为了研究致密化相关基因在水牛体外培养早期胚胎中的mRNA表达情况,采用Taqman探针法和SYBR GreenⅠ染料法分别分析了缝隙连结蛋白43和31(Cx43、Cx31)、上皮钙调素蛋白(E-cad)3个基因在水牛体外成熟卵母细胞及培养的早期胚胎中的mRNA表达。结果发现,Cx43基因在水牛成熟卵母细胞中表达量最高,显著高于后3个发育阶段(P〈0.05);Cx31基因mRNA表达趋势与Cx43相反,成熟卵母细胞中表达量最低,囊胚期最高,随体外胚胎发育Cx31mRNA表达量逐渐增高;E-cad基因在体外培养的各阶段胚胎中mRNA表达无显著差异(P〉0.05)。以上结果表明,3个致密化相关基因在水牛体外培养早期胚胎中均有表达,但是具有明显不同的mRNA表达方式。
In order to investigate the mRNA expression of compact-related genes in buffalo in vitro matured and fertilized embryos,the Taqman probe and SYBR-Green Ⅰ quantitative real-time PCR(Q-PCR)method was used to analyze mRNA expression of Cx43,Cx31 and E-cad genes in pooled baffalo in vitro cultured embryos.Cx43 gene showed significantly higher expression in matured oocytes than in other stage embryos(P0.05),reverse to Cx43 gene,Cx31 gene had the lowest expression in matured oocytes compared with other three stages,and gradually increased its expression along with the development of embryos.The level of E-cad mRNA expression did not vary among the tested embryos(P0.05).The present study demonstrated that the three compact-related genes had expression in all stages of baffalo IVF embryos,but showed definitely different mRNA expression style.