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Related Experiment Videos

Beta-galactosidase activity in single differentiating bacterial cells

F Russo-Marie1, M Roederer, B Sager

  • 1Department of Biochemistry, Beckman Center for Molecular and Genetic Medicine, Stanford University School of Medicine, CA 94305.

Proceedings of the National Academy of Sciences of the United States of America
|September 1, 1993
PubMed
Summary

Researchers developed a method to measure beta-galactosidase activity in Myxococcus xanthus using a fluorogenic substrate. This technique allows for the physical separation of cells based on gene expression levels, aiding in the study of cellular differentiation.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • Myxococcus xanthus exhibits complex developmental processes.
  • Understanding gene expression dynamics is crucial for deciphering cellular differentiation.

Purpose of the Study:

  • To develop and validate a method for analyzing beta-galactosidase expression in M. xanthus.
  • To investigate heterogeneity in gene expression during M. xanthus development.

Main Methods:

  • Utilized transcriptional fusions to lacZ in M. xanthus strains.
  • Employed the fluorogenic substrate fluorescein di-beta-galactopyranoside for beta-galactosidase detection.
  • Applied cell fractionation based on fluorescence levels.

Main Results:

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  • Fluorescein product was retained intracellularly, preserving cell viability.
  • Fluorescence intensity correlated linearly with beta-galactosidase activity.
  • Demonstrated cell separation based on high vs. no beta-galactosidase expression in a specific strain (Tn5 lac omega 4473) at 9 hours of development.

Conclusions:

  • The developed fluorogenic assay is effective for quantifying beta-galactosidase activity in M. xanthus.
  • This method enables the physical separation of cells with differential gene expression.
  • The approach has potential applications for studying differentiation in other organisms.