北京大学学报自然科学版 ›› 2019, Vol. 55 ›› Issue (4): 609-614.DOI: 10.13209/j.0479-8023.2019.050

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芽殖酵母细胞周期过程不同时期的定量观测

张志温, 李方廷†   

  1. 北京大学物理学院, 北京大学定量生物学中心, 北京 100871
  • 收稿日期:2018-06-13 修回日期:2018-12-25 出版日期:2019-07-20 发布日期:2019-07-20
  • 通讯作者: 李方廷, E-mail: lft(at)pku.edu.cn
  • 基金资助:
    国家自然科学基金(11174011)和国家重点研发计划(2015CB910300)资助

Time Marker and Quantitative Measurement of Budding Yeast Cell Cycle

ZHANG Zhiwen, LI Fangting†   

  1. Center for Quantitative biology, School of Physics, Peking University, Beijing 100871
  • Received:2018-06-13 Revised:2018-12-25 Online:2019-07-20 Published:2019-07-20
  • Contact: LI Fangting, E-mail: lft(at)pku.edu.cn

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摘要：

在芽殖酵母中构建CDC10-mCherry和SPC42-mCherry荧光蛋白, 作为酵母细胞周期不同时期的报告系统。根据酵母细胞中芽环(CDC10-mCherry)的产生和消失以及纺锤体极体(SPC42-mCherry)的复制和分离, 可以测量细胞周期中 G1期、S期、Early M期和Late M期的时间长度。在此报告系统的基础上, 分别构建S期细胞周期蛋白CLB5-GFP和M 期细胞周期蛋白CLB2-GFP菌株, 实现单细胞观测, 检验报告系统工作的稳定性和准确性。该报告系统可作为研究酵母细胞周期不同时期时间长度等定量生物学问题的背景菌株。

关键词: 芽殖酵母, 单细胞观测, 细胞周期, 时间标记

Abstract:

The budding yeast strains containing the CDC10-mCherry and SPC42-mCherry were constructed as a reporter system for different phases in the yeast cell cycle process. According to the generation and disappearance of bud neck (CDC10-mCherry) and the duplication and separation of spindle polar body (SPC42-mCherry), the durations of G1 phase, S phase, Early M phase and Late M phase in the yeast cell cycle can be quantitatively
measured. Based on the reporter system, CLB5 (S-phase cyclin)-GFP and CLB2 (M-phase cyclin)-GFP strains were constructed and single-cell observations were performed to verify the stability and accuracy of the reporter system. The reporter system can be used as a background strain to study the time length of different phase in the yeast cell cycle.

Key words: budding yeast, single-cell level observation, cell cycle, time marker

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