| Project/Area Number |
10670293
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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, 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, SCHOOL OF MEDICINE, ASSISTANT PROFESSOR, 医学部, 助教授 (30168794)
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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 |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2000: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1999: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1998: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | measles virus / hemagglutinin / baculovirus / 立体構造解析 / 麻疹IgM抗体 / 赤血球凝集素 / バキュロウイルス / 組換え体バキュロウイルス / ファクターXa |
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| Research Abstract |
2 kinds of recombination baculoviruses that manifested the H protein were produced in order to carry out three-dimensional structure analysis of the important hemagglutinin protein (H protein) as a protective antigen of measles virus. Except for cytoplasm domain of the H gene, recombination baculovirus which connected the HA protein signal sequence of the influenza virus A/Japan strain (H3 subtype) was produced so that the H protein may secrete it to culture medium from the infected insect cells. The second viruses produced recombinant baculovirus which introduced cutting arrangement of factor-Xa into cell membrane attachment protein of the H protein. The protein was refined by affinity chromatography, ion exchangechromatography and gel filtration chromatography using specific antibody in order to large recover these 2 kinds of H proteins. With the goof the stage of the purification, the reactivity with the monoclonal antibody for measles virus lowered through the H protein. That H pro
… More
tein which reacts with the monoclonal antibody which shows only neutralizing antibody existed only for the fractionation in which electrophoresis and gel filtration chromatography recognize the large aggregation clarified. And, the activity which also hemagglutination of the green monkey red blood cells in either protein could not be recognized. The isolated purification of the protein from the recombinant baculovirus for structure analysis seems to be unsuitable from the above result. Therefore, it is concluded that the isolated purification of the protein from the virion is important in order to carry out three-dimensional structure analysis of the virus.
And, we had already sensitized the measles virus in the gelatin particle, and the new measles antibody measuring methodwas developed. This was applied this time, and the measles IgM antibody measuring method was established. It was adopted for the measles eradication plan of WHO, because this method is convenient, and it would be examined in developing countries in Africa and South America, etc. Less
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