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Infection and Immunity, April 2002, p. 1761-1771, Vol. 70, No. 4
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.4.1761-1771.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification of Motility and Autoagglutination Campylobacter jejuni Mutants by Random Transposon Mutagenesis

Neal J. Golden1 and David W. K. Acheson2*

Department of Epidemiology and Preventative Medicine, University of Maryland School of Medicine, Baltimore, Maryland 21201,2 Department of Immunology and Pathology, Tufts University, Boston, Massachusetts 021111

Received 23 July 2001/ Returned for modification 17 September 2001/ Accepted 15 December 2001

Campylobacter jejuni has been identified as the leading cause of acute bacterial diarrhea in the United States, yet compared with other enteric pathogens, considerably less is understood concerning the virulence factors of this human pathogen. A random in vivo transposon mutagenesis system was recently developed for the purpose of creating a library of C. jejuni transformants. A total of 1,065 C. jejuni transposon mutants were screened for their ability to swarm on motility agar plates and autoagglutinate in liquid cultures; 28 mutants were subsequently identified. The transposon insertion sites were obtained by using random-primed PCR, and the putative genes responsible for these phenotypes were identified. Of these mutants, all 28 were found to have diminished motility (0 to 86% that of the control). Seventeen motility mutants had insertions in genes with strong homology to functionally known motility and chemotaxis genes; however, 11 insertions were in genes of unknown function. Twenty motility mutants were unable to autoagglutinate, suggesting that the expression of flagella is correlated with autoagglutination (AAG). However, four mutants expressed wild-type levels of surface FlaA, as indicated by Western blot analysis, yet were unable to autoagglutinate (Cj1318, Cj1333, Cj1340c, and Cj1062). These results suggest that FlaA is necessary but not sufficient to mediate the AAG phenotype. Furthermore, two of the four AAG mutants (Cj1333 and Cj1062) were unable to invade INT-407 intestinal epithelial cells, as determined by a gentamicin treatment assay. These data identify novel genes important for motility, chemotaxis, and AAG and demonstrate their potential role in virulence.

* Corresponding author. Mailing address: Department of Epidemiology and Preventative Medicine, University of Maryland School of Medicine, 10 S. Pine St., MSTF Building, Baltimore, MD 21201. Phone: (410) 706-4583. Fax: (410) 706-4581. E-mail: dacheson{at}epi.umaryland.edu.

Editor: A. D. O'Brien

Infection and Immunity, April 2002, p. 1761-1771, Vol. 70, No. 4
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.4.1761-1771.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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