番茄MIR398基因的克隆及其在烟草中的表达分析

Cloning of Tomato MIR398 Gene and Its Expression Analysis in Tobacco

miRNA作为真核生物的一类调控因子,在植物胁迫应答中发挥着重要的作用,成熟的miRNAs能降解或者阻遏靶mRNA的翻译。为从不同角度揭示miR398的生物学功能,我们从番茄中克隆到MIR398基因并构建了该基因的过表达载体pBI121-MIR398,通过农杆菌介导的叶盘法将重组质粒导入异源植物烟草中,利用psRNATarget预测miR398在烟草中的靶基因有PR5和CSD等。采用实时定量PCR检测基因表达,发现转基因烟草中MIR398的表达量增加,靶基因的表达量减少,为阐明miR398的异源调控功能奠定了基础。

As a kind of regulatory factor in eukaryote, miRNA plays an important role in the plant stress response.The mature miRNAs can degrade or repress the translation of the target mRNA. To explore the biological function of miR398, we cloned the MIR398 gene from tomato and constructed the over-expression vector pBI121-MIR398. Then we introduced this over-expression vector into tobacco and predicted the target genes of miR398 in tobacco, which were CSD, PR5 and so on. Then we detected the expression of MIR398 and its target genes with real-time PCR and found that the expression level of MIR398 was increased and its target was contrary,which provide a basis on elucidating the different source control of miR398.