生长分化因子9(GDF9)是TGF-β超家族的一个成员,在早期卵泡的生长和分化过程中起着重要的调节作用。试验采用RT-PCR和实时荧光定量PCR对湖羊GDF9的组织表达、发情期单羔和三羔湖羊卵巢组织mRNA表达水平进行了研究;用PCR-RFLP法对湖羊群体GDF9外显子1第152位点的多态性进行了分析。组织表达谱分析发现GDF9基因在湖羊的下丘脑、垂体、卵巢、子宫、输卵管、心、肝、脾、肺、肾等组织中均有表达;实时荧光定量PCR结果表明,三羔湖羊卵巢组织中GDF9基因mRNA表达水平显著高于单羔湖羊(P〈0.05);PCR-RFLP法没有检测到湖羊GDF9基因cDNA第152处发生的单碱基突变(A→G)。研究提示GDF9基因mRNA表达水平与湖羊的繁殖性状有一定的关系,在湖羊群体中的多态性还有待于进一步研究。
Growth differentiation factor 9 (GDF9) is one of the members of transforming growth factor β (TGF-β), which is essential for growth and differentiation of early ovarian follicles. The tissue expression profile and the mRNA expression levels in the ovary between single fetuses and triplets Hu sheep during oestrum were analysed using RT-PCR and real time PCR; The polymorphism at 152 on Exon1 of GDF9 gene was identified using PCR-RFLP. The tissue expression results indicated that GDF9 gene expresses in hypothalamus, pituitary, ovary, uterus, fallopian tube, heart, liver, spleen, lung and kidney of Hu sheep. Real time PCR results indicated that GDF9 mRNA expression levels in the ovary of triplets Hu sheep is prominent higher than single fetuses Hu sheep (P0.05); The single nucleotide mutation: A→G at GDF9 cDNA 152 in Hu sheep was not found using PCR-RFLP. The result indicated that GDF9 mRNA expression level have some relation with the reproductive traits in Hu sheep, and wait for the further analyses of the polymorphism of GDF9 gene in Hu sheep.