| Project/Area Number |
12306003
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
蚕糸・昆虫利用学
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| Research Institution | RIKEN |
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Principal Investigator |
MATSUMOTO Shogo RIKEN, Molecular Entomology Laboratory, Chief Scientist, 分子昆虫学研究室, 主任研究員 (60134516)
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| Co-Investigator(Kenkyū-buntansha) |
NAGAMINE Toshihiro RIKEN, Molecular Entomology Laboratory, Senior Research Scientist, 分子昆虫学研究室, 先任研究員 (90237553)
SUZUKI Masataka RIKEN, Molecular Entomology Laboratory, Research Scientist, 分子昆虫学研究室, 研究員 (30360572)
KAN Wonkyung RIKEN, Molecular Entomology Laboratory, Senior Research Scientist, 分子昆虫学研究室, 先任研究員 (30291917)
KURIHARA Masaaki RIKEN, Molecular Entomology Laboratory, Senior Technical Scientist, 分子昆虫学研究室, 先任技師(研究職)
MOTO Ken'ichi RIKEN, Molecular Entomology Laboratory, Special Postdoctral Researcher, 分子昆虫学研究室, 基礎科学特別研究員 (90333335)
三田 和英 農業生物資源研究所, 昆虫ゲノム研究チーム, チーム長 (30159165)
岡野 和広 理化学研究所, 分子昆虫学研究室, 基礎科学特別研究員 (70322691)
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| Project Period (FY) |
2000 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥47,600,000 (Direct Cost: ¥40,100,000、Indirect Cost: ¥7,500,000)
Fiscal Year 2003: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000)
Fiscal Year 2002: ¥10,140,000 (Direct Cost: ¥7,800,000、Indirect Cost: ¥2,340,000)
Fiscal Year 2001: ¥15,470,000 (Direct Cost: ¥11,900,000、Indirect Cost: ¥3,570,000)
Fiscal Year 2000: ¥15,100,000 (Direct Cost: ¥15,100,000)
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| Keywords | Silkworm / EST database / Normalized cDNA library / Neurohormone / Gene targeting / Pheromone / バキュロウイルス / 遺伝子導入 / GFP |
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| Research Abstract |
One of our research topics highlighted here is on the molecular mechanisms underlying sex pheromone production in B.mori. Production of the sex pheromone bombykol by the pheromone gland (PG) is under the control of a neurohormone termed PBAN. Following identification and morphological characterization of the bombykol-producing cells, we have examined the cellular dynamics during pheromonogenesis and discovered that cytoplasmic lipid droplets accumulate and morphometrically fluctuate in accordance with bombykol production (Fonagy et al.2000, 2001). Chemical characterization of the lipid droplet contents using mass spectrometry and analysis using experimental morphological techniques have confirmed that the role of the lipid droplets is to store the bombykol precursor fatty acid in the form of triacylglycerols (TGs) and then to release it for pheromone production in response to PBAN (Matsumoto et al., 2002). Further biochemical characterization has led us to propose a mechanism in which
… More
PBAN stimulates both lipolysis of TGs in the lipid droplets and fatty-acyl reduction, the latter of which consists of two consecutive enzyme reactions catalyzed by acyl-CoA synthetase and fatty-acyl reductase (FAR).
In order to understand the molecular basis of the B.mori PG and to further elucidate the molecular events occurring in the PG cells, we have constructed a PG expressed-sequence tag (EST) database from a normalized PG cDNA library. From 2,000 catalogued EST clones, we have identified and characterized several PG-specific genes encoding functional proteins including acyl-CoA desaturases, two acyl-CoA-binding proteins (pgACBP, mgACBP), and the heterosubunits of calcineurin (Yoshiga et al., 2000 ; Matsumoto et al., 2001 ; Yoshiga et al., 2002). Recently, we have successfully characterized a gene encoding a PG-specific FAR that functions to convert the bombykol precursor fatty acid to its corresponding alcohol (Moto et al., 2003).
Gene transformation has emerged as a powerful methodological tool for the elucidation of gene function and regulation. Despite the successes of dipteran insect systems, an efficient and practical germ-line transformation protocol for lepidopteran insects has not yet been achieved in spite of considerable efforts. After examining a number of baculovirus based approaches for in vivo gene analysis, we were able to achieve cell-specific expression of the enhanced green fluorescence protein (EGFP) gene in brain cells under the control of specific gene promoters of the neuropeptides, bombyxin B3 and B.mori prothoracicotropic hormone (Moto et al., 2003). Our results demonstrate that in vivo gene-expression systems mediated by baculoviruses can be utilized to elucidate molecular mechanisms underlying various cellular events even those occurring in the neural tissues/organs of insects. Less
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