Identification, expression and bioactivity of Paramisgurnus dabryanus β-defensin that might be involved in immune defense against bacterial infection

Abstract

β-defensins are a large family of multi-disulfide-bonded peptides with broad-spectrum antimicrobial activities that contribute to innate host defense in many organisms, but little information is available about β-defensins produced by freshwater fish lacking scales. We therefore cloned and identified a β-defensin gene from Chinese loach (Paramisgurnus dabryanus) by designing degenerate primers and using thermal asymmetric interlaced PCR. This gene is the first defensin gene ever identified in a non-scaled freshwater fish. Annotation of the protein domain architecture showed that the putative Chinese loach β-defensin contains the signature motif of six conserved cysteines within the mature peptide, an aspect similar to β-defensins of other marine fish. We also used quantitative real-time PCR to investigate the expression pattern of the Chinese loach β-defensin gene, mRNA of which could be observed in various tissues. After challenge with the pathogenic bacterium Aeromonas hydrophila, β-defensin expression was induced in the eye, gill, skin, and spleen of the adult loach. The bioactivity of the recombinant P. dabryanus β-defensin was examined against pathogenic bacteria, and the results suggest that this class 2 β-defensin has potential applications for treatment of bacterial infections.