Cloning, molecular characterization and expression analysis of heat shock cognate 70 (Hsc70) cDNA from turbot (Scophthalmus maximus).

As an essential member of the HSP70 family, heat shock cognate 70 (Hsc70) is a constitutively expressed molecular chaperone involved in protein metabolism. In this paper, turbot Hsc70 was cloned and the expression profile was also analyzed. The full-length cDNA of the turbot Hsc70 was 2,292 bp in length, including a 113-bp 5' UTR, a 223-bp 3' UTR and a 1,956-bp open reading frame coding a protein with 651 amino acid residues. Comparison of amino acid sequence revealed the existence of three classical HSP70 family signature motifs, a signature nonapeptide and one repeat of tetrapeptide in turbot Hsc70. The turbot Hsc70-deduced amino acids sequence exhibited 75.4-96.8 % homology with Hsp70s/Hsc70s of 24 other known sequences. In particular, the strongest homology was found with the cognate members of Hsc70 subfamily and the highest identity was found with Japanese flounder Hsc70. Semi-quantitative RT-PCR revealed that turbot Hsc70 transcripts were stably expressed in all tested tissues under normal physiological condition, while the expression levels also increased (~1.5-fold to ~threefold) after heat shock and bacterial infection. In addition, Hsc70 transcripts were detected throughout embryonic development and in turbot embryonic cell line (TEC) in the absence of any stress. Meanwhile, it was also heat inducible, but not cold inducible in TEC. These results suggest that Hsc70 gene may be involved in embryogenesis and cellular protection events under normal and stress condition.