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Novel short peptide tag from a bacterial toxin for versatile applications

Title
Novel short peptide tag from a bacterial toxin for versatile applications
Authors
Lee, Tae HeeKim, Kwang SooKim, Jin HeeJeong, Jae-HoWoo, Hye RyunPark, So RaSohn, Myung-HoLee, Hyeon JuRhee, Joon HaengCha, Sun-ShinHwang, Joo-HeeChung, Kyung Min
DGIST Authors
Woo, Hye Ryun
Issue Date
2020-04
Citation
Journal of Immunological Methods, 479, 112750
Type
Article
Article Type
Article
Author Keywords
Epitope tagMonoclonal antibodyImmunodetectionAffinity purification
Keywords
RECOMBINANT PROTEIN EXPRESSIONMYC ANTIBODY 9E10MONOCLONAL-ANTIBODIESESCHERICHIA-COLISUBCELLULAR-LOCALIZATIONSTEP PURIFICATIONAFFINITY PEPTIDEHNRNP A1BINDINGSYSTEM
ISSN
0022-1759
Abstract
The specific recognition between a monoclonal antibody (mAb) and its epitope can be used in a tag system that has proved valuable in a wide range of biological applications. Herein, we describe a novel tag called RA-tag that is composed of a seven amino acid sequence (DIDLSRI) and recognized by a highly specific mAb, 47RA, against the bacterial toxin Vibrio vulnificus RtxA1/MARTXVv. By using recombinant proteins with the RA-tag at the N-terminal, C-terminal, or an internal site, we demonstrated that the tag system could be an excellent biological system for both protein purification and protein detection in enzyme-linked immunosorbent, Western blot, flow cytometry, and immunofluorescence staining analyses in Escherichia coli, mammalian cell lines, yeast, and plant. In addition, our RA-tag/47RA mAb combination showed high sensitivity and reliable affinity (KD = 5.90 × 10−8 M) when compared with conventional tags. Overall, our results suggest that the RA-tag system could facilitate the development of a broadly applicable tag system for biological research. © 2020
URI
http://hdl.handle.net/20.500.11750/11614
DOI
10.1016/j.jim.2020.112750
Publisher
Elsevier BV
Related Researcher
  • Author Woo, Hye Ryun Lab of Plant Molecular Communication
  • Research Interests Genome biology; Epigenomics; Plant senescence
Files:
There are no files associated with this item.
Collection:
Department of New BiologyLab of Plant Molecular Communication1. Journal Articles


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