Frame shuffling: a novel method for in vitro protein evolution.
We describe 'frame shuffling', a novel method for preparing artificial protein libraries. With this method, a Y-family DNA polymerase known to introduce frame shift mutations at high rates is utilized to scramble the reading frames of a parental gene. The resultant progeny produce mutant proteins having segmental sequence changes. Such frame-shuffled mutant proteins exhibit physicochemical properties that differ from those of proteins obtained using conventional mutagenesis.
Kashiwagi
Kenji
K
Department of Protein Engineering, Cancer Institute, Japanese Foundation for Cancer Research, Ariake, Koto-ku, Tokyo, Japan.
Isogai
Yasuhiro
Y
Nishiguchi
Kei-ichi
K
Shiba
Kiyotaka
K
D000595:Amino Acid Sequence; D002499:Centrifugation, Density Gradient; D002942:Circular Dichroism; D018388:Codon, Terminator; D019951:DNA Polymerase beta / Q000235:genetics / Q000302:isolation & purification; D032621:DNA Shuffling / Q000379:methods*; D019020:Directed Molecular Evolution; D004926:Escherichia coli / Q000235:genetics; D016368:Frameshift Mutation; D066298:In Vitro Techniques; D008027:Light; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010455:Peptides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D015202:Protein Engineering / Q000379:methods*; D012542:Scattering, Radiation; D017386:Sequence Homology, Amino Acid
D018388:Codon, Terminator; D010455:Peptides; D019951:DNA Polymerase beta