ABSTRACT
Escherichia coli O25 strains that produce heat-stable toxin (ST) have been recently isolated in Japan, and epidemiological study of this type of enterotoxigenic E. coli is required. In this study the heterogeneity of 16 ST-producing and non-producing strains of E. coli O25 was investigated. All eight ST-producing strains were shown to have STIb gene, and seven of them had similar profiles of plasmids, ladder-banding of LPS in SDS-polyacrylamide gel electrophoresis, and chromosomal DNA digestions in pulsed-field gel electrophoresis (PFGE). In contrast, ST-non-producing strains were more heterogeneous in all parameters examined. PFGE of the digested chromosomal DNA with several restriction enzymes was proved to be an effective procedure to compare the closely related strains of E. coli O25.
Subject(s)
Bacterial Proteins/metabolism , Bacterial Toxins/metabolism , Enterotoxins/metabolism , Escherichia coli/genetics , Blotting, Southern , DNA, Bacterial/analysis , Electrophoresis, Gel, Pulsed-Field , Electrophoresis, Polyacrylamide Gel , Escherichia coli/chemistry , Escherichia coli/metabolism , Escherichia coli Proteins , Humans , Japan , Lipopolysaccharides/analysis , Plasmids/analysisABSTRACT
Mutants of Sphingomonaspaucimobilis defective in a part of the carbohydrate moiety of the glycosphingolipid (GSL) were constructed by transposon (Tn5)-insertional mutagenesis. Defective mutants were selected by ELISA using the antibody recognizing the tetrasaccharide-type GSL (GSL-4A) of S. paucimobilis. Eight defective mutants were selected from about 8,000 kanamycin-resistant strains, and seven of them were found to lack the terminal mannose of GSL-4A. The chemical structure of the mutant GSL was investigated, and proved that the rest of the structure was not changed by the mutation.
