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DREB1A转录因子蛋白的原核表达

Prokaryotic Expression of DREB1A Transcription Factor

  • 摘要: DREB1A是DREB转录因子的一员,它们都含有一段与DNA结合的保守区域,由58个氨基酸组成,称为EREBP/AP2结构域(EREBP/AP2 domain)。一个DREB转录因子可以调控多个与植物干旱、高盐及低温耐性有关的功能基因表达。从拟南芥中克隆了DREB1A转录因子基因,成功构建了DREB1A基因的原核表达载体,转化E.coliBL21(DE3),经1 mmol/L IPTG诱导表达获得了DREB1A蛋白,但以包涵体形式存在。为以后深入研究DREB转录因子与DRE顺式作用元件相互作用的具体机制奠定了基础。

     

    Abstract: DREB1A is a member of the DREB transcription factors.Each DREB protein contains 58 conserved amino acids named as EREBP/AP2 domain,which binds cis-acting element named DRE(for dehydration-responsive element).A DREB transcription factor could regulate multiple functional genes related to dehydration,low temperature and high salt tolerance of plant.We cloned the DREB1A gene from Arabidopsis thaliana(Columbia ecotype) genome by PCR and constructed the prokaryotic express vector,which was transferred into E.coli BL21(DE3).Expression of the DREB1A protein was induced by 1 mmol/L IPTG,but the target protein obtained in the form of inclusion body.This work provides a foundation for further understanding of the interaction mechanism between DREB transcription factors and DRE cis-acting elements.

     

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