Intracellular space-time distribution of fluorescent protein of luminous bacterium and the molecular link with cell cycle
| Project/Area Number |
17310127
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Living organism molecular science
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| Research Institution | Kyoto Institute of Technology |
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Principal Investigator |
KARATANI Hajime Kyoto Institute of Technology, Graduate School of Science and Technology, Professor (10169659)
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| Co-Investigator(Kenkyū-buntansha) |
WADA Minoru The University of Tokyo, Ocean Resemrh Institute, Assistant Professor (70292860)
SASAKI Ken Kyoto Institute of Technology, Graduate School of Science and Technology, Associate Professor (20205842)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥8,580,000 (Direct Cost: ¥8,100,000、Indirect Cost: ¥480,000)
Fiscal Year 2007: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2006: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2005: ¥5,000,000 (Direct Cost: ¥5,000,000)
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| Keywords | Bioluminescence / Bacterial luciferase / Fluorescent protein / Fusion protein / Fluorescence imaging / Bioluminescence imaging / Gene expression / cell division cycle / 生細胞イメージング / 遺伝子クローニング / 呼吸 |
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| Research Abstract |
In this study, we have investigated into the intracellular space-time distribution of fluorescent protein of luminous bacterium and the molecular link with cell cycle mainly using Vibrio fischeri Y1 cells. The fluorescence images of the cells have proven that the fluorescent proteins responsible for bioluminescence (BL) color change is scattered in the vicinity of the cell membrane together with luciferase.
To clarify the reversible BL color change, the relationship between the BL color and the redox state of yellow fluorescent protein (YFP) has been studied in vitro. The results indicated that the YFP fluorescence ability depends on the redox states of YFP, and that the reduced YFP no longer has ability to cause yellow BL. It was also clarified that once the reduced YFP is reoxidized, the BL color changes from blue to yellow reversibly.
To characterize the space-time distribution of fluorescent proteins, the genes encoding YFP and another blue fluorescent protein (BFP) were cloned and their Escherichia coli expression systems were established. The fluorescence properties of the recombinant proteins were in good agreement with those of the authentic ones.
Subsequently, several fusion fluorescent proteins consisting of YFP and BFP have been designed and constructed. The fusion protein, of which N-terminus is YFP, was found to exhibit the bimodal fluorescence characteristic of both YFP and BFP. The appearance of bimodal fluorescences depended on the cellular conditions. Both fluorescences observed as small spots at the same site in growing cells, whereas the cells in the stationary phase appeared to emit mainly blue fluorescence. From the results, the fused fluorescent proteins were evaluated to be useful for collecting the intracellular information including the cell cycle. Furthermore, BL imaging of the luminous colony suggested that the bacterial colonies grow exhibiting the non-linear pattern.
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Report
(4 results)
Research Products
(63 results)
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[Journal Article] Bioluminescence color modulation of Vibrio fischeri strain Y1 coupled with alterable levels of endogenous yellow fluorescent protein and its fluorescence imaging.2006
Author(s)
Karatani, H.*, Matsumoto, S., Miyata, K., Yoshizawa, S., Suhama, Y., Hirayama, S.
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Journal Title
Photochemistry and Photobiology 82
Pages: 587-592
Description
「研究成果報告書概要(和文)」より
Related Report
Peer Reviewed
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[Journal Article] LuxA gene of light organ symbionts of the bioluminescent fish Acropoma japonicum(Acropomatidae)and Siphamia versicolor(Apogonnidae)forms a lineage closely related to that of Photobacterium leiognathi ssp. Mandapamensis.2006
Author(s)
Wada, M.*, Kamiya, A., Uchiyama, N., Yoshizawa, S., Kita-Tsukamoto, K., Ikejima, K., Yu, R., Imada, C., Karatani, H., Mizuno, N., Suzuki, Y., Nishida, M., Kogure, K.
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Journal Title
FEMS Microbiology Letters 260
Pages: 186-192
Description
「研究成果報告書概要(和文)」より
Related Report
Peer Reviewed
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[Journal Article] Bioluminescence color modulation of Vibrio fischeri strain YI coupled with alterable levels of endogenous yellow fluorescent protein and its fluorescence imaging2006
Author(s)
Karatani, H., Matsumoto, S., Miyata, K., Yoshizawa, S., Suhama, Y., Hirayama, S
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Journal Title
Photochemistry and Photobiology 82
Pages: 587-592
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Journal Article] LuxA gene of light organ symbionts of the bioluminescent fish Acropomajaponicum(Acropomatidae) and Siphamia versicolor(Apogonnidae) forms a lineage closely related to that of Photobacterium leiognathi ssp. Mandapamensis.2006
Author(s)
Wada, M., Kamiya, A., Uchiyama, N., Yoshizawa, S., Kita-Tsukamoto, K., Ikejima, K., Yu, R., Imada, C., Karatani. H., Mizuno, N., Suzuki, Y., Nishida, M., Kogure, K
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Journal Title
FEMS Microbiology Letters 260
Pages: 186-192
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Presentation] Studies on the alteration in color of bioluminescence arising from Vibrio fischeri Y1.2007
Author(s)
Karatani, H., Osaki, T., Yasui, M., Matsumoto, S., Ohta, S.
Organizer
12th Congress European Society for Photobiology,
Place of Presentation
Bath, England
Description
「研究成果報告書概要(和文)」より
Related Report
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[Presentation] Bioluminescence color modulation of luminous bacteria and its application to bioimaging.2006
Author(s)
Karatani, H., Yasui, M., Heguri, A., Matsumoto, S., Ohta, S., Osaki, T.
Organizer
KIT-NIST
Place of Presentation
Kyoto
Year and Date
2006-12-05
Description
「研究成果報告書概要(和文)」より
Related Report
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[Presentation] Bioluminescence color modulation of luminous bacteria and its application to bioimaging2006
Author(s)
Karatani, H., Yasui, M., Heguri, A., Matsumoto, S., Ohta, S., Osaki, T
Organizer
KIT-NIST
Place of Presentation
Kyoto
Year and Date
2006-12-05
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Presentation] Bioluminescence image analysis of luminous colony growing on agar plate.2005
Author(s)
Karatani, H., Heguri, A., Fumiyama, M., Hirayama, S., Matsumura-Inoue, T
Organizer
PACIFCHEM
Place of Presentation
Honolulu, USA
Description
「研究成果報告書概要(和文)」より
Related Report
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[Presentation] Expression of a gene encoding yellow fluorescent protein of marine luminous bacterium in Escherichia coli with a view to practical application.2005
Author(s)
Karatani, H., Yasui, M., Osaki, T., Izuta, T., Hirayama, S.
Organizer
PACIFCHEM
Place of Presentation
Honolulu, USA
Description
「研究成果報告書概要(和文)」より
Related Report
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[Presentation] Bioluminescence image analysis of luminous colony growing on agar plate2005
Author(s)
Karatani, H., Heguri, A., Fumiyama, M., Hirayama, S., Matsumura-Inoue, T
Organizer
PACIFCHEM
Place of Presentation
Honolulu, USA
Description
「研究成果報告書概要(欧文)」より
Related Report
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