基于前期分型基础,为了更进一步提高分型的效率和准确性,本试验建立了Taqman探针和SNaPshot两种高通量FecB突变分型方法。结果显示,与传统的PCR-RFLP和PCR-SSCP分型方法相比,这两种高通量分型方法在节省检测成本的同时,大大缩短了检测周期,并在检测效率上也有了大幅度的提高,对利用FecB进行分子育种具有潜在的应用价值。自2003年至今,本实验室应用这两种方法对10个中国本地绵羊品种、3个培育品种以及一些杂交群体的23 264只绵羊的FecB频率进行了检测,结果显示,湖羊和小尾寒羊携带FecB基因的频率最高,小尾寒羊FecB基因的B等位基因频率为0.432-0.833。跟踪以小尾寒羊作为母本和杜泊羊级进杂交后代群体发现,随着杂交的进行FecB基因在培育的鲁西黑头肉羊中的B等位基因频率提升到了0.432,但在横交固定阶段有所下降。本研究所得结果可为今后在肉用绵羊品种多羔改良和多羔肉用新品种培育过程中通过引入FecB基因提高繁殖性能提供参考数据。
Based on the previous studies,the aim of this study was to further improve the efficiency and accuracy of genotyping.Two high-throughput genotyping methods(Taqman and SNaPshot)for FecBdetection in Chinese sheep breeds had been established in the present study.The results showed that the two methods not only reduced the cost but also saved the time and increased efficiency dramatically.The two methods had potential application value in large-scale molecular breeding introducing FecB.The detection results of 23 264 sheep including 10 Chineseindigenous sheep breeds,3crossbred breeds and some hybridized sheep populations collected since 2003 showed that the allele B frequencies in Small Tail Han and Hu sheep were much higher than other breeds.The allele B frequency in Small Tail Han sheep was 0.432-0.833.The allele B frequency in Luxi Blackhead sheep had been improved(up to 0.432)during upgrading and selection stage,while decreased during intercross stage.These results can provide reference data for improving sheep fecundity in sheep breeding program by introducing FecBgene.