玉米PLATZ基因GRMZM2G006585启动子的克隆及表达分析  

Cloning and Expression Analysis of Maize PLATZ Gene GRMZM2G006585 Promoter

作　　者：王亚丽 魏琦超 李成伟 WANG Yali;WEI Qichao;LI Chengwei(Henan Engineering Research Center of Crop Genome Editing,College of Life Science and Technology,Henan Institute of Science and Technology,Xinxiang 453003,China;College of Biological Engineering,Henan University of Technology,Zhengzhou 450001,China)

机构地区：[1]河南科技学院生命科技学院,河南省粮食作物基因组编辑工程技术研究中心,河南新乡453003 [2]河南工业大学生物工程学院,河南郑州450001

出　　处：《华北农学报》2023年第2期21-30,共10页Acta Agriculturae Boreali-Sinica

基　　金：河南省科技攻关项目(202102110148,212102110477);河南科技学院高层次人才科研启动项目(201010616004)。

摘　　要：高转录活性籽粒特异性启动子可调控目的基因在植物籽粒中特异性、高水平表达。为发掘玉米籽粒特异性启动子,以公开发表的玉米表达谱芯片数据为切入点,筛选出籽粒优势表达基因GRMZM2G006585,克隆其编码区上游约2000 bp的DNA序列,命名为PZm2G006585。利用在线网站New PLACE和PlantCARE对其进行启动子顺式作用元件分析,发现其含有E-box、P-box等多个籽粒特异性相关元件,初步认为所克隆编码区上游序列为玉米来源的籽粒特异性启动子。为验证其功能,构建该启动子驱动β-葡萄糖苷酸酶基因(GUS)的表达载体并进行植物遗传转化。转基因水稻的GUS组织化学染色结果表明,该启动子驱动外源基因表达模式为籽粒特异、胚优势表达;转基因拟南芥单拷贝株系T_(3)种子中GUS活性检测结果显示,PZm2G006585驱动的GUS活性为909.52 nmol/(min·mg)。籽粒特异性启动子PZm2G006585的发掘和功能验证为驱动目标基因在玉米、水稻等单子叶植物籽粒中特异性表达提供了候选启动子资源。Grain specific promoter with high transcription activity can regulate the specific and high-level expression of target genes in plant grains.In order to explore specific promoters of maize grain,the dominant expression gene GRMZM2G006585 was screened from the published data of maize expression profile microarray.The DNA sequence about 2000 bp upstream of its coding region was cloned and named PZm2G006585.By using the online websites New PLACE and PlantCARE to analyze its promoter elements,we found that it contained multiple grain specific related elements such as E-box and P-box.It was preliminarily considered that the upstream sequence of the cloned coding region was a grain specific promoter from maize.In order to verify its function,the expression vector of GUS gene was constructed and transformed into plants.GUS histochemical staining results of transgenic rice showed that the expression pattern of exogenous genes driven by the promoter was grain specific and embryo dominant.The results of GUS activity detection in T_(3) seeds of transgenic Arabidopsis thaliana single copy line showed that the GUS activity driven by PZm2G006585 was 909.52 nmol/(min·mg).The discovery and functional verification of the grain-specific promoter PZm2G006585 can provide candidate promoter resource for specific expression of target genes in maize,rice and other monocotyledons.

关 键 词：玉米 籽粒特异性启动子 植物生物反应器 PZm2G006585 遗传转化 

分 类 号：S513.03[农业科学—作物学] Q78[生物学—分子生物学]