Schmidt H, Karch H, Beutin L
Institut für Hygiene und Mikrobiolgie, Universität Würzburg, FRG.
FEMS Microbiol Lett. 1994 Apr 1;117(2):189-96. doi: 10.1111/j.1574-6968.1994.tb06763.x.
Most enterohemorrhagic Escherichia coli O157:H7 strains harbor a large-sized (90 kb) plasmid designated pO157 and show an enterohemolytic phenotype. In this study the hemolytic activity of E. coli O157:H7 strain EDL933 was investigated. Curing of strain EDL933 from pO157 resulted in loss of its hemolytic activity. By transformation with Tn801-tagged pO157 (pSK3), the hemolysin-negative E. coli K-12 strains C600 and DH5 alpha became positive for hemolysin production. By transformation of recombinant plasmids carrying a 11.9 kb BamHI fragment and a 5.3 kb SalI fragment of pSK3 hemolytic activity is revealed when transformed in E. coli C600 or DH5 alpha DNA-hybridization of pO157 and subclones with the alpha-hemolysin specific DNA probe was only found under conditions of low stringency. No hybridization was found with enterohemolysin I (EHly1) and enterohemolysin II (EHly2) probes. Our results indicate that a hitherto not described hemolysin belonging to the alpha-hemolysin family is encoded by the 90 kb plasmid of E. coli O157 strains.
大多数肠出血性大肠杆菌O157:H7菌株携带一个名为pO157的大型(90 kb)质粒,并表现出肠溶血表型。在本研究中,对大肠杆菌O157:H7菌株EDL933的溶血活性进行了研究。从pO157中去除EDL933菌株导致其溶血活性丧失。通过用Tn801标记的pO157(pSK3)进行转化,溶血素阴性的大肠杆菌K-12菌株C600和DH5α溶血素产生变为阳性。通过转化携带pSK3的11.9 kb BamHI片段和5.3 kb SalI片段的重组质粒,当在大肠杆菌C600或DH5α中转化时显示出溶血活性。pO157及其亚克隆与α-溶血素特异性DNA探针的DNA杂交仅在低严谨条件下发现。未发现与肠溶血素I(EHly1)和肠溶血素II(EHly2)探针杂交。我们的结果表明,大肠杆菌O157菌株的90 kb质粒编码一种迄今未描述的属于α-溶血素家族的溶血素。