Cloning and expression analysis of Keap1 of golden pompano (Trachinotus ovatus) and response to oxidized fish oil and LPS administration

Keap1 is an important component of Nrf2-Keap1 signaling pathways. However, information about Keap1 in T. ovatus is quiet limit. Thus, the full length of Keap1 was cloned from T. ovatus in this study. The full-length cDNA sequence was 3373 bp in length and comprised of a 1878 bp ORF preceded by 1205bp 5′-UTR, and followed a 290 bp 3′- UTR. The putative amino acid sequence of the Keap1 contains 625 residues. The Keap1 amino acid sequence contained a BTB domain, an IVR domain and a DGR domain. The phylogenetic tree showed that Seriola dumerili had the highest identity with T. ovatus in Keap1 amino acid. The results show that Keap1 gene was predominantly expressed in liver, kidney, spleen and gill, significantly higher than that of other tissues. The relative expression of Keap1 in foregut and heart were moderately expressed. After feeding with oxidized fish oil for 8 weeks, the head kidney expression levels of Keap1, Nrf2, HO-1, SOD and CAT in T. ovatus were significantly higher than those in control group. Administration of LPS for 72 h in T. ovatus activated the Nrf2-Keap1 pathways and upregulated the expression level of HO-1, GSH-PX, SOD and CAT. In conclusion, Keap1 is evolutionarily and functionally conserved in T. ovatus. Keap1 expression level was significantly higher in oxidative stress caused by oxidized fish oil and LPS administration, suggesting that Keap1 plays a pivotal role in Nrf2-Keap1 pathway of T. ovatus.

Abstract licence: CC BY-NC

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