| Project/Area Number |
10660044
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
植物保護
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| Research Institution | Utsunomiya University |
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Principal Investigator |
OKUDA Seiichi Faculty of Agriculture, Professor, 農学部, 教授 (90091941)
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| Project Period (FY) |
1998 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2000: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1999: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1998: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Plant Virus / Satellite RNA / Virus Replication / Template Switching / テンペレートスイッチング / キュウリモザイクウイルス / タバコモザイクウイルス / ウイルスRNA |
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| Research Abstract |
Previously we have showed that multimers of satellite RNA are present in cucumber mosaic virus (CMV) infecting tobacco leaves. First of all in this research we assumed that the hetero-joined RNA molecules were synthesized in virus-infected plants, for example CMV satellite RNA joined with CMV genomic RNA, oligonucleotide primers were designed on the basis of the nucleotide sequence of CMV genome- and satellite RNAs to amplify the junction region in head-to-tail hetero-joined molecules. The cDNA was synthesized from total RNA extracted from CMV-infecting tobacco leaves and amplified by PCR with the primers. Sequence analysis of RT-PCR products revealed that they were plus-sense or minus-sense head-to-tail joined RNAs between CMV genomic RNA1-3 but not plus-minus joined one. The junction sequences indicated that most molecules lacked sequence of 3' components and a few have insertion between 5' and 3' components. However, no head-to-tail joined RNA was detected from purified CMV preparation. We have also detected plus-sense or minus-sense head-to-tail joined RNAs from samples infected with cucumber green mottle mosaic virus (CGMMV). These data indicate that hetero-joined vital RNA molecules are generated by a replicase-driven copy-choice mechanism. In this research we have succeeded to develop the simple direct tube RT-PCR method that can detect virus RNA.Moreover, to confirm the head-to-tail joined RNAs from virus-infecting samples, the infectious full-length cDNA clone of CMV was constructed.
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