High-fidelity polymerase with preference for gapped dna and use thereof
AssigneeZhejiang University
InventorsYuejin HUA, Xingru ZHOU, Liangyan WANG, Xuanyi CHEN
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Abstract
The disclosure provides a high-fidelity polymerase with preference for gapped DNA and use thereof. The Klenow fragment (KlenDr) derived from Deinococcus radiodurans DNA polymerase I, which has the high-fidelity polymerization characteristics, is independent of 3′-5′ proofreading exonuclease activity, has the preference for binding gapped DNA, and is different from the existing commercial high-fidelity polymerase. Due to the specific affinity of KlenDr to gapped DNA substrate, the 3′ end of the forward primer will not be cut off, and the downstream nucleotide chain is rarely replaced.
Claims7
A high-fidelity polymerase with preference for gapped DNA, wherein having a high-fidelity polymerization characteristic independent of 3′-5′ proofreading exonuclease activity, and having a preference for binding gapped DNA, the polymerase is derived from Deinococcus radiodurans R1, purchased from American Type Culture Collection, with a collection number of ATCC 13939, and containing the amino acids 289-921 with protein sequence ID ANC71194.1, which is set forth in SEQ ID NO:
- 1.
A method for amplifying the nucleic acid, wherein the method comprises filling the DNA gap with high-fidelity polymerase with preference for gapped DNA.
Dependencies are inferred from the claim text.