水稻雄性不育及其育性恢复表达载体的构建

将ＰＣＲ扩增获得的水稻花药绒毡层特异表达基因Ｏｓｇ６Ｂ启动子（简写成６Ｂ）与来自质粒ｐＲＯＫⅡ上的ＮＯＳ终止子相连，产生了ｐＧＥＭ７Ｚ－６ＢＮＯＳ。然后将来自解淀粉芽孢杆菌中的Ｂａｒｎａｓｅ和Ｂａｒｓｔａｒ基因（简写成ＢＮ和ＢＳ）分别插入到ｐＧＥＭ７Ｚ－６ＢＮＯＳ上的ＢａｍＨ１和ＮｃｏＩ位点上，再用ＳａｌＩ和ＸｈｏＩ切下２．３ｋｂ的６ＢＢＮＮＯＳ和２．２ｋｂ的６ＢＢＳＮＯＳ片段，并分别将其插入到ｐ

Construction of Expression Vectors of Male Sterility and Its Fertility Restoration in Rice (Oryza sativa L. )

The 1.7kb promoter 6B of tapetum-specific genes from Oryza sativa L. byPCR was connected to the 270 bp NOS terminator from pROK II to generate pGEM7Z-6BNOS.The genes for Barnase and Barstar from Bacillus amylolique faciens were inserted between theBamH I and Nco I sites of pGEM7Z-6BNOS to give pGEM7Z-6BBNNOS and pGEM7Z-6BB-NOS, respectively. The fragments of 2.3kb 6BBNNOS and 2. 2kb 6BBSNOS were separatedfrom pGEM7Z-6BBNNOS and pGEM7Z-6BBSNOS with the Sal I and Xho I digestion, andcloned into the Xho I sites of pDM302 conferred resistance to phosphinothricin(PPT) , respectively. The orientation of the 6BBNNOS and 6BBSNOS fragments was diverse in pDM302.Therefore, the plant expression vectors of male sterility and its fertility restoration were successfully constructed. The immatured embryos from rice were transformed by particle bombardment. The rice transgcnic plants with resistance to PPT were generated. The superior male sterile line and its restorer are being expected.