METODO PARA A EVOLUCAO MOLECULAR IN VITRO DA FUNCAO DE PROTEINA

摘要

The invention provides a method for generating a polmucleotide sequence or population of sequences fromparent single stranded polynucleotide sequences encoding one or more protein motifs, comprising the steps of a) providing single stranded DNA constituting plus and minus strands of parent polynucleotide sequences; b) digesting the single stranded polynucleotide sequences with an exonuclease to generate populations of single stranded fragments; c) contacting said fragments generated from the plus strands with fragments generated from the minus strands and optionally, adding primer sequences that anneal to the 3' and 5' ends of at least one of the parent polynucleotides under annealing conditions; d) amplifying the fragments that anneal to each other to generate at least one polynucleotide sequence encoding one or more protein motifs having altered characteristics as compared to theone or more protein motifs encoded by said parent polynucleotides. Preferably, the exonuclease is BAL 31.