高光效基因植物表达载体的构建

Construction of Plant Expression Vectors for High Efficiency Photosynthesis Genes

通过一系列中间载体的构建 ,将来自C4作物 (甘蔗 )光合作用中的关键酶基因 (高光效基因 )RbcS和PEPCase基因插入到植物表达质粒中 ,获得了具有卡那霉素抗性的RbcS植物表达载体 pC2 0SNR和具有PPT除草剂抗性的PEPCase基因的正义 (pC30SNP)和反义 (pC30SNPr)植物表达载体。再将这 3个载体分别转入农杆菌LBA4 4 0 4和A2 81或EHA10 5中 ,获得了适于C3 植物转化的农杆菌重组工程菌株。为进一步利用C4植物高光效基因转化C3 植物 ,以期提高光合效率提供基础。

RbcS and PEPCase gene come from C 4 plant sugarcane, which are the key enzyme genes of photosynthesis. Three plant expression vectors pC20SNR (containing RbcS gene), pC30SNP (sense vector) and pC30SNPr (antisense vector) containg PEPCase gene were obtained by constructing a series of middle vectors. Vector pC20SNR has nptⅡ selective marker gene resistant to kanamycin, pC30SNP and pC30SNPr have bar selective marker gene resistant to herbicide PPT. The three vectors were introduced into different stains of Agrobacterium tumefacien LBA4404 and A281 or EHA105 by direct transformation, which could be used for C3 plants genetic transformation with high efficiency photosynthesis gene of C 4 plant to increase photosynthesis efficiency, and investigating expression and regulation of PEPCase gene.