| Project/Area Number |
16K19144
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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 |
Virology
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| Research Institution | Jichi Medical University |
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Principal Investigator |
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| Project Period (FY) |
2016-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2016: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
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| Keywords | E型肝炎ウイルス / レプリコン / リバースジェネティクス法 / 細胞培養 / ウイルス複製 / PSAPモチーフ / HEV / RNA複製 / PSAP / リアルタイムRT-PCR法 |
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| Outline of Final Research Achievements |
In this study, in order to elucidate the replication mechanism of hepatitis E virus (HEV), we constructed an HEV replicon that can multiplicate full-length genomic RNA but cannot produce virions in transfected cells. We could obtain evidence that a 2.2-kb subgenomic RNA was transcribed, from which ORF3 proteins were translated. Obtained results suggested that we could establish a replicon-based method to analyze the replication mechanism of the HEV genomic RNA in cultured cells. In addition, since rat HEV ORF3 protein lacks an amino acid sequence motif of proline, serine, alanine and proline (PSAP) that has been demonstrated to be essential for release of human HEV particles from infected cells, various infectious rat HEV cDNA clones with mutations in proline residues were constructed. Obtained results indicated that two proline residues of the PxYP sequence play a pivotal role for egress of rat HEV particles.
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