Tn5 transposase mutants and the use thereof
AssigneeWisconsin Alumni Research Foundation
InventorsWilliam S. Reznikoff, Mindy M. Steiniger-White, Jeremy D. Metzler
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Abstract
Tn5 transposase (Tnp) mutants that have higher transposase activities than the wild-type Tnp are disclosed. The Tn5 Tnp mutants differ from the wild-type Tnp at amino acid positions 54, 242, and 372 and have greater avidity than the wild-type Tnp for at least one of a wild-type Tn5 outside end sequence as defined by SEQ ID NO:3 and a modified Tn5 outside end sequence as defined by SEQ ID NO:5. Also disclosed are various systems and methods of using the Tnp mutants for in vitro or in vivo transposition.
Claims20
A polypeptide comprising a Tn5 transposase mutant modified relative to SEQ ID NO:2, the transposase mutant comprising a mutation at position 54 of SEQ ID NO:2, a mutation at position 242of SEQ ID NO:2, and a mutation at position 372of SEQ ID NO:2, wherein the transposase mutant has greater avidity than wild-type Tn5 transposase for at least one of a Tn5 outside end sequence as defined by SEQ ID NO:3 and a modified Tn5 outside end sequence as defined by SEQ ID NO:5.
A Tn5 transposase mutant modified relative to SEQ ID NO:2, the transposase mutant comprising a mutation at position 54of SEQ ID NO:2, a mutation at position 242of SEQ ID NO:2, and a mutation at position 372of SEQ ID NO:2, wherein the transposase mutant has greater avidity than wild-type Tn5 transposase for at least one of a Tn5 outside end sequence as defined by SEQ ID NO:3 and a modified Tn5 outside end sequence as defined by SEQ ID NO:5.
Dependencies are inferred from the claim text.