烟草花药特异启动子的克隆、活性测定及雄性不育基因和恢复基因的构建  

Biological Activity Assay of Tobacco Anther-specific Promoter and Construction of Chimeric Genes Confering Plant Male-sterility and Male-restoration

作　　者：李胜国[1] 刘玉乐[1] 康良仪[1] 田波[1] 

机构地区：[1]中国科学院微生物研究所

出　　处：《农业生物技术学报》1995年第3期25-32,共8页Journal of Agricultural Biotechnology

摘　　要：采用聚合酶链式反应方法（ＰＣＲ），以烟草（ＮｉｃｏｒｉａｎａｔａｂａｃｕｍｃｖＳａｍｓｕｎ）叶片ＤＮＡ为模板扩增得到１．５ｋｂ花药特异启动子ＴＡ２９的基因片段。然后克隆到ｐＧＥＭ７Ｚｆ（＋）的ＳｍａⅠ位点上。序列分析结果表明，该基因片段两端２８０ｂｐ区段的碱基序列与资料报道的完全相同。经转化离体组织测定，该启动子具有花药特异启动活性。该启动子经与核酸酶Ｂａｒｎａｓｅ及其相应的抑制剂Ｂａｒｓｔａｒ基因分别融合后构建成雄性不育基因和恢复基因。A tobacco anther-specifici promoter TA29 was obtained from tobacco genomic DNA by PCR and cloned into the site of Smal of pGEM7Zf(+).Partial DNA sequencing showed that about 280 bpsequences of two flanking regions of the gene were identical with the reported previously,and the result of in vitro assay in detached anthers indicated that the cloned promoter has anther specific prornoter activity.Two chimeric genes were constructed by fusing TA29 with Barnase and Barstar,respectively,in a binaryvector containing bromoxynil-resistai、tnitrilase(bxn)gene. TA29BN/bxn ha;been proved to confer plantmale sterility,and TA29BS/bxn to confer male restoration.

关 键 词：烟草 花药特异启动子 三系配套 

分 类 号：S572.035.3[农业科学—烟草工业]