二氯异氰尿酸钠胁迫下菲牛蛭GST基因分子克隆及表达规律研究

Cloning and Expression of GST Gene in Poecilobdella manillensis under Sodium Dichloroisocyanurate Stress

摘要: 谷胱甘肽转移酶(GST)是一种重要的解毒酶，在抗氧化和细胞解毒中起着重要作用。二氯异氰尿酸钠属于含氯消毒剂，常用于养殖行业、自来水、医院等杀灭细菌。为了研究菲牛蛭GST基因在应对二氯异氰尿酸钠胁迫中的作用，本研究对菲牛蛭GST基因进行了克隆和表达规律研究。结果表明，菲牛蛭GST基因编码区全长是654 bp，编辑217个氨基酸，其蛋白质序列与红鲍螺，红鲍，绿叶海蛞蝓，角突臂围轮虫，紫贻贝相应序列相似性分别为97%，97%，97%，96%，96%。实时荧光定量PCR结果显示GST基因在胁迫后表达量先下降后升高，在12 h表达量达到最低，48 h表达量最高，是12 h表达量的93倍。研究结果显示菲牛蛭GST基因参与了二氯异氰尿酸钠胁迫的解毒过程，为今后菲牛蛭人工养殖过程中含氯消毒剂的合理使用，及解析GST在生物体内排毒机制提供了参考。

Abstract: Glutathione transferase (GST) is an important detoxification enzyme, which plays an important role in antioxidant and cellular detoxification. Sodium dichloroisocyanurate is a chlorine-containing disinfectant, which is often used to kill bacteria in aquaculture industry, tap water, hospitals, etc. In order to study the role of GST gene in coping with sodium dichloroisocyanurate stress, the cloning and espression of GST gene in Poecilobdella manillensis were studied. The results showed that the total length of the GST gene coding region of the Poecilobdella manillensis was 654 bp, and the protein sequences were edited with 217 amino acids, and the protein sequences were 97%, 97%, 97%, 96% and 96% of the corresponding sequences of Haliotis rufescens, Haliotis rubra, Plakobranchus ocellatus, Brachionus angularis and Mytilus galloprovincialis. The real-time PCR results showed that the expression of GST gene decreased first and then increased after stress, reaching the lowest expression at 12 h and the highest expression at 48 h, which was 93 times the expression at 12 h. The results showed that the Poecilobdella manillensis GST gene participated in the detoxification process of sodium dichloroisocyanurate stress, which provided a reference for the rational use of chlorine-containing disinfectants in the artificial breeding process of Fifei bull leech in the future, and the analysis of the detoxification mechanism of GST in organisms.