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Fc-III 标记蛋白表达系统的开发用于蛋白纯化和检测。

Development of the Fc-III tagged protein expression system for protein purification and detection.

机构信息

School of Life Sciences, Tsinghua University, Beijing, China.

出版信息

PLoS One. 2012;7(8):e44208. doi: 10.1371/journal.pone.0044208. Epub 2012 Aug 29.

Abstract

In the present work, we developed the Fc-III tagged protein expression system for protein purification and detection. The Fc-III sequence encodes for a 13 residue peptide and this peptide is cyclized by disulfide bond formation when the fusion protein is expressed. The Fc-III-fusion proteins selectively bind to immunoglobulin Fc domains (IgG-Fc) expressed from E. coli. We showed the efficient purification of Fc-III tagged proteins by immobilized non-native IgG-Fc and the detection of the cellular locations of fusion proteins by fluorescent-conjugated IgG-Fc. Our results prove that Fc-III tagged protein expression system is a simple and efficient tool for protein purification and detection and is a useful addition to the biochemistry and proteomics toolbox.

摘要

在本工作中,我们开发了 Fc-III 标签蛋白表达系统,用于蛋白纯化和检测。Fc-III 序列编码一个 13 个氨基酸的肽段,当融合蛋白表达时,该肽段通过二硫键形成环化。Fc-III 融合蛋白特异性结合大肠杆菌表达的免疫球蛋白 Fc 结构域(IgG-Fc)。我们展示了通过固定化非天然 IgG-Fc 进行 Fc-III 标签蛋白的高效纯化,以及通过荧光标记 IgG-Fc 进行融合蛋白细胞定位的检测。我们的结果证明,Fc-III 标签蛋白表达系统是一种简单有效的蛋白纯化和检测工具,是生物化学和蛋白质组学工具包的有益补充。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/428d/3430635/3639ca31de7c/pone.0044208.g001.jpg

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