Shuttle mutagenesis: A mini-transposon for producing PhoA fusions with exported proteins in neisseria gonorrhoeae

Susan Boyle-Vavra, H. Steven Seifert*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

Shuttle mutagenesis is a method for producing stable mini-transposon (mTn) insertions into the genome of Neisseria gonorrhoeae (gonococcus, Gc) and other microbes. Using an mTn3 derivative, we have produced an mTn (mTnCmPhoA) which contains a phoA' gene lacking its N-terminal signal sequence useful for isolating genes which encode exported proteins, mTnCmPhoA was characterized in Gc and Escherichia coli using a cloned target containing the Gc genes, opaE1, pilA and piIB. PhoA+ Gc containing pi1B::mTnCmPhoA insertions confirm that Pi1B is an exported protein in Gc. This system will be useful for isolating and characterizing extracytoplasmic virulence factors from Gc and other bacterial pathogens.

Original languageEnglish (US)
Pages (from-to)101-106
Number of pages6
JournalGene
Volume155
Issue number1
DOIs
StatePublished - Mar 21 1995

Keywords

  • Exported protein
  • opaE1
  • pathogen
  • pilA
  • pilB
  • virulence factor

ASJC Scopus subject areas

  • Genetics

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