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Genetics and Molecular Biology |
Centers for Disease Control and Prevention, Respiratory Diseases Branch, 1600 Clifton Rd, Mailstop C02, Atlanta, GA 30333, USA1
Author for correspondence: Bernard Beall. Tel: +1 404 639 1237. Fax: +1 404 639 3123. e-mail: beb0{at}cdc.gov
The Bordetella bronchiseptica tonB gene was cloned by detection of a chromosomal restriction fragment hybridizing with each of two degenerate oligonucleotides that corresponded to Pro-Glu and Pro-Lys repeats characteristic of known TonB proteins. The tonBBb gene was situated upstream of exbB and exbD homologues and downstream of a putative Fur-regulated promoter. Hybridization results indicated that the tonB operon and flanking regions were highly conserved between B. bronchiseptica, Bordetella pertussis and Bordetella parapertussis. Disruption of tonB in B. bronchiseptica resulted in inability to grow in iron-limiting media, and inability to utilize alcaligin, enterobactin, ferrichrome, desferroxamine B, haemin and haemoglobin. Although it was not possible to inactivate tonB in a clinical B. pertussis isolate, tonB was disrupted in a laboratory B. pertussis strain previously selected for the ability to grow on Luria–Bertani medium. This B. pertussis tonB mutant shared a similar iron complex utilization deficient phenotype with the B. bronchiseptica tonB mutant. The B. bronchiseptica tonB operon present on a plasmid did not complement an Escherichia coli tonB mutant, but inefficient reconstitution of enterobactin utilization was observed in one fepA mutant harbouring plasmid copies of the B. pertussis fepA homologue and tonBBb operon.
Keywords: Bordetella, iron sources, TonB-dependent receptors
Abbreviations: DP, 2,2’-dipyridyl; EDDA, ethylenediamine-di-(o-hydroxyphenylacetic acid); LB, Luria–Bertani; SS, Stainer–Scholte minimal broth lacking added iron; SS-EDDA, SS agar containing 45µg EDDA ml-1
The GenBank accession number for the sequence reported in this paper is AF087669.
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