Green fluorescent protein as a quantitative reporter of relative promoter activity in E. coli.
Green fluorescent protein (GFP) has become a valuable tool for the detection of gene expression in prokaryotes and eukaryotes. To evaluate its potential for quantitation of relative promoter activity in E. coli, we have compared GFP with the commonly used reporter gene lacZ, encoding beta-galactosidase. We cloned a series of previously characterized synthetic E. coli promoters into GFP and beta-galactosidase reporter vectors. Qualitative and quantitative assessments of these constructs show that (a) both reporters display similar sensitivities in cells grown on solid or liquid media and (b) GFP is especially well suited for quantitation of promoter activity in cells grown on agar. Thus, GFP provides a simple, rapid and sensitive tool for measuring relative promoter activity in intact E. coli cells.
Lissemore
J L
JL
Department of Biology, John Carroll University, University Heights, OH 44118, USA. jlissemore@jcu.edu
Jankowski
J T
JT
Thomas
C B
CB
Mascotti
D P
DP
deHaseth
P L
PL
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D007763:Lac Operon / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D001616:beta-Galactosidase / Q000378:metabolism
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase