Cloning of catalase gene and antioxidant genes in Scophthalmus maximus response to metalloprotease of Vibrio anguillarum stress  

作　　者：Hai REN Jian LI Ping LIU Xianyun REN Tao SONG Guisheng GAO Duwen LI Shuaiting LIU 

机构地区：[1]Hebei Key Laboratory of Preventive Veterinary Medicine,Hebei Normal University of Science and Technology,Qinhuangdao 066004,China [2]Key Laboratory of Sustainable Development of Marine Fisheries,Ministry of Agriculture,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao 266071,China

出　　处：《Journal of Oceanology and Limnology》2022年第1期322-335,共14页海洋湖沼学报（英文）

基　　金：Supported by the National Natural Science Foundation of China(No.31502187);the Natural Science Foundation of Hebei Province(No.C2018407049);the Hebei Provincial Department of Science and Technology(Nos.20286701Z,20567621H);the Talent Engineering Training Funding Project of Hebei Province(No.A201901057)。

摘　　要：Metalloproteases represent a class of extracellular proteases found in Vibrio anguillarum that can generate toxic and pathogenic eff ects in turbot(Scophthalmus maximus).The toxicological eff ect partly results from oxidative damage due to the production of excessive reactive oxygen species(ROS).Catalase(CAT),superoxide dismutase(SOD),and glutathione peroxidase(GPx)are major antioxidant enzymes induced by various oxidative stresses and can scavenge peroxides generated in cells.To evaluate the eff ects of metalloprotease-induced ROS on the antioxidation defense mechanism of S.maximus head kidney cells,the cDNA of CAT gene(designated as SmCAT)was cloned and characterized.SmCAT comprises a 1584-bp coding sequence that encodes a protein containing 527 amino acids with a poly(A)tail.Bioinformatics analysis revealed an active site signature sequence,a heme-ligand signature sequence,and three catalytic amino acid residues.The deduced SmCAT amino acid sequence shares a sequence similarity of 66.1%-92.4%with those of other species.Phylogenetic analysis revealed that SmCAT is classifi ed with CAT of other fi shes.Quantitative real-time PCR analysis showed that SmCAT was extensively expressed in all tested tissues,especially in blood.The expression of SmCAT,SmMnSOD,and SmGPx were inhibited signifi cantly in head kidney cells treated with metalloprotease from 12 to 24 h.In 6 to 24 h metalloprotease-treated groups compared to that of the untreated group,it was found that the production of ROS was markedly increased,and the mitochondrial membrane potential was decreased considerably.Hoechst 33342 staining revealed the presence of apoptotic bodies when the cells were incubated with 8.0 or 40.0μg/mL metalloprotease for 12 and 24 h.Hence,the toxic eff ects of metalloprotease are associated with the down-regulation of antioxidant enzyme expression and increased ROS levels,which trigger the activation of apoptosis in the head kidney cells of turbot.Our fi ndings provide a better understanding on the mechanism of metalloprotease-in

关 键 词：gene cloning EXPRESSION reactive oxygen species METALLOPROTEASE head kidney cells 

分 类 号：S943[农业科学—水产养殖]