Simultaneous determination of gene expression and enzymatic activity in individual bacterial cells in microdroplet compartments

J Am Chem Soc. 2009 Oct 28;131(42):15251-6. doi: 10.1021/ja904823z.

Abstract

A microfluidic device capable of storing picoliter droplets containing single bacteria at constant volumes has been fabricated in PDMS. Once captured in droplets that remain static in the device, bacteria express both a red fluorescent protein (mRFP1) and the enzyme, alkaline phosphatase (AP), from a biscistronic construct. By measuring the fluorescence intensity of both the mRFP1 inside the cells and a fluorescent product formed as a result of the enzymatic activity outside the cells, gene expression and enzymatic activity can be simultaneously and continuously monitored. By collecting data from many individual cells, the distribution of activities in a cell is quantified and the difference in activity between two AP mutants is measured.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / analysis*
  • Escherichia coli / chemistry*
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Gene Expression*
  • Kinetics
  • Luminescent Proteins / analysis*
  • Microfluidic Analytical Techniques / instrumentation
  • Microfluidic Analytical Techniques / methods*
  • Red Fluorescent Protein

Substances

  • Luminescent Proteins
  • Alkaline Phosphatase