扩展莫尼茨绦虫不同体节电压门控性钙通道β亚基和胶原纤维mRNA转录水平的研究

Expression of mRNA transcription of voltage gated calcium channel β subunits and collagen fibers in different segments of Moniezia expansa

为了研究电压门控性钙通道β亚基和胶原纤维基因在扩展莫尼茨绦虫不同体节的mRNA转录水平,以beta-Tubulin基因作为内参基因,采用SYBR Green real-time RT-PCR方法检测该基因在扩展莫尼茨绦虫4个不同发育阶段即头节、幼节、成节、孕节中的表达差异。结果显示,电压门控性钙通道β亚基、胶原纤维基因和beta-Tubulin基因的Ct值与阳性质粒的浓度均呈良好的线性关系,相关系数均大于0.99;熔解曲线分析表明,产物为特异的单峰,具有较高的特异性。试验结果表明,电压门控性钙通道β亚基以及胶原纤维基因在虫体各发育阶段中的表达丰度存在差异。电压门控性钙通道β亚基在幼节的表达水平较高,而胶原纤维基因在孕节的表达水平较高,提示这2个基因可能在幼节和孕节的发育中有重要作用。

In the present study,we developed a SYBR Green real-time fluorescent quantitative PCR assay to explore the mRNA transcription of voltage gated calcium channel fl subunits and collagen fibers in different segments of Moniezia expansa, with beta-Tubulin as an interna control. In result, a good linear relationship（〉0. 99） between the Ct value and the concentration of positive plasmid for each gene were found. The melting curve analysis showed the product was specific to a single peak, with high specificity. Real-time PCR results showed that the expression abundance of voltage gated calcium channel β subunits and collagen fibers were different, the voltage gated calcium channel β subunits was high in immature segment,but the collagen fibers was high in gravid segment. This phenomenon showed that the voltage gated calcium channel β subunits and collagen fibers may play a key role in the development of immature segment.