Journal Article

Codon based mutagenesis using trimer phosphoramidites

John Randolph, Andrey Yagodkin, Marc Lamaitre, Alex Azhayev and Hugh Mackie

in Nucleic Acids Symposium Series

Volume 52, issue 1, pages 479-479
Published in print September 2008 | ISSN: 0261-3166
Published online September 2008 | e-ISSN: 1746-8272 | DOI: http://dx.doi.org/10.1093/nass/nrn243
Codon based mutagenesis using trimer phosphoramidites

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  • Chemistry
  • Bioinformatics and Computational Biology
  • Genetics and Genomics
  • Molecular and Cell Biology

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A set of Trimer phosphoramidites was synthesized covering all 20 amino acid codons. These trimer phosphoramidites can be added during synthesis using standard DNA chemistry. A Reaction Factor (RF) was determined empirically for each trimer to compensate for differences in their relative rate of reaction during coupling. It is therefore possible to introduce an equimolar mix of all 20 amino acid codons, or subsets thereof, at any location within a sequence. By mutating a gene at the codon level rather than at individual bases, it is possible to avoid codon bias, frame-shift mutations and the introduction of stop codons, making Trimer phosphoramidites a highly efficient tool for the exploration of sequence space in proteins.

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Subjects: Chemistry ; Bioinformatics and Computational Biology ; Genetics and Genomics ; Molecular and Cell Biology

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