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The process for generating specific amino acid changes by changing the coding sequence at a targeted site in a gene is called sitedirected mutagenesis.
However, for many proteins, such detailed information is often lacking, so sitedirected mutagenesis becomes a trial anderror strategy in which changes are made to those nucleotides that are most likely to yield a particular change in a protein property.
A straightforward sitedirected mutagenesis protocol introduces defined nucleotide substitutions into a gene by overlap extension PCR.
Figure 3.47 Sitedirected mutagenesis by overlap extension PCR.
Essentially any protein can be altered by substituting one amino acid for another using sitedirected mutagenesis.
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