Histochemical techniques for locating Escherichia coli beta-galactosidase activity in transgenic organisms

Genet Anal Tech Appl. Oct-Dec 1992;9(5-6):151-8. doi: 10.1016/1050-3862(92)90042-4.

Abstract

Escherichia coli beta-galactosidase is a commonly used reporter molecule for analyzing gene expression. Recently, beta-galactosidase fusions have been applied to a variety of eukaryotic systems. The techniques for constructing and introducing beta-galactosidase fusion constructs as well as soluble assays for total enzyme function have been described in detail elsewhere. This article describes histochemical techniques for analyzing organisms that contain a functional beta-galactosidase fusion construct. The object is to determine semiquantitatively which cells are expressing the beta-galactosidase fusion protein, as well as the subcellular localization of the protein. Due to its prevalence in the author's laboratory, Caenorhabditis elegans is used as a canonical organism for the detailed methods described.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Caenorhabditis elegans / enzymology*
  • Chromogenic Compounds
  • Escherichia coli / enzymology*
  • Galactosides
  • Histocytological Preparation Techniques*
  • Indoles
  • Recombinant Fusion Proteins / isolation & purification*
  • Substrate Specificity
  • beta-Galactosidase / isolation & purification*
  • beta-Galactosidase / metabolism

Substances

  • Chromogenic Compounds
  • Galactosides
  • Indoles
  • Recombinant Fusion Proteins
  • beta-Galactosidase
  • 5-bromo-4-chloro-3-indolyl beta-galactoside