PCR扩增妊娠奶牛牙釉蛋白基因进行胚胎性别鉴定的研究

Study of Embryo Sex Identification by PCR Amplification for AML Gene in Pregnant Dairy Cows

牙釉蛋白(amelogenin,简写为AML)基因是牙齿发育过程中丰富表达的多拷贝基因,AML基因的同源基因分别定位在XY染色体上。本试验利用X-Y同源的牙釉蛋白基因序列设计一对特异性引物(牛AML基因序列的扩增片段长度:雌性为只有467bp的特异性扩增片段;雄性为同时具有341bp和467bp的两条特异性扩增片段),应用PCR技术同时扩增X和Y染色体上的特异性片段,扩增产物用PAGE电泳分离技术,经硝酸银溶液染色及扫描分析进行妊娠奶牛早期胚胎的性别鉴定。结果显示,从X染色体上扩增出467bp的片段,从Y染色体上扩增出341bp的特异性片段。由此可知,PCR扩增妊娠奶牛牙釉蛋白基因可以进行胚胎的性别鉴定。

Amelogenin （AML） was a multi-copy gene expressed in abundance in the process of tooth development, and its homologous genes were located in XY-chromosome respectively. In this experiment, a pair of specific primers was designed to amplify the bovine AML gene in X and Y chromosome by PCR （The amplified fragment length of AML gene for female was 467bp, and the male had two fragment of 341bp and 467bp）, PCR product was separated and analyzed by PAGE electrophoresis and silver nitrate staining for the sex identification of pregnant dairy early embryo. The results showed that a specific fragment of 467bp was amplified in X chromosome, while the 341bp specific fragment was in Y chromosome. It was concluded that AML gene by PCR amplification can be used for the early embryo sex identification of pregnant dairy cows.