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目的克隆并分析刺五加甲羟戊酸焦磷酸脱羧酶(mevalonate diphosphate decarboxylase,MDD)基因的启动子序列。方法根据刺五加MDD基因的c DNA序列,采用PCR扩增和热不对称交错PCR(TAIL-PCR)技术,克隆MDD基因5’端的DNA序列及启动子序列。利用Plant CARE等软件对其进行生物信息学分析。结果克隆得到长1 423 bp的刺五加MDD基因启动子序列及长1 024 bp的5’端DNA序列。该启动子序列含有49个TATA-box、25个CAAT-box。还含有脱落酸响应元件、茉莉酸甲酯响应元件、胚乳表达必须顺式作用元件、干旱胁迫响应元件、光响应元件等多种顺式作用元件,以及2个MYBHv1与2个MBY结合位点。结论首次克隆并分析了刺五加MDD基因的启动子序列,为该基因的表达调控奠定了基础。
Objective To clone and analyze the promoter sequence of mevalonate diphosphate decarboxylase (MDD) gene. Methods According to the c DNA sequence of MDD gene of Acanthopanax senticosus, the DNA sequence and promoter sequence of the 5 ’end of MDD gene were cloned by PCR amplification and thermal asymmetric interlaced PCR (TAIL-PCR). Bioinformatics analysis using Plant CARE software. Results A 1 423 bp long promoter sequence of Acanthopanax senticosus MDD gene and a 5 ’end DNA sequence of 1 024 bp were obtained. The promoter sequence contains 49 TATA-box, 25 CAAT-box. Also contains abscisic acid response element, methyl jasmonate response element, the endosperm expression cis-acting element, drought stress response element, light-responsive elements and other cis-acting elements, and two MYBHv1 with two MBY binding sites. Conclusion The promoter sequence of MDD gene of Acanthopanax senticosus was cloned and analyzed for the first time, which laid the foundation for the regulation of the gene expression.