COMBINATORIAL LIBRARIES OF PROTEINS - ANALYSIS OF EFFICIENCY OF MUTAGENESIS TECHNIQUES

A number of considerations are made on the efficiency of mutagenesis t echniques used in protein engineering, particularly those that include a random component. The expected outcome of different protocols is an alyzed using computer programs. Special emphasis is made on the effect that the degeneracy of the genetic code has on the bias of the repres entation of amino acid replacements. The consequences of using alterna tive methods is analyzed in terms of the likelihood of obtaining under represented amino acid substitutions in mutant libraries. A considerat ion is also made of the outcome of combinatorial mutagenesis experimen ts with regard to the size of the amino acid window and the multiplici ty of replacements that could be sampled with different methods. Optim al mutagenesis rates for specific conditions could be derived from the presented data and the computer program made available with this pape r.