Technique - (1) Genetic code expansion and site-specific fluorophore labeling

Type: Experimental

Description: Orthogonal tRNA/synthetase pairs install non-canonical amino acids for click-chemistry attachment of dyes at single defined residues, enabling smFRET and SMLM in cells.

Department(s)/lab(s): Biology / Institute of Molecular Biology (IMB) | Lemke Lab - Synthetic Biophysics @ JGU
Summary:

Lemke holds the chair of Synthetic Biophysics at JGU and is adjunct director at the Institute of Molecular Biology. The group's signature is combining genetic code expansion -- installing non-canonical amino acids so a dye can be clicked onto one chosen residue -- with single-molecule fluorescence: smFRET on intrinsically disordered proteins, super-resolution imaging of the nuclear pore complex and its FG-nucleoporin permeability barrier, and engineered membraneless organelles used as designer compartments in living cells. The result is single-molecule-resolution measurement of conformational dynamics and phase behaviour inside cells rather than in vitro. Relative to the established NV-ensemble quantum-sensing playbook (DEER, nanoscale NMR, T1 relaxometry at pT/sqrt(Hz) ensemble sensitivity), this is the strongest biosensing/advanced-microscopy host in Mainz: the labelling chemistry is precisely what a quantum-sensing postdoc would need to attach nanodiamonds or spin labels to a defined protein site, and the group already operates at the single-molecule sensitivity limit optically. Large, well-funded, internationally recruiting group.