中国草食动物科学2024，Vol.44Issue(3)：33-38,6.DOI:10.3969/j.issn.2095-3887.2024.00.035
绵羊精液中X、Y精子比率快速评价方法的建立
Establishment of Rapid Method for Evaluating the Ratio of X and Y Spermatozoa in Sheep Semen
摘要
Abstract
The aim of this experiment was to establish a TaqMan fluorescence PCR-based method for rapid calculation of X and Y sperm ratio in sheep,and to provide technical support for the development and application of X and Y sperm separation technology in sheep.In this study,specific primers and probes were designed for the X and Y chromosome-specific genes F9 and SRY,respectively,and sheep genomic DNA was used as the template for fragment amplification,plasmid ligation,and standard curve was constructed by mixing at different ratios after amplification and purification,and the fitting equations were calculated.The accuracy,stability and reliability of the method were tested by measuring commercial X-sex-controlled semen(n=3),Y-sex-controlled semen(n=3)and fresh semen(n=2).The results showed that the ratios of X spermatozoa in commercial X-sex-controlled semen were(89.46±3.37)%,(89.86±2.85)%,(90.02±2.19)%,respectively;the ratios of Y spermatozoa in commercial Y-sex-controlled semen were(89.02±2.68)%,(88.21±2.63)%,(88.72±3.93)%;and there was no significant difference between the results of the assay and the ratios of X and Y spermatozoa as labelled in the quality control report(P<0.05).The assay results of fresh sperm showed that the X and Y sperm ratios were(49.86±2.90)%and(49.30±2.10)%,respectively,which were not signifi-cantly different from the theoretical value of 50%(P<0.05).In summary,the method established in this study based on TaqMan probe fluorescence quantitative PCR for rapid calculation of X and Y sperm ratio in sheep is fast,the volume of semen required for the test is small,and the accuracy,stability and reliability are high,which can provide technical support for the detection of X and Y sperm separation in sheep.关键词
绵羊/性控精液/X、Y精子/TaqMan探针/荧光定量PCRKey words
sheep/sexually controlled semen/X and Y spermatozoa/TaqMan probe/fluorescence quantitative PCR分类
农业科技引用本文复制引用
古丽孜巴·艾力,李伟建,艾力亚木·土尔衮,张慧,王旭光..绵羊精液中X、Y精子比率快速评价方法的建立[J].中国草食动物科学,2024,44(3):33-38,6.基金项目
新疆维吾尔自治区自然科学基金项目(2020D01A56) （2020D01A56）
