In Vitro Scanning-Saturation Mutagenesis
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Scanning mutagenesis strategies have proven to be a useful approach to structure-function and protein evolution studies (1
), yet great effort is required to generate comprehensive data for a given protein. In these experiments, one or several amino
acid residues are changed (i.e., to alanine), and the resulting mutant proteins analyzed for changes in function. Successful
applications include the mapping of functional binding epitopes (2
,3
), confirmation of low-resolution X-ray or computer-generated structural models (4
), and investigation of the energetic contributions of contact residues (5
,6
). However, a stumbling block to the wider application of these techniques is that each desired mutation must be introduced
into a gene, cloned into bacteria, and the protein produced then purified (7
). This is a resource and time intensive process that precludes routine use.
