A method for fractionating rat liver cell nuclei on equilibrium density gradients of CS2SO4.
A method for fractionating rat liver cell nuclei on equilibrium density gradients of CS2SO4.
复制标题
一种在 CS2SO4 平衡密度梯度上分离大鼠肝细胞核的方法。
DOI：
10.1016/0003-2697(75)90295-x
复制
发表时间：
1975
影响因子：
2.9
通讯作者：
W. Kaufmann
中科院分区：
文献类型：
--
作者：
V. Genta;D. Kaufman;W. Kaufmann
超声破碎的细胞核从增殖的肝细胞在Cs2 SO 4平衡密度梯度分级。核成分主要集中在三条带中，分别为轻带（LB，1.21 g/cm ~ 3）、中带（MB，1.26 g/cm ~ 3）和重带（HB，1.32 g/cm ~ 3）。蛋白质和核酸的梯度分布分析表明，一些大分子相互作用的保存。研究比较放射性标记的DNA的分布后，1-或120分钟的时间间隔后，氚标记的胸苷注射指示富集新生的DNA在LB和MB。这种富集对超声处理的时间和压力敏感。此外，DNA聚合酶活性证明在梯度馏分中去除Cs2 SO 4后，与LB和MB中的大部分活动再次。这些结果表明，该程序作为分离酶活性DNA复制复合物的初始步骤。
Sonically disrupted nuclei from proliferating liver cells were fractionated in Cs2SO4equilibrium density gradients. Nuclear constituents were concentrated in three bands designated as light band (LB, 1.21 g/cm3), middle band (MB, 1.26 g/cm3), and heavy band (HB, 1.32 g/cm3). Analysis of protein and nucleic acid distribution in gradients suggests preservation of some macromolecular interactions. Studies comparing distributions of radioactively labeled DNA after 1- or 120-min intervals following tritiated thymidine injection indicate enrichment of nascent DNA in LB and MB. This enrichment is sensitive to time and pressure of sonication. Furthermore, DNA-polymerase activity was demonstrated in the gradient fractions following removal of Cs2SO4, with most activity once again in the LB and MB. These results suggest this procedure as an initial step in the isolation of an enzymatically active DNA replication complex.