烟草维管束发育相关基因Nvas启动子顺式作用元件鉴定

Identification for Promoter Cis-acting Element of Vascular Bundle Development Related Gene Nvas in Nicotiana tobacum

为研究烟草维管束发育相关基因Nvas启动子顺式作用元件,将克隆得到的Nvas启动子ATG+1^-463区域分为10个不同长度的片段与GFP报告基因融合构建了植物表达载体并完成了5'端缺失分析.利用根癌农杆菌介导转化W38型烟草叶盘,得到了含有不同长度Nvas启动子片段的转基因烟草植株.体视显微镜下观察到含Nvas启动子-216^-463区域的转基因烟草均有绿色荧光蛋白的表达,表达部位集中在维管束组织.结果表明Nvas启动子能使外源基因在维管束组织中特异表达.该启动子活性高于Ca MV35S,在ATG+1^-216区域存在核心启动子元件,在-337^-379区域存在能增强启动子活性的元件.

In order to study the promoter cis-acting element of vascular bundle related gene Nvas in Nicotiana tobacum,Nvas＇ s promoter sequence（ ATG ＋ 1 --463） was divided into ten fragments of different lengths and fused with GFP to constract plant expresion vector and complete the deletion analysis at 5＇ end. Transgenic plants were obtained by Agrobacterium tumefaciens mediated transformation. The results of the stereomicroscope showed that the green fluorescence were observed in vascular bundle in-216 --463 transgenic plants. The expression results revealed the Nvas promoter could make the genes express in vascular bundle specifically,which had a higher promoter activity than Ca MV35 S. A key promoter element exist in ATG＋1 --216 region and an element could enhancing promoter activity may exist in the-337--379 region.