Histochemical staining of clonal mammalian cell lines expressing E. coli beta galactosidase indicates heterogeneous expression of the bacterial gene.
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.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001358:Avian Sarcoma Viruses / Q000235:genetics; D001374:Azacitidine / Q000494:pharmacology; D001426:Bacterial Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000276:immunology; D002478:Cells, Cultured; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005696:Galactosidases / Q000096:biosynthesis*; D005697:Galactosides; D005786:Gene Expression Regulation / Q000187:drug effects; D005814:Genes, Viral / Q000187:drug effects; D006027:Glycosides; D007211:Indoles; D011401:Promoter Regions, Genetic / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013194:Staining and Labeling / Q000379:methods*; D001616:beta-Galactosidase / Q000032:analysis / Q000096:biosynthesis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D005697:Galactosides; D006027:Glycosides; D007211:Indoles; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D005696:Galactosidases; D001616:beta-Galactosidase; D001374:Azacitidine; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside