Simultaneous detection of DsRed2-tagged and EGFP-tagged human β-interferons in the same single cells

Masato Maruyama, Teruko Nishio, Toyokazu Yoshida, Chisaki Ishida, Kayo Ishida, Yoshihiko Watanabe, Makiya Nishikawa, Yoshinobu Takakura

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)497-502
Number of pages6
JournalJournal of Cellular Biochemistry
Volume93
Issue number3
DOIs
Publication statusPublished - Dec 1 2004
Externally publishedYes

Keywords

  • Epithelial cell
  • Green fluorescent protein
  • Interferon-β
  • Red fluorescent protein
  • Secretory protein

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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  • Cite this

    Maruyama, M., Nishio, T., Yoshida, T., Ishida, C., Ishida, K., Watanabe, Y., Nishikawa, M., & Takakura, Y. (2004). Simultaneous detection of DsRed2-tagged and EGFP-tagged human β-interferons in the same single cells. Journal of Cellular Biochemistry, 93(3), 497-502. https://doi.org/10.1002/jcb.20203