牛冷冻试管胚胎性别鉴定的应用

Sexing of Frozen IVF Bovine Embryos by PCR during Embryo Transfer

利用双引物PCR技术对优质良种牛体外受精的冷冻胚胎进行性别鉴定 ,并对分割后的半胚分别进行PCR扩增和染色体分析 ,两种方法的判定结果相吻合。在本实验条件下 ,利用减少PCR体系中细胞的数量来确立最低的模板DNA的检出量 ,2 细胞不能检测 ,4 细胞检出率为 6 2 .5 % ,8 细胞为 90 % ,1 6 细胞以上为 1 0 0 %。对 2 3枚经过性别鉴定后的胚胎进行移植 ,获得 5头纯种试管牛 ,产犊率为 2 1 7% ,与正常的胚胎移植结果 2 7 3% ( 3 1 1 )无显著差异 ,预测性比为 1 0 0 %符合。

Sex of early embryos was determined by PCR using double primers (Y-specific DNA primer BOV-97,157 bp; Bovine α-lactalbumin primer in male and female,109 bp).The sex determination accuracy of biopsied half embryos by PCR with BOV97 primer was 100% in accordance with chromosome analysis. Under our conditions, detection rate of embryonic DNA were zero with 2 cells, 62.5% with 4 cells, 90% with 8 cells and 100% with 16 cells. A total of 23 embryos that had been biopsied and sexed were transferred into the donors, and 5 calves were born. The success rate is 21.7%, with no significant difference from the normal IVF-ET (27.3%, 3/11).