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Applied and Environmental Microbiology, May 1999, p. 1843-1848, Vol. 65, No. 5
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Export of Virulence Genes and Shiga Toxin by Membrane Vesicles of Escherichia coli O157:H7

Glynis L. Kolling and Karl R. Matthews*

Department of Food Science, Rutgers University, New Brunswick, New Jersey 08901

Received 2 November 1998/Accepted 15 February 1999

Membrane vesicles released by Escherichia coli O157:H7 into culture medium were purified and analyzed for protein and DNA content. Electron micrographs revealed vesicles that are spherical, range in size from 20 to 100 nm, and have a complete bilayer. Analysis of vesicle protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrates vesicles that contain many proteins with molecular sizes similar to outer membrane proteins and a number of cellular proteins. Immunoblot (Western) analysis of vesicles suggests the presence of cell antigens. Treatment of vesicles with exogenous DNase hydrolyzed surface-associated DNA; PCR demonstrated that vesicles contain DNA encoding the virulence genes eae, stx1 and stx2, and uidA, which encodes for beta -galactosidase. Immunoblot analysis of intact and lysed, proteinase K-treated vesicles demonstrate that Shiga toxins 1 and 2 are contained within vesicles. These results suggest that vesicles contain toxic material and transfer experiments demonstrate that vesicles can deliver genetic material to other gram-negative organisms.

* Corresponding author. Mailing address: Cook College, Department of Food Science, Rutgers, The State University of New Jersey, 65 Dudley Rd., New Brunswick, NJ 08901-8520. Phone: (732) 932-9611. Fax: (732) 932-6776. E-mail: matthews{at}aesop.rutgers.edu.

Applied and Environmental Microbiology, May 1999, p. 1843-1848, Vol. 65, No. 5
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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