Klimczak L J, Grummt F, Burger K J
Nucleic Acids Res. 1985 Jul 25;13(14):5269-82. doi: 10.1093/nar/13.14.5269.
DNA polymerase has been purified about 25,000-fold from the thermoacidophilic archaebacterium Sulfolobus acidocaldarius. On SDS-PAGE the enzyme was observed to have a molecular weight of 100 kDa and to be about 90% pure. The native molecular weight was 108 kDa indicating that the enzyme is composed of a single polypeptide. Activity gel analysis showed an active polypeptide of about 100 kDa. Under conditions promoting proteolysis this polypeptide was degraded to a slightly smaller form of 98 kDa. The enzyme has been characterized in respect to optimal assay conditions, template specificity, sensitivity to inhibitors and associated nuclease activities. The high temperature optimum of 65 degrees C should be emphasized. No substantial similarities have been found with other prokaryotic and eukaryotic DNA polymerases, although the enzyme bears certain resemblances to prokaryotic non-replicative polymerases.
已从嗜热嗜酸古细菌嗜酸热硫化叶菌中纯化出约25000倍的DNA聚合酶。在SDS - 聚丙烯酰胺凝胶电泳上观察到该酶的分子量为100 kDa，纯度约为90%。天然分子量为108 kDa，表明该酶由单一多肽组成。活性凝胶分析显示有一条约100 kDa的活性多肽。在促进蛋白水解的条件下，该多肽降解为稍小的98 kDa形式。已对该酶的最佳测定条件、模板特异性、对抑制剂的敏感性及相关核酸酶活性进行了表征。应强调其65摄氏度的高温最佳温度。与其他原核和真核DNA聚合酶未发现实质性相似之处，尽管该酶与原核非复制性聚合酶有一定相似性。
