Patent ID: 9593326
Date: 2017-03-14
CPC Classifications: C12N,C12P

Claim:
1. A method of introducing diversity into a reference protein sequence, the method comprising: (a) providing a first plurality of mutant proteins derived from the reference protein sequence, wherein the first plurality of mutant proteins each have one or more established beneficial properties and mutations in a first region of the reference protein sequence but not in substantially any other region of the reference protein sequence; (b) providing a second plurality of mutant proteins derived from the reference protein sequence, wherein the second plurality of mutant proteins each have one or more established beneficial properties and mutations in a second region of the reference protein sequence but not in substantially any other region of the reference protein sequence; (c) isolating a first group of nucleic acid segments from nucleic acids encoding the first plurality of mutant proteins, each nucleic acid segment in the first group encoding the first region of the reference protein sequence but not encoding substantially any other region of the reference protein sequence, wherein isolating the first group of nucleic acid segments is performed without first sequencing the first group of nucleic acid segments to identify the mutations in the first region; (d) isolating a second group of nucleic acid segments from nucleic acids encoding the second plurality of mutant proteins, each nucleic acid segment in the second group encoding the second region of the reference protein sequence but not encoding substantially any of the first region of the reference protein sequence, wherein isolating the second group of nucleic acid segments is performed without first sequencing the second group of nucleic acid segments to identify the mutations in the second region; and (e) assembling at least the isolated nucleic acid segments from (c) and (d) into full length nucleic acid sequences encoding new mutant proteins.