ChsA启动子驱动的IPT基因植物表达载体的构建

The Construction of a Plant Expression Vector pBI121-PchsA-IPT Using ChsA Gene Promoter

为研究异戊烯腺苷类细胞分裂素与花衰老进程的相关性,本研究通过PCR方法从矮牵牛中克隆到花瓣特异性表达的矮牵牛查尔酮合成酶基因A(ChsA)的启动子PchsA,并以其取代植物表达载体pBI121中的组成型启动子CaMV35S,然后将IPT基因插入到PchsA启动子下游,测序结果显示花瓣特异性启动子驱动的IPT植物表达载体构建成功.

In order to further investigate the correlation between isopentenyl adenosine cytokinin and flowerage senescence development,we constructed the binary vector pBI121-PchsA-IPT.Firstly PchsA fragment was obtained from petunia hybrida genome DNA by PCR,then PchsA was used to substitute constitutive promoter CaMV35S in plant expression vector pBI121.Secondly,isopentenyl pyrophosphate transferase gene（IPT） was inserted into pBI121-PchsA using XbaⅠ and Sac Ⅰ restriction enzymes and T4 ligase.The sequence results showed that plant expression vector that flower petal specificity promoter PchsA drive IPT was constructed successfully.