Optimization and in situ detection of Escherichia coli beta-galactosidase gene expression in Dictyostelium discoideum

Gene. 1989 Dec 28;85(2):353-62. doi: 10.1016/0378-1119(89)90428-9.

Abstract

We show that a fusion gene, containing the promoter and 5'-noncoding region of a Dictyostelium discoideum actin 6 gene linked to the Escherichia coli beta-galactosidase (beta Gal) gene (lacZ), directs the production of functionally active beta Gal in D. discoideum and that the enzyme can be detected by staining in situ; a procedure which will be of great value in analyzing cell-type-specific gene expression. We illustrate this by fusing lacZ to the promoter of the prespore-specific gene, D19, and localizing expressing cells in migrating slugs. Optimal expression requires the inclusion of termination and polyadenylylation signals and we describe pDDlac, a vector containing a multiple cloning site upstream from a lacZ-Dictyostelium terminator fusion, which can be used to analyze regulated promoters.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • Dictyostelium / genetics*
  • Escherichia coli / enzymology
  • Escherichia coli / genetics*
  • Galactosidases / genetics*
  • Gene Expression
  • Genes, Bacterial*
  • Histocytochemistry
  • Molecular Sequence Data
  • Plasmids
  • Recombinant Proteins / metabolism
  • beta-Galactosidase / genetics*
  • beta-Galactosidase / metabolism

Substances

  • Recombinant Proteins
  • Galactosidases
  • beta-Galactosidase