Hirono I, Masuda T, Aoki T
Laboratory of Genetics and Biochemistry, Tokyo University of Fisheries, Japan.
Microb Pathog. 1996 Sep;21(3):173-82. doi: 10.1006/mpat.1996.0052.
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.
使用黏粒载体charomid9 - 36从鱼类病原菌鳗弧菌中克隆出一个编码溶血素的5 kb DNA片段。溶血素基因（VAH1）的一个开放阅读框为2253 bp，对应一个由751个氨基酸残基组成的蛋白质。VAH1基因推导的氨基酸序列与先前报道的霍乱弧菌EI Tor溶血素、创伤弧菌细胞毒素 - 溶血素、嗜水气单胞菌AHH1溶血素和杀鲑气单胞菌ASH4溶血素显示出显著程度的序列同源性，总体氨基酸同一性分别为57.3%、25.8%、46.2%和43.7%。使用VAH1作为探针在高严格条件下进行DNA杂交分析表明，VAH1与28株鳗弧菌中的25株杂交，包括A至I血清型，但不与其他弧菌属、嗜水气单胞菌或杀鲑气单胞菌杂交。通过PCR成功地从鳗弧菌感染的鱼类组织中扩增出VAH1的目标DNA片段。
