Chimeric DNA polymerase and application thereof
A chimeric DNA polymerase includes: a first fragment having at least 80% homology to at least part of an N-end domain of KOD DNA polymerase; a second fragment having at least 80% homology to at least part of an exonucleolytic domain of Pab DNA polymerase; a third fragment having at least 80% homology to at least part of the N-end domain of KOD DNA polymerase; a fourth fragment having at least 80% homology to at least part of a palm domain of Pfu DNA polymerase; a fifth fragment having at least 80% homology to at least part of a finger domain of Pab DNA polymerase; a sixth fragment having at least 80% homology to at least part of the palm domain of Pfu DNA polymerase; and a seventh fragment having at least 80% homology to at least part of a thumb domain of KOD DNA polymerase.
1 . A chimeric DNA polymerase, comprising:
a first peptide fragment comprising an amino acid sequence of sites 1 to 130 of a KOD DNA polymerase;
a second peptide fragment comprising an amino acid sequence of sites 131 to 337 of a Pab DNA polymerase, an N-terminal end of the second peptide fragment being connected to a C-terminal end of the first peptide fragment;
a third peptide fragment comprising an amino acid sequence of sites 338 to 373 of the KOD DNA polymerase, an N-terminal end of the third peptide fragment being connected to a C-terminal end of the second peptide fragment;
a fourth peptide fragment comprising an amino acid sequence of sites 374 to 448 of a Pfu DNA polymerase, an N-terminal end of the fourth peptide fragment being connected to a C-terminal end of the third peptide fragment;
a fifth peptide fragment comprising an amino acid sequence of sites 449 to 500 of the Pab DNA polymerase, an N-terminal end of the fifth peptide fragment being connected to a C-terminal end of the fourth peptide fragment;
a sixth peptide fragment comprising an amino acid sequence of sites 501 to 591 of the Pfu DNA polymerase, an N-terminal end of the sixth peptide fragment being connected to a C-terminal end of the fifth peptide fragment; and
a seventh peptide fragment comprising an amino acid sequence of sites 591 to 774 of the KOD DNA polymerase, an N-terminal end of the seventh peptide fragment being connected to a C-terminal end of the sixth peptide fragment.
2 . The chimeric DNA polymerase according to claim 1 , wherein the chimeric DNA polymerase has an amino acid sequence set forth as SEQ ID NO: 1.
3 . An isolated nucleic acid, encoding the chimeric DNA polymerase according to claim 1 .
4 . The isolated nucleic acid according to claim 3 , wherein the isolated nucleic acid has a nucleotide sequence set forth as SEQ ID NO: 2.
5 . A construct, comprising the isolated nucleic acid according to claim 3 .
6 . A recombinant cell or recombinant microorganism, containing the isolated nucleic acid according to claim 3 .
7 . A method for obtaining the chimeric DNA polymerase according to claim 1 , comprising:
culturing a recombinant cell or recombinant microorganism containing an isolated nucleic acid encoding the chimeric DNA polymerase under conditions suitable for expressing the chimeric DNA polymerase to obtain the chimeric DNA polymerase.
8 . A kit, containing the chimeric DNA polymerase according to claim 1 .
9 . The chimeric DNA polymerase according to claim 1 , wherein the chimeric DNA polymerase is used in DNA amplification, genetic screening, sequencing, or mutation detection.
10 . The isolated nucleic acid according to claim 3 , wherein the isolated nucleic acid is used in DNA amplification, genetic screening, sequencing, or mutation detection.
11 . The construct according to claim 5 , wherein the construct is used in DNA amplification, genetic screening, sequencing, or mutation detection.
12 . The recombinant cell or recombinant microorganism according to claim 6 , wherein the recombinant cell or recombinant microorganism is used in DNA amplification, genetic screening, sequencing, or mutation detection.
13 . The kit according to claim 8 , wherein the kit is used in DNA amplification, genetic screening, sequencing, or mutation detection.