Site-directed mutagenesis
(Redirected from In vitro mutagenesis)
Site-directed mutagenesis is a molecular biology method that is used to make specific and intentional changes to the DNA sequence of a gene and any gene products. Also known as site-specific mutagenesis or oligonucleotide-directed mutagenesis, it is used for a variety of applications, but is most commonly used in the study of protein function.
History[edit | edit source]
The method of site-directed mutagenesis was first developed in the late 1970s. The original method used single-stranded DNA as a template for the synthesis of a complementary strand containing the desired mutation. This method, however, was time-consuming and required large amounts of DNA. In the 1980s, a more efficient method was developed using polymerase chain reaction (PCR), which is now the most commonly used method for site-directed mutagenesis.
Method[edit | edit source]
Site-directed mutagenesis involves the following steps:
- Design of a primer that carries the desired mutation.
- Amplification of the gene of interest using PCR.
- Transformation of the PCR product into suitable bacteria.
- Selection of bacteria that carry the mutated gene.
- Verification of the mutation by DNA sequencing.
Applications[edit | edit source]
Site-directed mutagenesis is used in a variety of applications, including:
- Studying the function of specific proteins.
- Creating proteins with improved properties.
- Creating proteins with novel functions.
- Studying the effects of specific mutations on protein function.
See also[edit | edit source]
References[edit | edit source]
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