柽柳 ThbHLH1上游表达调控基因1）

摘要

通过柽柳转录组分析克隆得到了一条bHLH基因（ ThbHLH1）。在此基础上，本研究通过TAIL－PCR的方法克隆ThbHLH1转录因子的启动子序列，长2061 bp。经PLACE等软件分析发现，ThbHLH1启动子中存在多个DOF元件，利用酵母单杂交和染色质免疫共沉淀试验（ ChIP 试验）研究发现，柽柳的ThDof16基因可识别Th-bHLH1基因启动子中的DOF元件，说明ThDof16基因是调控ThbHLH1基因表达的上游调控因子。另外，通过基因枪技术瞬时转化烟草叶片，进行GUS染色和GUS酶活测定的试验，得到进一步验证。%A bHLH gene, ThbHLH1 was cloned from transcriptome of Tamarix hispdi a.The promoter region of ThbHLH1with 2061 bp in length was cloned using TAIL-PCR.There were many DOF motifs in the promoter of ThbHLH1, and yeast one hybrid and ChIP analysis were performed to determine the upstream of ThbHLH 1.The results showed that a DOF protein from T.hispida, ThDof16 can bind to these DOF motifs in the promoter of ThbHLH 1, and is the upstream regulator of Th-bHLH1.The GUS staining and GUS enzyme activity determination assays of the transient-transformed tobacco leaves , which were obtained by gene-gun bombardment technology , were performed .