High-Throughput Scanning Mutagenesis by Recombination Polymerase Chain Reaction
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Site-directed in vitro mutagenesis is a powerful tool for investigating the structure-function relationships of proteins and
the expression of genes. Often, it is desirable to subject the protein or gene of interest to scanning mutagenesis. The genetic
analysis of scores of single or multiple point mutations contributes to a better understanding of cis
elements in gene expression (1
). In the case of proteins, cysteine (Cys) (2
,3
) and alanine scanning mutagenesis (4
) are popular means for investigating the structure and function of proteins and the interactions between proteins. Cys-scanning
mutagenesis is particularly useful, since the introduction of Cys allows targeted chemical modification with a wide variety
of sulfhydryl-reactive reagents (5
,6
).
