SpringerProtocols: Abstract: 9. Random Oligonucleotide Mutagenesis

9. Random Oligonucleotide Mutagenesis

By: Jessica L. Sneeden³, Lawrence A. Loeb³

Abstract

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Random oligonucleotide mutagenesis is a method to generate diversity that consists of incorporating random mutations, encoded on a synthetic oligonucleotide, into a specific region of a gene. The number of mutations in individual enzymes within the population can be controlled by varying the length of the target sequence and the degree of randomization during synthesis of the oligonucleotides.

Affiliation(s): (3) Department of Biochemistry, University of Washington, Seattle, WA

Book Title: Directed Evolution Library Creation: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 231 | Pub. Date: Apr-25-2003 | Page Range: 65-73 | DOI: 10.1385/1-59259-395-X:65

Subject: Genetics/Genomics

References

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9.	Encell, L. P. and Loeb, L. A. (1999) Redesigning the substrate specficity of human O⁶-alylguanine-DNA alkyltransferase. Mutants with enhanced repair of O⁴-methylthymine. Biochemistry 38, 12,097–12,103.

10.	Landis, D. M. and Loeb, L. A. (1998) Random sequence mutagenesis and resistance to 5-fluorouridine in human thymidylate synthases. J. Biol. Chem. 273, 25,809–25,817.

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13.	Sambrook, J. and Russell D. W. (2001) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

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