Single-cell measurement of plasmid copy number and promoter activity

Nat Commun. 2021 Mar 5;12(1):1475. doi: 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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacteria / genetics*
  • DNA Copy Number Variations*
  • Escherichia coli / genetics
  • Gene Expression Regulation, Bacterial
  • Plasmids*
  • Promoter Regions, Genetic
  • RNA, Messenger / metabolism
  • Transcription, Genetic

Substances

  • RNA, Messenger