1989 Fiscal Year Final Research Report Summary
Research on the expression of dengue virus gene.
| Project/Area Number |
63570213
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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 | University of the Ryukyus |
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Principal Investigator |
MAKINO Yoshihiro Department of Virology, University of the Ryukyus Associate Professor, 医学部, 助教授 (60039930)
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| Co-Investigator(Kenkyū-buntansha) |
TADANO Masayuki Department of Virology, University of the Ryukyus Research Associate, 医学部, 助手 (80179712)
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| Project Period (FY) |
1988 – 1989
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| Keywords | Dengue 4 virus / Gene expression / Baculovirus / dengue core gene / dengue envelope gene |
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| Research Abstract |
1. The dengue 4 virus (DEN-4) core gene was inserted into the baculovirus polyhedrin gene region and obtained a recombinant baculovirus (373-core), which directed the synthesis of a DEN-4 core fusion protein. The Sf-9 cells infected with the recombinant virus and incubated for 24-48hr were fixed and stained by peroxidase anti-peroxidase(PAP)staining method using DEN patient sera. The results showed that the patient sera did not react with the recombinant DEN-4 core protein, It can be explained as follows: (1) Only a portion of the epitopes on the authentic DEN-4 core protein may be present in a recombinant core protein. Thus only a part of the core antibodies in the patient sera reacted with the recombinant core protein. (2) The DEN core protein does not migrate to the surface of the infected cells. Therefore, the core protein is not recognized by immune mechanism, thus little DEN core antibody are produced.
2. The recombinant baculovirus which contained DEN-4 envelope(E) gene was constructed. This recombinant baculovirus (401E) directed the synthesis of DEN-4 envelope fusion protein with a molecular weight of about 43K dalton. The recombinant virus infected cells reacted with DEN patient sera as well as DEN specific polyclonal and monoclonal antibody as examined by PAP staining method. The staining titers of the patient sera were well correlated with ELISA titers. Thus the usefulness of the recombinant E protein as a diagnostic antigen has been suggested.
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