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Related Experiment Video

Updated: Dec 3, 2025

Saccharomyces cerevisiae Metabolic Labeling with 4-thiouracil and the Quantification of Newly Synthesized mRNA As a Proxy for RNA Polymerase II Activity
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Protocol for Titrating Gene Expression Levels in Budding Yeast.

Yimiao Qu1, Jun Jiang1, Xiaojing Yang1

  • 1Center for Quantitative Biology and Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871, China.

STAR Protocols
|October 28, 2020
PubMed
Summary
This summary is machine-generated.

This study presents a method to precisely control and measure gene expression levels in budding yeast by altering gene copy numbers. This technique allows for stable gene expression control, using fluorescent intensity as a readout.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Gene expression levels significantly influence an organism's biological traits and functions.
  • Precise control and measurement of gene expression are crucial for understanding cellular processes and developing biotechnological applications.

Purpose of the Study:

  • To develop a robust protocol for quantitative observation and titration of target gene expression in budding yeast.
  • To establish a method for stable, inherent control of gene expression by manipulating genomic copy number.

Main Methods:

  • The protocol involves altering the copy number of a target gene within the budding yeast genome.
  • Various yeast strains were constructed with differing copy numbers of the cell cycle inhibitor Whi5.
  • Fluorescent intensity was utilized as a quantitative readout to measure gene expression levels.

Main Results:

  • The developed protocol allows for precise titration of gene expression.
  • Stable and predictable control of gene expression at desired levels was achieved.
  • Fluorescent intensity accurately reflects the quantitative gene expression levels.

Conclusions:

  • This method provides a reliable way to manipulate and quantify gene expression in budding yeast.
  • The protocol facilitates the study of gene function and regulation by enabling fine-tuning of expression levels.
  • The technique is valuable for research in genetics, molecular biology, and synthetic biology.