青花菜谷胱甘肽-S-转移酶基因克隆及其表达分析

为探讨青花菜在模拟酸雨胁迫下谷胱甘肽-S-转移酶的表达变化,克隆了青花菜谷胱甘肽-S-转移酶基因(glutathione-S-transferase,GST)的cDNA序列全长,并进行了生物信息学和表达分析。结果表明:青花菜GST基因cDNA全长为915bp,开放阅读框为642bp,编码213个氨基酸,推测分子式为C1091H1719N289O306S5,分子量为23 940.7,没有跨膜螺旋区域和信号肽。系统进化树分析表明,该青花菜基因GST与芥菜的GST聚类关系最近。实时荧光定量PCR结果显示,在模拟酸雨胁迫下,GST基因的表达量在胁迫初期显著增大,随时间延长开始下降,表明其参与了青花菜抗酸雨的应答反应。

Cloning of GST and Its Expression in Broccoli (Brassica oleracea var.italica)

The full-length cDNA of GST was cloned in order to explore molecular response mechanism in broccoli under acid rain stress,which was analyzed by bioinformatics method and real-time PCR.The results showed that the full-length DNA was of 915 bp,opening reading frame(ORF) was of 642 bp.And the ORF encoded 213 amino acids,suggesting a formula of C₁₀₉₁H₁₇₁₉N₂₈₉O₃₀₆S₅,a molecular weight of 23 940.7,had no transmembrane helix region or signal peptide.Phylogenetic tree analysis revealed that this gene of broccoli had closer relationship with that from Brassica juncea.The analysis by real-time PCR suggested that the expression of GST genes in broccoli under acid rain stress increased significantly at first,and then declined due to longer duration of acid rain stress,which was involved in signaling pathways in response to the acid rain stress.