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用聚乙烯亚胺沉淀核酸。

Precipitation of nucleic acids with poly(ethyleneimine).

作者信息

Cordes R M, Sims W B, Glatz C E

机构信息

Department of Chemical Engineering, Iowa State University, Ames 50011.

出版信息

Biotechnol Prog. 1990 Jul-Aug;6(4):283-5. doi: 10.1021/bp00004a009.

Abstract

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.

摘要

从细胞提取物中去除核酸是蛋白质回收下游处理中常见的早期步骤。我们报道了通过添加阳离子聚电解质聚乙烯亚胺(PEI)从酿酒酵母匀浆中沉淀核酸的方法,重点研究了PEI用量对分批和连续模式下颗粒大小、蛋白质损失以及核酸去除程度的影响。通过PEI沉淀法,酵母匀浆中核酸的去除率超过95%,无论是否预先去除细胞碎片,蛋白质损失约为15%。共沉淀的蛋白质主要是大分子物质,等电点有高有低。这种处理不会使细胞碎片聚集;连续沉淀器中沉淀颗粒的大小分布与蛋白质沉淀的大小分布非常相似。

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