1. New and old reagents for fluorescent protein tagging of microtubules in fission yeast; experimental and critical evaluation.
- Author
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Snaith HA, Anders A, Samejima I, and Sawin KE
- Subjects
- Clinical Laboratory Techniques, Indicators and Reagents pharmacology, Microtubule-Associated Proteins analysis, Microtubule-Associated Proteins metabolism, Microtubules chemistry, Models, Biological, Schizosaccharomyces pombe Proteins analysis, Schizosaccharomyces pombe Proteins metabolism, Fluorescent Dyes pharmacology, Microtubules metabolism, Schizosaccharomyces cytology, Schizosaccharomyces drug effects, Schizosaccharomyces metabolism, Staining and Labeling methods
- Abstract
The green fluorescent protein (GFP) has become a mainstay of in vivo imaging in many experimental systems. In this chapter, we first discuss and evaluate reagents currently available to image GFP-labeled microtubules in the fission yeast Schizosaccharomyces pombe, with particular reference to time-lapse applications. We then describe recent progress in the development of robust monomeric and tandem dimer red fluorescent proteins (RFPs), including mCherry, TagRFP-T, mOrange2, mKate, and tdTomato, and we present data assessing their suitability as tags in S. pombe. As part of this analysis, we introduce new PCR tagging cassettes for several RFPs, new pDUAL-based plasmids for RFP-tagging, and new RFP-tubulin strains. These reagents should improve and extend the study of microtubules and microtubule-associated proteins in S. pombe., (2010 Elsevier Inc. All rights reserved.)
- Published
- 2010
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