Abstract
DNA colony hybridization with three 18- to 20-base-long synthetic oligonucleotide probes for cholera toxin (CT) was used to screen 12 clinical isolates of Aeromonas hydrophila. Under stringent hybridizing (overnight at 40 degrees C) and washing (1 h at 50 degrees C) conditions, nine strains reacted with the 32P-labeled CT probes. Concentrated (10x) cell-free supernatants or lysates from eight cultures, heated at 56 degrees C for 20 min, produced cytotonic effects in Y-1 mouse adrenal cells and Chinese hamster ovary (CHO) cells and caused a 1.5- to 22-fold increase in production of cyclic AMP in CHO cells. Preincubation with anti-CT reduced the CHO cell titer of cell lysates by 10-fold. In the GM1 ganglioside enzyme-linked immunosorbent assay, heated supernatants and lysates gave readings equivalent to 3.5 to 100 ng of CT. Three proteins with molecular weights of 89,900, 37,000, and 11,000 reacted with anti-CT on immunoblots of cell lysates from sodium dodecyl sulfate-polyacrylamide gels. These results suggest that there is DNA homology and immunological cross-reactivity between CT and the A. hydrophila cytotonic toxin.
MeSH Terms
Aeromonas
Bacterial Toxins/genetics,immunology,toxicity
Cholera Toxin/genetics,immunology,toxicity
Cross Reactions
Cyclic AMP/metabolism
Cytotoxins/genetics,immunology,toxicity
DNA, Bacterial/genetics
Nucleic Acid Hybridization
Sequence Homology, Nucleic Acid
Chemicals
Bacterial Toxins
Cytotoxins
DNA, Bacterial
Cholera Toxin
Cyclic AMP
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Schultz A J
Division of Microbiology, Food and Drug Administration, Washington, D.C. 20204.
McCardell B A
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