Project/Area Number |
05454132
|
Research Category |
Grant-in-Aid for General Scientific Research (B)
|
Allocation Type | Single-year Grants |
Research Field |
Applied molecular and cellular biology
|
Research Institution | Faculty of Biotechnology, Fukui Prefectural University |
Principal Investigator |
ASAHI Tadashi Fukui Prefectural University, Faculty of Biotechnology, Professor, 生物資源学部, 教授 (10023392)
|
Co-Investigator(Kenkyū-buntansha) |
IWASAKI Yukimoto Fukui Prefectural University, Faculty of Biotechnology, Associate Professor, 生物資源学部, 助教授 (20193732)
|
Project Period (FY) |
1993 – 1995
|
Project Status |
Completed (Fiscal Year 1995)
|
Budget Amount *help |
¥6,700,000 (Direct Cost: ¥6,700,000)
Fiscal Year 1995: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1994: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1993: ¥4,500,000 (Direct Cost: ¥4,500,000)
|
Keywords | catalase genes / cytocrome c oxidase / heterotrimeric G protein / F_1ATPase / microbody / mitochondria / seed germination / ヘテロ三重体Gタンパク質 / 遺伝子発現調整 / F_0F_1ATPase / カタラーゼ / シトクロムcオキシダーゼ / 植物遺伝子 |
Research Abstract |
The biogenesis of mitochondria is induced during germination of seeds, such as castor bean seeds, and in cut-wounded blant tissues. In order to elucidate the molecular mechanisms of the induction of biogenesis, we first isolated genes for mitochondrial proteins including some subunits of cytochome c oxidase and F_1 ATPase and analyzed the structure of the genes. Next, we examined how the genes were expressed in germinating seeds and wounded tissues but obtained no valuable data. Therefore, we turned our attention to microbodies, of which the biogenesisi was induced in response to seed germination and wounding of tissues just like mitochondria and investigated the bio-synthesis, induced in response to the stresses, of catalase, a marker enzyme of micro-bodies. We isolated two catalase genes from castor bean. The genes were located in a chromosome at a short distance and had similar structures but were expressed in different organ-specific manners. Both genes were, however, expressed in similar manners during seed germination ; namely, the expression was induced in response to germination. Then, we prepared fusion genes of 5' upstream regions of one of the catalase genes with E.coli beta-glucuronidase (GUS) gene, which were transferred to tobacco plant, and analyzed the regulatory functions of the regions through assay of GUS activity. The results showed the presence of a cis-element for the expression during seed germination in the regions, in addition to the elements for tissue-specific expression and the expression during seed formation. We are now conducting a series of experiments for identification of the cis-element. We also studied heterotrimeric G proteins in the plasma membrane in order to elucidate the signal transduction of tissue-wounding stress to nuclear gene, and obtained clones carrying cDNAs for the alpha and beta subunits from rice. We are now investigating the structure and function of the G protein.
|