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Visualizing Yeast Organelles with Fluorescent Protein Markers
Published on: April 20, 2022
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Visualizing Yeast Organelles with Fluorescent Protein Markers
Chang-Yue Liu1, Jing Zhu1, Zhiping Xie2
1State Key Laboratory of Microbial Metabolism & Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University.
Journal of Visualized Experiments : Jove
|May 9, 2022
Summary
This study provides a protocol for using fluorescent protein markers to visualize organelles in Saccharomyces cerevisiae (yeast). Researchers can now easily track organelle dynamics and protein localization in live yeast cells.
Area of Science:
- Cell Biology
- Microscopy
- Yeast Genetics
Background:
- Saccharomyces cerevisiae is a key model organism for organelle research.
- Previous work established fluorescent markers for major yeast organelles and endomembrane structures.
Purpose of the Study:
- To provide a detailed protocol for utilizing fluorescent protein markers in yeast.
- To enable researchers to study organelle function and dynamics in vivo.
- To serve as a starting point for live-cell fluorescent microscopy of yeast subcellular entities.
Main Methods:
- DNA preparation for yeast transformation.
- Selection and evaluation of transformants.
- Fluorescent microscopic observation and live-cell imaging.
Main Results:
- A comprehensive protocol for yeast organelle visualization is presented.
- The protocol covers essential steps from DNA preparation to microscopic observation.
- Technical notes on microscope hardware and common pitfalls are included.
Conclusions:
- The provided tools and methods facilitate the study of protein subcellular localization in yeast.
- Researchers can effectively track organelles of interest using time-lapse imaging.
- This protocol empowers newcomers to the field of yeast organelle research.
