Optimization and in situ detection of Escherichia coli beta-galactosidase gene expression in Dictyostelium discoideum.
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.
Dingermann
T
T
Institut für Biochemie, Medizinischen Fakultät, Erlangen, F.R.G.
Reindl
N
N
Werner
H
H
Hildebrandt
M
M
Nellen
W
W
Harwood
A
A
Williams
J
J
Nerke
K
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004023:Dictyostelium / Q000235:genetics*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005696:Galactosidases / Q000235:genetics*; D015870:Gene Expression; D005798:Genes, Bacterial; D006651:Histocytochemistry; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism
D011994:Recombinant Proteins; D005696:Galactosidases; D001616:beta-Galactosidase