Construction of mini-Tn 5cyaA′ and its utilization for the identification of genes encoding surface-exposed and secreted proteins in Bordetella bronchiseptica

Abstract

A mini-Tn 5 transposon derivative, mini-Tn5 cyaA′, 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.