Visualization of fluorescence-tagged proteins in fission yeast: the analysis of mitotic spindle dynamics using GFP-tubulin under the native promoter.
Visualization of fluorescence-tagged proteins in fission yeast: the analysis of mitotic spindle dynamics using GFP-tubulin under the native promoter.
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裂殖酵母中荧光标记蛋白的可视化：使用天然启动子下的 GFP-微管蛋白分析有丝分裂纺锤体动力学。
DOI：
10.1007/978-1-60327-993-2_11
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发表时间：
2009
影响因子：
--
通讯作者：
T. Toda
中科院分区：
文献类型：
--
作者：
Masamitsu Sato;M. Toya;T. Toda
有丝分裂纺锤体微管将染色体拉向两极，产生两个子细胞。正确的纺锤体形成和功能是防止肿瘤发生和细胞死亡所必需的。荧光蛋白系统和显微镜的发展使我们能够利用一株在其天然启动子下带有荧光标记基因的菌株来研究活裂变酵母细胞中蛋白质的“真实”行为。通过这种方式，标记蛋白的表达水平与野生型非标记蛋白的表达水平相似。在本章中，我们说明了标准的方法来产生表达荧光标记蛋白的菌株，并在显微镜下观察它们。具体来说，我们引入gfp微管蛋白应变来分析纺锤体微管的动态行为。gfp微管蛋白在其原生启动子下的观察，揭示了裂变酵母中着丝点与微管的附着过程。
Mitotic spindle microtubules pull chromosomes toward each pole to generate two daughter cells. Proper spindle formation and function are required to prevent tumorigenesis and cell death. The fission yeast Schizosaccharomyces pombe has been widely used as a model organism to understand the molecular mechanism of mitosis due to its convenience in genetics, molecular biology, and cell biology. The development of fluorescent protein systems and microscopy enables us to investigate the "true" behavior of proteins in living fission yeast cells using a strain with a fluorescence-tagged gene under its native promoter. In this way the level of expression of tagged protein is similar to the level of wild-type nontagged protein. In this chapter we illustrate standard methods to generate strains expressing fluorescently tagged proteins and to observe them under the microscope. Specifically, we introduce a GFP-tubulin strain to analyze the dynamic behavior of spindle microtubules. Observation of GFP-tubulin under its native promoter has illuminated the process of kinetochore-microtubule attachment process in fission yeast.
影响因子：
4
作者：
I. Hagan;J. Hyams
通讯作者：
I. Hagan;J. Hyams
影响因子：
10.5
作者：
Bridge, AJ;Morphew, M;Hagan, IM
通讯作者：
Hagan, IM
影响因子：
3.3
作者：
West, RR;Vaisberg, EV;McIntosh, JR
通讯作者：
McIntosh, JR