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从玉米自交系9801基因组DNA中克隆玉米醇溶蛋白基因ze19启动子,将其克隆到pMD18-T载体上。该序列与发表序列同源性为94%,包括TATAbox、CAATbox启动子基本元件以及多种参与调节醇溶蛋白表达的顺式元件。将ze19启动子取代质粒PBI121-gus中的CaMV35S启动子,构建由ze19启动子驱动GUS的植物表达载体pBIze19-gus,利用土壤杆菌介导法将重组载体pBIze19-gus转入烟草中。对转基因烟草植株GUS活性的定性与定量分析结果表明,ze19启动子驱动GUS基因在转基因烟草种子中表达活性最高,在叶片中活性很弱,在茎中没有活性。所克隆的ze19启动子具有种子特异表达特性,为玉米种子生物反应器的研究提供借鉴。
The ze19 promoter of zein gene was cloned from the maize inbred line 9801 genomic DNA and cloned into pMD18-T vector. This sequence shares 94% sequence homology with the published sequence, including the TATAbox, the basic elements of the CAATbox promoter, and various cis-elements involved in the regulation of gliadin expression. The ze19 promoter was substituted for the CaMV35S promoter in the plasmid PBI121-gus to construct the plant expression vector pBIze19-gus driven by the ze19 promoter and the recombinant vector pBIze19-gus was transformed into tobacco by Agrobacterium-mediated transformation. The qualitative and quantitative analysis of the GUS activity of the transgenic tobacco plants showed that the ze19 promoter-driven GUS gene had the highest expression activity in the transgenic tobacco seeds, the weak activity in the leaves, and no activity in the stem. The cloned ze19 promoter has seed-specific expression characteristics, which can provide reference for the research of corn seed bioreactor.