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Quantifying E. coli proteome and transcriptome with single-molecule sensitivity in single cells.

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TLDR
System-wide analyses of protein and mRNA expression in individual cells with single-molecule sensitivity using a newly constructed yellow fluorescent protein fusion library for Escherichia coli found that almost all protein number distributions can be described by the gamma distribution with two fitting parameters which, at low expression levels, have clear physical interpretations as the transcription rate and protein burst size.
Abstract
Protein and messenger RNA (mRNA) copy numbers vary from cell to cell in isogenic bacterial populations. However, these molecules often exist in low copy numbers and are difficult to detect in single cells. We carried out quantitative system-wide analyses of protein and mRNA expression in individual cells with single-molecule sensitivity using a newly constructed yellow fluorescent protein fusion library for Escherichia coli. We found that almost all protein number distributions can be described by the gamma distribution with two fitting parameters which, at low expression levels, have clear physical interpretations as the transcription rate and protein burst size. At high expression levels, the distributions are dominated by extrinsic noise. We found that a single cell's protein and mRNA copy numbers for any given gene are uncorrelated.

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A Kinetic Platform to Determine the Fate of Hydrogen Peroxide in Escherichia coli.

TL;DR: A quantitative, systems-level model of H2O2 detoxification in Escherichia coli that includes detoxification enzymes, H2o2-dependent transcriptional regulation, enzyme degradation, the Fenton reaction and damage caused by •OH, oxidation of biomolecules by H2 O2, and repair processes is constructed.
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Micro-patterned agarose gel devices for single-cell high-throughput microscopy of E. coli cells

TL;DR: Micro-patterning of agarose pads is implemented into multi-pad devices called CapsuleHotel and LineHotel for high-throughput imaging, which will accelerate discoveries in diverse fields ranging from microbiology to systems and synthetic biology.
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Protein complexes, big data, machine learning and integrative proteomics: lessons learned over a decade of systematic analysis of protein interaction networks

TL;DR: Key lessons learned are recounted, with an emphasis on successful workflows, and challenges, arising from the own and other groups’ ongoing efforts to generate, interpret and report proteome-scale interaction networks in increasingly diverse biological contexts.
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Single-cell transcriptomics of small microbial eukaryotes: limitations and potential

TL;DR: It is found that the reason of such observation is that the smaller organisms had much lower mRNA copy numbers, so the application of single-cell RNA-seq in studying smaller microbial eukaryotes in the context of these limitations is discussed.
Journal ArticleDOI

Dual transcript and protein quantification in a massive single cell array

TL;DR: A microwell-based cytometric method for simultaneous measurements of gene and protein expression dynamics in thousands of single cells and believes this platform is applicable for interrogating the dynamics of gene expression, protein expression, and translational kinetics at the single-cell level.
References
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疟原虫var基因转换速率变化导致抗原变异[英]/Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A

宁北芳, +1 more
TL;DR: PfPMP1)与感染红细胞、树突状组胞以及胎盘的单个或多个受体作用,在黏附及免疫逃避中起关键的作�ly.
Journal ArticleDOI

Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection.

TL;DR: These mutants—the ‘Keio collection’—provide a new resource not only for systematic analyses of unknown gene functions and gene regulatory networks but also for genome‐wide testing of mutational effects in a common strain background, E. coli K‐12 BW25113.
Journal ArticleDOI

Stochastic Gene Expression in a Single Cell

TL;DR: This work constructed strains of Escherichia coli that enable detection of noise and discrimination between the two mechanisms by which it is generated and reveals how low intracellular copy numbers of molecules can fundamentally limit the precision of gene regulation.
Journal ArticleDOI

Global analysis of protein localization in budding yeast

TL;DR: The construction and analysis of a collection of yeast strains expressing full-length, chromosomally tagged green fluorescent protein fusion proteins helps reveal the logic of transcriptional co-regulation, and provides a comprehensive view of interactions within and between organelles in eukaryotic cells.
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Fabrication of microfluidic systems in poly(dimethylsiloxane)

TL;DR: Fabrication of microfluidic devices in poly(dimethylsiloxane) (PDMS) by soft lithography provides faster, less expensive routes to devices that handle aqueous solutions.
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