| Project/Area Number |
02556011
|
|---|
| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
応用生物化学・栄養化学
|
|---|
| Research Institution | Nagoya University |
|---|
Principal Investigator |
ASAHI Tadashi Nagoya University, School of Agriculture, Professor, 農学部, 教授 (10023392)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
NAKAGAWA Tsuyoshi Shimane University, Research Institute of Molecular Genetics, Instructor, 遺伝子実験施設, 講師 (30202211)
MORIKAMI Atsushi Nagoya University, School of Agriculture, Research Associate, 農学部, 助手 (10211608)
|
|---|
| Project Period (FY) |
1990 – 1991
|
| Project Status |
Completed (Fiscal Year 1991)
|
| Budget Amount *help |
¥12,600,000 (Direct Cost: ¥12,600,000)
Fiscal Year 1991: ¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 1990: ¥8,500,000 (Direct Cost: ¥8,500,000)
|
|---|
| Keywords | castor bean / catalase / catalase gene / cis-element / promoter / seed germination / transgenic plant / 植物遺伝子 |
|---|
| Research Abstract |
Two catalase genes, CAT1 and CAT3 genes, in the castor bean genome had been cloned. The promoter of CAT3 gene was found to have a much higher activity than cauliflower mosaic virus 35S RNA (CaMV35S) promoter, which suggests that the promoter of transgenic plants. In order to analyze the promoter of CAT3 gene, we constructed fused genes of 5' -upstream regions with different lengths (4300, 1241, 816, 535 and 380 base pairs, bp) of CAT3 gene and bacterial beta-glucuronidase (GUS) gene, and analyzed the expression of the fuse genes (CAT3-GUS fused genes) in transgenic tobacco cells. Analysis of the transient expression in cultured cells and the stable expression in callus cellus cells indicated the presence of many cis-elements enhancing the promoter activity in the 4300-bp 5'-upstream region. It might be due to the presence of such cis-elements in the region that the CAT3 promoter showed a much higher activity than the CaMV35S promoter. ' Next, we harvested seeds from transgenic tobacco plants carrying the CAT3-GUS fused genes and analyzed the expression of the genes during germination. GUS activity in dry seeds may reflect the expression of the fused genes during seed formation.
Analysis on this assumption suggested that there seemed to be a cis-element(s) concerning gene expression during seed formation in a region of -535 to -816 (the position numbers were counted from the initiation codon). Analysis on changes in GUS activity during germination suggested that a cis-element(s) concerning gene expression during germination might also exist in a region of -535 to -816 ; The cis-element(s) seemed to concern gene expression in boundary regions between the cotyledons and hypocotyl and between the hypocotyl and root and in the procambium of cotyledons. There seemed to be a cis-elements concerning gene expression in central parts of the hypocotyl at later stages of germination in a region of -816 to -1241.
|
