Abstract
An evaluation has been made of the E. coli beta-galactosidase (beta-gal) gene for use as a reporter gene in mammalian cells in culture. We have adopted a histochemical procedure which enables identification of those cells within a population that express the introduced bacterial gene. Data is presented concerning the sensitivity of the histochemical method relative to an immunological method of detection. It has been found that several clonal cell lines generated after transfection of human 293 cells with a Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter-beta-gal construction are mosaic for expression of the introduced mini-gene. Furthermore, after treatment of these clonal cell lines with the nucleoside analog 5-aza-cytidine (5-aza-C), an increase in production of beta-gal under control of this promoter element was observed.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antibodies, Monoclonal / immunology
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Avian Sarcoma Viruses / genetics
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Azacitidine / pharmacology
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Bacterial Proteins / analysis
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Bacterial Proteins / biosynthesis*
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Bacterial Proteins / immunology
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Cells, Cultured
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Escherichia coli / enzymology
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Escherichia coli / genetics
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Galactosidases / biosynthesis*
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Galactosides*
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Gene Expression Regulation* / drug effects
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Genes, Viral / drug effects
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Glycosides*
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Indoles*
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Promoter Regions, Genetic / drug effects
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Recombinant Fusion Proteins / analysis
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Recombinant Fusion Proteins / biosynthesis*
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Recombinant Proteins / biosynthesis*
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Staining and Labeling / methods*
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beta-Galactosidase / analysis
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beta-Galactosidase / biosynthesis*
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beta-Galactosidase / immunology
Substances
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Antibodies, Monoclonal
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Bacterial Proteins
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Galactosides
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Glycosides
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Indoles
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Recombinant Fusion Proteins
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Recombinant Proteins
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Galactosidases
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beta-Galactosidase
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Azacitidine
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5-bromo-4-chloro-3-indolyl beta-galactoside