期刊名称：Journal of Clinical Biochemistry and Nutrition
印刷版ISSN：0912-0009
电子版ISSN：1880-5086
出版年度：2001
卷号：30
页码：33-42
DOI：10.3164/jcbn.30.33
出版社：The Society for Free Radical Research Japan
摘要：We constructed isogenic Shiga (-like) toxin (Stx-1 and/or Stx-2) gene-deletion mutants of enterohemorrhagic Escherichia coli (EHEC) GPU96MM (O157:H7). A vector with temperature-sensitive replication origin was used for the construction. The parts of stx-1 and stx-2 on the GPU96MM genome were replaced with kanamycin and chloramphenicol resistance genes, respectively. The mutants deficient in stx-1 , stx-2 , and both of them were named GPU993, GPU994, and GPU995, respectively. Each mutation was confirmed by the polymerase chain reaction, enzyme-linked immunosorbent assay using antibodies to B subunits of Stx-1 and Stx-2, and the cytotoxic activity of the bacterial culture supernatants toward HeLa cells was detected for GPU96MM and the mutants except for GPU995. These results indicate that GPU993 and GPU994 lack productivity for the respective toxins and GPU995, for both of them.
