| Project/Area Number |
61870025
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Virology
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| Research Institution | Nagasaki University |
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Principal Investigator |
IGARASHI Akira Nagasaki University, Institute of Tropical Medicine, 熱帯医学研究所, 教授 (40029773)
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| Co-Investigator(Kenkyū-buntansha) |
FUJITA Hiroyuki Research Foundation for Microbial Diseases of Osaka University, Kanonji Institut, 観音寺研究所, 生物工学センター主任 (30156880)
TAKAGI Mitsuo Research Foundation for Microbial Diseases of Osaka University, Kanonji Institut, 観音寺研究所, 技術部次長 (60113453)
YOSHIDA Iwao Research Foundation for Microbial Diseases of Osaka University, Kanonji Institut, 観音寺研究所, 品質管理部長 (60072782)
MORITA Kouichi Nagasaki University, Institute of Tropical Medicine, 熱帯医学研究所, 講師 (40182240)
SAKAKI Yoshiyuki Kyushu University, Institute of Genetic Information, 遺伝情報実験施設, 教授 (10112327)
住吉 日出夫 長崎大学, 熱帯医学研究所, 助手 (80179308)
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| Project Period (FY) |
1986 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥10,000,000 (Direct Cost: ¥10,000,000)
Fiscal Year 1988: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1987: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1986: ¥6,000,000 (Direct Cost: ¥6,000,000)
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| Keywords | Japanese encephalitis virus / recombinant DNA / 第二世代ワクチン / Eタンパク / 組換DNA / ワクチン / 遺伝子RNA / 塩基配列解析 |
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| Research Abstract |
Studies were made to develop the second generation Japanese encephalitis (JE) vaccine by recombinant DNA technology. In order to obtain basic information for this objective, complete nucleotide sequence of JE virus was analyzed. The virus genome consisted of 10,976 nucleotides, of which 95 bases existed in 5' noncoding region, 585 bases at 3' noncoding region, and 10,296 bases formed a long open reading frame (ORT) between both noncoding region. The N-terminal amino acid sequences of the structural proteins of JE virus showed that the structural protein gene existed in the 5' 1/4 of the ORF in the order of C, PreM (precursor of M), and E. Sicne E protein is known to produce neutralizing antibodies which play most important roles in the protection of je, e protein gene was inserted into various recombinant plasmids or viruses in order to express the E protein in the cells transformed or infected with these plasmids or viruses. The expression of E protein was detected in recombinant yeast, mammalian cells, vaccinia virus and bombix mori nuclear polyhedrosis virus, respectively, however, the immunogenicity of these expressed.E protein was much lower than that of the current vaccine in terms of eliciting neutralizing antibodies. The reason of the low immunogenicity could be (1) amount of the expressed E protein, (2) stability of the E protein in the expressed cells, (3) dependence of immunogenicity on the highly conformational structure of the E protein, (4) purity of the e protein in the expressed cells used for immunization. These problems should be overcome by further studies to improve the levels of expression, purification of the expressed E protein, and trying to express the neutralizing epitope which is resistant to protein denaturation and exists near C-terminal of the E protein.
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