A mini-Tn5 transposon derivative, mini-Tn5cyaA′, has been constructed. It contains a promoter-less and ribosome binding site-deficient reporter gene, encoding the catalytic domain of Bordetella pertussis adenylate cyclase toxin (CyaA′). We used this system to mutagenize B. bronchiseptica and we developed a screen for identification of mutants containing cyaA′ translational fusions. This system was used to identify B. bronchiseptica genes that encode surface-exposed and secreted proteins.
Bibliographical noteFunding Information:
This work was supported by the European Union Fifth Framework Quality of Life Program (Grant 30600, I.R.), the Israel Science Foundation (Grant 223/99, I.R.), the Max Deutsch Foundation for Applied Research (E.H. and I.R.), NIH (Grant AI38417, J.F.M.) and USDA (Grant 1999-02298, J.F.M.).
- Bordetella bronchiseptica
- Protein secretion
- Reporter gene
- Transposon mutagenesis