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Project

Photophysics and application of novel fluorescent proteins (FPs) and FP based calcium indicators.

Fluorescent proteins (FPs) are a class of proteins that possess the remarkable feature that they can autocatalytically form the fluorophore after expression from their respective gene, so the fluorescence is genetically encoded. This property, in combination with the existing knowledge of DNA manipulation, allows one to label whichever protein in the genomewith intrinsic fluorescence by making a fusion of the gene coding for the FP with the one coding for the protein. The field of biolabeling was revolutionized because of this. All FPs display a complex photophysical behavior which complicates quantitative data-analysis form these often used markers. In this project attention is focused on the photophysical characterization of new FPs (the fast folding protein Venus) or FP based indicators (a so-called chameleon-protein based on a green donor and a red acceptor for energy transfer (FRET)). This characterization will occur on bulk level (advanced ultrafast measurements) but also on the singlemolecule level (fluorescence microscopy). The acquired insights will beused on the one hand to improve FPs and on the other hand to obtain quantitative information in selected biophysical issues. For Venus a concrete application is the research for persistence in bacterial cells.
Date:1 Oct 2009 →  18 Sep 2016
Keywords:Single molecule fluorescence microscopy, Photochemistry, Molecular biophysics, Fluorescence spectroscopy
Disciplines:Medicinal and biomolecular chemistry, Molecular and cell biology, Plant biology, Systems biology, Biophysics, Physical chemistry, Sustainable chemistry
Project type:PhD project