title: Site-directed mutagenesis.
abstract: Site-directed mutagenesis is a PCR-based method to mutate specified nucleotides of a sequence within a plasmid vector. This technique allows one to study the relative importance of a particular amino acid for protein structure and function. Typical mutations are designed to disrupt or map protein-protein interactions, mimic or block posttranslational modifications, or to silence enzymatic activity. Alternatively, noncoding changes are often used to generate rescue constructs that are resistant to knockdown via RNAi.
authors.affiliation: Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: jbachma9@gmail.com.
authors.forename: Julia
authors.initials: J
authors.lastname: Bachman
mesh_terms: D017931:DNA Primers / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli; D005822:Genetic Vectors; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D009154:Mutation; D016133:Polymerase Chain Reaction
chemical_list: D017931:DNA Primers
keywords: Chemically competent E. coli; Colony screening; DNA polymerase; PCR amplification; Plasmid DNA; Site-directed mutagenesis