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鉴定霍乱弧菌中抗 FLAG 抗体结合蛋白。

Characterization of an anti-FLAG antibody binding protein in V. cholerae.

机构信息

Weill Institute for Cell and Molecular Biology, Cornell, University, Ithaca, NY, 14853, USA; Department of Microbiology, Cornell University, Ithaca, NY, 14853, USA.

Weill Institute for Cell and Molecular Biology, Cornell, University, Ithaca, NY, 14853, USA; Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY, 14853, USA.

出版信息

Biochem Biophys Res Commun. 2020 Jul 30;528(3):493-498. doi: 10.1016/j.bbrc.2020.05.169. Epub 2020 Jun 3.

Abstract

FLAG-tags are commonly used for protein abundance measurements and for identification of protein-protein interactions in living cells. We have observed that the cholera pathogen Vibrio cholerae encodes a FLAG-antibody-reactive protein and identified this protein as an outer membrane porin, Porin4, which contains a sequence very similar to the 3xFLAG epitope tag. We have demonstrated the binding affinity of the conserved peptide sequence (called Porin 4 tag) in Porin4 against monoclonal anti-FLAG M2 antibody. In addition, we created a porin4 deletion mutant, which can be used for background-less FLAG antibody detection experiments.

摘要

FLAG 标签常用于蛋白质丰度的测量和活细胞中蛋白质-蛋白质相互作用的鉴定。我们观察到霍乱病原体霍乱弧菌编码一种 FLAG 抗体反应蛋白,并将该蛋白鉴定为外膜孔蛋白 Porin4,其包含与 3xFLAG 表位标签非常相似的序列。我们已经证明了 Porin4 中保守肽序列(称为 Porin4 标签)对单克隆抗 FLAG M2 抗体的结合亲和力。此外,我们创建了一个缺失突变体,可用于无背景的 FLAG 抗体检测实验。

相似文献

1
Characterization of an anti-FLAG antibody binding protein in V. cholerae.鉴定霍乱弧菌中抗 FLAG 抗体结合蛋白。
Biochem Biophys Res Commun. 2020 Jul 30;528(3):493-498. doi: 10.1016/j.bbrc.2020.05.169. Epub 2020 Jun 3.

本文引用的文献

1
Overview of affinity tags for protein purification.蛋白质纯化亲和标签概述。
Curr Protoc Protein Sci. 2013 Sep 24;73:9.9.1-9.9.23. doi: 10.1002/0471140864.ps0909s73.
2
Several affinity tags commonly used in chromatographic purification.几种常用于色谱纯化的亲和标签。
J Anal Methods Chem. 2013;2013:581093. doi: 10.1155/2013/581093. Epub 2013 Dec 26.
7
The Vibrio cholerae chitin utilization program.霍乱弧菌几丁质利用程序
Proc Natl Acad Sci U S A. 2004 Feb 24;101(8):2524-9. doi: 10.1073/pnas.0308707101.

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