Green Fluorescent Protein as a Quantitative Reporter of Relative Promoter Activity in E. coli
Author(s) -
James L. Lissemore,
Jakub Jankowski,
C. Thomas,
David P. Mascotti,
P L deHaseth
Publication year - 2000
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/00281st02
Subject(s) - green fluorescent protein , reporter gene , promoter , escherichia coli , beta galactosidase , microbiology and biotechnology , lac operon , biology , gene , fluorescence , gene expression , biochemistry , physics , quantum mechanics
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 β-galactosidase. We cloned a series of previously characterized synthetic E. coli promoters into GFP and β-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.
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