CORE--PROTEIN

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Support is requested for Protein Production. While small-scale preparations of recombinant proteins using standard affinity purifications will be performed in the individual laboratories of the project, all projects require mg-amounts of highly purified recombinant and untagged proteins. The Core will provide technical and scientific support in two ways. First, it will optimize expression of proteins that either do not express at all or only at very low levels, including change of vectors, bacterial strains, co-expression of chaperones, or switching to an eukaryotic expression system. Second, the core will express and purify medium to large scale amounts of proteins using an array of conventional and affinity-based chromatographic methods. The core is equipped with a bacterial fermenter for large-scale expression, two state-of-the-art automated FPLC-systems, and a miniaturaized FPLC (SMART). An array of physico-chemical techniques will be used for initial characterization with respect to folding, oligomerization/aggregation and purity including CD-spectroscopy, multi-angle laser light scattering, and mass spectrometry. It is directd jointly by Drs. Fasshauer and Jahn, who will be assisted by M. Boeddener, an experienced Research Associate with several years of experience in all of the required techniques.