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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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Effects of σ factor competition are promoter initiation kinetics dependent.

TL;DR: A promoter's responsiveness to indirect regulation by σ factor competition is determined by the sequence-dependent kinetics of the rate limiting steps of transcription initiation, and it is concluded that, in E. coli, the smaller is the time-scale of the closed complex formation relative to the open complex formation, the weaker is a promoters' responsiveness to changes in σ(38) numbers.
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Bacterial responses to antibiotics and their combinations.

TL;DR: Recent progress in the understanding of bacterial responses to antibiotics and their combinations is reviewed, focusing on effects at the levels of growth rate and gene expression.
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Quantification of very low-abundant proteins in bacteria using the HaloTag and epi-fluorescence microscopy

TL;DR: The use of the self-labelling protein tag called HaloTag, in conjunction with an organic fluorescent dye, to label and accurately count endogenous proteins present in very low numbers (<7) in individual Escherichia coli cells is characterized.
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Method for physiologic phenotype characterization at the single-cell level in non-interacting and interacting cells.

TL;DR: A measurement platform that features the capability to quantify oxygen consumption rates of individual, non-interacting and interacting cells under normoxic and hypoxic conditions is presented and the effects of cell-to-cell interactions by using the system's capability to isolate two and three cells in a single well are measured.
Journal ArticleDOI

Kinetic Foundation of the Zero-Inflated Negative Binomial Model for Single-Cell RNA Sequencing Data

TL;DR: Single-cell RNA sequencing data have complex features such as dropout events, overdispersion, and high-magnitude outliers, resulting in complicated probability distributions of mRNA abundances that...
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

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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.
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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.
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