Green fluorescent protein is a quantitative reporter of gene expression in individual eukaryotic cells.
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.
Soboleski
Mark R
MR
Department of Microbiology and Immunology, Tulane University Medical School, New Orleans, Louisiana, USA.
Oaks
Jason
J
Halford
William P
WP
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015152:Blotting, Northern; D002522:Chlorocebus aethiops; D004318:Doxycycline / Q000494:pharmacology; D005057:Eukaryotic Cells / Q000737:chemistry / Q000378:metabolism*; D005434:Flow Cytometry; D005453:Fluorescence; D015870:Gene Expression / Q000187:drug effects; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics*; D017405:Herpes Simplex Virus Protein Vmw65 / Q000235:genetics / Q000502:physiology; D018259:Herpesvirus 1, Human / Q000235:genetics; D008856:Microscopy, Fluorescence; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D014162:Transfection; D014709:Vero Cells
D017405:Herpes Simplex Virus Protein Vmw65; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D004318:Doxycycline