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【目的】探讨Ac SERK1启动子的活性,为进一步研究Ac SERK1转录调控的分子机制提供依据。【方法】利用hi TAIL-PCR法从菠萝基因组DNA中分离Ac SERK1启动子序列,并利用Plant Care和PLACE分析该序列的顺式作用元件。将启动子与GUS融合,农杆菌真空渗透法浸染烟草叶片后,光照处理、暗处理和0.5 mg·L~(-1)2,4-D处理36 h后利用组织化学染色法检测GUS活性。【结果】克隆了Ac SERK1上游2097bp启动子序列,命名为P_(Ac SERK1)。预测结果显示,该序列除含有TATA-box、CAAT-box等真核生物启动子核心元件外,还含有生长素、赤霉素和茉莉酸甲酯等激素响应元件,高温和低温胁迫响应元件以及光响应元件。瞬时表达结果表明,在光和2,4-D的诱导下P_(Ac SERK1)具有启动基因表达的功能。【结论】分离获得了Ac SERK1启动子序列且该启动子在光和2,4-D诱导下能驱动GUS的表达。
【Objective】 The purpose of this study was to investigate the activity of Ac SERK1 promoter and provide the basis for further study on the molecular mechanism of Ac SERK1 transcriptional regulation. 【Method】 Ac SERK1 promoter sequence was isolated from pineapple genomic DNA by hi TAIL-PCR method. The cis-acting element of this sequence was analyzed by Plant Care and PLACE. The promoter was fused with GUS, and the Agrobacterium tumefaciens vacuolization method was used to detect the GUS activity after the tobacco leaves were infiltrated by the light infiltration method, and the light treatment, dark treatment and 0.5 mg · L -1 2,4-D treatment for 36 h. 【Result】 The 2097bp promoter upstream of Ac SERK1 was cloned and named P_ (Ac SERK1). The results showed that the sequence contained hormonal response elements such as ghrelin, gibberellin and methyl jasmonate, high temperature and low temperature stress response elements, in addition to eukaryotic promoter elements such as TATA-box and CAAT-box, and Light response element. The transient expression results showed that P_ (Ac SERK1) has the function of initiating gene expression under the induction of light and 2,4-D. 【Conclusion】 The Ac SERK1 promoter was isolated and the promoter was able to drive GUS expression under light and 2,4-D induction.