title: The glpD gene is a novel reporter gene for E. coli that is superior to established reporter genes like lacZ and gusA.
abstract: Reporter genes facilitate the characterization of promoter activities, transcript stabilities, translational efficiencies, or intracellular localization. Various reporter genes for Escherichia coli have been established, however, most of them have drawbacks like transcript instability or the inability to be used in genetic selections. Therefore, the glpD gene encoding glycerol-3-phosphate dehydrogenase was introduced as a novel reporter gene for E. coli. The enzymatic assay was optimized, and it was verified that growth on glycerol strictly depends on the presence of GlpD. The 5'-UTRs of three E. coli genes were chosen and cloned upstream of the new reporter gene glpD as well as the established reporter genes lacZ and gusA. Protein and transcript levels were quantified and translational efficiencies were calculated. The lacZ transcript was very unstable and its level highly depended on its translation, compromising its use as a reporter. The results obtained with gusA and glpD were similar, however, only glpD can be used for genetic selections. Therefore, glpD was found to be a superior novel reporter gene compared to the established reporter genes lacZ and gusA.
authors: Goethe-University, Biocentre, Institute for Molecular Biosciences, Max-von-Laue-Str. 9, D-60438 Frankfurt, Germany. Electronic address: marius-wegener@gmx.de.
authors: Marius
authors: M
authors: Wegener
authors: Goethe-University, Biocentre, Institute for Molecular Biosciences, Max-von-Laue-Str. 9, D-60438 Frankfurt, Germany. Electronic address: Vogtmann_K@ukw.de.
authors: Kristina
authors: K
authors: Vogtmann
authors: Goethe-University, Biocentre, Institute for Molecular Biosciences, Max-von-Laue-Str. 9, D-60438 Frankfurt, Germany. Electronic address: s4282343@stud.uni-frankfurt.de.
authors: Madeleine
authors: M
authors: Huber
authors: Goethe-University, Biocentre, Institute for Molecular Biosciences, Max-von-Laue-Str. 9, D-60438 Frankfurt, Germany. Electronic address: s.laass@bio.uni-frankfurt.de.
authors: Sebastian
authors: S
authors: Laass
authors: Goethe-University, Biocentre, Institute for Molecular Biosciences, Max-von-Laue-Str. 9, D-60438 Frankfurt, Germany. Electronic address: soppa@bio.uni-frankfurt.de.
authors: Jörg
authors: J
authors: Soppa
mesh_terms: D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D057075:Enzyme Assays / Q000379:methods; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D015964:Gene Expression Regulation, Bacterial; D050939:Gene Fusion; D005798:Genes, Bacterial / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics*; D005990:Glycerol / Q000378:metabolism; D005993:Glycerolphosphate Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D007763:Lac Operon / Q000235:genetics*; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic
chemical_list: D004269:DNA, Bacterial; D005993:Glycerolphosphate Dehydrogenase; C098831:glycerol-3-phosphate oxidase; D005990:Glycerol
keywords: Escherichia coli; In vivo selection; Reporter gene; Transcript stability; Translation efficiency; glpD; gusA; lacZ