奶牛性别鉴定的研究与应用

Research and Application of Sex Identification in Dairy Cows

选取牛雄性性别决定基因SRY (sex region of Y chromosome),根据基因序列设计特异引物,应用PCR技术对5头荷斯坦奶牛DNA样品进行扩增,鉴定其性别;并对设计的引物灵敏度进行检测;对已有的公、母各20头荷斯坦奶牛DNA样品进行PCR盲检,获取奶牛高灵敏度特异性引物,用于奶牛性别鉴定。结果表明,4头公牛DNA样品可以扩增出目标条带(66 bp),1头母牛DNA样品无法扩增出条带,阴性对照扩增无条带;最佳引物灵敏度为1.6 pg/μL,可以很好地满足性别鉴定需要。40头个体中,20头个体DNA样品可以扩增出条带,其余20头个体DNA样品无法扩增出条带,检测结果与实际性别对比准确率为100%。试验结果表明,设计的引物灵敏度比较好,能够满足奶牛性别鉴定的需要。

To obtain high sensitivity specific primers of dairy cattle to determine the their sex, the experiment selected sex determining gene SRY(sex region of Y chromosome) of male cattle to design soecific primers according to gene sequence. PCR was used to amplify the DNA samples from 5 Holstein dairy cattle to determine their sex, and the sensitivity of designed primers was detected. Finally, DNA samples from existing 20 male and 20 female Holstein dairy cattle were taken to conduct PCR blind test. The results show 4 bulls’ DNA samples could be amplified the target band(66 bp), 1 cow’s DNA could not. The negative control had no target band after amplification. The best primer’s sensitivity was 1.6 pg/μL, which could better satisfy the need of sex identification. Results also show among 40 cattle, the DNA samples of 20 of which could be amplified target band, while the other 20 could not. The test results were 100% accurate compared with the reality. The primer designed in this experiment has better sensitivity, which can satisfy the demand of sex determination.