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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
Reducing codon redundancy and screening effort of combinatorial protein libraries created by saturation mutagenesis
ACS Synthetic Biology, Volume 2, No. 2, Year 2013
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Description
Saturation mutagenesis probes define sections of the vast protein sequence space. However, even if randomization is limited this way, the combinatorial numbers problem is severe. Because diversity is created at the codon level, codon redundancy is a crucial factor determining the necessary effort for library screening. Additionally, due to the probabilistic nature of the sampling process, oversampling is required to ensure library completeness as well as a high probability to encounter all unique variants. Our trick employs a special mixture of three primers, creating a degeneracy of 22 unique codons coding for the 20 canonical amino acids. Therefore, codon redundancy and subsequent screening effort is significantly reduced, and a balanced distribution of codon per amino acid is achieved, as demonstrated exemplarily for a library of cyclohexanone monooxygenase. We show that this strategy is suitable for any saturation mutagenesis methodology to generate less-redundant libraries. © 2012 American Chemical Society.
Authors & Co-Authors
Opperman, Diederik Johannes
Germany, Mulheim an Der Ruhr
Max Planck Institute for Coal Research
Reetz, Manfred T.
Germany, Mulheim an Der Ruhr
Max Planck Institute for Coal Research
Germany, Marburg
Philipps-universität Marburg
Acevedo, Juan Pablo
Chile, Santiago
Universidad de Los Andes, Chile
Statistics
Citations: 189
Authors: 3
Affiliations: 3
Identifiers
Doi:
10.1021/sb300037w
ISSN:
21615063
Research Areas
Genetics And Genomics
Study Design
Randomised Control Trial
Study Approach
Qualitative