Brown M, Bollum F J, Chang L M
Proc Natl Acad Sci U S A. 1981 May;78(5):3049-52. doi: 10.1073/pnas.78.5.3049.
Immunoglobulin (IgG) and the F(ab')2 fragment of IgG were prepared from serum of a rabbit immunized with purified calf thymus DNA polymerase alpha (deoxynucleosidetriphosphate:DNA deoxynucleotidyltransferase, EC 2.7.7.7). An indirect immunofluorescent method based on these reagents was used to detect the intracellular localization of DNA polymerase alpha in primary fetal bovine fibroblasts. The results show that the bulk of DNA polymerase alpha is located in the perinuclear region of the cytoplasm. Immunofluorescent staining of cytoplast and Ficoll-Paque gradient-purified karyoplast fragments resulting from cytochalasin enucleation show the presence of DNA polymerase alpha in cytoplasts and the virtual absence of the enzyme in the nucleus of the karyoplast itself. The implication of this unusual intracellular location for DNA polymerase alpha is discussed.
用纯化的小牛胸腺DNA聚合酶α(脱氧核苷三磷酸:DNA脱氧核苷酸转移酶,EC 2.7.7.7)免疫兔子,从其血清中制备免疫球蛋白(IgG)和IgG的F(ab')2片段。基于这些试剂建立了间接免疫荧光法,用于检测原代胎牛成纤维细胞中DNA聚合酶α的细胞内定位。结果表明,大部分DNA聚合酶α位于细胞质的核周区域。对经细胞松弛素去核产生的胞质体和经Ficoll-Paque梯度纯化的核质体片段进行免疫荧光染色,结果显示胞质体中存在DNA聚合酶α,而核质体本身的细胞核中几乎没有该酶。本文讨论了DNA聚合酶α这种不寻常的细胞内定位的意义。