Terent'ev L L, Terent'eva N A, Rassazov V A
Biokhimiia. 1980 Sep;45(9):1603-8.
Using DEAE-cellulose chromatography, three peaks of the DNA-polymerase activity were found in a homogenate of the sea urchin Strongylocentrotus intermedius embryos at stage 32 of the blastomer. The isolation and purification of DNA-polymerase making up the bulk of he DNA-synthesizing activity of the sea urchin embryo cells included fractionation by ammonium sulfate, chromatography on DEAE-cellulose, hydroxyapatite and DNA-cellulose, resulting in a 2750-fold purification of the enzyme. The enzyme activity requires the presence of two-chain DNA activated by pancreatic DNAse, four dNTP and Mg2+. The enzyme is inhibited by a high ionic strength (150 mM KCl or NaCL) and the sulfhydryl reagent--N-ethylmaleimide; the pH optimum is 8.0. The molecular weight of the enzyme as determined by gel-filtration is about 150 000. It is assumed that the enzyme under study can be related to DNA-polymerases of the alpha-type.
利用二乙氨基乙基纤维素色谱法，在间型强刺海胆胚胎处于32细胞期的卵裂球匀浆中发现了三个DNA聚合酶活性峰。对构成海胆胚胎细胞大部分DNA合成活性的DNA聚合酶进行分离和纯化，包括硫酸铵分级分离、二乙氨基乙基纤维素色谱法、羟基磷灰石柱色谱法和DNA纤维素柱色谱法，最终该酶得到了2750倍的纯化。该酶活性需要经胰脱氧核糖核酸酶激活的双链DNA、四种脱氧核苷三磷酸和镁离子。该酶会受到高离子强度（150 mM氯化钾或氯化钠）和巯基试剂N - 乙基马来酰亚胺的抑制；最适pH为8.0。通过凝胶过滤法测定该酶的分子量约为150 000。据推测，所研究的这种酶可能属于α型DNA聚合酶。
