| 研究課題/領域番号 |
17K15053
|
|---|
| 研究機関 | 総合研究大学院大学 |
|---|
研究代表者 |
QUINTIN LAU 総合研究大学院大学, 先導科学研究科, 特別研究員 (60794518)
|
|---|
| 研究期間 (年度) |
2017-04-01 – 2020-03-31
|
| キーワード | Rana / MHC / chytrid / frog |
|---|
| 研究実績の概要 |
Study A- Expression of MHC class I and II in tadpoles during development: Completed qPCR experiments to measure MHC expression in Rana ornativentris tadpoles, and also conducted normal PCR to detect presence of MHC amplification from Xenopus tadpoles. Prepared and submitted manuscript.
Study B-Transcriptome of numerous Japanese frog species to study immune genes: isolated MHC class I and II sequences and validated using cloning/sequencing. Completed phylogenetic and supertyping analyses of the MHC sequences. Found that all Japanese frogs share some common physiochemical properties in MHC II based on supertyping anaylsis. Conducted preliminary computer-based prediction of binding affinity between frog MHC sequence and chytrid fungus sequence. Other immune genes were also isolated from transcriptome data including innate immune genes (Toll-like receptors and antimicrobial peptides).
|
| 現在までの達成度 (区分) |
現在までの達成度 (区分)
3: やや遅れている
理由
Study A: During the end of FY2018, we received heavy reviewer criticism related to our study of MHC expression in tadpoles, which drove us to the decision to continue this project with more robust and in-depth approach (i.e. transcriptome sequencing).
Study B: The isolation of MHC sequences from transcriptome data was complex. Since MHC is the most polymorphic gene in the genome, it was challenging to validate allele sequences based on transcriptome data alone (because each individual may have multiple MHC loci due to gene duplication). Thus, I had to conduct cloning and sequencing to confirm presence of true MHC alleles. This additional experimental work caused delay of subsequent analyses.
|
| 今後の研究の推進方策 |
Study A- conduct cDNA library sequencing of Rana and Xenopus tadpole samples, isolate differentially expressed genes (DEGs) as well as expression levels of MHC class I and II. Update the former manuscript for submission
Study B- complete MHC supertyping analyses. Conduct further analyses of predicting MHC binding, utilizing more diverse sequences from the chytrid fungus after consulting with Australian collaborators.
|
| 次年度使用額が生じた理由 |
Due to time constraints and delays in experimental work, the project has taken longer than expected to finish
|
