| Project/Area Number |
16406009
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 海外学術 |
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| Research Field |
Parasitology (including Sanitary zoology)
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| Research Institution | Ehime University |
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Principal Investigator |
TSUBOI Takafumi Ehime University, Cell-free Science and Technology Research Center, Professor, 無細胞生命科学工学研究センター, 教授 (00188616)
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| Co-Investigator(Kenkyū-buntansha) |
TAKEO Satoru Ehime University, Cell-free Science and Technology Tesearch Center, Lecturer, 無細胞生命科学工学研究センター, 講師 (40302666)
IRIKO Hideyuki Tottori University, Faculty of Medicine, Instructor, 医学部, 助手 (60346674)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥13,200,000 (Direct Cost: ¥13,200,000)
Fiscal Year 2006: ¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2005: ¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2004: ¥5,400,000 (Direct Cost: ¥5,400,000)
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| Keywords | malaria / vaccine / genome / biotechnology / protein / serum / 感染症 / 国際研究者交流 / タイ / 無細胞タンパク質合成 |
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| Research Abstract |
Malaria remains one of the leading causes of both morbidity and mortality in humans residing in the tropical countries. The evidence of increasing resistance of the Plasmodium falciparum parasite to chemotherapeutic agents highlights the critical need for an effective vaccine. After the establishment of malaria genome database, we have now free access to the genome data to search for novel vaccine candidates. However, one of the bottlenecks for the malaria vaccine research is the difficulty of the recombinant protein expression using conventional methods. We applied a high-throughput protein production method based on the wheat germ cell-free system to the malaria vaccine research. To identify novel malaria vaccine candidates, we selected and cloned more than 180 genes which are expected to be expressed in merozoite stage, then prepared transcription templates through PCR-based high throughput procedures, followed by protein synthesis by wheat germ cell-free system. Using this approach, we succeeded in obtaining 149 recombinant merozoite proteins. At the same time, we successfully obtained approximately 30 human immune sera from asymptomatic villagers living along the Thai-Myanmar border, and approximately 30 symptomatic malaria patient sera from hospital in Bangkok, Thailand. All human serum samples used in this study were reviewed and approved by the Institutional Ethics Committee of the Thai Ministry of Public Health and of the Ehime University, Japan. Finally, we tried to screen these recombinant proteins for identifying novel malaria vaccine candidates with five cases of the malaria immune sera by ELISA. As a result of this preliminary screening, we identified ten merozoite proteins as antigens. Accordingly, this strategy using wheat germ cell-free system will be a major breakthrough in the novel malaria vaccine candidate discovery research.
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