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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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Journal ArticleDOI

Leveraging genome-wide datasets to quantify the functional role of the anti-Shine-Dalgarno sequence in regulating translation efficiency.

TL;DR: It is shown that—after accounting for predicted mRNA structure—aSD sequence complementarity increases the translation of endogenous mRNAs by roughly 50%, and it is observed that this relationship is nonlinear, with translation efficiency maximized for m RNAs with intermediate levels of aSD sequences complementarity.
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Barriers to transmission of transcriptional noise in a c-fos c-jun pathway

TL;DR: It is found that in individual cells the expression of c‐fos and c‐jun is noisy and uncorrelated with each other, andquestration of promoters of the downstream genes within compact chromatin is a likely cause of this insensitivity.
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Fluorescent Reporter Libraries as Useful Tools for Optimizing Microbial Cell Factories: A Review of the Current Methods and Applications.

TL;DR: The use of genetically encoded fluorescent reporters for determining the expression level of a particular promoter, not only the synthesis of a specific protein but also the content of intracellular metabolites is used, allowing the rapid identification of the best producers.
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29-Color Flow Cytometry: Unraveling Human Liver NK Cell Repertoire Diversity

TL;DR: A robust approach for diversity profiling of tissue-resident NK cells that can be applied in various homeostatic and pathological conditions such as reproduction, infection, and cancer is presented.
Journal ArticleDOI

Protein Expression Analyses at the Single Cell Level

TL;DR: Key recent progresses in fluorescence imaging-based methods are reviewed and discussed and their application to proteome analysis at the single cell level is discussed.
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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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.
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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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