Site-saturation Mutagenesis Library Construction to characterize structure-function relationships (CAT#: STEM-MB-0023-WXH)

Introduction

Site-saturation mutagenesis (SSM) is a simple, semi-rational approach to protein engineering that occupies a useful middle ground between fully rational site-directed mutagenesis strategies and non-rational mutagenesis strategies. In SSM, the semi-rational designation derives from the fact that target residues are identified in a specific, knowledge-guided manner, but are then mutated randomly, in either an individual or combinatorial fashion. Through the site-saturation mutagenesis, it is possible to create a library of mutants containing all possible combinations of 20 different amino acids at one or more predetermined positions in a gene sequence. SSM is a powerful approach in protein engineering to characterize structure-function relationships, as well as to create improved protein variants.