Analysis of mammalian cell genetic regulation in situ by using retrovirus-derived "portable exons" carrying the Escherichia coli lacZ gene.
Self-inactivating derivatives of Moloney murine leukemia retrovirus containing the Escherichia coli lacZ gene were used to detect and study the regulation of transcription initiated at chromosomally located promoters in mouse fibroblasts. The introduction of splice acceptor sites in all three translational reading frames relative to lacZ and the inclusion of an in-frame ATG translation start codon in one construct allowed synthesis of beta-galactosidase fusion proteins upon insertion of retrovirus vectors containing lacZ into introns 3' to either protein-coding or noncoding exons. Selection of lacZ-expressing cells by fluorescence-activated cell sorting and the analysis of beta-galactosidase production after serum deprivation has yielded lines in which lacZ was fused to genes induced by growth arrest in the G0 state.
Brenner
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DG
Department of Genetics, Stanford University School of Medicine, CA 94305-5120.
Lin-Chao
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S
Cohen
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SN
D000818:Animals; D015139:Blotting, Southern; D002478:Cells, Cultured; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005091:Exons; D005696:Galactosidases / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D005809:Genes, Regulator; D005822:Genetic Vectors; D051379:Mice; D008979:Moloney murine leukemia virus / Q000235:genetics*; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000235:genetics*
D005696:Galactosidases; D001616:beta-Galactosidase