SIMPLE approach for isolating mutants expressing fimbriae

Sean Paul Nuccio, Daniela Chessa, Eric H. Weening, Manuela Raffatellu, Steven Clegg, Andreas J Baumler

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Genomes of members of the family Enterobacteriaceae contain large repertoires of putative fimbrial operons. Since many of these operons are poorly expressed in vitro, a convenient method for inducing elaboration of the encoded fimbriae would greatly facilitate their functional characterization. Here we describe a new technique for identifying fimbriated bacteria from a library of transposon mutants by screening with immunomagnetic particles for ligand expression (SIMPLE). The SIMPLE method was applied to identify the T-POP mutants of Salmonella enterica serotype Typhimurium carrying on their surfaces filaments composed of PefA, the major subunit product of a fimbrial operon (pef) that is not expressed during growth in Luria-Bertani broth. Four such mutants were identified from a library of 24,000 mutants, each of which carried a T-POP insertion within the hns gene, which encodes a global silencer of horizontally acquired genes. Our data suggest that the SIMPLE method is an effective approach for isolating fimbriated bacteria, which can be readily applied to fimbrial operons identified by whole-genome sequencing.

Original languageEnglish (US)
Pages (from-to)4455-4462
Number of pages8
JournalApplied and Environmental Microbiology
Volume73
Issue number14
DOIs
StatePublished - Jul 2007

Fingerprint

fimbriae
operon
Operon
ligand
Ligands
screening
mutants
Libraries
genome
Genome
Bacteria
bacterium
Salmonella enterica
gene
bacteria
Enterobacteriaceae
methodology
Salmonella Typhimurium
transposons
Genes

ASJC Scopus subject areas

  • Environmental Science(all)
  • Biotechnology
  • Microbiology

Cite this

Nuccio, S. P., Chessa, D., Weening, E. H., Raffatellu, M., Clegg, S., & Baumler, A. J. (2007). SIMPLE approach for isolating mutants expressing fimbriae. Applied and Environmental Microbiology, 73(14), 4455-4462. https://doi.org/10.1128/AEM.00148-07

SIMPLE approach for isolating mutants expressing fimbriae. / Nuccio, Sean Paul; Chessa, Daniela; Weening, Eric H.; Raffatellu, Manuela; Clegg, Steven; Baumler, Andreas J.

In: Applied and Environmental Microbiology, Vol. 73, No. 14, 07.2007, p. 4455-4462.

Research output: Contribution to journalArticle

Nuccio, SP, Chessa, D, Weening, EH, Raffatellu, M, Clegg, S & Baumler, AJ 2007, 'SIMPLE approach for isolating mutants expressing fimbriae', Applied and Environmental Microbiology, vol. 73, no. 14, pp. 4455-4462. https://doi.org/10.1128/AEM.00148-07
Nuccio, Sean Paul ; Chessa, Daniela ; Weening, Eric H. ; Raffatellu, Manuela ; Clegg, Steven ; Baumler, Andreas J. / SIMPLE approach for isolating mutants expressing fimbriae. In: Applied and Environmental Microbiology. 2007 ; Vol. 73, No. 14. pp. 4455-4462.
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