扩展莫尼茨绦虫不同体节CREB和PICK基因mRNA转录水平研究

mRNA Transcription Levels of CREBand PICK Genes in Different Segments of Moniezia expansa

为研究CREB和PICK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。标准曲线分析显示,CREB、PICK基因和beta-Tubulin基因标准质粒实时定量扩增Ct值与标准质粒的质量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。同时结果还显示,CREB以及PICK基因在虫体各发育阶段中的表达丰度存在差异。CREB基因在扩展莫尼茨绦虫头颈节、幼节、成节和孕节的表达水平依次呈上升趋势,而PICK基因在扩展莫尼茨绦虫成节以及孕节的表达水平较高。

To explore the mRNA transcription levels of CREB and PICK in different segments of Moniezia expansa, this study developed a SYBR Green Real time fluorescent quantitative PCR assay for detection of Moniezia expansa mRNA with Moniezia expansa beta Tubulin as an internal control. The results showed a good linear relationships （R2〉0.99 ） between the Ct value and the concentration of positive plasmid for each gene. The melting curve analysis showed the product was a specific amplification. Real time PCR results showed that the expression abundance of CREB and PICK were different. The expression level of CREB was from high to low in gravid segment, mature segment, immature segment and scolex and the expression level of PICK was high in mature segment and gravid segment.