Gene expression and cell fusion analyzed by lacZ complementation in mammalian cells.
Complementing reporter genes provide biological indicators of coincident expression of proteins in cells. We have adapted intracistronic complementation of the Escherichia coli lacZ gene for use in mammalian cells. Enzymatic activity detectable by quantitative biochemical assay, flow cytometry, or microscopy is produced upon convergent expression of two distinct mutant lacZ peptides within single cells, or upon fusion of cells expressing such mutants. A novel fluorescent substrate for beta-galactosidase (Fluor-X-Gal) increases detection and permits simultaneous microscopic visualization of other fluorescent markers. The enzymatic complementation described here should facilitate studies of cell fusion, cell lineage, and signal transduction, by producing activity only when two proteins are expressed at the same time and place in intact cells.
Mohler
W A
WA
Department of Molecular Pharmacology, Stanford University Medical Center, CA 94305-5322, USA.
Blau
H M
HM
D000818:Animals; D002459:Cell Fusion; D002478:Cells, Cultured; D005697:Galactosides / Q000378:metabolism; D005786:Gene Expression Regulation; D005816:Genetic Complementation Test; D007211:Indoles / Q000378:metabolism; D007763:Lac Operon; D001616:beta-Galactosidase / Q000235:genetics
D005697:Galactosides; D007211:Indoles; D001616:beta-Galactosidase; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside