Identification and expression of cathepsin B from the freshwater mussel Cristaria plicata

Cathepsin B plays crucial roles in host immune defense against pathogen infection. In present study, a cathepsin B gene from the freshwater mussel, Cristaria plicata (CpCathB) was cloned and characterized. The full-length cDNA of CpCathB was 1825 bp, and contained a 5′ untranslated region (UTR) of 36 nucleotides, an open reading frame (ORF) of 1044 bp and a 3′ UTR of 745 bp with a poly (A) tail. The deduced CpCathB protein was encoded as a preproenzyme with 347 amino acid residues and predicted molecular weight of 38.55 kDa. Sequence alignment revealed that CpCathB protein shared 56% - 60.7% identity comparison with other species. The predicted tertiary structure of CpCathB protein was highly similar to that of human. The CpCathB mRNA was expressed in hemocytes, hepatopancreas, adductor muscle, gills and mantle tissues of healthy mussels, and the highest expression level was in hepatopancreas. The transcripts of CpCathB were increased in hemocytes and hepatopancreas from mussels after Aeromonas hydrophila challenge. Moreover, the recombinant CpCathB was expressed in the Escherichia coli Rosetta-gami (DE3) strain. The maximum titer of the anti-CpCathB polyclonal antibodies was 1:640,000.The CpCathB protein had a higher expression in hepatopancreas and mantle and a lower level in hemocytes.