Methods to perform the selective evolution of proteins in vitro.

摘要

Methods to perform the selective evolution of proteu00ecnas in vitro.The present invention relates to the production of variants of a protein in a method of evolution in vitro, comprising the steps of: (a) providing an in vitro expression system comprising (I) a nucleic acid sequence coding for a protein S Y to be varied, (II) a The target molecule X that is able to bind to the protein y, and / or at least one variant y ',(III) an RNA polymerase (POL) that is capable of transcribing the sequ00fau00eancia nucleic acid s, (IV) a reverse transcriptase (RT) that is capable of reverse transcription of transcripts of the sequ00fau00eancia of u00e1ci nucleic s, where the target molecule x is coupled to Pol and the protein y is acoplad The RT, or target molecule x is coupled to RT and the protein y is acopiada was,(b) incubation of the expression system in vu00edtro of (the) under conditions that enable transcription, reverse transcription and transu00edau00e7u00e3o to form variants y 'y sequ00fau00eancias protein and nucleic acid s' encoding for these, and for the formation of variants with propr y' Iedades link improved for the target molecule x, (the) isolation and, where appropriate,Characterization of the variants y 'which exhibit improved bonding properties connected to x, and / or isolation of variants s' of sequ00fau00eancia nucleic acid coding for y'.