2017 Fiscal Year Research-status Report
Saving the frog: Understanding how Japanese frogs are resistant to a deadly worldwide fungal disease through in silico and in vitro assays of MHC and related immune genes
| Project/Area Number |
17K15053
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| Research Institution | The Graduate University for Advanced Studies |
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Principal Investigator |
QUINTIN LAU 総合研究大学院大学, 先導科学研究科, 特別研究員 (60794518)
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| Project Period (FY) |
2017-04-01 – 2019-03-31
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| Keywords | transcriptome / anuran / frog / chytridiomycosis |
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| Outline of Annual Research Achievements |
-Completed study of transcriptome and MHC class II characterization of three Ranidae species, now published in BMC Genomics.
-Conducted and completed study characterizing TLR2 and TLR4 genes in the three Ranidae species, manuscript is now in review.
-Studied expression levels of MHC class I and II in tadpoles during development.
-Collected RNA samples from nine other Japanese frog species. From RNA, generated cDNA libraries for Illumina sequencing, conducted assembly and blast search. Collated MHC sequences from the nine species.
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| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
During the first half of FY2017, I was continuing and completing related research in three Ranidae species. This research is directly related to the currently-funded research. For example, I completed a transcriptome study of the three Ranidae species, and characterized MHC class II genes (including analyses of sequences and linking it to resistance to chytridiomycosis). These approaches will be repeated and applied to nine other Japanese frog species in the currently-funded research project.
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| Strategy for Future Research Activity |
-Analyze MHC class I and II sequences from the newly acquired transcriptome data of nine Japanese frog species. This will include supertype analyses, to compare MHC sequences across disease resistant and susceptible species across the world. We will also begin analyses to test predicted binding of chytrid-related peptides by MHC receptors.
-With assistance of Australian collaborators listed in the application, we will collect RNA from two Australian species of frogs, one that is susceptible and one that is resistant. We will then construct cDNA libraries for Illumina sequencing, and compare sequences of MHC and other immune genes with Japanese frogs.
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| Causes of Carryover |
-Costs for field sampling as well as consumables for sample preparation and genotyping were lower than expected. This was partly because we purchased several consumables using remaining funds prior to this project.
-For the next fiscal year we plan to purchase new kits for transcriptome library, and conduct Illumina sequencing of additional samples from Australian frogs species. I will also attend an International conference to present our results. In addition, we plan to purchase more consumables, and have local research meetings, and purchase a computer to assist in computational analyses.
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