Synchronization of HeLa cell cultures by inhibition of DNA polymerase alpha with aphidicolin.
Synchronization of HeLa cell cultures by inhibition of DNA polymerase alpha with aphidicolin.
复制标题
通过用 aphidicolin 抑制 DNA 聚合酶 α 来同步 HeLa 细胞培养物。
DOI：
10.1093/nar/8.2.377
复制
发表时间：
1980
影响因子：
14.9
通讯作者：
G. Koch
中科院分区：
文献类型：
--
作者：
G. Pedrali;Silvio Spadari;A. Miller;A.O.A. Miller;J. Kruppa;G. Koch
阿非迪霉素对复制型α-聚合酶的抑制作用和其体内作用的可逆性（Pedrali-Noy & Spadari，1979，Biochem.Biophys.通信资源88，1194-2002）允许培养物中细胞的同步化。Aphidicolin阻止G1期细胞进入DNA合成期，阻滞细胞于“S”期，使G2、M和G1期细胞继续进行细胞周期，并在G1/S边界聚集。Aphidicolin是比羟基脲和胸苷更有用的试剂，因为它不影响细胞活力或“S”期持续时间，并且不干扰dNTP或DNA聚合酶的合成。事实上，暴露于药物的细胞继续合成所有三种DNA聚合酶α，β和γ以及所有dNTP，当阻断被移除时，它们以DNA起始和复制的最佳水平存在。该技术很简单，可以应用于悬浮或单层生长的细胞，并允许收获大量的同步细胞。
Both the inhibitory effect of aphidicolin on the replicative alpha-polymerase and the reversibility of its action in vivo (Pedrali-Noy & Spadari, 1979, Biochem. Biophys. Res. Commun. 88, 1194-2002) allow the synchronization of cells in culture. Aphidicolin prevents G1 cells from entering the DNA synthetic period, blocks cells in "S" phase, allows G2, M and G1 cells to continue the cell cycle and to accumulate at the G1/S border. Aphidicolin is a more useful reagent than hydroxyurea and thymidine because it does not affect cell viability or "S" phase duration and does not interfere with the synthesis of dNTPs or DNA polymerases. In fact cells exposed to the drug continue to synthesize all three DNA polymerases alpha, beta and gamma as well as all dNTPs which, when the block is removed, are present at levels optimal for DNA initiation and replication. The technique is simple and can be applied to cells growing in suspension or monolayers and allows one to harvest large quantities of synchronized cells.