The invention relates to thermostable polymerases that have polymerase activity temperatures in the range from 90.degree. C. up to 113.degree. C., such as those derived from Pyrolobus fumaria, and to polynucleotides encoding the polymerases In addition, methods of designing new thermostable DNA polymerases and methods of use thereof are also provided. The polymerases have increased activity and stability at increased pH and temperature.
1. An isolated, synthetic, or recombinant polypeptide comprising:
a) an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2, having polymerase activity at a temperature range from about 95° C. to 113° C. or
b) a fragment of a), having polymerase activity at a temperature range from about 95° C. to 113° C.
2. The isolated, synthetic, or recombinant polypeptide of claim 1 , comprising a fragment of a), having polymerase activity at a temperature range from about 95° C. to 113° C.
3. The polypeptide of claim 1 encoded by the nucleic acid sequence as set forth in SEQ ID NO: 1.
4. The polypeptide of claim 1 , having at least 97% sequence identity to SEQ ID NO: 2.
5. The polypeptide of claim 1 , having at least 99% sequence identity to SEQ ID NO: 2.