| Project/Area Number |
17K15410
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Applied molecular and cellular biology
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| Research Institution | University of Tsukuba |
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Principal Investigator |
Utada Andrew 筑波大学, 生命環境系, 准教授 (90776626)
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| Project Period (FY) |
2017-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2018: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2017: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
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| Keywords | Pseudomonas aeruginosa / mucoid phenotype / spontaneous mutation / phenotypic switching / reversion phenotype / localization / Single-cell tracking / Applied microbiology / Microfluidics / mucoid / 応用微生物 / マイクロ・ナノデバイス |
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| Outline of Final Research Achievements |
Pseudomonas aeruginosa (PA), an opportunistic human pathogen, is often used as a model organism to understand biofilms. The PA mucoid mutant produces copious amounts of the polysaccharide alginate and is a leading cause of mortality in cystic fibrosis patients.
We found that the mucoid strain rapidly acquires spontaneous mutations, resulting in a loss of the alginate overexpression phenotype; this new strain is called the “revertant”. To distinguish and track these different mutants, we generated a reporter strain in the mucoid mutant background to report the phenotype through expression of fluorescent proteins. We are characterizing the transition from mucoid (green) to revertant (red) using flow cytometry, wide-field and confocal microscopy. Moreover, we are beginning to quantify the frequency and location of appearance of the reversion mutant within mucoid colonies. We are still working to clarify the major factors that induce spontaneous mutations in the mucoid mutant.
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| Academic Significance and Societal Importance of the Research Achievements |
We have generated a reporter strain through this work that has the potential to enable the clarification of the biofilm forming properties of the mucoid mutant strain. This research has potential to clarify the ecological significance of polysaccharide overproduction.
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