RApid Parallel Protein EvaluatoR (RAPPER), from gene to enzyme function in one day.
Cell-free transcription-translation systems offer an effective and versatile platform to explore the impact of genetic variations on protein function. We have developed a protocol for preparing linear, mutagenic DNA templates for direct use in the PURE system, enabling the fast and semi-quantitative evaluation of amino acid variations on catalytic activity and stereo-selectivity in native and engineered variants of Old Yellow Enzyme.
Department of Chemistry, Emory University, 1515 Dickey Drive, Atlanta, GA 30322, USA. sal2@emory.edu.
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D001547:Benzaldehydes / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D020134:Catalytic Domain; D004926:Escherichia coli / Q000378:metabolism; D005486:Flavin Mononucleotide / Q000737:chemistry / Q000378:metabolism; D015723:Gene Library; D005822:Genetic Vectors / Q000378:metabolism; D062105:Molecular Docking Simulation; D016296:Mutagenesis; D009252:NADPH Dehydrogenase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012440:Saccharomyces / Q000201:enzymology
D001547:Benzaldehydes; D005486:Flavin Mononucleotide; D009252:NADPH Dehydrogenase; C011483:4-hydroxybenzaldehyde