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MicroRNAs在植物发育与逆境响应等多个方面具有重要的调节作用。本研究采用生物信息学方法分析gmamiR1514b的靶基因和启动子中的顺式作用元件。PLACE和Plant CARE启动子在线预测工具分析表明gma-miR1514b启动子序列具有TATA-box、CAAT-box基本的顺式作用元件和一些参与光、逆境和植物激素应答相关的顺式作用元件。PMRD软件预测获得5个gma-miR1514b靶基因,分别与拟南芥CA2、NTL9和ANAC014基因同源。通过重叠PCR方法替换拟南芥At-MIR319的成熟链和互补链序列,构建含有amiR1514b前体的植物表达载体amiR1514b-p CAMBIA3301。
MicroRNAs play an important regulatory role in many aspects of plant development and stress response. In this study, bioinformatics methods were used to analyze the target genes of gmamiR1514b and cis-acting elements in the promoter. Online prediction tools analysis of PLACE and Plant CARE promoters showed that the gma-miR1514b promoter sequence has the TATA-box, the basic cis-acting element of CAAT-box and some cis-acting elements involved in light, stress and plant hormone response. PMRD software predicted five gma-miR1514b target genes, respectively, and Arabidopsis CA2, NTL9 and ANAC014 gene homology. The mature and the complementary strand sequences of Arabidopsis At-MIR319 were replaced by overlapping PCR methods to construct the plant expression vector amiR1514b-p CAMBIA3301 containing the amiR1514b precursor.