Bagshaw J C
Nucleic Acids Res. 1976 Jun;3(6):1449-61. doi: 10.1093/nar/3.6.1449.
RNA polymerase II from larvae of the brine shrimp, Artemia salina, was highly purified by two cycles of DEAE-cellulose chromatography followed by centrifugation through discontinuous sucrose gradients. Gradient fractions were subjected to elctrophoresis is polyacrylamide gels containing sodium dodecyl sulfate. The subunit structure of RNA polymerase II was determined by quantitative comparison of the polypeptides and enzyme activity present in each gradient fraction. The enzyme contains one copy of each of four subunits with estimated molecular weights of 170,000, 130,000, 36,000 and 24,000. The total molecular weight agrees well with the molecular weight estimated for the native enzyme by density gradient centrifugation.
通过两轮DEAE - 纤维素层析,随后经不连续蔗糖梯度离心,对卤虫(Artemia salina)幼虫的RNA聚合酶II进行了高度纯化。将梯度级分在含十二烷基硫酸钠的聚丙烯酰胺凝胶中进行电泳。通过对每个梯度级分中存在的多肽和酶活性进行定量比较,确定了RNA聚合酶II的亚基结构。该酶包含四个亚基各一个拷贝,其估计分子量分别为170,000、130,000、36,000和24,000。总分子量与通过密度梯度离心法对天然酶估计的分子量非常吻合。