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Applied and Environmental Microbiology, July 2000, p. 3098-3101, Vol. 66, No. 7
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Absence of a Putative Mannose-Specific Phosphotransferase System Enzyme IIAB Component in a Leucocin A-Resistant Strain of Listeria monocytogenes, as Shown by Two-Dimensional Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis

M. Ramnath,1 M. Beukes,1 K. Tamura,2 and J. W. Hastings1,*

School of Molecular and Cellular Biosciences, University of Natal, Scottsville, Pietermaritzburg, South Africa,1 and Institute for Protein Research, Osaka University, 3-2 Yamadaoka Suita, Osaka 565-0871, Japan2

Received 13 January 2000/Accepted 17 April 2000

Leucocin A is a class IIa bacteriocin produced by Leuconostoc spp. that has previously been shown to inhibit the growth of Listeria monocytogenes. A spontaneous resistant mutant of L. monocytogenes was isolated and found to be resistant to leucocin A at levels in excess of 2 mg/ml. The mutant showed no significant cross-resistance to nontype IIa bacteriocins including nisaplin and ESF1-7GR. However, there were no inhibition zones found on a lawn of the mutant when challenged with an extract containing 51,200 AU of pediocin PA-2 per ml as determined by a simultaneous assay on the sensitive wild-type strain. DNA and protein analysis of the resistant and susceptible strains were carried out using silver-stained amplified fragment length polymorphism (ssAFLP) and one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), respectively. Two-dimensional SDS-PAGE clearly showed a 35-kDa protein which was present in the sensitive but absent from the resistant strain. The N-terminal end of the 35-kDa protein was sequenced and found to have an 83% homology to the mannose-specific phosphotransferase system enzyme IIAB of Streptococcus salivarius.

* Corresponding author. Mailing address: School of Molecular and Cellular Biosciences, University of Natal, P.O. Box X01, Scottsville, Pietermaritzburg 3209, South Africa. Phone: 27 33 260 5434. Fax: 27 33 260 5435. E-mail: Hastings{at}gene.unp.ac.za.

Applied and Environmental Microbiology, July 2000, p. 3098-3101, Vol. 66, No. 7
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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