摘要
水稻种子萌发后,新生器官分化和生长所需磷素主要来自于植酸酶对种子中贮存植酸及其盐类的降解。针对水稻植酸酶基因OsPHY2在萌发种子中优势表达、但其转录调控机制尚不明确的现状,本研究克隆了该基因的启动子。利用生物信息学工具对该启动子中含有的顺式调控元件分析表明,该启动子中除含有RNA聚合酶结合的重要保守元件TATA盒和调控转录效率的保守元件CAAT盒外,还含有调控OsPHY2呈组织特异表达、参与碳代谢和应答光照等环境信号的调控元件。利用DNA重组技术,构建了OsPHY2启动子不同长度片段驱动报告基因GUS表达的系列双元表达载体。本试验为鉴定OsPHY2启动子中含有的重要调控元件及其在启动子中的精细定位,进而阐明水稻植酸酶基因OsPHY2的转录调控机制奠定了基础。
During seed germination process,phosphorus is to be hydrolyzed by phytase for the differentiation and growth of novel tissues.The phosphorus is derived from the degradation of phytic acid and its derivatives stored in the seeds.OsPHY2,a rice phytase gene identified previously in our group,was shown to be predominantly expressed in the germinating seeds.However,how OsPHY2 expression is regulated is still largely unknown.In this study,the OsPHY2 promoter was cloned and several important cis-regulatory elements located in the promoter were identified based on bioinformatic analysis.It was found that the OsPHY2 promoter contains the conserved motiofs such as TATA box and CAAT box which play critical roles on binding of RNA polymerase and in regulation of transcriptional efficiency,respectively.The OsPHY2 promoter also harbors other putative key cis-regulatory elements such as those involved in tiussue-specific expression,carbon metabolism,and response of light signal,etc..Using DNA recombinant technique,a series of binary expression vectors were generated,in which various lengths of truncated OsPHY2 promoter fragments were fused to GUS reporter.These results set a base for experimentally identifying the important regulatory elements and their fine locations in the promoter,and for systematically elucidating the transcriptional mechanism of OsPHY2.
出处
《河北农业大学学报》
CAS
CSCD
北大核心
2011年第6期10-15,29,共7页
Journal of Hebei Agricultural University
基金
国家自然科学基金(No.30871466)
河北省作物生长发育调控重点实验室资助
