Cloning and expression analysis of hif-2? gene in cobia (Rachycentron canadum) under hypoxia stress
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Hypoxia-inducible factor 2? (hif-2?) is a critical regulator of hypoxia response and plays a vital role in hypoxia stress in the organism. To understand the regulatory role of hif-2? in response to hypoxia stress in juvenile cobia (Rachycentron canadum), hif-2? was cloned using Rapid Amplification of cDNA Ends (RACE) technology. The full length of hif-2? is 4021 bp, with 2634 bp open reading frame (ORF), 5' non-coding region (5'UTR), 285 bp, 3' non-coding region (3'-UTR), 1102 bp, and encoding 877 amino acids. The encoded protein contains the HLH (Helix-loop-helix) domain (amino acids 20-75), the PAS (PER-ARNT-SIM) domain (amino acids 91-157 and 237-303), and the PAC (PAS Associated C-terminal) domain (amino acids 309-352). The results of phylogenetic tree analysis showed that hif-2? in cobia clustered with hif-2? in Echeneis naucrates and were closely related. Real-time fluorescence quantitative PCR (qRT-PCR) was used to analyze the expression of hif-2? in nine different tissues of cobia and the expression of hif-2? mRNA in the liver and gill under hypoxia stress. The results suggested that the hif-2? was expressed in all tissues of the cobia, with higher expression in the liver and gill. Under hypoxia stress, the expression of the hif-2? in the liver and gill was tissue-specific. In liver tissues, hif-2? expression was significantly higher than that of the control at 14 and 28 days. In gill tissues, hif-2? expression decreased at 7, 14, and 28 days and was lowest at 14 day. The results suggest that hif-2? plays a vital role in hypoxic stress in cobia and may provide basic information for studying the molecular genetic mechanism of hypoxia tolerance in cobia.
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