Define site‑directed mutagenesis and give one application of this technique in biotechnology.
Answer
Site‑directed mutagenesis is a technique to introduce a specific nucleotide change at a predetermined position in a DNA sequence; it is used to engineer enzymes with altered substrate specificity.
Step-by-step solution
- State the definition: site‑directed mutagenesis allows precise alteration of a chosen base pair in a gene using designed primers and a DNA polymerase.
- Explain the typical workflow: design mutagenic primers containing the desired change, amplify the plasmid by PCR, digest the parental template with DpnI, transform the product into competent cells, and screen colonies for the mutation.
- Provide an example application: engineering a protease to improve its activity on a new substrate, or creating a vaccine strain with a specific antigenic epitope mutation.
- Conclude with the concise answer linking definition and application.
Common mistake: Confusing it with random mutagenesis, which introduces uncontrolled changes
Concept tested: Site‑directed mutagenesis
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