普通野生稻绿色组织特异表达启动子的克隆与鉴定

摘要

Green tissue-specific expression promoter,which makes the exogenous gene express efficiently in green tissues of receptor crop. The green tissue-specific promoter OrGSP was cloned from Oryza rufipogon,and the fusion vector of OrGSP and GUS gene was constructed and transferred into Arabidopsis thaliana in order to identify the function of the promoter. Bioinformatics analysis shows that the length of 825 bp of OrGSP,contains the basic transcription initiation elements TATA-box and CAAT-box,and light responsive elements TCCC-motif,Sp1,G-box,I-box,GA-motif and as-2-box,etc. The GUS histochemical staining of transgenic Arabidopsis thaliana shows that OrGSP regulates the expression of GUS gene only in green tissue,and the GUS activity in leaf and stem is significantly higher than in roots. The promoter of Oryza rufipogon,OrGSP,is a green tissue-specific promoter and the results can provide new regulatory elements for crop molecular breeding.%绿色组织特异表达启动子可调控外源基因只在受体作物的绿色组织中定点、高效地表达.以普通野生稻为实验材料,克隆了绿色组织特异表达启动子OrGSP,构建OrGSP和GUS基因融合的表达载体,转入拟南芥中鉴定功能.启动子OrGSP长度为825 bp,含有基本的转录起始元件TATA-box和CAAT-box,以及光响应元件TCCC-motif、Sp1、G-box、I-box、GA-motif和as-2-box等.转基因拟南芥GUS组织化学染色结果表明,启动子OrGSP调控GUS基因只在绿色组织中特异表达.GUS活性测定结果显示,叶和茎中的GUS活性比根中明显提高.普通野生稻中克隆的启动子OrGSP为绿色组织特异表达启动子,可为作物分子育种提供新的调控元件.