两温度梯度多重PCR鉴别牛早期胚胎性别的技术

The Technique of Sexing Bovine Pre-implantation Embryos with Two-temperature Gradient Multiplex PCR

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。

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 clone 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.