Construction of mini-Tn5cyaA' and its utilization for the identification of genes encoding surface-exposed and secreted proteins in Bordetella bronchiseptica

FEMS Microbiol Lett. 2001 Nov 27;205(1):119-23. doi: 10.1111/j.1574-6968.2001.tb10934.x.

Abstract

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.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenylate Cyclase Toxin
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Base Sequence
  • Bordetella bronchiseptica / genetics*
  • Bordetella bronchiseptica / metabolism
  • DNA Transposable Elements*
  • Genes, Reporter
  • Membrane Proteins / genetics*
  • Membrane Proteins / metabolism*
  • Molecular Sequence Data
  • Mutagenesis, Insertional*
  • Protein Biosynthesis
  • Protein Precursors / genetics
  • Protein Precursors / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • Adenylate Cyclase Toxin
  • Bacterial Proteins
  • DNA Transposable Elements
  • Membrane Proteins
  • Protein Precursors
  • Recombinant Fusion Proteins