邻单胞菌邻苯二酚1,2-双加氧酶基因(tfdC)在拟南芥中表达的初步研究

A Preliminary Study of Expression of a New Catechol-1,2-Dioxygenase Gene ( tfd C) from Plesiomanas in Arabidopsis thaliana

将从一株邻单胞菌中克隆到的一个新的邻苯二酚 1,2 双加氧酶基因 (tfdC)的起始密码子由GTG突变成ATG ,并克隆到农杆菌双元载体 pPZPY12 2中 ,利用农杆菌介导转化模式植物拟南芥 ,获得了转化植株 .经过PCR ,PCR Southern和Southerndotblot方法检测证实 ,tfdC基因已经整合到拟南芥基因组中 .邻苯二酚 1,2 双加氧酶酶活性检测表明 ,转基因植株具有一定的酶活性 。

A new catechol 1,2 dioxygenase gene ( tfd C) was cloned from a strain of Plesiomonas in a previous study. The start codon of the tfd C gene was changed from GTG to ATG using PCR method. The modified tfd C gene was constructed into a binary vector, pPZPY122, and the construct was transferred into Arabidopsis thaliana mediated by Agrobacterium tumefaciens LBA4404 by the leaf discs transformation method.Regenerated plants were obtained from transformed calli. The result of PCR, PCR southern and Southern dot blot assays showed that the tfd C gene was incorporated into the genome of Arabidopsis thaliana, and the results of catechol 1,2 dioxygenase activity assay indicated that the gene was expressed in one of the transgenic plants.