I have question regarding Site-directed mutagenesis and Mutagenesis mapping. (See copied from Wikipedia below)
For site-directed mutagenesis how do you introduce systematic mutation of amino acids into a protein sequence? Is it through genetic manipulation through PCR? In Mutagenesis mapping where comprehensive mutation library is used. How is this comprehensive mutation library is made?
What's the difference between Site-directed mutagenesis and mutagenesis mapping.
Site-directed mutagenesis: Using this approach, systematic mutations of amino acids are introduced into a protein sequence followed by measurement of antibody binding in order to identify amino acids that comprise an epitope. This techniquedo can be used to map both linear and conformational epitopes, but is labor-intensive and slow, typically limiting analysis to a small number of amino acid residues.
Mutagenesis Mapping.[8] This approach utilizes a comprehensive mutation library, with each clone containing a unique amino acid mutation and the entire library covering every amino acid in the target protein. Amino acids that are required for antibody binding can be identified by a loss of reactivity and mapped onto protein structures to visualize epitopes.[9] This approach has recently been used to epitope map a panel of antibodies against human CCR5, a GPCR coreceptor for HIV entry.[10]
Thank you!
