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Applied and Environmental Microbiology, November 2002, p. 5480-5487, Vol. 68, No. 11
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.11.5480-5487.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Expression of Duplicate msa Genes in the Salmonid Pathogen Renibacterium salmoninarum

Northwest Fisheries Science Center, National Marine Fisheries Service, National Oceanic and Atmospheric Administration, United States Department of Commerce, Seattle, Washington 98112

Received 8 April 2002/ Accepted 24 August 2002

Renibacterium salmoninarum is a gram-positive bacterium responsible for bacterial kidney disease of salmon and trout. R. salmoninarum has two identical copies of the gene encoding major soluble antigen (MSA), an immunodominant, extracellular protein. To determine whether one or both copies of msa are expressed, reporter plasmids encoding a fusion of MSA and green fluorescent protein controlled by 0.6 kb of promoter region from msa1 or msa2 were constructed and introduced into R. salmoninarum. Single copies of the reporter plasmids integrated into the chromosome by homologous recombination. Expression of mRNA and protein from the integrated plasmids was detected, and transformed cells were fluorescent, demonstrating that both msa1 and msa2 are expressed under in vitro conditions. This is the first report of successful transformation and homologous recombination in R. salmoninarum.

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