Expression pattern of heat-shock cognate 70 gene of humphead snapper, Lutjanus sanguineus (Cuvier), infected by Vibrio harveyi.

ABSTRACT

The heat-shock cognate 70 (HSC70) gene of humphead snapper, Lutjanus sanguineus, designated as ByHSC70, was cloned by rapid amplification of cDNA ends (RACE) with the primers designed from the known expressed sequence tag (EST) identified from the subtracted cDNA library of the head kidney of humphead snapper. The full-length cDNA of ByHSC70 is 2313 bp, containing a 5' terminal untranslated region (UTR) of 96 bp, a 3' terminal UTR of 267 bp, and an open reading frame (ORF) of 1950 bp encoding a polypeptide of 650 amino acids with a theoretical molecular weight of 71.21 kDa and an estimated isoelectric point (pI) of 5.08. ByHSC70 contained three classical HSP70 family signatures. BLAST analysis showed that the amino acid sequence of ByHSC70 had the highest similarity of 99% when compared with other HSC70s. Fluorescent real-time quantitative RT-PCR was used to examine the expression of ByHSC70 gene in eight kinds of tissues/organs of humphead snapper after challenge with Vibrio harveyi. There was a clear time-dependent expression pattern of ByHSC70 in head kidney, spleen and thymus after bacterial challenge, and the expression of mRNA reached a maximum level at 9, 6 and 24 h post-infection and then returned to control levels after 15, 24 and 36 h, respectively. Our results suggest that HSC70 is an important component in the immune system of humphead snapper, its their rapid transcriptional upregulation in response to V. harveyi infection might be important for survival of humphead snapper.