Tn5 transposase mutants and the use thereof
First Claim
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1. 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.
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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.
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Citations
20 Claims
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1. 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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 16, 17, 18, 19, 20)
- 2, the transposase mutant comprising a mutation at position 54 of SEQ ID NO;
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12. 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. - View Dependent Claims (13, 14, 15)
- 2, the transposase mutant comprising a mutation at position 54of SEQ ID NO;
Specification
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Current AssigneeWisconsin Alumni Research Foundation (University of Wisconsin System)
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Original AssigneeWisconsin Alumni Research Foundation (University of Wisconsin System)
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InventorsMetzler, Jeremy D., Steiniger-White, Mindy M., Reznikoff, William S.
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Primary Examiner(s)Ketter, James
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Assistant Examiner(s)KATCHEVES, KONSTANTINA T
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Application NumberUS10/826,573Publication NumberTime in Patent Office837 DaysField of Search435/455, 435/461, 435/462US Class Current435/455CPC Class CodesC12N 9/22 Ribonucleases RNAses, DNAse...
