葡萄风信子谷胱甘肽S转移酶基因克隆与表达分析

Clone and Characterization of A Glutathione-s-transferase Gene in Muscari armeniacum

以亚美尼亚葡萄风信子为材料,利用本课题组前期获得的葡萄风信子转录组数据库,根据已经获得的葡萄风信子GST基因片段,通过PCR技术克隆得到葡萄风信子GST基因的c DNA序列,命名为Ma GST。Ma GST的c DNA全长为711 bp,开放阅读框为666 bp,编码221个氨基酸,推测蛋白质分子量为54.1 k D,理论等电点p I为5.13。利用生物信息分析软件对Ma GST基因进行同源性比对和系统进化分析表明,结果显示该基因编码的氨基酸具有谷胱甘肽S转移酶典型的C端与N端双结构域,属于GST Tau家族蛋白;与洋葱和小麦GST基因的一致性分别为76.72%和62.50%。实时定量PCR结果显示Ma GST在葡萄风信子各组织器官表达强度相似,属于组成型表达;水杨酸(SA)可以明显诱导Ma GST表达,Ma GST对氯化钠(Na Cl)应答不是很明显,甚至表达量有稍微的下调。

Based on the latex the transcriptome database, the full length cDNA of glutathione-S-transferase （GST） from Muscari armeniacum was cloned by reverse-PCR and PCR, designated as MaGST. The full length eDNA of MaGST was 711 bp, and the ORF （Open Reading Frame） length was 666 bp, encoding a protein polypeptide of 221 amino acids with a predicted molecular weight of 54.1 kD and pI of 5.13. By phylogenetic tree analysis, the putative MaGST protein displayed identities to the GSTs of Allium cepa and Triticum aestivum of 76.72% and 62.50%, respectively, and contained the Tau GST-specific N-terminal domain（ G site） and the C-terminal domain（ H site）, belonging to the family of GST Tau. By real-time PCR, the expression pattern of MaGST in different organs were similar, belonging to constitutive expression. The expression of MaGST was regulated by salicylic acid, but not by the NaCl.