title: Efficient procedure for site-directed mutagenesis mediated by PCR insertion of a novel restriction site.
abstract: Better understanding of proteins' structure/function relationship and dissecting their functional domains are still challenges yet to be mastered. Site-directed mutagenesis approaches that can alter bases at precise positions on the gene sequence can help to reach this goal. This article describes an efficient strategy that can be applied not only for both deletion and substitution of target amino acids, but also for insertion of point mutations in promoter regions to study cis-regulating elements. This method takes advantage of the plasticity of the genetic code and the use of compatible restriction sites.
authors.affiliation: Laboratoire de Biochimie et Physiologie Moléculaire des Plantes, Centre National de la recherche Scientifique/Institut National de la Recherche Agronomique/Agro-M/Université Montpellier II, Montpellier, France. hatem.rouached@gmail.com
authors.forename: Hatem
authors.initials: H
authors.lastname: Rouached
mesh_terms: D016297:Mutagenesis, Site-Directed / Q000379:methods*; D010940:Plant Proteins / Q000737:chemistry / Q000502:physiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D013329:Structure-Activity Relationship
chemical_list: D010940:Plant Proteins