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Journal of Bacteriology, April 2001, p. 2384-2388, Vol. 183, No. 7
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.7.2384-2388.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

In Vitro Transposition System for Efficient Generation of Random Mutants of Campylobacter jejuni

Oscar R. Colegio,1 Thomas J. Griffin IV,2 Nigel D. F. Grindley,2 and Jorge E. Galán1,*

Section of Microbial Pathogenesis, Boyer Center for Molecular Medicine, Yale School of Medicine, New Haven, Connecticut 06536-0812,1 and Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520-81142

Received 3 November 2000/Accepted 11 January 2001

Campylobacter jejuni is the most common cause of food-borne illnesses in the United States. Despite the fact that the entire nucleotide sequence of its genome has recently become available, its mechanisms of pathogenicity are poorly understood. This is in part due to the lack of an efficient mutagenesis system. Here we describe an in vitro transposon mutagenesis system based on the Staphylococcus aureus transposable element Tn552 that allows the efficient generation of insertion mutants of C. jejuni. Insertions occur randomly and throughout the entire bacterial genome. We have tested this system in the isolation of nonmotile mutants of C. jejuni. Demonstrating the utility of the system, six nonmotile mutants from a total of nine exhibited insertions in genes known to be associated with motility. An additional mutant had an inactivating insertion in sigma 54, implicating this transcription factor in flagellum regulation. The availability of this efficient system will greatly facilitate the study of the mechanisms of pathogenesis of this important pathogen.

* Corresponding author. Mailing address: Section of Microbial Pathogenesis, Boyer Center for Molecular Medicine, Yale School of Medicine, New Haven, CT 06536-0812. Phone: (203) 737-2404. Fax: (203) 737-2630. E-mail: jorge.galan{at}yale.edu.

Journal of Bacteriology, April 2001, p. 2384-2388, Vol. 183, No. 7
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.7.2384-2388.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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