Cloning and functional characterization of thioredoxin genes from large yellow croaker Larimichthys crocea. Issue 77 (June 2018)

• Record Type: Journal Article
• Authors: Chen, Mengnan; Zhang, Jianshe; Xie, Xiaoze; Wu, Changwen
• Is Part Of: Fish & shellfish immunology. Issue 77(2018)
• Page Start: 385
• Page End: 391
• Publication Date: 2018-06
• Subjects: Thioredoxin -- Larimichthys crocea -- Expression analysis -- Real-time quantitative PCR; Fishes -- Immunology -- Periodicals; Shellfish -- Immunology -- Periodicals; Poissons -- Immunologie -- Périodiques; Crustacés -- Immunologie -- Périodiques; 571.9617
• DOI: 10.1016/j.fsi.2018.03.045
• Languages: English
• ISSNs: 1050-4648

Abstract

Abstract: Thioredoxin(Trx)with a redox-active disulfide/dithiol in the active site, is an ubiquitous disulfide reductase majorly responsible for maintaining the balance of reactive oxygen species. In this study, the complete thioredoxin-like protein 1 (designated as LcTrx) was cloned from large yellow croaker Larimichthys crocea through rapid amplification of cDNA ends. The full-length cDNA of LcTrx was 1295 bp in length containing a 131 bp 5′ untranslated region (UTR), a 3′UTR of 294bp with a poly (A) tail, and an 870 bp open reading frame (ORF) encoding a polypeptide of 289 amino acids. Protein sequence analysis revealed that LcTrx contains the evolutionarily conserved redox motif CRPC (Cys-Arg-Pro-Cys-). Multiple alignments revealed that LcTrx is highly identical to Trx from other organisms, especially in the CRPC motifs. The recombinant LcTrx showed obvious insulin reduction activity in vitro. The LcTrx transcripts were constitutively expressed in all examined tissues with the highest levels found in the muscles and the lowest in the head kidney. Results of Vibrio parahaemolyticus infection experiment showed that the expression levels of LcTrx were tissue and time dependent. In the liver and kidney, LcTrx was down-regulated both at 12 h and 48 h post-infection. In contrast, LcTrx showed induced expression in the spleen and head kidney at same post-infection time points. The different responses to pathogen stimulation indicated the diversified physiological function of LcTrx in the four examined tissues. Highlights: Clone the full-length cDNA of Trx from large yellow croaker. Investigate the expression profile of thioredoxin-like protein 1 (LcTrx) transcript after a bacterial challenge. Recombinant LcTrx protein in vitro and test its antioxidant activity.