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将大麦β-1,3-葡聚糖酶同功酶基因(GIII)启动子(PGIII)与其报告基因gus(β-葡聚糖酸醛苷酶基因)耦联,构建植物表达载体,通过农杆菌介导法转化水稻。PCR、DNA印迹法结果显示,构建的pGIII-gus表达载体已整合到水稻基因组DNA中。GUS组织化学染色、RNA印迹法及荧光法结果显示,该启动子驱动的gus在水稻叶片中为低水平表达;而用水扬酸(SA)与稻瘟菌来源的激发子处理,可诱导gus的高水平表达。T1代种子的GUS组织化学染色结果也表明,SA与激发子可以诱导高水平的PGIII活性。这些结果表明PGIII是一种强诱导型启动子,并可能是一种病原菌诱导型的启动子。
The barley β-1,3-glucanase isoenzyme gene (GIII) promoter (PGIII) was coupled with its reporter gene gus (β-glucanase gene) to construct a plant expression vector. Bacillus mediated transformation of rice. PCR and Southern blotting showed that the constructed pGIII-gus expression vector had been integrated into rice genomic DNA. GUS histochemical staining, Northern blotting and fluorescence analysis showed that the promoter-driven gus was expressed at a low level in rice leaves, whereas salicylic acid (SA) and Magnaporthe grisea-derived excitons induced gus High level of expression. GUS histochemical staining of T1 seed also showed that SA and the elicitor can induce high levels of PGIII activity. These results indicate that PGIII is a strong inducible promoter and may be a pathogen-inducible promoter.