By Tadhg P Begley; Wiley-Blackwell (Firm); Wiley InterScience (Online service); et al
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Additional info for Wiley Encyclopedia of Chemical Biology [4 vols]
Second, the proteins on the surface are accessible to external molecules, such as substrates or other target molecules. Finally, the DNA is restricted inside the phage particle or microbial cells, enabling easy tracking of the genotype. With the need for enzyme engineering growing, researchers recognized the potential of display technologies and progressively adapted them for enzyme engineering. Phage display is the most commonly used technique for in vitro selection. , M13) are used for protein display for their ability to infect host cells without killing them (25).
Screening involves examining every mutant individually for the desired property, whereas selection is a method whereby only proteins with the desired property are carried through. Although various technologies have been developed in each category, a common principle underlying these assays exists: tagging the DNA (genotype) and the protein it encodes (phenotype) followed by screening/selection (phenotype analysis) that is compatible with the tagging. Physical linkage and spatial compartmentalization are two ways of tagging.
Nucleic acid evolution and minimization by nonhomologous random recombination. Nat. Biotechnol. 2002;20:1024–1029. Chockalingam K, Chen Z, Katzenellenbogen JA, Zhao H. Directed evolution of specific receptor-ligand pairs for use in the creation of gene switches. Proc. Natl. Acad. Sci. A. 2005;102:5691–5696. Reetz MT, Bocola M, Carballeira JD, Zha D, Vogel A. Expanding the range of substrate acceptance of enzymes: combinatorial active-site saturation test. Angew. Chem. Int. Ed. Engl. 2005;44:4192–4196.
Wiley Encyclopedia of Chemical Biology [4 vols] by Tadhg P Begley; Wiley-Blackwell (Firm); Wiley InterScience (Online service); et al