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KMID : 0613820180280060639
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Journal of Life Science
2018 Volume.28 No. 6 p.639 ~ p.647
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Molecular Cloning and Expression Analysis of Red-spotted Grouper, Epinephelus akaara Hsp70
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Min Byung-Hwa
Hur Jun-Wook
Park Hyung-Jun
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Abstract
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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℃) for sampled livers revealed that expression gradually increased at 12℃ compared to 21℃. 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.
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KEYWORD
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Epinephelus akaara, expression, Hsp70, RACE PCR, quantitative real-time PCR
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