| Project/Area Number |
04454203
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Virology
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| Research Institution | Jichi Medical School |
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Principal Investigator |
TOBITA Kiyotake Professor, Department of Virology Jichi Medical School, 医学部, 教授 (00077174)
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| Co-Investigator(Kenkyū-buntansha) |
ODAGIRI Takato Lecturer, Department of Virology, Jichi Medical School, 医学部, 講師 (80177237)
TANAKA Toshinori Lecturer, Department of Virology, Jichi Medical School, 医学部, 講師 (30146154)
TASHIRO Masato Assistant Professor, Department of Virology, Jichi Medical School, 医学部, 助教授 (90111343)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥4,800,000 (Direct Cost: ¥4,800,000)
Fiscal Year 1993: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1992: ¥3,600,000 (Direct Cost: ¥3,600,000)
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| Keywords | influevza virus / latent infection / polymerase chain reaction / copetitive PCR / viral genes / pUC19ベクター / 遺伝子の塩基配列 |
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| Research Abstract |
By repeated superinfections of MDBK cells with AWBY-140, a weakly cytolytic mutant of influenza virus A/WAN ( HINI ), at a high multiplicity and subsequent cultivations of the surviving cells, we established a latent infection of influenza virus in cell culture ( MDBK-RR ). Within MDBK-RR cells, we detected all the 8 genes of influenza virus by a polymerase chain reaction ( PCR ) using influenza viral gene-specific primer pairs.
At least some of them retained their full size, accompanied by a few point mutations.
In addition to viral RNA ( vRNA ), comlementary RNA ( cRNA ) and viral messenger RNA ( mRNA )were also detected within the cells.
Competitive PCR revesled that the copy number of the persisting viral genes was 10^5 to 10^6 per 8 x 10^6 cells, which did not alter significantry as the passage number of the culture increased.
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