PRECIPITATION OF NUCLEIC-ACIDS WITH POLY(ETHYLENEIMINE)
PRECIPITATION OF NUCLEIC-ACIDS WITH POLY(ETHYLENEIMINE)
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DOI：
10.1021/bp00004a009
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发表时间：
1990-07-01
影响因子：
2.9
通讯作者：
GLATZ, CE
中科院分区：
文献类型：
--
作者：
CORDES, RM;SIMS, WB;GLATZ, CE
从细胞提取物中去除核酸是蛋白质回收下游加工的常见早期步骤。我们报道了通过添加阳离子聚电解质聚亚胺（PEI）从酵母匀浆中沉淀核酸，重点研究了PEI用量对颗粒大小、蛋白质损失和核酸去除程度的影响，包括批处理和连续模式。通过PEI沉淀，酵母匀浆中核酸的去除率超过95%，无论是否事先去除细胞碎片，蛋白质损失约为15%。共沉淀蛋白主要为大分子量物质，具有高、低等电点。这种处理不会使细胞碎片聚集；从连续沉淀器中沉淀的颗粒的大小分布与蛋白质沉淀非常相似。
Removal of nucleic acids from cell extracts is a common early step in downstream processing for protein recovery. We report on the precipitation of nucleic acids from a homogenate of Saccharomyces cerevisiae by addition of the cationic polyelectrolyte poly(ethyleneimine) (PEI), focusing on the effect of PEI dosage on particle size, protein loss, and extent of nucleic acid removal in both batch and continuous mode. Better than 95% removal of nucleic acids from yeast homogenates was achieved by means of precipitation with PEI with protein losses of approximately 15% with or without previous removal of cell debris. The coprecipitated protein is predominately large molecular weight material and exhibits both low and high isoelectric points. Such treatment does not aggregate the cell debris; size distribution of the precipitated particles from a continuous precipitator is very similar to that for protein precipitation.