Molecular characterization of heat shock protein 70 gene transcripts during Vibrio harveyi infection of humphead snapper, Lutjanus sanguineus.

ABSTRACT

In the present study, heat shock cognate 70 (HSC70) and inducible heat shock protein 70 (HSP70) gene of humphead snapper, Lutjanus sanguineus, were cloned by rapid amplification of cDNA ends (RACE) technique with the primers designed from the known expressed sequence tags (ESTs) identified from the subtracted cDNA library of the head kidney of humphead snapper. BLAST program analysis indicated that both HSC70 and HSP70 shared high homology with their counterparts in other species. However, the homology between HSC70 and HSP70 is only 82.5% identity. Phylogenetic trees were constructed by the neighbor-joining method, and the results suggested that both HSC70 and HSP70 could be used for phylogenetic analysis at order levels. The expression profiles of HSC70 and HSP70 were measured by fluorescent real-time RT-PCR after Vibrio harveyi infection. Our results suggested that both HSC70 and HSP70 could be induced by V. harveyi challenge. However, the expression pattern of HSP70 showed some differences compared with that of HSC70. Original level of HSP70 in head kidney was lower than that of HSC70. The expression of HSP70 could increase faster and last longer than that of HSC70 and maintain a high level from the time point of 6-15 h. Our results suggested that the rapid transcriptional upregulation of HSC70 and HSP70 in response to V. harveyi infection might be important for the survival of humphead snapper.