JOURNAL ARTICLE

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

Jung-uk Shim, Luis F Olguin, Graeme Whyte, Duncan Scott, Ann Babtie, Chris Abell, Wilhelm T S Huck, Florian Hollfelder
Journal of the American Chemical Society 2009 October 28, 131 (42): 15251-6
19799429
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.

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