RRC ID 27196
Author Snaith HA, Anders A, Samejima I, Sawin KE.
Title New and old reagents for fluorescent protein tagging of microtubules in fission yeast; experimental and critical evaluation.
Journal Methods Cell Biol
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.
Volume 97
Pages 147-72
Published 2010-1-1
DOI 10.1016/S0091-679X(10)97009-X
PII S0091-679X(10)97009-X
PMID 20719270
MeSH Clinical Laboratory Techniques Fluorescent Dyes / pharmacology* Indicators and Reagents / pharmacology Microtubule-Associated Proteins / analysis Microtubule-Associated Proteins / metabolism Microtubules / chemistry Microtubules / metabolism* Models, Biological Schizosaccharomyces* / cytology Schizosaccharomyces* / drug effects Schizosaccharomyces* / metabolism Schizosaccharomyces pombe Proteins / analysis Schizosaccharomyces pombe Proteins / metabolism Staining and Labeling / methods*
IF 1.441
Times Cited 26
DNA material pDUAL-GFH1c (RDB06145) pDUAL-HFG1c (RDB06133) pDUAL-HFG41c (RDB06134) pDUAL-HFG81c (RDB06135) pDUAL-GFH1c (RDB06145) pDUAL-GFH41c (RDB06146) pDUAL-GFH81c (RDB06147) pDUAL (RDB06178) pDUAL-HFG31c (RDB06494) pDUAL-HFG51c (RDB06495) pDUAL-GFH31c (RDB06498) pDUAL-GFH51c (RDB06499) pHFF1 (RDB06580).