Ribosome-inactivation display system.
We present a novel strategy for the connection of phenotype and genotype in vitro that can be used for the selection of functional proteins. The strategy involves the generation of a stable complex among a ribosome, an messenger RNA and its translated protein, without removal of the termination codon, as a result of the action of the ricin A chain during translation. The technique requires no transfection, no chemical synthesis, no ligation, and no removal of the termination codon. Thus, our novel ribosome-inactivation display system should provide, without loss of the pool population, a reliable, simple, and robust selection system for the in vitro evolution of the properties of proteins in a predictable direction by a combination of randomization and appropriate selection strategies.
Fujita
Satoshi
S
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Japan.
Zhou
Jing-Min
JM
Taira
Kazunari
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D018388:Codon, Terminator; D019020:Directed Molecular Evolution; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D008954:Models, Biological; D008957:Models, Genetic; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D015202:Protein Engineering / Q000379:methods*; D012333:RNA, Messenger / Q000378:metabolism; D012270:Ribosomes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012276:Ricin / Q000378:metabolism; D019809:Streptavidin / Q000235:genetics / Q000378:metabolism
D018388:Codon, Terminator; D012333:RNA, Messenger; D012276:Ricin; D019809:Streptavidin; D005982:Glutathione Transferase