Cloning of the gene for the surface array protein of Aeromonas salmonicida and evidence linking loss of expression with genetic deletion

Abstract

A gene bank of DNA from the fish pathogenic bacterium Aeromonas salmonicida was constructed in the bacteriophage lambda gt11. Phage lambda gt11/10G, a recombinant carrying a 4.0-kilobase fragment of A. salmonicida DNA, was found to express the surface array protein (A protein) in Escherichia coli. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the protein expressed from the cloned gene had a subunit molecular weight of 49,000, which was identical to that of subunits in the native assembled A layer. Genomic Southern analysis showed that the gene coding for this predominant cellular protein was in a single copy on the chromosome and was conserved among a wide range of A. salmonicida strains with different phenotypic characteristics and isolated from diverse geographic locations, fish species, and means of pathogenesis. Results of genomic blotting experiments also showed that loss of expression of the A layer resulting from growth at 30 degrees C was accompanied by genetic rearrangement in which N-terminal sequences of the gene for A protein were lost by deletion.