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A rapid screening procedure to identify mini-Tn10 insertion mutants of Escherichia coli K-12 with altered adhesion properties
Authors:Pierre Genevaux  Stephane Muller  Pascale Bauda
Affiliation:Laboratoire de Microbiologie, Centre des Sciences de l'Environnement, BP 4025, 1 rue des Récollets, 57040 Metz Cedex 1, France
Abstract:Abstract A rapid screening procedure was developed for detection of Escherichia coli mutants with altered adhesion abilities using polystyrene 96-well microtiter plates as attachment surfaces. During this assay, bacterial strains grew and adhered simultaneously, and attached cells were measured after crystal violet staining. Starting with a total of 7000 W3110::Tn10 insertion mutants of E. coli K-12 W3110, 50 adhesion-deficient mutants were isolated which showed less than 40% attachment, and 22 mutants were found with an attachment of 40–75%. Motility assays were performed on these 72 mutants, and 34 displayed altered motility.
Keywords:Escherichia coli    Mini-Tn10 transposon mutagenesis    Bacterial adhesion    Attachment    Adhesion assay    Inanimate surface
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