用无启动子的GUS报告基因捕获水稻基因启动子

构建了嵌合质粒p13DGUTs，它是在Ds转座子中插入了无启动子的B．葡萄糖醛酸酶报告基因(GUS)，用于分离水稻基因启动子。将p13DGUTs转化粳稻品种中花11的胚性愈伤组织，获得了496个转基因植株。抗性愈伤组织与转基因植株的GUS染色与PCR分析表明整合在水稻染色体上的Ds因子都发生了随机跳跃。转基因植株T0代与部分T1代的GUS染色结果表明，M92转基因植株中Ds转座子整合位置上游的水稻基因启动子指导GUS基因的表达及表达的特性是可遗传的。文章对此方法在分离水稻基因启动子与基因上的应用进行了讨论。

Promoter Trapping by Using Promoterless GUS Reporter Gene in Rice

The left border of Ds element, coding region of GUS reporter gene, promoter region of Ubi , the right border of Ds element and transponase of Ac transposon were inserted into the T-DNA region of pCAMBIA1300 in turn to construct plasmid p13DGUTs (Fig.1). The plasmid p13DGUTs was introduced into calli of Oryza sativa subsp. japonica cv.Zhonghua 11 and then 496 transgenic plants were obtained. Histochemical staining and PCR analysis showed that the T-DNA region of plasmid p13DGUTs was integrated into rice chromosome and Ds element was excised randomly from the original insertion sites in some of transgenic calli and plants (Figs.2-4). The histochemical staining in T 0 generation of 496 transgenic plants and T 1 generation of 13 transgenic plants revealed that the expression of GUS gene was driven by an unknown promoter located upstream of the Ds re-integrating site in transgenic plant numbered M92 (Table 1). The expression of GUS reporter gene was found to be inheritable. These results indicated that the plasmid p13DGUTs was available in promoter trapping system.