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PMID: 2501787 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Analysis of mammalian cell genetic regulation in situ by using retrovirus-derived "portable exons" carrying the Escherichia coli lacZ gene.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 14 ·1989-07-00 ·Pages 5517-21

Brenner DG, Lin-Chao S, Cohen SN

Abstract

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.

MeSH Terms
Animals Blotting, Southern Cells, Cultured Escherichia coli/enzymology,genetics Exons Galactosidases/genetics Genes Genes, Bacterial Genes, Regulator Genetic Vectors Mice Moloney murine leukemia virus/genetics Promoter Regions, Genetic Restriction Mapping Transcription, Genetic beta-Galactosidase/genetics
Chemicals
Galactosidases beta-Galactosidase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Brenner D G
Department of Genetics, Stanford University School of Medicine, CA 94305-5120.
Lin-Chao S
Cohen S N
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.86.14.5517
Published
1989-07-00
Pages
5517-21
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC297654
Subset
IM
Grants
NIGMS NIH HHS · GM 27241 · United States
External Links
PubMed source DOI: 10.1073/pnas.86.14.5517 PMC Full Text View journal details
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