| Project/Area Number |
08404054
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (A)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
動物生理・代謝
|
|---|
| Research Institution | THE UNIVERSITY OF TOKYO |
|---|
Principal Investigator |
ISHIKAWA Hajime THE UNIVERSITY OF TOKYO,GRADUATE SCHOOL OF SCIENCE,PROFESSOR, 大学院・理学系研究科, 教授 (70012482)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
SATO Shigeharu THE UNIVERSITY OF TOKYO,GRADUATE SCHOOL OF SCIENCE,ASSISTANT, 大学院・理学系研究科, 助手 (80250215)
SASAKI Tetsuhiko THE UNIVERSITY OF TOKYO,GRADUATE SCHOOL OF SCIENCE,ASSISTANT, 大学院・理学系研究科, 助手 (60235257)
MORIOKA Mizue THE UNIVERSITY OF TOKYO,GRADUATE SCHOOL OF SCIENCE,ASSISTANT, 大学院・理学系研究科, 助手 (20272461)
|
|---|
| Project Period (FY) |
1996 – 1998
|
| Project Status |
Completed (Fiscal Year 1998)
|
| Budget Amount *help |
¥33,900,000 (Direct Cost: ¥33,900,000)
Fiscal Year 1998: ¥4,700,000 (Direct Cost: ¥4,700,000)
Fiscal Year 1997: ¥9,300,000 (Direct Cost: ¥9,300,000)
Fiscal Year 1996: ¥19,900,000 (Direct Cost: ¥19,900,000)
|
|---|
| Keywords | APHIDS / BACTERIOCYTE SYMBIOTIC SYSTEM / GENE EXPRESSION / RIBOFLAVIN / GENOME SIZE / GENOMIC COPY NUMBER / HISTONE H1 / URICASE / ヒストンHI / 菌細胞 / ディファレンシャルディスプレイ / シンビオニン / ボルバキア / 細胞質不和合 / rPOH / エネルギー共役的リン酸基転移 / ディファレンシャルディスプレー / 定量的RT-PCR |
|---|
| Research Abstract |
We studied on Buchnera, intracellular symbiotic bacteria of aphids and intracellular yeast-like symbionts of brown planthoppers, the results of which are summarized as follows :
1. Comparison of gene expression patterns of the bacteriocyte symbiotic systems from young and old aphids revealed that there are more than ten genes, of both host cell and Buchnera, that are specifically expressed only when the symbiotic system is kept functional. These genes include those for enzymes involved in the amino acid metabolism, confirming our previous results.
2. We attained convincing evidence that Buchnera produce riboflavin and provision the vitamin to hosts.
3. We estimated the genome size of Buchnera by digesting the genomic DNA with restriction enzymes whose restriction sites are rich in G/C, followed by pulse-field gel electrophoresis. It turned out that the genome size, 657kbp, is only one seventh of that of the genome of E.coli.
4. Dot-blot analyses of two different genes and direct fluorimetry of the cellular DNA content revealed that Buchnera are very extraordinary bacteria with more than 100 genomic copies in a single cell. In addition, the genomic copy number of Buchnera is variable to a great extent due to morph and developmental stage of the host insect.
5. Buchnera are exceptional as bacteria in that they contain a histone Hi -like protein, which is likely in interaction with the genomic DNA.
6. We successfully cloned the gene for uricase of the yeast-like symbiont that is essential for planthoppers to mobilize the uric acid stored in their tissue. Also, we constructed a large scale expression system of the uricase gene by transforming E.coil with the gene ligated into an expression vector.
|
