根据GenBank中植物病原物诱导型启动子碱基序列设计引物,以烟草基因组DNA为模板,扩增PPP3启动子。用PPP3替换pCAMBIA1301中与gus基因相连的35S启动子,构建重组质粒pCOMBIA+PPP3+gus后导入农杆菌GV3101。通过农杆菌介导的瞬时表达技术将受PPPs控制的gus基因导入烟草。分别接种青枯菌和水杨酸24 h后,烟草叶片中检测到gus基因转录效率分别提高了27.94,17.69倍。表明PPP3启动子具有本底表达水平低、诱导表达活性强的特点。
According to the sequences of pathogen- inducible plant promoters PPP3 at GenBank，PPP3 promot- ers was cloned from tobacco genome．It was used to replace cauliflower mosaic virus 35S promoter of pCAM- BIA1301．The recombinant plasmid was used to transform Agrobacterium tumefaciens GV3101．The inducibility of the PPP3 promoters in tobacco leaf was evaluated by Agrobacterium tumefaciens- transient genetic transformation as- sye．Real- time quantitative PCR was used to screen the PPP3 promoters with high inducible expression．The results showed that the gus transcript level under the control of PPP3 promoters increased respectively 27．94 fold and 17. 69 fold after inoculation with Ralstonia solanacearum and SA．The PPP3 promoter had the advantages such as low basal activity and high expression activity．
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