Cloning and detection of the hemolysin gene of Vibrio anguillarum.
A 5 kb DNA fragment encoding a hemolysin was cloned from the fish pathogenic bacterium Vibrio anguillarum using the cosmid vector charomid9-36. An open reading frame of the hemolysin gene (VAH1) was 2253 bp and corresponded to a protein of 751 amino acid residues. The deduced amino acid sequence of the VAH1 gene and the previously reported Vibrio cholerae EI Tor hemolysin, V. vulnificus cytolysin-hemolysin, Aeromonas hydrophila AHH1 hemolysin, and A. salmonicida ASH4 hemolysin showed a significant degree of sequence homology, and the overall amino acid identities were 57.3%, 25.8%, 46.2%, and 43.7%, respectively. DNA hybridization analysis under high-stringent conditions using VAH1 as a probe, demonstrated that VAH1 hybridized with 25 out of 28 strains of V. anguillarum including serotypes A to I, but did not hybridize with other species of Vibrio, A. hydrophila or A. salmonicida. The targeted DNA fragment of VAH1 was successfully amplified from V. anguillarum-infected fish tissues by PCR.
Hirono
I
I
Laboratory of Genetics and Biochemistry, Tokyo University of Fisheries, Japan.
Masuda
T
T
Aoki
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D002347:Carps / Q000382:microbiology; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D006460:Hemolysin Proteins; D006461:Hemolysis; D008969:Molecular Sequence Data; D017686:Oncorhynchus mykiss / Q000382:microbiology; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014733:Vibrio / Q000235:genetics*
D001426:Bacterial Proteins; D006460:Hemolysin Proteins; C103308:VAH1 protein, Listonella anguillarum