| Project/Area Number |
07670358
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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 |
Virology
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| Research Institution | NATIONAL INSTITUTE OF HEALTH |
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Principal Investigator |
SATO Takeshi NATIONAL INSTITUTE OF HEALTH,DEPARTMENT OF VIRUS DISEASE AND VACCINE CONTROL,SENIOR RESEARCHER, ウイルス製剤部, 主任研究官 (00221284)
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| Co-Investigator(Kenkyū-buntansha) |
ENAMI Masayoshi KANAZAWA UNIVERSITY OF SCHOOL OF MEDICINE,ASSISTANT PROFESSOR, 医学部, 助教授 (30168794)
KOHAMA Tomoaki NATIONAL INSTITUTE OF HEALTH,DEPARTMENT OF VIRUS DISEASE AND VACCINE CONTROL,LAB, ウイルス製剤部, 室長 (70150183)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1996: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1995: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | measles virus / hemagglutinin / baculovirus |
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| Research Abstract |
Genetically-manipulated measles virus hemagglutinin (H) protein was expressed via Baculovirus expression system, in which the signal sequence and the following two residues of the influenza virus hemagglutinin (HA) protein (MAIIYLILLFTAVRG/DQ) was attached to the amino-terminus of the ecto-domain of the H protein. The recombinant H protein was highly expressed in the Sf21 cells and it was efficiently secreted into the medium. Approximately 70% of the expressed protein was recovered in the medium.
The recombinant H protein was purified through affinity-column using anti-measles antibody and the following gel-filtration column. Typically, 1 mg of the purified H protein was obtained starting from 1 liter of the culture. The majority of the recombinant H protein was present in a monomer. It possessed antigenic conformation against monoclonal antibodies and retained receptor binding activity but lacked HA activity. This protocol may be useful for large scale isolation of the viral glycoproteins for the component vaccines as well as for crystallography.
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