香石竹ACC氧化酶重复基因植物表达载体的构建及转基因植株的获得

香石竹ACC氧化酶基因从核NDA中克隆之后,将其首先构建成正义及反义的单拷贝植物表达载体,在此基础上又同向插入一个ACO基因片段,从而获得ACO的正义重复基因和反义重复基因植物表达载体,所得载体已通过酶切分析和PCR鉴定。将该表达载体导入根癌农杆菌LBA4404菌株,并转化香石竹品种Master、爱卡迪幼叶,经PCR检测和Southem杂交鉴定,获得了8株转化植珠。

Construction of Repeated Carnation ACC Oxidase Gene PlantRecombinant Expression Vectors

The genomic fragment of carnation ACC oxidase,the key enzyme in ethylene biosynthesis,was cloned and single copy plant recombinant expression vectors were constructed first.The sense and antisense repeated plant recombinant expression vectors were constructed based on the single copy vectors.The repeated vectors were indentified by restriction enzyme and PCR analysis.The vectors were transformed into Agrobacterium tumefaciens ,which was confirmed by PCR analysis.Transgenic plants of carnation were obtained by using those vectors,which were confirmed by PCR analysis and Southern blotting.The transgenic plants were under further analysis for transgene silence effect.