Rasskazov V A, Shumilov Iu N, Pin'kevich E V
Mol Biol (Mosk). 1976 May-Jun;10(2):609-17.
In the sea urchin embryonic cells, all newly synthesized nuclear DNA (n-DNA) pulse-labeled by 3H-thymidine was found within DNA-membrane complex (DNA-mc) isolated by centrifugation of lysates of nuclei after their treatment with Sarkosyl, Brij-35, or sodium dodecylsulfate through neutral sucrose (10--30%) gradients. This attachment has been shown not to be an artifact due to the unspecific effect of the detergents or the destabilization of the secondary structure of n-DNA because the association of the exogenous 14C-DNA with nuclear membrane and chromatin did not occur during the isolation of the DNA-mc. n-DNA was not replaced from DNA-mc when the latter was isolated in the excess of unlabeled denatured DNA. n-DNA associated with DNA-mc behaved as a precursor of chromosomal DNA. It is suggested that in sea urchin embryonic cells the synthesis of nuclear DNA is carried out by the replicative complex attached to the nuclear membrane.
在海胆胚胎细胞中,通过用 Sarkosyl、Brij - 35 或十二烷基硫酸钠处理细胞核裂解物,然后通过中性蔗糖(10 - 30%)梯度离心分离得到的 DNA - 膜复合物(DNA - mc)中,发现了所有经 3H - 胸腺嘧啶脉冲标记的新合成核 DNA(n - DNA)。这种附着已被证明不是由于去污剂的非特异性作用或 n - DNA 二级结构的不稳定而产生的假象,因为在分离 DNA - mc 的过程中,外源 14C - DNA 并未与核膜和染色质发生结合。当在过量未标记的变性 DNA 存在下分离 DNA - mc 时,可以观察到 n - DNA 并未从 DNA - mc 中被取代。与 DNA - mc 相关联的 n - DNA 表现为染色体 DNA 的前体。这表明在海胆胚胎细胞中,核 DNA 的合成是由附着在核膜上的复制复合物进行的。