[24] Selecting and evolving functional proteins in vitro by ribosome display

摘要: Publisher Summary In vivo systems have a number of limitations. One important restriction is that the library size limited by transformation efficiency, step all these techniques in common. Another limitation -based selection methods becomes apparent if diversification must be included. It requires either repeatedly switching between and vitro or use mutator strains. The former approach quite laborious, as it makes new generation large-scale necessary for each cycle sequence selection. latter may disadvantage possible candidate molecules removed from mutations generated host genome plasmid regions expression replication. All limitations can simultaneously overcome ribosome display. Ribosome display an technology simultaneous evolution proteins diverse libraries. Because no necessary, large libraries prepared applied Diversification conveniently introduced this method, thereby making evolutionary approaches easily accessible. was first to short peptides has subsequently been improved work with folded proteins. This chapter describes detail methodology nature improvements provides information perform functional proteins, using E. coli rabbit reticulocyte translation system.