蔗茅野生种ErDREB1A基因的克隆与表达分析

Cloning and Expression Analysis of ErDREB1A Gene in the Wild Species of Erianthus fulvus

为了充分利用蔗茅野生资源,培育耐寒转基因甘蔗提供可靠的候选基因,本研究以电子克隆结合RT-PCR的方法,在蔗茅(Erianthus fulvus)野生种(昆明蔗茅99-1)中克隆到1个干旱应答元件结合蛋白(dehydration responsive element binding protein,DREB)基因。生物信息学分析表明:该基因的ORF长度为720 bp,编码239个氨基酸,具有1个保守的AP2结构域,编码蛋白以α-螺旋和β-折叠为主,无跨膜结构,没有信号肽,具有多个磷酸化位点,命名为ErDREB1A基因(GenBank ID:MK726363)。荧光定量PCR分析表明:在蔗茅野生种中,随着低温胁迫的延时,ErDREB1A基因的表达量逐渐增加,表明该基因表达受低温胁迫诱导。本研究为进一步研究蔗茅低温胁迫下的分子调控网络和培育耐寒转基因甘蔗提供一定科学依据。

In order to make full use of the wild resources of Erianthus fulvus,and provide reliable candidate genes for breeding cold-tolerant transgenic sugarcane,in this study,a dehydration responsive element binding protein(DREB)gene was cloned from the wild species of Erianthus fulvus(Kunming99-1)by electron cloning combined with RT-PCR.Bioinformatics analysis showed that the ORF length of this gene was 720 bp,encoded 239 amino acids and of a conserved AP2 domain.The encoded protein was mainlyα-helix andβ-sheet,of no transmembrane structure,no signal peptide,and multiple phosphorylation sites.The gene was named ErDREB1 A(GenBank ID:MK726363).Quantitative fluorescence PCR analysis showed that the expression of ErDREB1 A gene increased gradually with the delay of low temperature stress in wild species of Erianthus fulvus.This results showed that the expression of ErDREB1 A was induced by cold stress.This study laid a foundation for further study of the molecular regulation network under the low temperature stress of Erianthus fulvus and the cultivation of cold-tolerant transgenic sugarcane.