Abstract: A DNA polymerase (Neq DNA polymerase) derived from Nanoarchaeum equitans is split into Neq L and Neq S fragments, each of which contains inteins. A Neq hot-start (HS) DNA polymerase in which the inteins of the Neq L and Neq S fragments are linked with each other is provided in the form of a precursor of Neq DNA polymerase. A purification method can be significantly improved by inserting a His-tag sequence composed of six histidine residues between the inteins of the Neq L and Neq S fragments at a gene level. As a result of effort to enhance PCR efficiency of the Neq HS DNA polymerase, a gene coding for the Neq HS DNA polymerase is mutated at specific positions to screen mutant Neq HS polymerases (M1, M2, and M3) having a highly improved PCR amplification rate and amplification level.

Abstract: A DNA polymerase (Neq DNA polymerase) derived from Nanoarchaeum equitans is split into Neq L and Neq S fragments, each of which contains inteins. A Neq hot-start (HS) DNA polymerase in which the inteins of the Neq L and Neq S fragments are linked with each other is provided in the form of a precursor of Neq DNA polymerase. A purification method can be significantly improved by inserting a His-tag sequence composed of six histidine residues between the inteins of the Neq L and Neq S fragments at a gene level. As a result of effort to enhance PCR efficiency of the Neq HS DNA polymerase, a gene coding for the Neq HS DNA polymerase is mutated at specific positions to screen mutant Neq HS polymerases (M1, M2, and M3) having a highly improved PCR amplification rate and amplification level.

Abstract: The present invention relates to a hyperthermophilic DNA polymerase and a preparation method thereof. The invention provides a novel hyperthermophilic DNA polymerase isolated from a Thermococcus sp. strain, a functional equivalent thereof, a protein having the amino acid sequence thereof, and a preparation method thereof. The DNA polymerase according to the invention is a DNA polymerase, which is hyperthermophilic and has an elongation ability and fidelity higher than those of prior commercial DNA polymerases. Thus, the DNA polymerase according to the invention will be useful in precision analysis, precision diagnosis, identification and the like, which require accurate PCR.

Abstract: Disclosed is a hot-start PCR method, based on protein trans-splicing of intein-inserted large (Neq L) and small (Neq S) fragments of Neq DNA polymerase. The method comprises: preparing a PCR reaction mixture containing a sample DNA and primers; adding the Neq L fragment and the Neq S fragment together to the PCR reaction mixture, said Neq L fragment consisting of an amino acid sequence of SEQ ID NO: 2, with an intein amino acid sequence stretching from position 579 to 676 therein, said Neq S fragment consisting of an amino acid sequence of SEQ ID NO: 4 with an intein amino acid sequence stretching from position 1 to 30 therein; inducing the Neq L fragment and the Neq S fragment to undergo a protein trans-splicing process to form a polypeptide exhibiting Neq DNA polymerase activity; and performing a certain number of cycles of DNA denaturation, primer annealing and DNA extension.

Abstract: Disclosed herein is a mutant Neq A523R DNA polymerase consisting of an amino acid sequence of SEQ ID NO: 2 wherein alanine at amino acid position 523 in a Nanoarchaeum equitans DNA polymerase (Neq DNA polymerase) consisting of an amino acid sequence of SEQ ID NO: 1 has been substituted with arginine by site-directed mutagenesis. Also disclosed are a gene consisting of a nucleotide sequence encoding the mutant Neq A523R DNA polymerase, a recombinant vector comprising the gene, and a transformant transformed with the vector. In addition, disclosed are a PCR kit comprising the mutant Neq A523R DNA polymerase having excellent performance compared to the wild-type Neq DNA polymerase, and a method for preparing the Neq A523R DNA polymerase.

Abstract: The present invention relates to mutant DNA polymerases, their genes and their uses. More specifically, the present invention relates to mutant DNA polymerases which are originally isolated from Thermococcus sp NA1. strain and produced by site-specific mutagenesis, their amino acid sequences, genes encoding said mutant DNA polymerases, their nucleic acids and PCR methods by using thereof. As mutant DNA polymerases according to the present invention have decreased proofreading activity and changed function of inosine sensing effectively compared to wild type DNA polymerase, PCR using primers with specific nucleic acids has made rapid progress. Therefore, the present invention is broadly applicable for PCR in various molecular genetic technologies.

Abstract: Disclosed are a method of preparing an active Nanoarchaeum equitans B-type DNA polymerase (Neq DNA polymerase), an active Neq DNA polymerase prepared according to the method, and a polymerase chain reaction (PCR) using the active Neq DNA polymerase. The active Neq DNA polymerase may be used in various nucleic acid polymerization reactions, such as PCR.

Abstract: The present invention provides uracil-DNA glycosylase (UDG) gene originating from Psychrobacter sp. HJ147, and amino acid sequences deduced from the gene; expression and purification of Psp HJ147 UDG gene in Escherichia coli; and characterization of UDG obtained therefrom, and the use thereof in a polymerase chain reaction (PCR). The UDG according to the present invention has a specific activity of excising uracil bases in a uracil-containing DNA substrates at a low temperature, and is easily heat-inactivated. It thus can effectively eliminate cross contamination and carry-over contamination of PCR templates often occurring after a PCR process using dUTP. Therefore, it is useful for increasing preciseness (elimination of false positives), purity and amplification efficiency of PCR.

Abstract: The present invention relates to a hyperthermophilic DNA polymerase and a preparation method thereof. The invention provides a novel hyperthermophilic DNA polymerase isolated from a Thermococcus sp. strain, a functional equivalent thereof, a protein having the amino acid sequence thereof, and a preparation method thereof. The DNA polymerase according to the invention is a DNA polymerase, which is hyperthermophilic and has an elongation ability and fidelity higher than those of prior commercial DNA polymerases. Thus, the DNA polymerase according to the invention will be useful in precision analysis, precision diagnosis, identification and the like, which require accurate PCR.

Abstract: The present invention provides uracil-DNA glycosylase (UDG) gene originating from Psychrobacter sp. HJ147, and amino acid sequences deduced from the gene; expression and purification of Psp HJ147 UDG gene in Escherichia coli; and characterization of UDG obtained therefrom, and the use thereof in a polymerase chain reaction (PCR). The UDG according to the present invention has a specific activity of excising uracil bases in a uracil-containing DNA substrates at a low temperature, and is easily heat-inactivated. It thus can effectively eliminate cross contamination and carry-over contamination of PCR templates often occurring after a PCR process using dUTP. Therefore, it is useful for increasing preciseness (elimination of false positives), purity and amplification efficiency of PCR.

Abstract: Disclosed are a method of preparing an active Nanoarchaeum equitans B-type DNA polymerase (Neq DNA polymerase), an active Neq DNA polymerase prepared according to the method, and a polymerase chain reaction (PCR) using the active Neq DNA polymerase. The active Neq DNA polymerase may be used in various nucleic acid polymerization reactions, such as PCR.

Abstract: A gene encoding an enzyme aqualysin I is disclosed. The gene of the present invention comprises a region encoding a signal peptide; a pro-region located downstream the signal peptide-encoding region; a region encoding aqualysin I, which is located downstream the pro-region; and a tail region of aqualysin I, which is located downstream the aqualysin I-encoding region. This invention also provides a recombinant vector comprising the gene, Escherichia coli transformed with the recombinant vector and a process of producing aqualysin I utilizing the Escherichia coli.