小叶章DaGST基因克隆及启动子分析

Cloning and Analysis of the DaGST Gene and Its Promoter in Deyeuxia angustifolia Kom.

为了解小叶章(Deyeuxia angustifolia Kom.)中GST基因的生物学信息,为后续研究GST基因在小叶章中的功能提供理论基础。利用RT-PCR技术和染色体步移法克隆小叶章GST基因及其上游启动子序列,对其进行生物信息学分析和启动子的顺式作用元件分析。克隆得到小叶章GST基因的cDNA序列,命名为DaGST,其开放阅读框为703 bp,编码233个氨基酸,蛋白质相对分子量为25.45 kD。保守结构域预测小叶章DaGST蛋白含有GST-N-Tau和GST-C-Tau两个典型的结构域。系统进化分析表明小叶章DaGST蛋白与大麦(Hordeum vulgare)亲缘关系最近,其次为山羊草(Aegilops tauschii)。克隆得到1 184 bp启动子序列,启动子结构分析表明,小叶章DaGST基因启动子除具有典型的TATA-box、CAAT-box作用元件外,还包括激素响应元件(ABRE,MeJA,SA)等。通过小叶章DaGST基因及启动子的分析,为进一步阐明小叶章的胁迫应答机制提供理论依据。

In order to acquiring the biological information of GST gene and provided a reference basis for the next step in the study of gene function in Deyeuxia angustifolia Kom.The GST gene and its upstream promoter sequence were cloned by RT-PCR and genome walking,and the bioinformatics and promote cis-acting elements were analyzed.The DaGST gene contained a open reading frame(ORF) of 703 bp,which encoded 233 amino acid and its molecular weight was 25.45 k D.Conserved domain analysis showed that DaGST protein had two typical domains,GST-N-Tau and GST-C-Tau.Phylogenetic analysis showed that DaGST protein was the closest to the GST of Hordeum vulgare and Aegilops tauschii.The 1 184 bp DaGST promoter sequence was cloned,and the analysis of DaGST promoter showed that it had typical promoter core elements such as TATA-box and enhancer element CAAT-box,as well as hormone response elements(ABRE,MeJA,SA).Cloning and analysis of the DaGST gene and its promoter,these results provide a theoretical foundation for further clarifying the stress-response of Deyeuxia angustifolia.