1994 Fiscal Year Final Research Report Summary
Persistence of Viral genes in a variant MDCK cells after productive Replication of a mutant of influenza virus A/WSN
| Project/Area Number |
05670283
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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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, Dept.of Virology |
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Principal Investigator |
TANAKA Toshinori Jichi Med.Sch.Dept.of Virology Ass.Prof., 医学部, 講師 (30146154)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Toshinori Jichi Med.Sch.Dept.of Virology Ass.Prof. (30146154)
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| Project Period (FY) |
1993 – 1994
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| Keywords | Persist Infection / Influenza Virus / MDCK cells / NS gene / M gene / competive PCR / RT-PCR法 |
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| Research Abstract |
Maedin-Darby canine kidney (MDCK) cells, which received high multiplicity superinfections with a weakly cytolytic mutant of influenza A virus, could survive and be passaged stably, carrying the genes of the infected virus. PB1, PB2, PA,NP,HA,NA,M and NS genes was detected in the 25th passaged cells using polymerase chain reaction (PCR).
In the 8th passaged cells, cRNA and mRNA of M gene was detected. The quantitation of the viral NS gene persisting in these cultured cells using competive PCR revealed that the gene existed at a relatively constant level (approximately 10^5 to 10^6 copies per 8 x 10^6 cells) over a long range of cell generations without producing any detectable projeny virus, suggesting that the persisting NS gene was not silent, but was amplified and inherited to the daughter cells.
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