Berlin 2012 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 7: Posters: Proteins
BP 7.2: Poster
Montag, 26. März 2012, 17:30–19:30, Poster A
Characterization and Application of the redox-sensitive GFP-mutant roGFP — •Sebastien Peter1, Sebastian Wierer3, Kirstin Elgass4, Stefan Bieker1, Alfred Meixner2, Ulrike Zentgraf1, and Frank Schleifenbaum1 — 1Zentrum für Molekularbiologie der Pflanzen, Universität Tübingen, Deutschland — 2Institut für physikalische und theoretische Chemie, Universität Tübingen, Deutschland — 3Protein Crystallography and Molecular Bioinformatics, Universität Konstanz, Deutschland — 4Biochemistry Department, La Trobe University, Bundoora, Australia
For the quantitative analysis of molecular processes in living (plant) cells new combinatorial approaches in optical and spectroscopic technologies are required. The use of green fluorescent protein (GFP) and its variants has revolutionized the in vivo analysis of cell biological processes. More recently, fluorescent proteins have been designed and adapted for targeted sensing of cellular metabolites. Recent progress has been made in generating a redox- sensitive mutant of GFP (roGFP). Here, we characterize the optical properties of roGFP in dependence on the environmental redox state including a comprehensive comparison of different spectral analysis approaches in vitro. Furthermore, we demonstrate several applications of roGFP in living plants such as long-term redox potential readout in plant cells.