Effective elimination of nucleic acids from bacterial protein samples for optimized blue native polyacrylamide gel electrophoresis
Effective elimination of nucleic acids from bacterial protein samples for optimized blue native polyacrylamide gel electrophoresis
复制标题
有效消除细菌蛋白质样品中的核酸，以优化蓝色天然聚丙烯酰胺凝胶电泳
DOI：
10.1002/elps.200900026
复制
发表时间：
2009-07-01
期刊：
影响因子：
2.9
通讯作者：
Wang, Zhijun
中科院分区：
文献类型：
--
作者：
Liang, Jingdan;Niu, Qiule;Wang, Zhijun
从变铅青链霉菌和大肠杆菌的细菌蛋白质样品中残留的核酸被发现显着干扰蓝色天然聚丙烯酰胺凝胶电泳（BN-PAGE），一种经常用于分析蛋白质组学研究中的细菌蛋白质复合物的技术。我们已经使用超离心和/或用硫酸链霉素沉淀细胞裂解物来从总蛋白和/或膜蛋白样品中消除核酸。首先通过用硫酸链霉素沉淀来富集核酸结合蛋白，然后通过添加聚乙烯亚胺沉淀来消除污染核酸。发现通过这些样品制备步骤，BN-PAGE的性能显著提高。
Nucleic acids remaining within bacterial protein samples from Streptomyces lividans and Escherichia coli were found to interfere significantly with blue native polyacrylamide gel electrophoresis (BN-PAGE), a technique used frequently for analyzing bacterial protein complexes in proteomics studies. We have used ultracentrifugation and/or precipitation of cell lysates with streptomycin sulfate to eliminate nucleic acids from total and/or membrane protein samples. Nucleic acid-binding proteins were first enriched by precipitation with streptomycin sulfate, and contaminating nucleic acids were then eliminated by precipitation by adding polyethyleneimine. The performance of BN-PAGE was found to be dramatically improved by these sample preparation steps.