Patent ID: 7432083

Claim:
A method of generating one or more assembled polynucleotide sequences 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) and each pair binding adjacent to a variant polynucleotide sequence encoding a variant protein motif from the at least one polynucleotide sequence of step a); 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), performing PCR amplification on the at least one polynucleotide sequence of step a) and forming amplified polynucleotide sequences encoding the variant protein motifs; 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 from at least one polynucleotide sequence, wherein at least one polynucleotide sequence is different from the at least one polynucleotide sequence of step a); and f) annealing the polynucleotide sequences from step d) with the unmutated , specifically selected, scaffold sequences from step e) such that annealed polynucleotides with one or more gaps are formed, and filling the one or more gaps present in the annealed polynucleotides, thereby generating the one or more assembled polynucleotide sequences ecoding a protein of desired characteristics.