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Journal of Clinical Microbiology, September 1999, p. 2913-2919, Vol. 37, No. 9
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Isolation of Rifampin-Resistant Mutants of Listeria monocytogenes and Their Characterization by rpoB Gene Sequencing, Temperature Sensitivity for Growth, and Interaction with an Epithelial Cell Line

Robert Morse,1,* Karen O'Hanlon,1 Mumtaz Virji,2 and Matthew D. Collins1

Department of Food Science and Technology, The University of Reading, Reading RG6 6AP,1 and Department of Pathology and Microbiology, School of Medical Sciences, University of Bristol, Clifton, Bristol BS8 1TD,2 United Kingdom

Received 20 January 1999/Returned for modification 26 April 1999/Accepted 25 May 1999

The sequence of the rpoB gene from Listeria monocytogenes was determined. Rifampin-resistant (Rifr) mutants arising from L. monocytogenes cultures exposed to rifampin were isolated, and by partial sequencing of their rpoB genes, seven different point mutations affecting five different amino acids (473Aspright-arrowAsn or Gly, 479Glyright-arrowAsp, 483Hisright-arrowTyr or Leu, 528Ileright-arrowPhe, and 530Serright-arrowTyr), which led to MICs of 0.5 to 100 µg/ml for the organisms, were determined. These mutants showed various deficiencies for growth at 42°C, with only one being comparable to the wild-type strain. The interaction of these Rifr mutants with human Caco-2 cells was examined by using an immunofluorescence technique. Three mutants failed to interact, while three showed a reduced interaction compared to that of the wild type. It is believed that these pleiotropic phenotypes have arisen as a result of mutations within the DNA-dependent RNA polymerase holoenzyme.

* Corresponding author. Mailing address: Department of Food Science and Technology, The University of Reading, Whiteknights, P.O. Box 226, Reading RG6 6AP, United Kingdom. Phone: 44-118-9357228. Fax: 44-118-9267917. E-mail: robert.morse{at}bbsrc.ac.uk.

Journal of Clinical Microbiology, September 1999, p. 2913-2919, Vol. 37, No. 9
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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