Molecular cloning, expression and antioxidative activity of 2-cys-peroxiredoxin from freshwater mussel Cristaria plicata. Issue 66 (July 2017)

• Record Type: Journal Article
• Authors: Wang, Xiaobo; Hu, Baoqing; Wen, Chungen; Zhang, Ming; Jian, Shaoqing; Yang, Gang
• Is Part Of: Fish & shellfish immunology. Issue 66(2017)
• Page Start: 254
• Page End: 263
• Publication Date: 2017-07
• Subjects: Cristaria plicata -- Peroxiredoxin -- Molecular clone -- Recombinant protein -- Antioxidant activity; Fishes -- Immunology -- Periodicals; Shellfish -- Immunology -- Periodicals; Poissons -- Immunologie -- Périodiques; Crustacés -- Immunologie -- Périodiques; 571.9617
• DOI: 10.1016/j.fsi.2017.05.026
• Languages: English
• ISSNs: 1050-4648

Abstract

Abstract: Peroxiredoxins (Prxs) play an important role against various oxidative stresses by catalyzing the reduction of hydrogen peroxide (H2 O2 ) and organic hydroperoxides to less harmful form. A 2-cys peroxiredoxin, designated as CpPrx, was cloned from hemocytes of freshwater mussel Cristaria plicata . The full length cDNA of CpPrx is 1247 bp, which includes an open reading frame (ORF) of 591bp, encoding 196 amino acids. CpPrx possesses two conserved cysteine residues (Cys49, Cys170). The deduced amino acid sequence of CpPrx showed a high level (67–74%) of sequence similarity to 2-Cys Prxs from other species. The results of real-time quantitative PCR revealed that CpPrx mRNA was constitutively expressed in tissues, and the highest expression levels were in hepatopancreas and gills. After peptidoglycan (PGN) and Aeromonas hydrophila challenge, the expression levels of CpPrx mRNA were up-regulated in hemocytes and hepatopancreas. The cDNA of CpPrx was cloned into the plasmid pET-32, and the recombinant protein was expressed in Escherichia coli BL21(DE3). Comparison with DE3-pET-32 and DE3 strain, the cells of DE3-pET-32-CpPrx exhibited resistance to the concentration of 0.4, 0.8 and 1.2 mmoL/L H2 O2 in vivo. Highlights: The full cDNA sequences of CpPrx were cloned. The transcripts of CpPrx were up-regulated after stimulation. The recombinant had antioxidant activity.