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Single-cell measurement of plasmid copy number and promoter activity.
Shao, Bin; Rammohan, Jayan; Anderson, Daniel A; Alperovich, Nina; Ross, David; Voigt, Christopher A.
Afiliação
  • Shao B; Synthetic Biology Center, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
  • Rammohan J; Biosystems and Biomaterials Division, Material Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, MD, USA.
  • Anderson DA; Synthetic Biology Center, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
  • Alperovich N; Biosystems and Biomaterials Division, Material Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, MD, USA.
  • Ross D; Biosystems and Biomaterials Division, Material Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, MD, USA.
  • Voigt CA; Synthetic Biology Center, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA. cavoigt@gmail.com.
Nat Commun ; 12(1): 1475, 2021 03 05.
Article em En | MEDLINE | ID: mdl-33674569
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.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Plasmídeos / Bactérias / Variações do Número de Cópias de DNA Idioma: En Ano de publicação: 2021 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Plasmídeos / Bactérias / Variações do Número de Cópias de DNA Idioma: En Ano de publicação: 2021 Tipo de documento: Article