Single-cell measurement of plasmid copy number and promoter activity
Open Access
- 5 March 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Communications
- Vol. 12 (1), 1-9
- https://doi.org/10.1038/s41467-021-21734-y
Abstract
Accurate measurements of promoter activities are crucial for predictably building genetic systems. Here we report a method to simultaneously count plasmid DNA, RNA transcripts, and protein expression in single living bacteria. From these data, the activity of a promoter in units of RNAP/s can be inferred. This work facilitates the reporting of promoters in absolute units, the variability in their activity across a population, and their quantitative toll on cellular resources, all of which provide critical insights for cellular engineering.Funding Information
- United States Department of Commerce | National Institute of Standards and Technology (70-NANB16H164)
- U.S. Department of Energy (DE-FOA-0001650)
This publication has 67 references indexed in Scilit:
- Single-molecule dynamics of transcription of the lar promoterPhysical Biology, 2012
- Mechanism of Transcription Initiation at an Activator-Dependent Promoter Defined by Single-Molecule ObservationCell, 2012
- Central dogma at the single-molecule level in living cellsNature, 2011
- Transcript amplification from single bacterium for transcriptome analysisGenome Research, 2011
- General properties of transcriptional time series in Escherichia coliNature Genetics, 2011
- Synthetic biology: applications come of ageNature Reviews Genetics, 2010
- Structural basis for the coevolution of a viral RNA–protein complexNature Structural & Molecular Biology, 2007
- Real-Time Kinetics of Gene Activity in Individual BacteriaCell, 2005
- From Silencing to Gene Expression: Real-Time Analysis in Single CellsCell, 2004
- Markovian Modeling of Gene-Product SynthesisTheoretical Population Biology, 1995