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Today, already many transcription factors have been characterized, but this is only a fraction of the real diversity that exists in nature. One important family of transcription factors is the well-characterized and widespread Leucine-responsive Regulatory Protein (Lrp) family, which has typically an amino acid as ligand. It is interesting to further explore the diversity of these Lrp-type transcriptional regulators, and to use them in industrial biotechnology approaches to improve detection of amino acids or the bio-based production of certain metabolites. Therefore, I aim to perform an in vivo screening of Lrp-type transcription factors that exist in nature, but that have not yet been characterized. A set of 30 predicted Lrp-type transcription factors will be selected, and together with their corresponding promoter regions, an inducible promoter and fluorescent reporter gene, two plasmids will be constructed and transformed into Escherichia coli to build a set of biosensors. These biosensors will be screened by performing plate reader measurements in both the absence and the presence of different amino acids, to easily find candidates in which regulation and ligand-binding are occurring. Positive candidates will be further characterized with experiments to examine DNA-binding, oligomeric state and protein structure. This screening will result in the characterization of numerous transcription factors and will contribute to the development of efficient production strains.
|Publication status||Published - 2 Nov 2020|
|Event||5th Applied Synthetic Biology in Europe - Online, Delft, Netherlands|
Duration: 2 Nov 2020 → 4 Nov 2020
|Conference||5th Applied Synthetic Biology in Europe|
|Period||2/11/20 → 4/11/20|