Quantifying E. coli proteome and transcriptome with single-molecule sensitivity in single cells.
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.
Taniguchi
Yuichi
Y
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA.
Choi
Paul J
PJ
Li
Gene-Wei
GW
Chen
Huiyi
H
Babu
Mohan
M
Hearn
Jeremy
J
Emili
Andrew
A
Xie
X Sunney
XS
D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000032:analysis* / Q000378:metabolism; D015870:Gene Expression; D020869:Gene Expression Profiling; D015723:Gene Library; D017404:In Situ Hybridization, Fluorescence; D008164:Luminescent Proteins; D046210:Microfluidic Analytical Techniques; D008856:Microscopy, Fluorescence; D014176:Protein Biosynthesis; D020543:Proteome / Q000032:analysis*; D020871:RNA Stability; D012329:RNA, Bacterial / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
D029968:Escherichia coli Proteins; D008164:Luminescent Proteins; D020543:Proteome; D012329:RNA, Bacterial; D012333:RNA, Messenger