选育作物的雄性不育系是杂种优势利用的重要环节。1990年以后,基因工程雄性不育发展非常迅速,许多植物基因表达调控系统已经被建立。本论文采用热激启动子和铜诱导系统控制位点专一性重组酶CRE的表达,通过删除花药特异性启动子(水稻的Osg6B启动子)与RNA酶基因(Barnase基因)之间的阻断序列,从而控制基因工程雄性不育的产生,探索建立可诱导的、集不育系和保持系于一身的植物基因工程雄性不育系统。
1.从大豆中克隆热激蛋白启动子Gmhsp17.5C(Genbank Number AF544399),并以Gus基因为报告基因转化烟草,组织化学染色和荧光定量测定结果显示,经过连续两次热激处理(42℃ 2h)后,在转基因植株的叶和茎组织化学染色显蓝色,诱导后荧光强度是诱导前荧光强度的5倍左右。其次,我们构建的载体系统通过热激诱导表达的CRE重组酶删除靶载体上的两侧带有loxp位点的Gus基因片段(CaMV35S-Gus)。实验结果表明,在转基因烟草中热诱导的基因删除效率可达41%。
2.利用已经建立的热激诱导定位重组系统调控RNA酶基因(Barnase基因)在转基因烟...
【英文摘要】
Finding male sterility in crops is the key point of utilization of heterosis. Since 1990, transgenic male sterility was improving. Several inducible systems for transgene expression were constructed in plant. The promoter of heat shock protein and copper-inducible system were used to control the expression of site-specific recombination enzyme gene (Cre) in our research. By excision of blocking fragment between anther-specific promoter (Osg6B) and RNase gene (barnase), effects of induciable male sterility s...