烟草orf25基因植物表达载体的构建及遗传转化

摘要

为更好地研究烟草雄性不育的机理，构建orf25基因的植物表达载体。 orf25基因是ATP合成酶F（0）部分的4个亚基因之一。以雄性不育烟草MS革新3号为受体材料，利用组成型启动子CaMV35S构建orf25基因植物表达载体，将表达载体pBI121－orf25导入根癌农杆菌LBA4404。采用根癌农杆菌介导orf25基因遗传转化MS革新3号，结果证明载体上的目的基因orf25已整合到烟草基因组中，并获得了52株转pBI121－CaMV35S－orf25基因烟草植株，经PCR鉴定，转基因阳性植株有14株，转化率为26．9％。为下一步研究该基因的功能及其与烟草雄性不育性的关系提供依据。%To better research the mechanism of tobacco male sterility,the plant expression vector of orf25 gene was constructed.Orf25 gene is one of the four subunit genes of F(0) section in ATP synthetase.In this study,male sterility of tobacco named “MS Gexin 3” was used as the receptor materials,adopting constitutive promoter CaMV35S to construct the plant expression vector of orf25 gene, and then the expression vector pBI121⁃orf25 was imported into agrobacterium tumefaciens LBA4404.By agrobacterium tumefaciens orf25 gene was mediated to genetic transform “MS Gexin 3”. The experimental results indicated that the target gene orf25 on the vector had been integrated into tobacco genome,and 52 strains of pBI121⁃CaMV35S⁃orf25 transgenic tobacco plants were obtained.14 transgenic positive plants were identified by PCR,and the conversion rate was 26.9%.All these lays a foundation for further study of the gene function and its relationship with tobacco male sterility.