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Site Directed Mutagenesis

Information on site directed mutagenesis.

Site Directed Mutagenesis

In the past protein biochemists relied on chemical methods to alter certain amino acids in the proteins they study inorder to observe the effects of these changes on protein activites. The use of chemicals are a very crude way too manipulate proteins, since it is very difficult to be sure that only one amin acid or even one kinfd of amino acid has been altered. Cloned genes make this type of an investigation much more precise, allowing us to perform microsurgery on a protein. With the ability to change different bases in a gene we also change amino acids ar corresponding sites in the protein product, which we then can observe the effects of those changes on the protein's function.

Site directed mutagenesis requires a cloned gene whose base sequence is known. Then we need to obtain our gene is a single stranded form. This can be done by cloning it into M13 phage and collecting the single-stranded progeny phage DNA (plus strands) or by using an analogous procedure with a phagemid.

A popular class of site-directed mutagenesis techniques relies on PCR.

Learn more about PCR site directed mutagenesis.

DNA Cloning

Copyright Molecular Station 2012



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