Yamamoto K, Ichinose Y, Nakasone N, Tanabe M, Nagahama M, Sakurai J, Iwanaga M
Infect Immun. 1986 Mar;51(3):927-31. doi: 10.1128/iai.51.3.927-931.1986.
Hemolysins purified from non-O1 Vibrio cholerae (non-O1 hemolysin) and a Vibrio cholerae O1, biotype El Tor (El Tor hemolysin) were investigated for their homology. The hemolysins were isolated from the culture supernatant fluids by ammonium sulfate precipitation and gel filtration on Sephadex G-100 columns. The purified hemolysins gave single bands with an identical mobility on conventional polyacrylamide gel disc electrophoresis. The molecular weights of the non-O1 and El Tor hemolysins were estimated to be about 60,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the amino acid compositions of the hemolysins were very similar. The specific activities of the hemolysins were identical, and both hemolysins were neutralized to the same extent with antisera against the homologous and heterologous hemolysins. Ouchterlony double immunodiffusion tests with both hemolysins and antihemolysin serum gave a common (fused) precipitin line. These data indicate that the non-O1 hemolysin is biologically, physicochemically, and immunologically indistinguishable from the El Tor hemolysin.
对从非O1群霍乱弧菌中纯化的溶血素(非O1溶血素)和O1群霍乱弧菌埃尔托生物型(埃尔托溶血素)的同源性进行了研究。通过硫酸铵沉淀和在Sephadex G - 100柱上进行凝胶过滤,从培养上清液中分离出溶血素。纯化后的溶血素在常规聚丙烯酰胺凝胶圆盘电泳上呈现出具有相同迁移率的单一条带。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳估计,非O1溶血素和埃尔托溶血素的分子量约为60,000,并且溶血素的氨基酸组成非常相似。溶血素的比活性相同,两种溶血素都能被针对同源和异源溶血素的抗血清同等程度地中和。用两种溶血素和抗溶血素血清进行的双向免疫扩散试验产生了一条共同(融合)的沉淀线。这些数据表明,非O1溶血素在生物学、物理化学和免疫学上与埃尔托溶血素无法区分。