pREPORT: a multi-readout transcription reporter vector for fission yeast.
Transcription factors are prominent regulators of gene expression that execute responses to various intracellular and extracellular stimuli. Recombinant transcription reporter systems can be conveniently used to study the DNA binding preferences and regulatory activity of a transcription factor under a range of conditions. Several reporter genes have been used to study transcription regulation in the fission yeast Schizosaccharomyces pombe. Each of these reporters has distinct advantages, such as high sensitivity or ease of use, and limitations, such as prohibitive costs or use of hazardous substances. To combine the strengths and mitigate the weaknesses of individual reporter genes, we have created pREPORT, a flexible multi-readout transcription reporter vector for fission yeast that employs an enhanced GFP-lacZ fusion and a customizable minimal promoter. With pREPORT, gene expression driven by the transcription factor of interest can be quantified in a number of ways, both in live cells and in vitro, using a single reporter construct.
Department of Cell Biology, Faculty of Science, Charles University in Prague, Prague, Czech Republic.
Martin
M
Převorovský
D020869:Gene Expression Profiling / Q000379:methods; D015966:Gene Expression Regulation, Fungal; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000032:analysis; D012568:Schizosaccharomyces / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D001616:beta-Galactosidase / Q000032:analysis
C569834:Cbf11 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase
GFP; Schizosaccharomyces pombe; fission yeast; plasmid; transcription reporter; β-galactosidase