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

作　　者：王文东[1] 陈文峻[1] 罗如新[2] 蒯本科[1] 

机构地区：[1]复旦大学生命科学学院生物化学系 [2]复旦大学环境科学与工程系

出　　处：《复旦学报（自然科学版）》2002年第1期16-21,26,共7页Journal of Fudan University：Natural Science

基　　金：国家自然科学基金资助项目 (39770 4 78)

摘　　要：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.

关 键 词：邻苯二酚1 2-双加氧酶基因(tfd C) 遗传转化 外源基因表达 植物修复 

分 类 号：Q789[生物学—分子生物学]