This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Guerry, P Articles by Bourgeois, A L Search for Related Content PubMed PubMed Citation Articles by Guerry, P Articles by Bourgeois, A L

research-article

Development and characterization of recA mutants of Campylobacter jejuni for inclusion in attenuated vaccines.

Enterics Program, Naval Medical Research Institute, Bethesda, Maryland 20814.

ABSTRACT

Isogenic recA mutants of Campylobacter jejuni have been constructed for evaluation of their usefulness in attenuated vaccines against this major worldwide cause of diarrhea. The recA+ gene of C. jejuni 81-176 was cloned by using degenerate primers to conserved regions of other RecA proteins in a PCR. The C. jejuni recA+ gene encodes a predicted protein with an M(r) of 37,012 with high sequence similarity to other RecA proteins. The termination codon of the recA+ gene overlaps with the initiation codon of another open reading frame which encodes a predicted protein which has > 50% identity with the N terminus of the Escherichia coli enolase protein. A kanamycin resistance gene was inserted into the cloned recA+ gene in E. coli and returned to C. jejuni VC83 by natural transformation, resulting in allelic replacement of the wild-type recA gene. The resulting VC83 recA mutant displayed increased sensitivity to UV light and a defect in generalized recombination as determined by natural transformation frequencies. The mutated recA gene was amplified from VC83 recA by PCR, and the product was used to transfer the mutation by natural transformation into C. jejuni 81-176 and 81-116, resulting in isogenic recA mutants with phenotypes similar to VC83 recA. After oral feeding, strain 81-176 recA colonized rabbits at levels comparable to wild-type 81-176 and was capable of eliciting the same degree of protection as wild-type 81-176 against subsequent homologous challenge in the RITARD (removable intestinal tie adult rabbit diarrhea) model.

This article has been cited by other articles:

• Jeon, B., Zhang, Q. (2007). Cj0011c, a Periplasmic Single- and Double-Stranded DNA-Binding Protein, Contributes to Natural Transformation in Campylobacter jejuni. J. Bacteriol. 189: 7399-7407 [Abstract] [Full Text]  
• Larsen, J. C., Szymanski, C., Guerry, P. (2004). N-Linked Protein Glycosylation Is Required for Full Competence in Campylobacter jejuni 81-176. J. Bacteriol. 186: 6508-6514 [Abstract] [Full Text]  
• Batchelor, R. A., Pearson, B. M., Friis, L. M., Guerry, P., Wells, J. M. (2004). Nucleotide sequences and comparison of two large conjugative plasmids from different Campylobacter species. Microbiology 150: 3507-3517 [Abstract] [Full Text]  
• Wilson, D. L., Bell, J. A., Young, V. B., Wilder, S. R., Mansfield, L. S., Linz, J. E. (2003). Variation of the natural transformation frequency of Campylobacter jejuni in liquid shake culture. Microbiology 149: 3603-3615 [Abstract] [Full Text]  
• Sander, P., Papavinasasundaram, K. G., Dick, T., Stavropoulos, E., Ellrott, K., Springer, B., Colston, M. J., Bottger, E. C. (2001). Mycobacterium bovis BCG recA Deletion Mutant Shows Increased Susceptibility to DNA-Damaging Agents but Wild-Type Survival in a Mouse Infection Model. Infect. Immun. 69: 3562-3568 [Abstract] [Full Text]  
• Bacon, D. J., Alm, R. A., Burr, D. H., Hu, L., Kopecko, D. J., Ewing, C. P., Trust, T. J., Guerry, P. (2000). Involvement of a Plasmid in Virulence of Campylobacter jejuni 81-176. Infect. Immun. 68: 4384-4390 [Abstract] [Full Text]  
• Wösten, M. M. S. M., Boeve, M., Koot, M. G. A., van Nuenen, A. C., van der Zeijst, B. A. M. (1998). Identification of Campylobacter jejuni Promoter Sequences. J. Bacteriol. 180: 594-599 [Abstract] [Full Text]