Thermostable DNA polymerase from the archaebacterium Sulfolobus acidocaldarius
Open Access
- 1 January 1989
- journal article
- research article
- Published by Wiley in European Journal of Biochemistry
- Vol. 178 (3) , 619-626
- https://doi.org/10.1111/j.1432-1033.1989.tb14490.x
Abstract
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 enzymes 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.degree. C and is active between 55.degree. C in the presence of activated calf-thymus DNA.This publication has 26 references indexed in Scilit:
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