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PMID: 15640280 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Green fluorescent protein is a quantitative reporter of gene expression in individual eukaryotic cells.

FASEB journal : official publication of the Federation of American Societies for Experimental Biology ·Vol. 19 ·No. 3 ·2005-03-00 ·Pages 440-2

Soboleski MR, Oaks J, Halford WP

Abstract

Green fluorescent protein (GFP) has gained widespread use as a tool to visualize spatial and temporal patterns of gene expression in vivo. However, it is not generally accepted that GFP can also be used as a quantitative reporter of gene expression. We report that GFP is a reliable reporter of gene expression in individual eukaryotic cells when fluorescence is measured by flow cytometry. Two pieces of evidence support this conclusion: GFP fluorescence increases in direct proportion to the GFP gene copy number delivered to cells by a replication-defective adenovirus vector, Ad.CMV-GFP, and the intensity of GFP fluorescence is directly proportional to GFP mRNA abundance in cells. This conclusion is further supported by the fact that the induction of GFP gene expression from two inducible promoters (i.e., the TRE and ICP0 promoters) is readily detected by flow cytometric measurement of GFP fluorescent intensity. Collectively, the results presented herein indicate that GFP fluorescence is a reliable and quantitative reporter of underlying differences in gene expression.

MeSH Terms
Adenoviridae/genetics Animals Blotting, Northern Chlorocebus aethiops Doxycycline/pharmacology Eukaryotic Cells/chemistry,metabolism Flow Cytometry Fluorescence Gene Expression/drug effects Genetic Vectors Green Fluorescent Proteins/genetics Herpes Simplex Virus Protein Vmw65/genetics,physiology Herpesvirus 1, Human/genetics Microscopy, Fluorescence Promoter Regions, Genetic/genetics RNA, Messenger/analysis Transfection Vero Cells
Chemicals
Herpes Simplex Virus Protein Vmw65 RNA, Messenger Green Fluorescent Proteins Doxycycline
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Soboleski Mark R
Department of Microbiology and Immunology, Tulane University Medical School, New Orleans, Louisiana, USA.
Oaks Jason
Halford William P
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Article Info
Journal
FASEB journal : official publication of the Federation of American Societies for Experimental Biology
Abbr.
FASEB J
ISSN
1530-6860
DOI
10.1096/fj.04-3180fje
Published
2005-03-00
Epub
2005-00-07
Pages
440-2
Language
English
Region
United States
NLM ID
8804484
PMCID
PMC1242169
Subset
IM
Grants
NIAID NIH HHS · R01 AI051414 · United States
NIAID NIH HHS · R01 AI051414-03 · United States
NIAID NIH HHS · R01 AI51414 · United States
External Links
PubMed source DOI: 10.1096/fj.04-3180fje PMC Full Text View journal details
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