Patent ID: 6989250
Filing Date: 2006-01-24
Classification: C07K,C12N,Y10T

Abstract:
1. A method of generating an assembled polynucleotide sequence encoding a protein of desired characteristics comprising the steps of: a) providing at least one polynucleotide sequence comprising one or more variant polynucleotide sequences encoding one or more variant protein motifs; b) providing one or more pairs of defined oligonucleotides, each pair representing spaced apart locations on the at least one polynucleotide sequence of step a), each pair binding adjacent to a variant polynucleotide sequence encoding a variant protein motif; c) using the pairs of defined oligonucleotides as amplification primers for PCR to amplify the variant polynucleotide sequences encoding the variant protein motifs of the at least one polynucleotide sequence of step a) and performing PCR amplification on the at least one polynucleotide sequences of step a); d) obtaining one or more single-stranded polynucleotide sequences from the amplified polynucleotide sequences in step c); e) providing one or more unmutated, specifically selected, scaffold polynucleotide sequences encoding one or more unmutated peptide regions; and f) annealing said one or more single-stranded polynucleotide sequences from step d) with the unmutated, specifically selected, scaffold polynucleotide sequences from step e) such that annealed polynucleotides with one or more gaps is formed, and filling the one or more gaps present in the annealed polynucleotides, thereby generating one or more assembled polynucleotide sequences.