Thermostable DNA polymerase with enhanced thermostability and enhanced length and efficiency of primer extension
First Claim
1. A DNA polymerase comprising substantially the same amino acid sequence as that of Thermus aquaticus DNA polymerase, excluding the N-terminal 280 amino acid residues of Thermus aquaticus DNA polymerase.
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Abstract
A DNA polymerase having an amino acid sequence comprising substantially the same amino acid sequence as that of Thermus aquaticus or Thermus flavus DNA polymerase, excluding the N-terminal 280 amino acid residues of Thermus aquaticus DNA polymerase or the N-terminal 279 amino acid residues of Thermus flavus DNA polymerase, recombinant DNA sequences encoding said DNA polymerases, vectors comprising said DNA sequences, and host cells containing such vectors. A formulation of thermostable DNA polymerases comprising a majority component comprised of at least one thermostable DNA polymerase of the type described above, wherein the DNA polymerase lacks 3'"'"'-exonuclease activity, and a minority component comprised of at least one thermostable DNA polymerase exhibiting 3'"'"'-exonuclease activity, and an improved method for amplifying nucleic acid sequences by polymerase chain reaction wherein the above formulation is mixed and used to catalyze primer extension, are also provided.
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Citations
16 Claims
- 1. A DNA polymerase comprising substantially the same amino acid sequence as that of Thermus aquaticus DNA polymerase, excluding the N-terminal 280 amino acid residues of Thermus aquaticus DNA polymerase.
- 5. A DNA polymerase comprising substantially the same amino acid sequence as that of Thermus flavus DNA polymerase, excluding the N-terminal 279 amino acid residues of Thermus flavus DNA polymerase.
- 6. A formulation of thermostable DNA polymerases comprising at least one thermostable DNA polymerase lacking 3'"'"'-exonuclease activity and at least one thermostable DNA polymerase exhibiting 3'"'"'-exonuclease activity, wherein the DNA polymerases are present in a ratio of from about 4 units to about 2000 units of the DNA polymerase lacking 3'"'"'-exonuclease activity to 1 unit of the DNA polymerase exhibiting 3'"'"'-exonuclease activity.
Specification