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We have initiated a structural genomics project based on selected protein families, with the representatives for structural studies obtained from the genome of E. coli. An essential task is the development of effective methods to express, purify and crystallize large numbers of proteins efficiently. We are currently applying robotics methods allowing cloning in an automated manner. Over 100 genes have been cloned to date, and more than 75 overexpressed as soluble proteins in at least one fusion system. Target genes have been cloned as N-terminal fusions with GST and (His)8 affinity tags, which are proteolytically removed after affinity purification. Using …
