Thermostable DNA polymerase from the archaebacterium Sulfolobus acidocaldarius

• Published in: European journal of biochemistry Vol. 178; no. 3; pp. 619 - 626
• Main Authors: ELIE, Christiane, RECONDO, Anne Marie, FORTRRE, Patrick
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.01.1989
• ISSN: 0014-2956; 1432-1033
• DOI: 10.1111/j.1432-1033.1989.tb14490.x

We have purified to near homogeneity a DNA polymerase from the thermoacidophilic archaebacterium Sulfolobus acidocaldarius. Sodium dodecyl sulfate gel electrophoresis of the purified enzyme revealed a polypeptide of 100 kDa. On the basis of a Stokes radius of 4.2 nm and a sedimentation coefficient of 6 S, the purified enzyme has an estimated molecular mass of 109 kDa. These results are consistent with the enzyme being a monomer of 100 kDa. In addition a polyclonal antiserum, obtained by injection of the electroeluted 100‐kDa polypeptide into a rabbit, specifically neutralized the DNA‐polymerase activity. The enzyme is sensitive to both N‐ethylmaleimide and 2′,3′‐dideoxyribosylthymine triphosphate and resistant to aphidicolin. The purified DNA polymerase has neither exonuclease nor primase activities. In our in vitro conditions, the enzyme is thermostable up to 80°C and is active between 55°C and 85°C in the presence of activated calf‐thymus DNA.