1996 Fiscal Year Final Research Report Summary
Development of a influenza virus vector for practical use.
| Project/Area Number |
07557215
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
Virology
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| Research Institution | Kanazawa University |
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Principal Investigator |
ENAMI Masayoshi Kanazawa Univ.Sch.of Med., Associate Prof., 医学部, 助教授 (30168794)
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| Co-Investigator(Kenkyū-buntansha) |
NAKADA Susumu Yamanouchi Pharmaceutical Co., Ltd.Group Leader, 筑波研究センター, 主管研究員 (90129255)
A.SATO Takeshi National Institute of Health, Senior Scientist, ウイルス製剤部, 主任研究官 (00221284)
ODAGIRI Takato Jichi Med.Sch., Assistant Prof., 医学部, 講師 (80177237)
TOYODA Tetsuya Kurume Univ.Sch.of Med., Prof., 医学部, 教授 (00197972)
NAGATA Kyosuke Tokyo Institute of Technology, Associate Prof., 生命理工学部, 助教授 (40180492)
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| Project Period (FY) |
1995 – 1996
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| Keywords | Influenza Virus / Vector / Reverse genetics / Vaccine |
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| Research Abstract |
1) We have isolated and analyzed viruses containing foreign genes (100-1800bp) in the NA genome. Short inserts were stable but the expression was low. Insertion of the measles virus H gene lead to 70% deletion of the insert. Therefore, previously described protocol was shown to be insufficient for practical use.
2) Then we have developed a novel protocol to isolate influenza transfectant containing mutations in the NS genome. The RNP,which was isolated from influenza virion, was treated with RNase H in the presence of NS cDNA.This was co-transfected to the cells together with in vitro-reconstituted NS RNPs. We successfully isolated several mutants in which deletion of the C-terminal half of the NS1 protein attenuated the virus. The deletion of 12 residues in the N-terminal half read to the temperature-sensitive phenotype. This would be good protocol to generate live attenuate vaccine.
3) It was shown that noncoding region of the genome may contain segment-specific regulatory signals.
4) A host factor was isolated which stimulates influenza virus RNA transcription and replication. This factor may be useful for efficient expression of proteins from the virus genome in the future.
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