title: Rational-based protein engineering: tips and tools.
abstract: The rational engineering of proteins is driven by contemporary needs for new and altered biomolecular forms. Utilizing manipulative procedures of molecular biology, it is relatively straightforward to alter protein structure and function to create mutated or fused sequences. We here give an overview of procedures and strategies for site-directed mutagenesis, construction of fusion proteins, and insertion of tags. The design of new protein constructs as well as their over-expression as recombinant products is considered. We also summarize approaches for the engineering of protein complexes by co-expression, a valuable route to generate bioactive multicomponent systems.
authors.affiliation: Structural and Computational Biology Division, Victor Chang Cardiac Research Institute, Sydney, Australia.
authors.forename: Meghna
authors.initials: M
authors.lastname: Sobti
authors.forename: Bridget C
authors.initials: BC
authors.lastname: Mabbutt
mesh_terms: D003001:Cloning, Molecular; D003062:Codon; D004926:Escherichia coli; D005822:Genetic Vectors; D016297:Mutagenesis, Site-Directed / Q000379:methods; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction / Q000379:methods; D014176:Protein Biosynthesis; D015202:Protein Engineering / Q000379:methods*; D059748:Proteolysis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D003062:Codon; D011993:Recombinant Fusion Proteins