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本研究利用染色体步移法从大豆栽培品种东农L13基因组中分离得到了GmPLP1基因1.5 kb的启动子片段,利用PLACE和PlantCARE在线启动子预测工具发现了107个调控元件。将该片段定向替换载体pBI121的CaMV35S组成型启动子,构建表达载体pBI121-pGmPLP1,驱动下游报告基因GUS基因表达,并通过组织化学染色和GUS定量分析对预测结果进行验证,结果显示GmPLP1受黑暗、蓝光、低温、GA3和ABA诱导表达。
In this study, a 1.5 kb promoter fragment of GmPLP1 gene was isolated from soybean cultivar Dongnong L13 using chromosome walking method. 107 regulatory elements were found using PLACE and PlantCARE online promoter prediction tools. The fragment was directed to CaMV35S constitutive promoter of vector pBI121, and the expression vector pBI121-pGmPLP1 was constructed to drive the expression of downstream reporter GUS gene. The prediction results were verified by histochemical staining and GUS quantitative analysis. The results showed that GmPLP1 was affected by darkness, Blue light, low temperature, GA3 and ABA induced expression.