In vitro scanning saturation mutagenesis of proteins

Date
2001-01-30
Authors
Brent L. Iverson
George M. Georgiou
Elizabeth A. Burks
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United States Patent and Trademark Office
Abstract

The present invention combines PCR.TM. mutagenesis with in vitro transcription/translation and ELISA for the rapid generation and characterization of protein mutants. The PCR.TM. products are used directly as the template for the in vitro transcription/translation reactions and because no cloning steps are required, the in vitro saturation mutagenesis of one residue can be completed in duplicate within a week by a single investigator. This high throughput enables the saturation mutagenesis of numerous residues of interest, a process that can be described as in vitro scanning saturation mutagenesis. Compositions and methods of use of such a process are described herein.

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