烟草NtGCN2启动子的克隆及功能研究

目的]探究烟草蛋白激酶NtGCN2启动子驱动GUS基因在ABA、SA、AZA和MeJA四种处理下的表达模式.方法]克隆烟草NtGCN2上游2000bp的启动子序列,并采用Plant CARE和PLACE数据库进行顺式作用元件预测.构建NtGCN2启动子驱动β-葡萄糖苷酸酶基因(GUS)的融合表达载体并转化烟草K32...

Cloning and functional analysis of NtGCN2 promoter in Nicotiana tabacum

  Objective  This study aims to explore the expression pattern of GUS gene driven by NtGCN2 promoter of Nicotiana tabacum protein kinase under different phytohormone treatments in tobacco.  Methods  The NtGCN2 promoter was cloned and the cisacting elements were predicted by Plant CARE software and PLACE database for sequence analysis. The expression vector of NtGCN2 promoter linked to the β-glucuronidase (GUS) gene was constructed and transformed into Nicotiana tabacum K326. The gene expression of GUS and its enzymatic activity were assayed under the treatment of ABA, SA, AZA and MeJA using the transgenic lines in order to explore the function of NtGCN2 promoter.  Results  The prediction results of cis-acting elements showed that NtGCN2-Nsy-pro2000 promoter contains cis-acting elements, including abscisic acid response element (ABRE), methyl jasmonate response element (TGACGmotif) and so on. The expression and enzymatic activity GUS reporter gene driven by NtGCN2 promoter increased under ABA, SA and AZA treatments, but decreased under MeJA treatment.  Conclusion  The gene expression of NtGCN2-Nsy-pro2000 showed different response patterns under the treatment of ABA, SA, AZA and MeJA. The expression and enzymatic activity of GUS driven by NtGCN2 promoter increased significantly under ABA, SA and AZA treatment, but decreased under MeJA treatment.