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Simultaneous detection of DsRed2‐tagged and EGFP‐tagged human β‐interferons in the same single cells
Author(s) -
Maruyama Masato,
Nishio Teruko,
Yoshida Toyokazu,
Ishida Chisaki,
Ishida Kayo,
Watanabe Yoshihiko,
Nishikawa Makiya,
Takakura Yoshinobu
Publication year - 2004
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.20203
Subject(s) - green fluorescent protein , fusion protein , protein subcellular localization prediction , plasmid , confocal , protein tag , confocal microscopy , microbiology and biotechnology , fluorescence , fluorescent protein , cell , fluorescence microscope , chemistry , biology , gene , recombinant dna , biochemistry , physics , geometry , mathematics , quantum mechanics
The red fluorescent protein DsRed2 is a useful fusion tag for various proteins, together with the enhanced green fluorescent protein (EGFP). These chromoproteins have spectral properties that allow simultaneous distinctive detection of tagged proteins in the same single cells by dual color imaging. We used them for tagging a secretory protein, human interferon‐β (IFN‐β). Expression plasmids for human IFN‐β tagged with DsRed2 or with EGFP at the carboxyl terminal were constructed and their coexpression was examined in Mardin–Darby canine kidney epithelial cells. Although maturation of DsRed2 for coloration was slow and the color intensity was weak compared with EGFP, low temperature treatment (20°C) allowed DsRed2‐tagged human IFN‐β to be detected in the cells using color imaging. Consequently, the two chimeric proteins were shown to be colocalized in the same single cells by dual color confocal microscopy. This approach will be useful for investigating subcellular localization of not only cell resident proteins but also secretory proteins. © 2004 Wiley‐Liss, Inc.