| Budget Amount *help |
¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2012: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2011: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Research Abstract |
We established a new reverse genetics system driven by a mammalian promoter, that functions without helper virus. The complete genome of the HuNoV GII.3 U201, Saga1, TCH, NV68 and also MNV-S7 strain was cloned downstream of an EF-1 alpha mammalian promoter. These constructs produced progeny viruses produced from cells that contained the complete NoV genomic RNA, VP1, VP2 and VPg were detected in isopycnic gradients of CsCl at the same density as native infectious NoV particles from a patient's stool. A GFP reporter construct containing the GFP gene in ORF1 produced complete virions that contain VPg linked RNA. RNA from virions containing the encapsidated GFP genomic RNA was successfully transfected back into cells producing fluorescent puncta indicating that the encapsidated RNA is replication competent. Our results are useful to develop of an antiviral medicine which targeted these in future.
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