New and Old Reagents for Fluorescent Protein Tagging of Microtubules in Fission Yeast: Experimental and Critical Evaluation

被引：36

作者：

Snaith, Hilary A. ^{[1
]}

Anders, Andreas ^{[1
]}

Samejima, Itaru ^{[1
]}

Sawin, Kenneth E. ^{[1
]}

机构：

[1] Univ Edinburgh, Wellcome Trust Ctr Cell Biol, Sch Biol Sci, Edinburgh EH9 3JR, Midlothian, Scotland

来源：

MICROTUBULES: IN VIVO | 2010年 / 97卷

基金：

英国惠康基金;

关键词：

GAMMA-TUBULIN COMPLEX; SPINDLE-POLE BODY; SCHIZOSACCHAROMYCES-POMBE; MITOTIC SPINDLE; CELL-DIVISION; MONOMERIC RED; INTERPHASE MICROTUBULES; POLARIZED GROWTH; BETA-TUBULIN; GENE;

D O I：

10.1016/S0091-679X(10)97009-X

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

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.

引用

页码：147 / 172

页数：26

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