拟南芥冷诱导型启动子CBF3驱动IPT基因在烟草中的表达

用PCR的方法获得拟南芥冷诱导型启动子CBF3和农杆菌Ti质粒上的IPT基因,构建由启动子CBF3驱动IPT植物表达载体,并经农杆菌介导将基因整合到烟草的基因组中,利用Kan筛选、PCR和RT-PCR检测,最终获得转基因的植株。通过对烟草的植物学性状和农艺学性状的分析发现,随着启动子CBF3驱动的IPT基因在烟草中的表达,使得植株性状发生很大的改变,如株型变为塔型,株高增高,节距增长,且有很多的分枝和侧芽,叶片明显增多,表现出短窄且平滑的性状,茎围、腰叶长和腰叶宽都减小许多,相应的最大叶面积、顶叶面积和叶质量也都下降,但叶的密度却有所增加。

Expression of IPT Gene Linked with Cold induced Promoter CBF3 from Arabidopsis thaliana in Tobacco

The Arabidopsis thaliana cold-inducible promoter CBF3 and the IPT gene located on Ti plasmid of Agrobacterium were amplified by PCR,constructed a plant expression vector driven by the promoter CBF3,and then integrated into tobacco genome by Agrobacterium-mediated,screening with Kan,and confirmed by PCR and RT-PCR,and finally transgenic plants were obtained.Through the analysis of botany traits and agronomic traits,we found that,with the expressing of IPT gene drove by promoter CBF3 in tobacco,the characters of tobacco changed a lot,such as plant type changed to tower,plant height and pitch increased,and many branches and buds emerged,the number of leaves increased obviously,and leaves became narrow and smooth,circumference of stem,length and width of waist leaves had reduced a lot,as well as the corresponding maximum area of leaf,parietal area and leaf weight,however,the leaf density increased.