Schmidt H, Bielaszewska M, Karch H
Institut für Hygiene und Mikrobiologie der Universität Würzburg, 97080 Würzburg, Germany.
Appl Environ Microbiol. 1999 Sep;65(9):3855-61. doi: 10.1128/AEM.65.9.3855-3861.1999.
We investigated the ability of a detoxified derivative of a Shiga toxin 2 (Stx2)-encoding bacteriophage to infect and lysogenize enteric Escherichia coli strains and to develop infectious progeny from such lysogenized strains. The stx(2) gene of the patient E. coli O157:H7 isolate 3538/95 was replaced by the chloramphenicol acetyltransferase (cat) gene from plasmid pACYC184. Phage phi3538(Deltastx(2)::cat) was isolated after induction of E. coli O157:H7 strain 3538/95 with mitomycin. A variety of strains of enteropathogenic E. coli (EPEC), enteroinvasive E. coli (EIEC), Stx-producing E. coli (STEC), enterotoxigenic E. coli (ETEC), enteroaggregative E. coli (EAEC), and E. coli from the physiological stool microflora were infected with phi3538(Deltastx(2)::cat), and plaque formation and lysogenic conversion of wild-type E. coli strains were investigated. With the exception of one EIEC strain, none of the E. coli strains supported the formation of plaques when used as indicators for phi3538(Deltastx(2)::cat). However, 2 of 11 EPEC, 11 of 25 STEC, 2 of 7 EAEC, 1 of 3 EIEC, and 1 of 6 E. coli isolates from the stool microflora of healthy individuals integrated the phage in their chromosomes and expressed resistance to chloramphenicol. Following induction with mitomycin, these lysogenic strains released infectious particles of phi3538(Deltastx(2)::cat) that formed plaques on a lawn of E. coli laboratory strain C600. The results of our study demonstrate that phi3538(Deltastx(2)::cat) was able to infect and lysogenize particular enteric strains of pathogenic and nonpathogenic E. coli and that the lysogens produced infectious phage progeny. Stx-encoding bacteriophages are able to spread stx genes among enteric E. coli strains.
我们研究了一种编码志贺毒素2(Stx2)的噬菌体的解毒衍生物感染肠道大肠杆菌菌株并使其溶原化,以及从这些溶原化菌株产生感染性后代的能力。将患者大肠杆菌O157:H7分离株3538/95的stx(2)基因替换为质粒pACYC184中的氯霉素乙酰转移酶(cat)基因。用丝裂霉素诱导大肠杆菌O157:H7菌株3538/95后,分离出噬菌体phi3538(Deltastx(2)::cat)。用phi3538(Deltastx(2)::cat)感染多种致病性大肠杆菌(EPEC)、侵袭性大肠杆菌(EIEC)、产志贺毒素大肠杆菌(STEC)、产肠毒素大肠杆菌(ETEC)、聚集性大肠杆菌(EAEC)菌株以及来自生理性粪便微生物群的大肠杆菌,并研究野生型大肠杆菌菌株的噬菌斑形成和溶原性转化。除一株EIEC菌株外,当用作phi3538(Deltastx(2)::cat)的指示菌时,没有一株大肠杆菌菌株支持噬菌斑的形成。然而,11株EPEC中的2株、25株STEC中的11株、7株EAEC中的2株、3株EIEC中的1株以及来自健康个体粪便微生物群的6株大肠杆菌分离株中的1株将噬菌体整合到其染色体中,并表现出对氯霉素的抗性。用丝裂霉素诱导后,这些溶原性菌株释放出phi3538(Deltastx(2)::cat)的感染性颗粒,这些颗粒在大肠杆菌实验室菌株C600的菌苔上形成噬菌斑。我们的研究结果表明,phi3538(Deltastx(2)::cat)能够感染并溶原化致病性和非致病性大肠杆菌的特定肠道菌株,并且溶原菌产生了感染性噬菌体后代。编码Stx的噬菌体能够在肠道大肠杆菌菌株之间传播stx基因。