Mikhaĭlov V S, Guliamov D B, Peskin A V, Filatova L S
Biokhimiia. 1983 Jun;48(6):1012-9.
The homogenate of loach (Misgurnus fossilis) embryonic cells was fractionated by differential centrifugation into fractions, in which the activities of DNA polymerases alpha and gamma were measured. About half of the total alpha-polymerase activity was retained in the postmicrosomal supernatant. The activity of gamma-polymerase in the supernatant did not exceed 2% of the total activity. The specific activity of gamma-polymerase was maximal in the mitochondrial pellet. The enzyme activity in the nuclear pellet was proportional to the activity of mitochondrial enzymes, cytochrome oxidase and KCN-insensitive superoxide dismutase. The nuclei were separated from gamma-polymerase-containing fraction by ultracentrifugation in a 10-50% sucrose density gradient. The material of this fraction was identical to the mitochondria in terms of buoyant density, morphology and enzymatic properties. Thus, in loach embryos the fraction of gamma-polymerase associated with the nuclei is absent; the total gamma-polymerase activity is localized in the mitochondria.
将泥鳅(Misgurnus fossilis)胚胎细胞匀浆通过差速离心法分离成不同组分,测定其中DNA聚合酶α和γ的活性。总α聚合酶活性的约一半保留在微粒体后上清液中。上清液中γ聚合酶的活性不超过总活性的2%。γ聚合酶的比活性在线粒体沉淀中最高。核沉淀中的酶活性与线粒体酶、细胞色素氧化酶和对KCN不敏感的超氧化物歧化酶的活性成正比。通过在10 - 50%蔗糖密度梯度中进行超速离心,将细胞核与含γ聚合酶的组分分离。该组分的物质在浮力密度、形态和酶学性质方面与线粒体相同。因此,在泥鳅胚胎中,与细胞核相关的γ聚合酶组分不存在;总γ聚合酶活性定位于线粒体中。