Abstract: The present invention provides a GG-specific mismatch endonuclease variant, a TT-specific mismatch endonuclease variant, and a GT/TG-specific mismatch endonuclease variant. The present invention also provides a mismatch specific cleaving reaction using said variant, a method for removing errors in a nucleic acid amplification reaction using a mismatch nuclease, a method for suppressing amplification of a nucleic acid having a specific base sequence during a nucleic acid amplification reaction, and a method for detecting a nucleic acid having a single base polymorphic mutation using said suppression method.

Abstract: The present invention relates to a method for detecting a target nucleic acid, the method including cleaving a first flap of a first cleavage structure formed by a target nucleic acid, a first nucleic acid, and a second nucleic acid; cleaving a second flap of a second cleavage structure formed by a third nucleic acid, the cleaved first flap, and a fourth nucleic acid; and detecting the presence of the target nucleic acid by detecting the cleaved second flap, wherein cleaving the first flap and cleaving the second flap are carried out by cleaving the first flap and the second flap with a flap endonuclease, and the flap endonuclease has an amino acid sequence having a sequence identity of 65% or higher with an amino acid sequence of a flap endonuclease of a microbe selected from the group consisting of microbes belonging to the Order Thermococcales and microbes belonging to the Order Methanobacteriales.

Abstract: The present invention relates to a method for detecting a target nucleic acid, the method including cleaving a first flap of a first cleavage structure formed by a target nucleic acid, a first nucleic acid, and a second nucleic acid; cleaving a second flap of a second cleavage structure formed by a third nucleic acid, the cleaved first flap, and a fourth nucleic acid; and detecting the presence of the target nucleic acid by detecting the cleaved second flap, wherein cleaving the first flap and cleaving the second flap are carried out by cleaving the first flap and the second flap with a flap endonuclease, and the flap endonuclease has an amino acid sequence having a sequence identity of 96% or higher with an amino acid sequence of a flap endonuclease of Thermococcus kodakarensis strain KOD1, or the like.

Abstract: The present invention relates to a Thermus aquaticus (Taq) polymerase having a strand displacement activity in which an amino acid residue in a template DNA binding site of the DNA polymerase is substituted with an amino acid to increase a total charge in the site, a nucleic acid encoding the polymerase, a vector containing the nucleic acid, a transformant containing the vector containing the nucleic acid or the nucleic acid, a method for producing the polymerase, a method for amplifying nucleic acids utilizing the polymerase, and a kit containing the polymerase. According to the present invention, a DNA polymerase having a high thermostability, capable of efficiently replicating a long-strand of a template DNA, and having a strong strand displacement activity is provided.

Abstract: A polypeptide having a mismatch endonuclease activity of recognizing a mismatch and cleaving the mismatch; a mismatch-specific cleavage reaction using the polypeptide; a method for removing an error in a nucleic acid amplification reaction utilizing the polypeptide; a method for inhibiting the amplification of a nucleic acid comprising a specific nucleotide sequence during a nucleic acid amplification reaction; and a method for detecting a nucleic acid having a single-nucleotide polymorphism mutation utilizing the inhibition method.

Abstract: Provided are various novel DNA polymerases. Provided are: a DNA polymerase comprising: an amino acid sequence modified from the amino acid sequence of SEQ ID NO: 12, which has a substitution of arginine at position 651 by an amino acid residue having a negatively charged side chain, preferably by asparatic acid or glutamic acid, more preferably by glutamic acid; and a DNA polymerase comprising an amino acid sequence modified from the amino acid sequence of SEQ ID NO: 14, which has a substitution of proline at position 653 by an amino acid residue having a negatively charged side chain, preferably by asparatic acid or glutamic acid, more preferably by glutamic acid.

Abstract: The present invention relates to a fusion polypeptide containing, in a direction of from an N-terminal side to a C-terminal side, one or more peptides which bind to a PCNA, and a polypeptide having a DNA polymerase activity; a method for amplifying nucleic acids using the polypeptide; and a composition and a kit, containing the polypeptide. According to the present invention, it is made possible to amplify a long-strand DNA in a short time in amplifying nucleic acids in the presence of PCNA even with a Pol I-type DNA polymerase.

Abstract: Provided are various novel DNA polymerases. Provided are: a DNA polymerase comprising: an amino acid sequence modified from the amino acid sequence of SEQ ID NO: 12, which has a substitution of arginine at position 651 by an amino acid residue having a negatively charged side chain, preferably by asparatic acid or glutamic acid, more preferably by glutamic acid; and a DNA polymerase comprising an amino acid sequence modified from the amino acid sequence of SEQ ID NO: 14, which has a substitution of proline at position 653 by an amino acid residue having a negatively charged side chain, preferably by asparatic acid or glutamic acid, more preferably by glutamic acid.

Abstract: The present invention relates to a Thermus aquaticus (Taq) polymerase having a strand displacement activity in which an amino acid residue in a template DNA binding site of the DNA polymerase is substituted with an amino acid to increase a total charge in the site, a nucleic acid encoding the polymerase, a vector containing the nucleic acid, a transformant containing the vector containing the nucleic acid or the nucleic acid, a method for producing the polymerase, a method for amplifying nucleic acids utilizing the polymerase, and a kit containing the polymerase. According to the present invention, a DNA polymerase having a high thermostability, capable of efficiently replicating a long-strand of a template DNA, and having a strong strand displacement activity is provided.

Abstract: There is provided an enzyme which has an activity of cleaving a phosphodiester bond of deoxyribonucleotide having a damaged base and deoxyribonucleotide adjacent to the 5? side of the deoxyribonucleotide in DNA strands which contain the damaged base as a reagent or the like for manipulating a gene, and further provided a method of removing a damaged base from DNA strands using the enzyme.

