【摘要】 根据水稻碳酸酐酶基因5’端序列,从4处不同位置设计上游引物,在碳酸酐酶基因ATG前0位点处设计下游引物。通过PCR扩增基因5’端1600bp、964bp、585bp、313bp片段,将以上目的片段克隆到以GUS为报告基因的植物表达载体pBI121上,通过农杆菌介导法转化烟草。转化植株GUS活性检测结果,发现-1600～0bp片段启动GUS在烟草的叶、茎中有GUS活性,而其余3段区域(-964～0bp,-585～0bp,-313～0bp)未检测出GUS活性,这些暗示了-1600～0bp启动子区域在控制基因表达时,具有在叶、茎中表达,在根中不表达的组织特异性。更多还原

【Abstract】 Based on the 5’ flanking regulatory regions of carbonic anhyrase (CA) g ene in rice (Oryza sativa L.), we designed four primers in different positions o f the upstream of the translation initiation ATG codon. The fragments of -1 600b p, -964bp, -585bp and -313bp were amplified using PCR, and then cloned into pBI1 21 vector by fusing with the report gene encoding ?茁-glucuronidase (GUS). The t ransgenic tobaccos were obtained by Agrobacterium tumefaciens-mediated transform ation. GUS staining showed that 1 600bp of the promoter directed GUS expression in leaf and stem of the transgenic tobacco, and the others showed the opposite r esults. This result indicate that -1 600~0bp of the promoter regulated gene spec ific expression in leaf and stem, and not in root.更多还原