Selectable plasmid vectors with alternative and ultrasensitive histochemical marker genes.
Three different histochemical marker genes--E. coli beta-galactosidase gene (lacZ), Drosophila alcohol dehydrogenase gene (ADH) and human placenta alkaline phosphatase gene (ALP)--were cloned into a eukaryotic expression vector also containing the neomycin resistance gene. After calcium phosphate transfection and G418 sulfate selection of recipient BALB/c 3T3 cells, stable transfectants were pooled for histochemical staining. The lacZ-bearing cells produce aqua blue staining for beta-galactosidase; ADH-bearing cells, blue-black staining for alcohol dehydrogenase; and ALP-bearing cells, red staining for alkaline phosphatase. Cells carrying different marker genes can be easily differentiated by double-staining protocols. In addition, various photographic films can be used to enhance the colors of specific histochemically tagged cell classes. These plasmid vectors, providing selectability with the neomycin resistance gene and ultrasensitivity of alternative histochemical marker genes, will be very effective in virtually any biological system requiring analyses of multiple cell clones or classes in culture model systems or in situ.
Dept. of Molecular Biology & Microbiology, Case Western Reserve University School of Medicine, Cleveland, OH 44106.
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D016475:3T3 Cells; D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D003001:Cloning, Molecular; D004352:Drug Resistance, Microbial / Q000235:genetics; D005819:Genetic Markers; D005822:Genetic Vectors; D006651:Histocytochemistry; D006801:Humans; D007763:Lac Operon; D051379:Mice; D008807:Mice, Inbred BALB C; D009355:Neomycin / Q000494:pharmacology; D010784:Photomicrography; D010957:Plasmids; D014162:Transfection
D005819:Genetic Markers; D000426:Alcohol Dehydrogenase; D000469:Alkaline Phosphatase; D009355:Neomycin