Abstract: A polypeptide having a mismatch endonuclease activity of recognizing a mismatch and cleaving the mismatch; a mismatch-specific cleavage reaction using the polypeptide; a method for removing an error in a nucleic acid amplification reaction utilizing the polypeptide; a method for inhibiting the amplification of a nucleic acid comprising a specific nucleotide sequence during a nucleic acid amplification reaction; and a method for detecting a nucleic acid having a single-nucleotide polymorphism mutation utilizing the inhibition method.

Abstract: There is provided an enzyme which has an activity of cleaving a phosphodiester bond of deoxyribonucleotide having a damaged base and deoxyribonucleotide adjacent to the 5? side of the deoxyribonucleotide in DNA strands which contain the damaged base as a reagent or the like for manipulating a gene, and further provided a method of removing a damaged base from DNA strands using the enzyme.

Abstract: Provided are various novel DNA polymerases. Provided is a DNA polymerase comprising an amino acid sequence modified from the amino acid sequence of SEQ ID NO: 8 by inserting nine amino acids “-A737-A738-A739-A740-A741-A742-A743-A744-A745-” between the amino acid residue at position 736 and the amino acid residue at position 737, wherein: A737 is an amino acid residue having a non-polar aliphatic side chain; A738 is an amino acid residue having a non-polar aliphatic side chain; A739 is an amino acid residue having a positively charged side chain; A740 is an amino acid residue having a positively charged side chain; A741 is an amino acid residue having a non-polar aliphatic side chain; A742 is an amino acid residue having a non-polar aliphatic side chain; A743 is any given amino acid residue; A744 is an amino acid residue having a positively charged side chain; and A745 is an amino acid residue having a non-polar aliphatic side chain).

Abstract: Provided are various novel DNA polymerases. Provided are: a DNA polymerase comprising: an amino acid sequence modified from the amino acid sequence of SEQ ID NO: 12, which has a substitution of arginine at position 651 by an amino acid residue having a negatively charged side chain, preferably by asparatic acid or glutamic acid, more preferably by glutamic acid; and a DNA polymerase comprising an amino acid sequence modified from the amino acid sequence of SEQ ID NO: 14, which has a substitution of proline at position 653 by an amino acid residue having a negatively charged side chain, preferably by asparatic acid or glutamic acid, more preferably by glutamic acid.

Abstract: Provided are various novel DNA polymerases. Provided is a DNA polymerase comprising an amino acid sequence modified from the amino acid sequence of SEQ ID NO: 8 by inserting nine amino acids “-A737-A738-A739-A740-A741-A742-A743-A744-A745-” between the amino acid residue at position 736 and the amino acid residue at position 737, wherein: A737 is an amino acid residue having a non-polar aliphatic side chain; A738 is an amino acid residue having a non-polar aliphatic side chain; A739 is an amino acid residue having a positively charged side chain; A740 is an amino acid residue having a positively charged side chain; A741 is an amino acid residue having a non-polar aliphatic side chain; A742 is an amino acid residue having a non-polar aliphatic side chain; A743 is any given amino acid residue; A744 is an amino acid residue having a positively charged side chain; and A745 is an amino acid residue having a non-polar aliphatic side chain).

Abstract: The present invention is to construct a DNA replication reaction system which is excellent in versatility and is easily used. An amino acid sequence of a PCNA monomer which is one of factors involved in DNA replication is prepared so that amino acid residues causing mutual charge repulsion constitute a site which causes, when an N terminal region of the PCNA monomer and a C terminal region of another PCNA monomer act as an interface to form a multimeric complex, an intermolecular interaction of the monomers in an interface region of the monomers.

Abstract: It is intended to obtain DNA ligase improved in binding ability and reactivity with DNA, particularly thermostable DNA ligase improved in binding ability and reactivity with DNA. The present invention provides a DNA ligase mutant improved in binding ability and reactivity with DNA, which is obtained by partially or completely deleting a C-terminal helix portion of DNA ligase. Particularly, the mutant is derived from a thermostable bacterium.

Abstract: The present invention is to construct a DNA replication reaction system which is excellent in versatility and is easily used. An amino acid sequence of a PCNA monomer which is one of factors involved in DNA replication is prepared so that amino acid residues causing mutual charge repulsion constitute a site which causes, when an N terminal region of the PCNA monomer and a C terminal region of another PCNA monomer act as an interface to form a multimeric complex, an intermolecular interaction of the monomers in an interface region of the monomers.

Abstract: There is provided a method for the detection of a base sequence of interest when amount of a sample DNA or RNA is little and plural base sequences of interest to be detected are present in the sample DNA or RNA. The Problem is solved by a method for the detection of an base sequence of interest in a sample DNA or RNA comprising the steps of (1) contacting a sample DNA or RNA to a probe DNAs or RNAs in an aqueous solution to form a hybridization complex; (2) isolating the hybridization complex; (3) dissociating the complex to recover the probe DNAs or RNAs; and (4) identifying the said probe DNAs or RNAs to detect an base sequence of interest in the sample DNA or RNA.

Abstract: The object of the invention is to provide proteins that have both DNA primase activity and DNA polymerase activity. This subject is solved by a protein (p41) that has an amino acid sequence shown in SEQ ID NO: 1. This is for the first time that proteins that have both DNA primase activity and DNA polymerase activity were found. A protein (p46) that has amino acid sequence shown in SEQ ID NO: 2 forms a complex with p41, and enforces DNA synthesis activity that is independent and/or dependent from primer of p41.