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Journal of Bacteriology, October 2001, p. 5756-5761, Vol. 183, No. 19
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.19.5756-5761.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Genetic Analysis of a Pyocin-Resistant Lipooligosaccharide (LOS) Mutant of Haemophilus ducreyi: Restoration of Full-Length LOS Restores Pyocin Sensitivity

Melanie J. Filiatrault,1,2 Robert S. Munson Jr.,3,4 and Anthony A. Campagnari1,3,5,*

Department of Microbiology,1 Department of Medicine, Division of Infectious Diseases,5 and The Witebsky Center for Immunology and Microbial Pathogenesis,2 State University of New York at Buffalo, Buffalo, New York 14214, and Children's Research Institute3 and Department of Molecular Virology, Immunology, and Medical Genetics,4 Ohio State University, Columbus, Ohio 43205-2696

Received 29 March 2001/Accepted 29 June 2001

DNA sequence and Southern blot analyses were used to determine the genetic defect of a Haemophilus ducreyi pyocin-resistant lipooligosaccharide (LOS) mutant, HD35000R. The region of the HD35000R chromosome containing the suspected mutation was amplified, and sequence analysis detected a 3,189-bp deletion. This deletion resulted in the loss of the entire waaQ gene, another open reading frame that encodes a putative homolog to a hypothetical protein (HI0461) of H. influenzae, the gene encoding an argininosuccinate synthase homolog, and a change in the 3' sequence of the lgtF gene. Southern blot analysis confirmed that no genomic rearrangements had occurred. Isogenic LOS mutants and the respective complemented mutants were evaluated for susceptibility to pyocin C. The mutants expressing truncated LOS were resistant to lysis by pyocin C, and complementation restored sensitivity to the pyocin. We conclude that HD35000R is defective in both glycosyltransferase genes and that pyocin resistance is due to truncation of the full-length LOS molecule.

* Corresponding author. Mailing address: Department of Microbiology, State University of New York at Buffalo, Biomedical Research Bldg., Rm. 143, 3435 Main St., Buffalo, NY 14214. Phone: (716) 829-2673. Fax: (716) 829-3889. E-mail: AAC{at}acsu.buffalo.edu.

Journal of Bacteriology, October 2001, p. 5756-5761, Vol. 183, No. 19
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.19.5756-5761.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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