摘要
通过采取PCR-SSCP和限制性内切酶酶切的方法检测类固醇21-羟化酶(steroid 21-hydroxylase,CYP21)基因的10个外显子在高繁殖力绵羊品种(小尾寒羊)、中等繁殖力品种(湖羊)和低繁殖力品种(多赛特羊和特克塞尔羊)间的单核苷酸多态性,同时研究该基因对小尾寒羊高繁殖力的影响。结果显示,在所设计的10对引物中,仅引物P8扩增的片段存在多态性。对于P8扩增得到的片段,经HinPⅠ1酶切后,可在小尾寒羊、湖羊和多赛特羊中检测到AA和AB型,而在特克塞尔羊中只检测到AA型;经BcnⅠ酶切后,在小尾寒羊、湖羊、多赛特和特克塞尔羊中均检测到CC和CD型。测序结果显示AB型与AA型相比在2340位发生了A→G突变,但并未引起所编码的氨基酸发生改变;CD型与CC型相比在2376位发生了G→A突变,也未引起所编码氨基酸的变化。AB型小尾寒羊产羔数最小二乘均值比AA型的多0.96只(P<0.05),CC型和CD型小尾寒羊产羔数差异不显著(P>0.05)。研究结果初步表明CYP21基因的B等位基因是提高绵羊产羔数的一个候选DNA标记。
The steroid 21-hydroxylase (CYP21) gene was studied as a candidate gene for the high prolificacy in Small Tail Han sheep. According to the sequence of bovine CYP21 gene, ten pairs of primers were designed to detect single nucleotide polymorphisms of exons 1-10 of CYP21 gene in high prolificacy breed (Small Tail Han sheep), medium prolificacy breed (Hu sheep) and low prolificacy breeds (Dorset and Texel sheep) by PCR-SSCP and diges- tion with restriction endonucleases. Only the products amplified by primer P8 displayed polymorphisms. For prime- PS, two genotypes (AA and AB) were detected in Small Tail Han, Hu and Dorset sheep by digesting the PCR prod- ucts with HinP I 1, at the same time only one genotype (AA) was found in Texel sheep; as well, two genotypes (CC and CD) were detected in all the four sheep breeds by digesting the PCR products with Bcn I. Sequencing revealed one mutation (2340A--*G) of CYP21 gene in the genotype AB in comparison to the genotype AA; another mutation (2376G--*A) of CYP21 gene in the genotype CD in comparison to the genotype CC, and both of mutations did not cause any amino acid change. The Small Tail Han sheep with genotype AB had mean 0.96 (P〈0.05) lambs more than those with genotype AA; and there was no significant difference of the litter size between genotypes CC and CD (P〉0.05). These results preliminarily indicated that allele B of CYP21 gene is a potential DNA marker for improving the litter size of sheep.
出处
《安徽农业大学学报》
CAS
CSCD
北大核心
2013年第6期937-944,共8页
Journal of Anhui Agricultural University
基金
国家肉羊产业技术体系专项(CARS-39)
北京市自然科学基金项目(5042017)
中央级公益性科研院所基本科研业务费专项(2010jc-9)共同资助