旨在研究 Bna-miR1140的表达模式及其调控机理，依据本实验室前期的油菜 miRNA 芯片实验结果，从油菜栽培种Westar 中克隆了 Bna-miR1140前体序列上游1.5 kb 的片段，并进行了顺式作用元件分析，然后构建了 GUS 报告基因植物表达载体，通过农杆菌介导法将 miR1140 pro∷GUS 转化到甘蓝型油菜 Westar 品种中，经 PCR 法鉴定获得5株阳性株。对阳性株油菜 T1代各组织的 GUS 化学染色分析表明，miR1140前体序列上游1.5 kb 区域具有启动子的功能，能够驱动 GUS 在油菜中表达，并且 GUS基因仅在叶柄及叶腋中特异性表达，说明油菜 miR1140Pro 为特异性启动子。%In order to explore the expression pattern and regulation mechanism of Bna-miR1140,the upstream 1.5 kb fragment from Bna-miR1140 precursor was cloned in a cultivated variety Westar of rape by PCR approach on the basis of rapeseed miRNAs chip results in our laboratory,and their cis-acting elements were analyzed. Then plant expression vector of GUS reporter genes was constructed,the vector miR1140 pro∷GUS was transformed into variety Westar by Agrobacterium-mediated approach,and 5 positive transgenic lines were obtained. GUS chemically staining the varied tissues of T1 generation groups of positive rape strains,the results showed that the upstream 1.5 kb region of miR1140 precursor sequence presented the function of promoter,driving GUS expression in rapeseed,moreover,only specific expression in the petiole and leaf axil,indicating that miR1140Pro was a specific promoter.
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