Unit

Parallel High-Throughput Automated Assays to Measure Cell Growth and Beta-Galactosidase Reporter Gene Expression in the Yeast Saccharomyces cerevisiae

  1. Andrew D. Napper1,3,
  2. Nuzhat Motlekar1,
  3. Rogerio Alves de Almeida2,
  4. Graham D. Pavitt2

Published Online: 1 JAN 2011

DOI: 10.1002/9780470559277.ch100119

Current Protocols in Chemical Biology

Current Protocols in Chemical Biology

How to Cite

Napper, A. D., Motlekar, N., de Almeida, R. A. and Pavitt, G. D. 2011. Parallel High-Throughput Automated Assays to Measure Cell Growth and Beta-Galactosidase Reporter Gene Expression in the Yeast Saccharomyces cerevisiae. Current Protocols in Chemical Biology. 3:1–14.

Author Information

  1. 1

    Penn Center for Molecular Discovery, Institute for Medicine and Engineering, and Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, Pennsylvania

  2. 2

    Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom

  3. 3

    Nemours Center for Childhood Cancer Research, Wilmington, Delaware

Publication History

  1. Published Online: 1 JAN 2011
  2. Published Print: JAN 2011

Abstract

Parallel high-throughput automated assays are described for the measurement of cell growth and β-galactosidase reporter gene expression from a single culture of the yeast S. cerevisiae. The dual assay measures the effect of test compounds on expression of a specific gene of interest linked to the β-galactosidase reporter gene, and simultaneously tests for compound toxicity and other effects on cell growth. Examples of assay development and validation results are used to illustrate how this protocol may be used to screen two yeast cell lines in parallel. Yeast cells are grown overnight in V-bottom polypropylene 384-well plates, after which portions of the cell suspension are transferred to clear and to white flat-bottom 384-well plates for measurement of cell growth and reporter gene expression, respectively. Cell growth is determined by measurement of absorbance at 595 nm, and β-galactosidase expression is quantified by Beta-Glo, a commercially available luminescent β-galactosidase substrate. Curr. Protoc. Chem. Biol. 3:1-14 © 2011 by John Wiley & Sons, Inc.

Keywords:

  • cell growth;
  • yeast;
  • reporter gene;
  • luciferase;
  • cell-based assay;
  • model organism;
  • mutant gene;
  • β-galactosidase;
  • luminescence