Chapter 9 - New and old reagents for fluorescent protein tagging of microtubules in fission yeast; experimental and critical evaluation

Hilary A Snaith, Andreas Anders, Itaru Samejima, Kenneth E Sawin

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

Abstract / Description of output

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.
Original languageEnglish
Title of host publicationMicrotubules
Subtitle of host publicationin vivo
EditorsLynne Cassimeris, Phong Tran
PublisherElsevier
Pages147-172
Number of pages26
ISBN (Print)978-0-12-381349-7
DOIs
Publication statusPublished - 2010

Publication series

NameMethods in Cell Biology
PublisherAcademic Press Inc.
Volume97
ISSN (Print)0091-679X

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