scispace - formally typeset
Open AccessJournal ArticleDOI

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

Reads0
Chats0
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.

read more

Content maybe subject to copyright    Report

Citations
More filters
Journal ArticleDOI

Ubiquitin-mediated control of plant hormone signaling.

TL;DR: The ubiquitin-proteasome system plays a role in nearly every aspect of plant biology and rivals the well-studied area of transcriptional regulation for importance in cellular regulation.
Journal ArticleDOI

Promoter sequence determines the relationship between expression level and noise

TL;DR: A single transcription factor can activate or repress expression by three different mechanisms: one that increases cell-to-cell variability in target gene expression (noise) and two that decrease noise.
Journal ArticleDOI

Mapping the environmental fitness landscape of a synthetic gene circuit

TL;DR: It is found that knowing the fitness and nongenetic memory associated with specific gene expression states were necessary for predicting the overall fitness of cell populations in combined environments, and may have important implications for phenotypic variability of drug resistance in natural settings.
Journal ArticleDOI

Measurements of translation initiation from all 64 codons in E. coli

TL;DR: In this article, the authors quantified translation initiation of green fluorescent protein and nanoluciferase in E. coli from all 64 triplet codons and across a range of DNA copy number.
Journal ArticleDOI

Single cell-resolution western blotting

TL;DR: This protocol describes how to perform western blotting on individual cells to measure cell-to-cell variation in protein expression levels and protein state and describes an optional gel de-cross-linking protocol for more effective introduction of antibodies into the gel layer.
References
More filters

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.
Journal ArticleDOI

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.
Related Papers (5)