Cloning,characterization, hypoxia and heat shock response of hypoxia inducible factor-1 (HIF-1) from the small abalone Haliotis diversicolor |
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Authors: | Xiuhong Cai Yitao Huang Xin Zhang Shuhong Wang Zhihua Zou Guodong Wang Yilei Wang Ziping Zhang |
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Institution: | 1. Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture, Fisheries College, Jimei University, Xiamen 361021, China;2. Department of Natural Sciences and Mathematics, State University of New York at Cobleskill, NY 12043, USA |
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Abstract: | In this study, hypoxia inducible factor-1α (HIF-1α) and hypoxia inducible factor-1β (HIF-1β) from small abalone Haliotis diversicolor were cloned. The cDNA of H. diversicolor HIF-1α (HdHIF-1α) is 2833 bp encoding a protein of 711aa and H. diversicolor HIF-1β (HdHIF-1β) is 1919 bp encoding a protein of 590aa. Similar to other species' HIF-1, HdHIF-1 has one basic helix–loop–helix (bHLH) domain and two Per-Arnt-Sim (PAS) domains, and HdHIF-1α has a oxygen-dependent degradation domain (ODDD) with two proline hydroxylation motifs and a C-terminal transactivation domain (C-TAD) with an asparagine hydroxylation motif. Under normoxic conditions, HdHIF-1α and HdHIF-1β mRNAs were constitutively present in all examined tissues. Under hypoxia (2.0 mg/L DO at 25 °C) stress, HdHIF-1α expression was up-regulated in gills at 4 h, 24 h and 96 h, and in hemocytes at 24 h and 96 h, while HdHIF-1β remained relatively constant. Under thermal stress (31 °C), HdHIF-1α expression was significantly increased in gills at 4 h, and hemocytes at 0 h and 4 h, while HdHIF-1β expression still remained relatively constant. These results suggested that HIF-1α may play an important role in adaption to poor environment in H. diversicolor. |
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Keywords: | HIF-1α hypoxia inducible factor-1α HIF-1β hypoxia inducible factor-1β ARNT aryl hydrocarbon receptor nuclear translocator bHLH/PAS basic helix&ndash loop&ndash helix/Per-Arnt-Sim CBP CREB1-binding protein C-TAD C-terminal transactivation domain DO dissolved oxygen HRE hypoxia response element NRR neutral red retention NRU neutral red uptake N-TAD N-terminal transactivation domain ODDD oxygen dependent degradation domain ORF open reading frame PHDs proline hydroxylases Pro proline pVHL von Hippel&ndash Lindau protein qRT-PCR quantitative real-time PCR RACE rapid amplification of cDNA ends THC hemocyte count UTR untranslated region |
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