Quantifying E. coli proteome and transcriptome with single-molecule sensitivity in single cells.
Yuichi Taniguchi,Paul J. Choi,Gene-Wei Li,Huiyi Chen,Mohan Babu,Jeremy Hearn,Andrew Emili,X. Sunney Xie +7 more
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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.read more
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Lighting-up RNA aptamer transcription synchronization amplification for ultrasensitive and label-free imaging of microRNA in single cells
TL;DR: A new in situ rolling circle transcription synchronization machinery (RCTsm) of lighting-up RNA aptamer strategy for highly sensitive imaging and selective differentiation of miRNA expression levels in cells is reported, with great potential for early cancer diagnosis as well as biomedical research.
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Label-free nanofluidic scattering microscopy of size and mass of single diffusing molecules and nanoparticles
Barbora Špačková,Henrik Klein Moberg,Joachim Fritzsche,Johan Tenghamn,Gustaf Sjösten,Hana Šípová-Jungová,David Albinsson,Quentin Lubart,Daniel van Leeuwen,Fredrik Westerlund,Daniel Midtvedt,Elin K. Esbjörner,Mikael Käll,Giovanni Volpe,Christoph Langhammer +14 more
TL;DR: In this article , a nanofluidic scattering microscopy (NSM) was proposed to enable real-time imaging of single biomolecules diffusing inside a nano-fluidic channel.
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Nucleoid and cytoplasmic localization of small RNAs in Escherichia coli.
TL;DR: The intracellular localization of sRNAs transcribed from plasmids in Escherichia coli is investigated using RNA fluorescent in-situ hybridization and it is found that s RNAs have equal preference for the nucleoid and cytoplasm, and no preferential localization at the cell membrane.
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Single-cell analysis of ribonucleotide reductase transcriptional and translational response to DNA damage.
TL;DR: Surprisingly, RNR subunit mRNA levels were comparably low in both damaged and undamaged G1 cells and highly induced in damaged S/G2 cells, and the differential RNR response to DNA damage correlated with variable Mec1 kinase activity in the cell cycle in single cells.
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Exponential trajectories, cell size fluctuations, and the adder property in bacteria follow from simple chemical dynamics and division control.
TL;DR: A cell is modeled as a set of chemical populations undergoing nonlinear mass action kinetics in a container whose volume is a linear function of the chemical populations, which explains exponential balanced growth of bacterial cells without invoking any regulatory mechanisms.
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
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.
Tomoya Baba,Takeshi Ara,Miki Hasegawa,Yuki Takai,Yoshiko Okumura,Miki Baba,Kirill A. Datsenko,Masaru Tomita,Barry L. Wanner,Hirotada Mori,Hirotada Mori +10 more
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.
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Global analysis of protein localization in budding yeast
Won-Ki Huh,James V. Falvo,Luke C. Gerke,Adam S. Carroll,Russell W. Howson,Jonathan S. Weissman,Erin K. O'Shea +6 more
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)
J.C. McDonald,David C. Duffy,Janelle R. Anderson,Daniel T. Chiu,Hongkai Wu,Olivier Schueller,George M. Whitesides +6 more
TL;DR: Fabrication of microfluidic devices in poly(dimethylsiloxane) (PDMS) by soft lithography provides faster, less expensive routes to devices that handle aqueous solutions.