草鱼MYH mRNA表达量分析中采用的内参基因稳定性比较

本研究分别以β-actin、18S rRNA和GAPDH为内参基因,采用实时荧光定量PCR对草鱼早期发育时期肌球蛋白重链(myosin heavy light,MYH)基因的mRNA表达量进行分析,并比较不同内参基因对MYH基因mRNA表达水平检测结果的准确性.研究结果表明,以β-actin和GAPDH作为内参,MYH基因mRNA表达水平完全一致,其表达量从原肠到仔鱼阶段逐次递增,仔鱼与原肠期阶段相比表达量差异显著;当采用18S rRNA作为内参时,MYH基因mRNA在发育阶段的表达量呈不稳定状态.因此,β-actin和GAPDH均可作为内参基因,用于草鱼早期发育中MYH基因mRNA的相对定量研究:而18S rRNA作为内参时,可能会对检测结果造成偏差.本研究不仅准确的揭示了草鱼MYH基因mRNA的表达特征,并且为荧光定量PCR技术在鱼类基因表达研究方面提供了有价值的参考.

Comparative Research on the Stability of Internal Control Genes Based on mRNA Expression Analysis of MYH Gene in the Grass carp,Ctenopharyngodon idellus

In this study,β-actin,18S rRNA and GAPDH were used as internal control genes to study the MYH mRNA expression at early development stages of the grass carp Ctenopharyngodon ideUus by using real-time PCR technique,and to compare the accuracy of testing results for MYH mRNA expression levels.The results showed that when both β-actin and GA PDH were used as internal control the MYH mRNA expression was complete consistent,and its expression was increased from the gastrula stage to larval stage,which represented the statistic significant difference at the level of 0.05 possibility.However,the level of mRNA expression during the developmental stages was instable when 18S rRNA was used as internal control gene.Therefore,The evidences support a conclusion that both β-actin and GAPDH were suitable internal control genes for mRNA relative quantification study of MYH gene during the developmental stages.In contrary,The deviation was appeared in testing results when 18S rRNA was used as internal control genes.This research not only accurately revealed the expression characteristics of MYH mRNA in the grass carp,but also would provide valuable references for the gene expression analysis in grass carp using real-time PCR technique.