Recombinase mutants
AssigneeIllumina Cambridge Limited
InventorsErin Bomati, Matthew William Kellinger, Jonathan Mark Boutell
Life of this patent
Estimating the term…
Abstract
Presented herein are recombinases for improved recombinase-mediated amplification of nucleic acids, such as a PCR-library having single-stranded adapter regions, on a patterned flow cell surface for improved cluster amplification, as well as methods and kits using the same.
Claims24
A UvsX recombinase polypeptide comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:
- 1, which UvsX recombinase polypeptide comprises an amino acid substitution mutation at the position functionally equivalent to position 256 in the Enterobacteria phage RB49 UvsX of SEQ ID NO:
- 1, said UvsX recombinase polypeptide having increased recombinase activity compared to a T4 UvsX recombinase polypeptide having SEQ ID NO:
- 8 or a RB49 UvsX recombinase polypeptide having all of the amino acid sequence of SEQ ID NO:1 except for a His to Ser substitution at amino acid position 63 of SEQ ID NO:
- 1.
A UvsX recombinase polypeptide comprising the amino acid sequence of SEQ ID NO:
- 2.
A UvsX recombinase polypeptide comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:1 and comprising a semiconserved domain, said semiconserved domain comprising all of SEQ ID NO:
- 3, 4, or 5 except for a substitution at position 7 of SEQ ID NO:
- 3, 4, or 5 to any residue other than Phe, Pro, Asp, Glu or Asn, said UvsX recombinase polypeptide having increased recombinase activity compared to a T4 UvsX recombinase polypeptide having SEQ ID NO:
- 8 or a RB49 UvsX recombinase polypeptide having all of the amino acid sequence of SEQ ID NO:
- 1 except for a His to Ser substitution at amino acid position 63 of SEQ ID NO:
- 1.
A UvsX recombinase polypeptide comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:
- 1 and comprising a semiconserved domain, said semiconserved domain comprising all of SEQ ID NO:
- 6 or 7 except for a substitution at position 12 of SEQ ID NO:
- 6 or 7 to any residue other than Phe, Pro, Asp, Glu or Asn, said UvsX recombinase polypeptide having increased recombinase activity compared to a T4 UvsX recombinase polypeptide having SEQ ID NO:
- 8 or a RB49 UvsX recombinase polypeptide having all of the amino acid sequence of SEQ ID NO:1 except for a His to Ser substitution at amino acid position 63 of SEQ ID NO:
- 1.
A UvsX recombinase polypeptide comprising all of the amino acid sequence of SEQ ID NO:
- 1 except for a proline (Pro) to lysine (Lys) substitution at amino acid position 256.
Dependencies are inferred from the claim text.