【摘要】 选择合适的启动子是植物抗病基因工程的关键性因素,病原菌诱导型启动子的获得将为植物提供更多的启动子选择。将大麦β-1,3-葡聚糖酶同工酶GⅢ基因启动子的缺失体片段P3与报告基因gus(β-葡聚糖酸醛苷酶基因)偶联,构建植物表达载体,通过农杆菌介导法转化水稻。PCR结果表明,所获得的10株潮霉素抗性水稻植株均呈PCR阳性;DNA印迹法结果显示,9株含P3/gus的融合基因已整合到水稻基因组DNA中。GUS组织化学染色及荧光法结果显示,P3缺失体驱动的gus在激发子诱导后,获得了高水平表达。T1代种子的GUS组织化学染色结果也表明,激发子可以诱导高水平的P3活性。更多还原

【Abstract】 P3 deletion of β-1, 3-glucanase isoenzyme GIII gene promoter was ligated upstream into the gus report gene and P3/gus fusion fragment was then cloned into a binary vector pCAMBIA1300 for Agrobacte-rium-mediated transformation of rice (Oryza sativa L. cv. Taipei 309). PCR analysis indicated that the fusion gene was presented in 10 T0 transgenic plants. The integration of the genes in rice genomic DNA was further confirmed by Southern blot. The results showed that 9 plants contain the fusion gene. Histochemical staining and spectrofluorophotometric analysis of transgenic rice leaves showed that the GUS activity was increased after treatment with the fungal elicitor from Magnaporthe griea. Histochemical staining of T1 rice seeds displayed marked GUS activity after induction with the elicitor, while no GUS activity was detected in untreated T1 rice seeds.更多还原