刺五加MDD基因启动子的克隆与生物信息学分析

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。

Cloning and bioinformatic analysis of mevalonate diphosphate decarboxylase gene promoter in Eleutherococcus senticosus

Objective To clone and analyze the sequence of mevalonate diphosphate decarboxylase (MDD) gene promoter from Eleutherococcus senticosus.Methods According to the full length of MDD cDNA sequence, using PCR cloning and TAIL-PCR methods, we are cloning 5''DNA sequence and promoter sequence of MDD gene. And bioinformatic analysis is used. Such as PlantCARE.Results We have cloned the 5''DNA sequence successfully, with full length of 1 024 bp. And the promoter sequence is 1 423 bp. Bioinformatic analysis shows that the promoter sequence has 49 TATA-box and 25 CAAT-box. In addition, the promoter has some cis-acting elements responsive to abscisic acid, environmental stresses, MeJA, required for endosperm expression, light-regulated and so on. Last but not least, there have two MYBHv1 and two MYB binding sites.Conclusion We have firstly cloned and analyzed the E. senticosus MDD gene promoter sequence. It is important to it''s expression.