Site-directed mutagenesis is one of the most important laboratory techniques for creating DNA libraries by introducing mutations into DNA sequences. There are numerous methods for achieving site-directed mutagenesis, but with decreasing costs of oligonucleotide synthesis, artificial gene synthesis is now occasionally used as an alternative to site-directed mutagenesis.
We propose a new method of mutagenesis without prior use of DNA by bioinformatic methods.
In fact, we do not perform pure mutagenesis, but replace amino acid residues in the wild-type protein with any other amino acid residue. To complete this procedure, we will need three-dimensional structures of the protein complex. It is desirable that this be a structure obtained by X-ray diffraction analysis of good quality, otherwise you can use servers that reproduce the three-dimensional structure of proteins. We also perform this procedure.
The mutagenesis procedure completely repeats the mechanism of alanine scanning. The result of mutagenesis is the determination of changes in affinity when replacing the amino acid residue in one of the proteins.
You can order mutagenesis of any protein complexes, but the accuracy of the calculations will directly depend on the quality of the three-dimensional structure.