福建农业学报2023，Vol.38Issue(10)：1185-1194,10.DOI:10.19303/j.issn.1008-0384.2023.10.007
细菌刺激下克氏原螯虾prx4基因的免疫响应和抗氧化功能研究
Immune Response and Antioxidant Activities of Bacteria-stimulated Prx4 in Procambarus clarkii
摘要
Abstract
[Objective]Immune response of peroxiredoxin 4 gene of Procambarus clarki(Pc-prx4)after being stimulated by bacterial infection was studied.[Methods]Pc-prx4 were cloned before and after an artificial inoculation of Staphylococcus aureus or Edwardsiella ictaluri on the crayfish.Bioinformatic analysis was conducted with the aid of relevant online websites and software including Expasy,translate tool,SMART,Expasy ProtParam tool,BLASTx,and MEGA-X.Expressions of Pc-prx4 from the tissues of normal and pathogenically inoculated crayfish were detected by qRT-PCR.Antioxidant function of the constructed recombinant protein was determined.[Results]Pc-prx4 had an open reading frame of 744 bp encoding 247 amino acids with an alkyl hydroperoxide reductase(AhpC)structural domain,a sulfhydryl-specific antioxidant structural domain,and a C-terminal structural domain of 1-Cys prx peroxisome iron oxidase.The predicted protein molecular formula was C1249H1934N330O357S10 with a molecular weight of 27.61 kDa and a pI of 5.88.Its signal peptide region was located at the N-terminal end,and the 1-Cys peroxiredoxin domain at the C-terminal end.The gene expressed in all sampled tissues—most highly in the hemolymph and high in the hemocytes,hepatopancreas,gills,and intestines of the crayfish inoculated by S.aureus or E.catarrhalis.In the plasmid DNA protection assay,rPc-PRX4 displayed varying degrees of antioxidant activity,especially in high concentrations.[Conclusion]A kind of 1-Cys prx,Pc-prx4 was highly expressed in the hemolymph of P.clarkii stimulated by S.aureus or E.catarrhalis.The protein exhibited a concentration-dependent activity associated with the antioxidant homeostasis of the crayfish.关键词
克氏原螯虾/过氧化物还原酶4/金黄色葡萄球菌/鲶爱德华氏菌/表达模式/抗氧化Key words
Procambarus clarkii/peroxiredoxin 4/Staphylococcus aureus/Edwardsiella ictaluri/expression of target genes/antioxidant分类
农业科技引用本文复制引用
张明达,李倩倩,张文硕,秦仕宇,沈秀丽,杜志强..细菌刺激下克氏原螯虾prx4基因的免疫响应和抗氧化功能研究[J].福建农业学报,2023,38(10):1185-1194,10.基金项目
国家自然科学基金项目(32060834) （32060834）
内蒙古科技大学 2022 年基本科研业务费专项(028) （028）
