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Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level
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Quantifying E. coli proteome and transcriptome with single-molecule sensitivity in single cells.

Yuichi Taniguchi1, Paul J Choi, Gene-Wei Li

  • 1Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA.

Science (New York, N.Y.)
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PubMed
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Single bacterial cells show cell-to-cell variations in protein and messenger RNA (mRNA) levels. These molecular numbers are uncorrelated between protein and mRNA for any specific gene.

Area of Science:

  • Bacterial gene expression
  • Single-cell biology
  • Molecular biology

Background:

  • Cell-to-cell variability in protein and messenger RNA (mRNA) copy numbers is observed in bacterial populations.
  • Low molecule numbers make detection in single cells challenging.

Purpose of the Study:

  • To perform quantitative system-wide analyses of protein and mRNA expression in individual bacterial cells.
  • To achieve single-molecule sensitivity in analyzing gene expression.

Main Methods:

  • Utilized a newly constructed yellow fluorescent protein fusion library for Escherichia coli.
  • Conducted quantitative system-wide analyses of protein and mRNA expression.
  • Applied single-molecule sensitivity detection methods.

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Probing mRNA Kinetics in Space and Time in Escherichia coli using Two-Color Single-Molecule Fluorescence In Situ Hybridization
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Probing mRNA Kinetics in Space and Time in Escherichia coli using Two-Color Single-Molecule Fluorescence In Situ Hybridization

Published on: July 30, 2020

Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments
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Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments

Published on: December 21, 2017

Related Experiment Videos

Last Updated: Jun 10, 2026

Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level
08:29

Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level

Published on: April 19, 2019

Probing mRNA Kinetics in Space and Time in Escherichia coli using Two-Color Single-Molecule Fluorescence In Situ Hybridization
10:01

Probing mRNA Kinetics in Space and Time in Escherichia coli using Two-Color Single-Molecule Fluorescence In Situ Hybridization

Published on: July 30, 2020

Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments
07:51

Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments

Published on: December 21, 2017

Main Results:

  • Protein number distributions in single cells largely follow a gamma distribution.
  • At low expression, distribution parameters relate to transcription rate and protein burst size.
  • Extrinsic noise significantly influences distributions at high expression levels.
  • Protein and mRNA copy numbers for a given gene are uncorrelated within individual cells.

Conclusions:

  • The gamma distribution effectively models protein number variations in single bacterial cells.
  • Distinguishes contributions of intrinsic and extrinsic noise to gene expression variability.
  • Highlights the lack of correlation between mRNA and protein levels in single bacterial cells.