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PMID: 2440117 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Histochemical staining of clonal mammalian cell lines expressing E. coli beta galactosidase indicates heterogeneous expression of the bacterial gene.

Somatic cell and molecular genetics ·Vol. 13 ·No. 3 ·1987-05-00 ·Pages 253-65

MacGregor GR, Mogg AE, Burke JF, Caskey CT

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.

MeSH Terms
Animals Antibodies, Monoclonal/immunology Avian Sarcoma Viruses/genetics Azacitidine/pharmacology Bacterial Proteins/analysis,biosynthesis,immunology Cells, Cultured Escherichia coli/enzymology,genetics Galactosidases/biosynthesis Galactosides Gene Expression Regulation/drug effects Genes, Viral/drug effects Glycosides Indoles Promoter Regions, Genetic/drug effects Recombinant Fusion Proteins/analysis,biosynthesis Recombinant Proteins/biosynthesis Staining and Labeling/methods beta-Galactosidase/analysis,biosynthesis,immunology
Chemicals
Antibodies, Monoclonal Bacterial Proteins Galactosides Glycosides Indoles Recombinant Fusion Proteins Recombinant Proteins Galactosidases beta-Galactosidase Azacitidine 5-bromo-4-chloro-3-indolyl beta-galactoside
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
MacGregor G R
Mogg A E
Burke J F
Caskey C T
Article Info
Journal
Somatic cell and molecular genetics
Abbr.
Somat Cell Mol Genet
ISSN
0740-7750
Published
1987-05-00
Pages
253-65
Language
English
Region
United States
NLM ID
8403568
Subset
IM
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