水曲柳SnRK2B基因和启动子的克隆及分析

Cloning and Analysis of SnRK2B Gene and Promoter in Fraxinus mandshurica

为了研究水曲柳中同源SnRK2转录因子的功能,在水曲柳干旱响应转录组中寻得SnRK2B的同源序列并克隆得到SnRK2B转录因子基因全长,将其命名为FmSnRK2B。使用SiteFinding-PCR法扩增获得SnRK2B启动子序列,并对FmSnRK2B基因编码区及其启动子进行生物信息学分析和启动子顺式调控元件分析。结果表明,克隆得到的全长序列包含完整的开放阅读框1074bp,编码357个氨基酸。启动子顺式作用元件分析结果表明,其启动子区包含脱落酸应答元件ABRE及胁迫相关元件HSE、MBS等。干旱胁迫下,FmSnRK2B基因的表达随胁迫时间的延长先增加后降低,表明其参与植株的抗逆过程。本研究对进一步揭示水曲柳的抗逆机制具有重要意义。

In order to study the function of homologous SnRK2 transcription factor gene in Fraxinus mandshurica, the homologous of the SnRK2B gene was find in the transcriptome responsed to drought from Fraxinus mandshurica and a complete transcription factor gene was cloned by Homology-based cloning method, and it was named by FmSnRK2B. The SnRK2B promoter fragment was isolated and identified from the genomic DNA of Fraxinus mandshurica using the method of SiteFinding-PCR. The gene sequence and promoter sequence was analyzed by bioinformatics analysis and cis- acting element analysis respectively. The results show that the complete transcription factor gene contain a complete open reading frame of 1 074bp which encoded 357 amino acids. The cis-acting elements analysis indicate that the promoter contain abscisic acid response component ABRE and other stress-related components as HSE, MBS etc. Under drought stress, the expression level of FmSnRK2B changed with time extension, which was first rised and then dropped, that means the gene was related to stress tolerance in plants. This study has a great significance to reveal the mechanism of resistance of Fraxinus mandshurica.