The 70-kDa heat shock proteins (HSP70) play crucial roles in protecting cells against environmental stresses, such as heat shock, heavy metals and pathogenic bacteria. The full-length HSP70 cDNA of Sepiella maindroni (designated as SmHSP70, GenBank accession no. KJ739788) was 2109bp, including an ORF of 1950bp encoding a polypeptide of 649 amino acids with predicted pI/MW 5.24/71.30kDa, a 62bp-5′-UTR and a 97bp-3′-UTR. BLASTp analysis and phylogenetic relationship strongly suggested that the amino acid sequence was a member of HSP70 family. Multiple sequence alignment revealed that SmHSP70 and other known HSP70 were highly conserved, especially in the regions of HSP70 family signatures, the bipartite nuclear targeting sequence, ATP/GTP-binding site motif and ‘EEVD’ motif. Time-dependent mRNA expression of SmHSP70 in the liver was recorded by quantitative real-time RT-PCR after Vibrio harveyi injection and Cd2+ exposure. The results indicated that SmHSP70 played a significant role in mediating the environmental stress and immune response against pathogens.