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Mannose-resistant Proteus-like (MR/P) fimbriae, produced by the uropathogen Proteus mirabilis, are major contributors to urinary tract infection. Expression of mrp genes is controlled by an invertible element in the mrp operon promoter, and promoter orientation is controlled by a single recombinase, MrpI, which determines whether the element is in the on or off orientation. Detailed here is a simple assay to determine the orientation of the invertible element in a population of P. mirabilis, a rapid screen to detect element-locked mrpI mutants, and quantification of mixtures of on and off bacteria.

Proteus mirabilis is generally easy to culture, but its tendency to swarm on a wide variety of media can interfere with isolation of single colonies or identification of other species in a sample. Therefore, specialized media may be needed to control swarming or to study the bacteria under chemically defined conditions. Here, methods are described for routine culture of P. mirabilis, isolation of P. mirabilis from mixed cultures, and culture of P. mirabilis on physiologically relevant media.

Genetic mutation enables the study of the function of specific genes, particularly when a mutant is compared against its isogenic parent. In Proteus mirabilis bacteria, traditional allelic exchange mutation is labor-intensive and has a high failure rate in some strains. Likewise, there is no working protocol for lambda red recombinase-based mutation in P. mirabilis. Here we describe an alternative method of insertional mutagenesis based on retargeting of group II introns. The protocol includes steps to generate single or multiple mutations, with the possibility to delete intervening sequences of DNA.

A critical first step in bacterial virulence and colonization is adherence to mucosal surfaces, often mediated by fimbriae and other protein adhesins. Here are described three short methods for studying these surface proteins and their behaviors, using protocols developed for the opportunistic pathogen Proteus mirabilis. Unlike the mannose-binding type 1 fimbriae produced by Escherichia coli, most P. mirabilis strains produce mannose-resistant/Proteus-like (MR/P) fimbriae. Both types of fimbrial production and adhesion can be easily demonstrated by a simple and economical hemagglutination assay which uses a model system of erythrocytes. The second and third fimbrial methods presented here show how to shear surface-exposed proteins and use acid treatment to separate interlocked fimbrial subunits into monomers.

Proteus mirabilis is well known for using its flagella to swim through liquids or swarm across solid surfaces. Both phenomena are easy to observe. Described here are two agar-based assays for studying both swimming and swarming behavior, and considerations that affect the outcome.