Purified thermostable
Purified thermostable Pyrococcus furiosus DNA polymerase that migrates on a non-denaturing polyacrylamide gel faster than phosphorylase B and Taq polymerase and more slowly than bovine serum albumin and has an estimated molecular weight of 90,000–93,000 daltons when compared with a Taq polymerase standard assigned a molecular weight of 94,000 daltons.
1. An isolated polynucleotide encoding a thermostable DNA polymerase, wherein the isolated polynucleotide is capable of hybridizing to a polynucleotide complementary to a polynucleotide that encodes amino acids 1 to 775 of SEQ ID NO: 1 under hybridization conditions comprising incubating at 42° C. in a hybridization buffer comprising 0.9 M NaCl and 50% formamide, and under wash conditions comprising incubating at 68° C. in a wash buffer comprising 0.015 M NaCl and 0.5% sodium dodecyl sulfate.
2. The isolated polynucleotide of claim 1 , wherein the thermostable DNA polymerase is a fusion protein.
3. A vector comprising the isolated polynucleotide of claim 1 .
4. A host cell comprising the isolated polynucleotide of claim 1 .
5. A method of preparing a recombinant thermostable DNA polymerase, comprising culturing the host cell of claim 4 under conditions suitable for the expression of the thermostable DNA polymerase.