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AssigneeLarge Scale Biology Corporation
InventorsHal S. Padgett, John A. Lindbo, Wayne P. Fitzmaurice
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We describe here an in vitro method of increasing complementarity in a heteroduplex polynucleotide sequence. The method uses annealing of opposite strands to form a polynucleotide duplex with mismatches. The heteroduplex polynucleotide is combined with an effective amount of enzymes having strand cleavage activity, 3′ to 5′ exonuclease activity, and polymerase activity, and allowing sufficient time for the percentage of complementarity to be increased within the heteroduplex. Not all heteroduplex polynucleotides will necessarily have all mismatches resolved to complementarity. The resulting polynucleotide is optionally ligated. Several variant polynucleotides result. At sites where either of the opposite strands has templated recoding in the other strand, the resulting percent complementarity of the heteroduplex polynucleotide sequence is increased. The parent polynucleotides need not be cleaved into fragments prior to annealing heterologous strands. Therefore, no reassembly is required.
An in vitro method of making sequence variants from at least one heteroduplex polynucleotide where said heteroduplex has at least two non-complementary nucleotide base pairs, said method comprising:
An in vitro method of making sequence variants from at least one heteroduplex polynucleotide wherein said heteroduplex has at least two non-complementary nucleotide base pairs, said method comprising:
An in vitro method of increasing diversity in a population of sequences, comprising:
An in vitro method of increasing diversity in a population of sequences, comprising:
A method of obtaining a polynucleotide encoding a desired functional property, comprising:
A method of obtaining a polynucleotide encoding a desired functional property, comprising:
A method of obtaining a polypeptide having a desired functional property, comprising:
A method of obtaining a polynucleotide encoding a desired functional property, comprising:
A kit used for increasing diversity in a population of sequences, comprising:
Dependencies are inferred from the claim text.