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http://dx.doi.org/10.5352/JLS.2018.28.6.639

Molecular Cloning and Expression Analysis of Red-spotted Grouper, Epinephelus akaara Hsp70  

Min, Byung Hwa (Aquaculture Industry Research Division, East Sea Fisheries Research Institute)
Hur, Jun Wook (Bio-Monitoring Center)
Park, Hyung Jun (Aquaculture Management Division, National Institute of Fisheries Science)
Publication Information
Journal of Life Science / v.28, no.6, 2018 , pp. 639-647 More about this Journal
Abstract
A new heat shock protein 70 was identified in red-spotted grouper (Epinephelus akaara) based on an expression analysis. The cDNA of red-spotted grouper Hsp70 (designated RgHsp70) was cloned by the rapid amplification of cDNA ends (RACE) techniques. The full-length of RgHsp70 cDNA was 2,152 bp, consisting of a 5'-terminal untranslated region (UTR) of 105 bp, a 3'-terminal UTR of 274 bp, and an open reading frame (ORF) of 1,773 bp that encode a polypeptide of 590 amino acids with a theoretical molecular weight of 64.9 kDa and an estimated isoelectric point of 5.2. Multiple alignment and phylogenetic analyses revealed that the RgHsp70 gene shares a high similarity with other Hsp70 fish genes. RgHsp70 contained all three classical Hsp70 family signatures. The results indicated the RgHsp70 is a member of the heat shock protein 70 family. RgHsp70 mRNA was predominately expressed in the liver, with reduced expression noted in the head-kidney tissues. The expression analysis of different water temperatures (21, 18, 15 and $12^{\circ}C$) for sampled livers revealed that expression gradually increased at $12^{\circ}C$ compared to $21^{\circ}C$. In this study, the effects of water temperature lowering on the physiological conditions were investigated, and the results revealed that novel RgHsp70 may be an important molecule involved in stress responses.
Keywords
Epinephelus akaara; expression; Hsp70; RACE PCR; quantitative real-time PCR;
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