| Project/Area Number |
10556078
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 展開研究 |
|---|
| Research Field |
Applied molecular and cellular biology
|
|---|
| Research Institution | The University of Tokyo |
|---|
Principal Investigator |
ABE Keiko Graduate School of Agricultural and life Sciences, The University of Tokyo, Professor, 大学院・農学生命科学研究科, 教授 (10151094)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MATSUMOTO Ichiro Graduate School of Agricultural and Life Sciences, The University of Tokyo, Assistant Professor, 大学院・農学生命科学研究科, 助手 (00291328)
KURODA Masaharu Hokuriku National Agricultural Experiment Station, Researcher, 試験場, 研究員
EMORI Yasufumi Graduate School of Science, The University of Tokyo, Associate Professor, 大学院・理学系研究科, 助教授 (60160389)
|
|---|
| Project Period (FY) |
1998 – 1999
|
| Project Status |
Completed (Fiscal Year 1999)
|
| Budget Amount *help |
¥7,800,000 (Direct Cost: ¥7,800,000)
Fiscal Year 1999: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 1998: ¥4,200,000 (Direct Cost: ¥4,200,000)
|
|---|
| Keywords | Coleoptera / insect gut proteinase / cysteine proteinases / Sitophilus cysteine proteinase / Oryzacystatin / 立体構造 |
|---|
| Research Abstract |
Detailed analysis were conducted of cysteine proteinases of Sitophilus zeamais (Coleoptera), a rice-seed insect pest, to elucidate their properties and physiological functions at the molecular level. It was found that the cysteine proteinases comprise multiple molecular species originating from a gene family and also that one of the species is a central enzyme. involved in damaging the seeds. The result made us try to develop a transgenic rice cultivar with oryzacystatin or corn cystatin expressed at a higher level. For the trial, we first fused a cystatin cDNA and a hyglomycin-resistant gene to Ca MV35S promoter and then introduced the fusion gene to rice protoplasts by electroporation or using an agrobacterium-mediated system. The consequently regenerated transformant was found to express 10-15 times more cystatin than the non-transformant (normal rice) used as control. The cystatin expressed in the transformant seeds, when purified, showed a strong inhibitory activity (ICィイD250ィエD2 : 〜10ィイD1-8ィエD1M) toward cathepsin H as well as toward the Sitphilus cysteine proteinase. We then produced transgenic rice of this type at a field scale and harvested the seeds. An investigation to find out the antipest and even lethal effects of the seeds themselves to successfully progressing.
|
