2018 Fiscal Year Annual Research Report
A multidisciplinary research program to enable the discovery of inhibitors of colanic acid excretion in bacteria
| Project/Area Number |
18H02109
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| Research Institution | Kagawa University |
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Principal Investigator |
Kong Lingbing 香川大学, 国際希少糖研究教育機構, 助教 (20798384)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Keywords | Colanic acid |
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| Outline of Annual Research Achievements |
The three pathogenic bacterial strains have been purchased, delivered, successfully cultured and stored at Kagawa University, according to the relevant regulations. Genomic DNA has been successfully extracted from the strains. WzaCA is transporter of colonic acid. Polymerase chain reaction has allowed the WzaCA gene copied from the genomic DNA and subsequent cloning onto desired vectors. Expression of the WzaCA is currently in progress. On the other hand, various genomic mutants are being created with a scarless genome editing technique. Single-molecule setup for the Wza pores has been established. An invited book chapter related to this project is in press:
Lingbing Kong, Revelation of function and inhibition of Wza through single channel studies, Methods Mol. Biol., 2019, in press.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
There was delay in the delivery of some equipment, strains and reagents, which led to the delay of some experiments. However, most of the scheduled goals have been achieved. The implementation of the following scheduled research plans will therefore be fine.
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| Strategy for Future Research Activity |
Single-molecule screening of blockers will be performed with wild-type and mutant WzaCA protein pores. Once a promising blocker is found, chemical synthesis of stronger blockers will be carried out. Effects of the blockers on colanic acid production in live pathogenic bacteria will be evaluated. Imaging of bacterial cells in the presence and absence of blockers will be performed by using a fluorescence microscope to be purchased with the grant. Wild-type and mutant bacterial strains will be used to validate the target specificity of the blockers.
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