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本研究用来自烟草的具有高度病原物特异性的两个诱导启动子EAS4和hsr203J,串联驱动GUS基因的表达,同时考虑到转基因植物的安全性,引入双边界序列,构建了含双病原物诱导启动子的植物安全表达载体。将表达载体转化烟草获得转基因植株。分析显示,在正常生长情况下转基因烟草检测不到GUS活性,或活性极低;而受疫霉激发子parasiticein、Phytophthora nicotianea[0]的孢子悬浮液和Ralstonia solanacarum的菌悬液诱导后,转基因烟草叶片中可检测到明显的GUS活性。结果表明,构建的植物表达载体所含双病原物诱导启动子均具有良好的诱导活性,可用于植物遗传转化。
In this study, two highly inducible promoters EAS4 and hsr203J from tobacco were used to drive the expression of GUS gene in tandem, taking into account the safety of transgenic plants and the introduction of double border sequences to construct a two-pathogen-inducing Promoter plant safety expression vector. The expression vector was transformed into tobacco to obtain transgenic plants. The results showed that GUS activity or low activity was not detected in the transgenic tobacco under normal growth conditions. After induced by the spore suspension of parasiticein, Phytophthora nicotianea [0] and the bacterial suspension of Ralstonia solanacarum, the transgenic tobacco Significant GUS activity was detectable in leaves. The results showed that the constructed two-pathogen-inducible promoter contained in the plant expression vector has good inducing activity and can be used for plant genetic transformation.