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© Research
Publication : Methods in molecular biology (Clifton, N.J.)

Analysis of Bacterial Pilus Assembly by Shearing and Immunofluorescence Microscopy

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Methods in molecular biology (Clifton, N.J.) - 01 Jan 2018

Luna-Rico A, Thomassin JL, Francetic O

Link to Pubmed [PMID] – 29605922

Methods Mol. Biol. 2018;1764:291-305

Bacterial surface appendages of the type 4 pilus superfamily play diverse roles in adherence, aggregation, motility, signaling, and macromolecular transport. Here we describe two analytical approaches to study assembly of type 4 pili and of pseudopili produced by type 2 protein secretion systems: the shearing assay and immunofluorescence microscopy. These complementary antibody-based methods allow for semiquantitative analysis of fiber assembly. The shearing assay can be scaled up to yield crude extracts of pili that can be further analyzed by electron and atomic force microscopy or by mass spectrometry.

https://www.ncbi.nlm.nih.gov/pubmed/29605922