Single Cell Reporter Assay Using Cell Surface Displayed Vargula Luciferase

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概要

• 論文の詳細を見る

• Reporter genes such as firefly luciferase are common tools to monitor gene expression in various systems. As reporter gene represents the expression level of the gene of interest with its enzyme activity, firefly luciferase is most frequently used because its luminescent activity is highly sensitive and less time consumable for assay. However, since firefly luciferase is expressed internally in the cell, lysis of the cell is a critical step, and thus it is difficult to monitor the gene expression level continuously. In this report, we utilized secretive Vargula hilgendorfii luciferase modified to cell surface displayed one by fusing with human EGFR transmembrane sequence. This modified Vargula luciferase was expressed on cell surface without losing its bioluminescent activity. Co-transfection with secretive alkaline phosphatase showed that the behaviors of cell surface displayed Vargula luciferase and secretive alkaline phosphatase are comparable to each other. Furthermore, the luminescence of a single cell expressing cell surface displayed Vargula luciferase can be monitored by using photon counting CCD camera, which indicates that this reporter gene can monitor gene expression in a single cell without cell lysis.

• 社団法人日本生物工学会の論文
• 2001-12-25

著者

• Nagamune T

  Univ. Tokyo Tokyo Jpn

• Nagamune T

  Department Of Chemistry And Biotechnology School Of Engineering The University Of Tokyo

• Nagamune Teruyuki

  Dep. Of Chemistry And Biotechnology Graduate School Of Engineering The Univ. Of Tokyo 7-3-1 Hongo Bu

• Nagamune Teruyuki

  Department Of Bioengineering Graduate School Of Engineering The University Of Tokyo:department Of Ch

• Kazami Jun

  Chemicals Research Laboratories Toray Industries Inc.

• Kawano Genji

  Tsukuba Laboratories Aquas Corporation

• Nagamune Teruyuki

  Department Of Chemistry And Biotechnology Graduate School Of Engineering The University Of Tokyo:dep

• URA SEIJI

  Institute of Molecular and Cellular Biosciences, The University of Tokyo

• UEDA HIROSHI

  Graduate School of Frontier Science, The University of Tokyo

• NAGAMUNE TERUYUKI

  Graduate School of Engineering, The University of Tokyo

• Ura Seiji

  Institute Of Molecular And Cellular Biosciences The University Of Tokyo

• Ueda Hiroshi

  Graduate School Of Engineering Science Osaka University

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