireprosci.2007.11.004. Epub 2007 Nov 19.
The objective of this study was to obtain a fast, accurate and reliable method of determining the sex of goat embryos prior to implantation through amplification of the high-motility-group (HMG) box of the sex-determining region of the Y chromosome (SRY) gene of the goats. Goat specific primers were designed for duplex polymerase chain reaction (PCR). As an internal control gene, the goat beta-action gene sequence was simultaneously amplified together with the HMG box of goat SRY gene. Males showed both 1 SRY band and 1 beta-action band, but only 1 beta-action band was present in the agarose gel electrophoresis of females. The result indicated that the goat HMG-box sequence motif of SRY was male specific. Afterward, the optimized PCR procedure was applied in 30 embryo biopsies and the biopsied embryos were transferred into 30 recipient female goats. The sex of the 13 kids proved anatomically corresponded to the sex determined by PCR (100% accuracy). Thus, this study showed that this duplex PCR method can be applied to sex the goat pre-implantation embryos and to manipulate the sex ratio of offspring in goat breeding programs.
本研究的目的是通过扩增山羊Y染色体性别决定区域（SRY）基因的高迁移率族（HMG）盒，获得一种在植入前快速、准确且可靠地确定山羊胚胎性别的方法。设计了山羊特异性引物用于双重聚合酶链反应（PCR）。作为内对照基因，山羊β-肌动蛋白基因序列与山羊SRY基因的HMG盒同时进行扩增。雄性在琼脂糖凝胶电泳中显示出1条SRY带和1条β-肌动蛋白带，而雌性仅出现1条β-肌动蛋白带。结果表明，山羊SRY的HMG盒序列基序具有雄性特异性。随后，将优化后的PCR程序应用于30个胚胎活检样本，并将活检后的胚胎移植到30只受体母山羊体内。经解剖证实，13只羔羊的性别与PCR确定的性别相符（准确率100%）。因此，本研究表明，这种双重PCR方法可用于山羊植入前胚胎的性别鉴定，并可在山羊育种计划中控制后代的性别比例。
