11/18/2023 0 Comments Klenow fragment mutagenesisD., Morin, C., Itakura, K., and Richards, J. (1992) Randomization of genes by PCR mutagenesis. (ed.) (2001) In Vitro Mutagenesis Protocols, Second Edition, Humana, Totowa, NJ.Ĭadwell, R. (2001) Discovery of superior enzymes by directed molecular evolution. (2001) How enzymes adapt: lessons from directed evolution. This process is experimental and the keywords may be updated as the learning algorithm improves.Īrnold, F. These keywords were added by machine and not by the authors. These synthetic methods may be applicable only to introduce mutations within a narrow range of the target gene. To introduce codon-based mutations at various positions, the split-and-mix method ( 8) or other synthetic methods of constructing DNAs using dinucleotide or trinucleotide units have been attempted ( 9– 11). These methods are limited to a defined region of the gene and can not introduce mutations at random positions. To achieve a non-biased random replacement on the amino acid level, oligonucleotide-directed mutagenesis ( 5) and cassette mutagenesis ( 6, 7) have been carried out. For example, mutation from AUG (Met) to UGG (Trp) is unlikely to take place. However, the error-prone PCR method has an inherent drawback of biased occurrence of amino acids as the result of single base replacements in the triplet codons. Among these, error-prone PCR mutagenesis, based on inaccurate copying by DNA polymerase, is the most commonly used technique to introduce random point mutations ( 4). Several methods for introducing random mutations in vitro have been reported ( 3). Random mutagenesis combined with high-throughput screening is a versatile strategy for improving protein functions or creating artificial enzymes ( 1, 2).
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |