两温度梯度多重PCR鉴别牛早期胚胎性别的技术研究

稳定、可靠和快速的牛胚胎性别鉴定方法在生产应用中具有重要意义。通过两温度梯度PCR方法对牛基因组、克隆胚胎、胚胎样品进行性别鉴别实验研究，建立了稳定、简便、快速的牛早期胚胎性别鉴别两温度梯度PCR方法，鉴定时间仅为57分钟。采用两温度梯度PCR方法对30枚奶牛胚胎进行了早期性别鉴别，并将鉴别的15枚胚胎（11枚为雌性，4枚为雄性）移植到同期处理的15头受体母牛体内。60天后妊娠检查，有7个受体成功受孕，5头受体怀孕晚期流产，流产犊牛全部为母犊。结果产下1公1母两头犊牛，流产个体与出生个体的性别与PCR鉴别结果完全相符。

The Study on the Technique of Sexing Bovine Pre-Implantation Embryos with Two-Temperature Gradient Multiplex PCR

For the stable,reliable,fast method of sexing bovine pre-implantation embryos is still play very important role in husbandry production,the amplification experiment under the condition of the two-temperature gradient PCR was done with bovine samples such as genome,cloning embryos,embryos respectively.As a result,the stable,simple,fast method of two-temperature gradient PCR for sexing bovine pre-implantation embryos was obtained,which only took 57 minutes to identify the embryo sex.A total of 30 dairy embryos were sexed with the two-temperature gradient PCR method in the study.15 identified embryos(11 female,4 male) were transferred into 15 recipient cows,among them 7 were pregnant after 60 days.In the end,5 female calves were aborted in late pregnancy,and 1 female and 1 male dairy calf were born.The sexes of aborted and born calves were fully in accordance with the embryo sex predetermination with PCR method.