反义PSY基因植物表达载体的构建及其对中国水仙的转化

实验从中国水仙花瓣中分离得到该基因的572bp的片段，将该片段反向连接到pCAMBIA1301双元载体质粒上，得到CaMV 35S组成型启动子的表达载体pCAM35S-PSY，用“冻融法”将pCAM35S-PSY质粒转入农杆菌LBA4404和EHA105两个菌株中。PCR扩增和酶切鉴定结果表明，所构建的反义表达载体pCAM35S-PSY是正确的并已经成功导入农杆菌中。随后，按照业已建立的遗传转化体系对中国水仙进行了遗传转化，得到了转基因抗性芽。

Construction of Plant Expression Plasmid Vector Containing the Anti Sense PSY Gene and its Transformation to Narcissus tazetta var. chinensis

Phytoene synthase catalyzes one of the key steps of the carotenoid biosynthnic pathway. 572 bp fragment of Narcissus tazetta var. chinensis petal were isolated and introduced its anti sense fragment to pCAMBIA1301 generating pCAM35S-PSY vector which contained CaMV35S promoter, and then the pCAM35S-PSY was transferred into Agrobacterium tumefaciens strain LBA4404. Result of PCR amplification and restriction digestion showed that the fragment of anti sense gene PSY was correctly introduced into pCAMBIA1301 plasmid and the expression vector pCAM35S-PSY was successfully transferred into Agrobacterium. Then, the anti sense gene fragment was transferred into N. tazetta var. chinensis and many HygR shoots were obtained.