Services offered to members of the Helmholtz Zentrum München
The PEPF supports the following cloning technologies
- Cloning using restriction enzymes
- Gateway cloning technology
- Ligase independent cloning (LIC)
- Sequence and ligase independent cloning (SLIC)
For all these cloning technologies appropriate vectors are available (see Materials).
Small-scale bacterial expression screening
For the small-scale expression screening in Escherichia coli we have developed the so-called 'Fast Soluble Expression Screen'.
Large-scale bacterial protein production
Target proteins will be expressed on a large-scale (1-10L) in the best E. coli expression host and under optimal expression conditions. The expressed proteins will be subsequently purified using affinity, ion exchange and/or size exclusion chromatography.
Protein characterization of targets proteins using the following techniques:
- Dynamic Light Scattering (DLS) to determine the size and the monodispersity of a target protein.
- Static Light Scattering (SLS) to determine the oligomeric state of a target protein and the stoichiometry of protein-protein and protein-ligand complexes.
- Isothermal Titration Calorimetry to study ligand binding.
- Thermal shift (ThermoFluor) Assay to determine protein stabiliy and study ligand binding and protein function.