Mismatched end DNA ligase
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsChun Tsai, Seonhi Kim, Gilbert Chu
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Abstract
A mismatched end DNA ligase is provided, which ligates two single strands to each other at a high efficiency, even if the other two single strands are not compatible. In one embodiment, the polypeptides of the ligase are Ku, Cernunnos, and XRCC4/Ligase4 (XL). This association can ligate DNA ends with a 3′ overhang to a recessed 5′ end, to a blunt end, or to a compatible end. In another embodiment, the proteins are Ku, Cernunnos, XRCC4/Ligase4 (XL) and DNA-PK.
Claims11
A method of efficiently catalyzing direct ligation of noncompatible DNA ends in vitro, the method comprising:
- contacting in a reaction mix a purified mismatched end DNA ligase complex comprising the polypeptides Ku, Cernunnos, and XRCC4/Ligase4, and DNA comprising non-compatible ends, in the presence of ATP for a period of time sufficient to permit direct ligation and preservation of DNA sequence, wherein the efficiency of ligation is at least 10% of a comparable reaction with DNA comprising compatible ends.
A method of efficiently catalyzing direct ligation of noncompatible DNA ends in vitro, the method comprising:
- contacting in a reaction mix a mismatched end DNA ligase complex consisting of substantially pure Ku, Cernunnos, XRCC4/Ligase4 and DNA-PK, and DNA comprising non-compatible ends, in the presence of ATP for a period of time sufficient to permit direct ligation and preservation of DNA sequence, wherein the efficiency of ligation is at least 10% of a comparable reaction with DNA comprising compatible ends.
A method of catalyzing direct ligation of DNA ends, the method comprising:
- contacting in a reaction mix a purified mismatched end DNA ligase complex consisting of substantially pure Ku, Cernunnos, XRCC4/Ligase4 and DNA-PK, and DNA, in the presence of ATP for a period of time sufficient to permit direct ligation and preservation of DNA sequence.
Dependencies are inferred from the claim text.