应用牙釉质基因鉴定绒山羊早期胚胎的性别

Embryo sexing identified by amplification of Amelogenin gene in Nanjiang cashmere goats

采用酚-氯仿法和煮沸法从南疆绒山羊血液和早期胚胎中提取基因组DNA,分别以雄羊和雌羊的血液基因组DNA和早期胚胎基因组DNA(约20～30胚胎细胞)为模板,以AMEL引物进行PCR扩增和电泳分析.结果表明:绒山羊5ng量和10pg量血液基因组DNA经扩增后雌性只得到1条非特异性带,雄性得到1条非特异性带和1条特异性带;超微量血液基因组DNA样本(10pg)经巢式扩增和电泳分析能够鉴定绒山羊性别;32枚绒山羊胚胎鉴定结果,雄雌性别比17/15;移植胚胎产羔雄雌比15/14.采用牙釉质基因(AMEL),经巢式扩增和电泳分析能够鉴定绒山羊性别,并对胚胎无损伤;南疆绒山羊早期胚胎性别鉴定结果与胚胎移植后产羔性别结果对比,雌雄性别比率差异不显著(P>0.05).

Genomic DNA was extracted from blood and early embryo（20~30 embryo） of Nanjiang cashmere goats by using phenol-choroform and boiled method.The genomic DNA from male and female goats was employed as model for PCR amplification with AMEL premier.The results showed that 5 ng blood genomic DNA following amplify displayed a single non-specificity band in the female goat sample,and a non-specificity band and a specificity band in the male goat sample,and 10 pg blood genomic DNA following amplify displayed a single non-specificity band in the female goat sample and a non-specificity band and a specificity band in the male goat sample,dram blood genomic DNA（10 pg）can be used to evaluate embryo sexing by nested PCR and agarose gel electrophoresis.The sex ratio of the embryo was the 17/15（male/female）.The sex ratio of offspring was the 15/14（male/female）.It is suggested that there is no significantly deviated for the sex ratio of embryo using PCR and offspring（P0.05） by applying Amelogenin（AMEL） gene in the sex identification of embryos from superovulated Nanjiang cashmere goats.