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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。
The selection of a suitable promoter is a key factor in plant resistance to genetic engineering. The acquisition of pathogen-inducible promoters will provide more promoter options for plants. Coupling the deletion fragment P3 of promoter of GIII gene of barley β-1,3-glucanase isozyme with gus (β-glucanase gene), a plant expression vector was constructed, Bacillus mediated transformation of rice. PCR results showed that the 10 strains of hygromycin-resistant rice plants were PCR positive; Southern blot results showed that nine fusion genes containing P3 / gus have been integrated into rice genomic DNA. GUS histochemical staining and fluorescence results showed that the P3-deficient gus-driven gus obtained high level of expression after induced by the exciton. GUS histochemical staining of T1 seed also showed that the elicitor can induce a high level of P3 activity.