Labeling of Peroxisomes for Live Cell Imaging in the Filamentous Fungus Ustilago maydis

This is the author accepted manuscript. The final version is available from Humana Press via the DOI in this record.The basidiomycete fungus Ustilago maydis has emerged as a powerful model organism to study fundamental biological processes. U. maydis shares many important features with human cells but provides the technical advantages of yeast. Recently, U. maydis has also been used to investigate fundamental processes in peroxisome biology. Here, we present an efficient yeast recombination-based cloning method to construct and express fluorescent fusion proteins (or conditional mutant protein alleles) which target peroxisomes in the fungus U. maydis. In vivo analysis is pivotal for understanding the underlying mechanisms of organelle motility. We focus on the in vivo labelling of peroxisomes in U. maydis and present approaches to analyze peroxisomal motility.We would like to thank G. Steinberg for his support and the opportunity to publish this method chapter. This work was supported by the Portuguese Foundation for Science and Technology and FEDER/COMPETE (SFRH/BD/73532/2010 to S.C. Guimarães) and CRUP/Treaty of Windsor (ACÇÕES INTEGRADAS 2009, B-33/09 to G. Steinberg and M. Schrader). M. Schrader acknowledges support from the Marie Curie Initial Training Network (ITN) action (FP7-2012 PERFUME-316723)