大豆抗逆相关基因GmERF10克隆及生物信息学分析

Cloning and Bioinformatics of Resistance Related Gene GmERF10 in Soybean

为分离克隆大豆抗旱功能重要基因,前期运用RNA-Seq技术对M18苗期根系转录组进行了分析,筛选与苗期大豆根系生长发育相关的显著差异表达基因,选取差异较大的基因进行克隆,提取M18叶片基因组,设计引物,得到目标片段,命名为GmERF10。为了解GmERF10的结构和功能特点,利用生物信息学工具和软件对GmERF10的理化性质和蛋白质结构等进行了系统分析,并探讨了同物种及不同物种间乙烯应答转录因子(ERF)基因的亲缘关系。结果表明:GmERF10为不具有信号肽和跨膜结构域的亲水性蛋白。GmERF10含有38个潜在磷酸化位点。GmERF10蛋白的二级结构主要以α-螺旋、β-折叠和无规则卷曲组成。GmERF10亚细胞主要定位于细胞核中,结构域为AP2/ERF结构域,预测其功能为乙烯应答转录因子(ERF),主要通过结合GCC顺式作用元件来调节抗性基因的表达。

In order to isolate and clone the important genes with drought resistance function in soybean,the transcriptome of M18 root system at seedling stage was analyzed by RNA-Seq technique in the early stage of this experiment.The significant differentially expressed genes related to the growth and development of soybean roots at seedling stage were screened,and the genes with greater differences were selected and cloned.The M18 leaf genome was extracted and designed,and primers were designed to obtain a 633 bp target fragment,which was named GmERF10.In order to clarify the structure and functional characteristics of GmERF10,this paper systematically analyzed the physicochemical properties and protein structure of GmERF10 using bioinformatics tools and software,and explored the genetic relationship of ERF genes between the same species and different species.The results show that GmERF10 does not have a hydrophilic protein of a signal peptide and a transmembrane domain.GmERF10 contains 38 potential phosphorylation sites,and the secondary structure of GmERF10 protein consists of α-helix β-sheet and random coil.The GmERF10 sub-cell is localized in the nucleus and its domain is the AP2/ERF domain.Its function is predicted to be an ethylene-responsive transcription factor,which regulates the expression of resistance genes mainly by binding to GCC cis-acting elements.