携带DNA的亲和乳胶的制备以及用该乳胶纯化转录因子

Preparation of DNA-carrying affinity latex and purification of transcription factors with the latex.

作者信息

Inomata Y, Wada T, Handa H, Fujimoto K, Kawaguchi H

机构信息

Department of Applied Chemistry, Faculty of Science and Technology, Keio University, Yokohama, Japan.

出版信息

J Biomater Sci Polym Ed. 1994;5(4):293-302. doi: 10.1163/156856294x00031.

DOI:10.1163/156856294x00031

PMID:8025028

Abstract

We have developed DNA-carrying latex particles for the separation and purification of transcription factors. These particles consist of styrene (St), glycidyl methacrylate (GMA) and divinylbenzene (DVB). It was confirmed that the ethanolamine-treated surface of these particles suffered no nonspecific adsorption of proteins. To the latex particles sequence-specific DNA oligomers were immobilized via covalent coupling. A transcription factor, E4TF3, was efficiently purified to homogeneity using the latext particles. In contrast, the purification using DNA-carrying Sepharose gel yielded poor results. Compared to DNA-carrying Sepharose gel, the latex particles exhibited several times higher efficiency in the purification of E4TF3 from the crude nuclear extract.

摘要

我们已开发出用于分离和纯化转录因子的携带DNA的乳胶颗粒。这些颗粒由苯乙烯（St）、甲基丙烯酸缩水甘油酯（GMA）和二乙烯基苯（DVB）组成。已证实这些颗粒经乙醇胺处理的表面不会发生蛋白质的非特异性吸附。通过共价偶联将序列特异性DNA寡聚物固定到乳胶颗粒上。使用乳胶颗粒可将转录因子E4TF3高效纯化至均一状态。相比之下，使用携带DNA的琼脂糖凝胶进行纯化的效果较差。与携带DNA的琼脂糖凝胶相比，乳胶颗粒从粗核提取物中纯化E4TF3的效率要高出几倍。