| Project/Area Number |
07558226
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 試験 |
|---|
| Research Field |
Biophysics
|
|---|
| Research Institution | The University of Tokyo |
|---|
Principal Investigator |
SUTOH Kazuo The University of Tokyo, Graduate School of Arts and Sciences, Professor, 大学院・総合文化研究科, 教授 (20111453)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
ADACHI Hiroyuki The University of Tokyo, Graduate School of Arts and Sciences, Research Associat, 大学院・総合文化研究科, 助手 (00211699)
|
|---|
| Project Period (FY) |
1995 – 1996
|
| Project Status |
Completed (Fiscal Year 1996)
|
| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1996: ¥2,300,000 (Direct Cost: ¥2,300,000)
|
|---|
| Keywords | genetic engineering / protein expression / selection marker / oligomeric proteins / Dictyostelium discoideum / 細胞性粘菌 / 蛋白質結晶 / 大量発現系 / ミオシン / 蛋白質多量体 |
|---|
| Research Abstract |
We have constructed a protein expression system by using Dictyostelium discoideum, eukaryotic organisms living as single cells. For expressing oligomeric proteins in Dictyostelium cells, it is necessary to use multiple selection markers since one marker is required for expressing a single gene. We have used the blasticidin S deaminase for the new selection marker in Dictyostelium cells. The actin 15 promoter and the actin 6 terminator were fused to the Bsr gene in frame to drive its expression. Since the neomycin-resistance gene has been already in use, we can now use two selection markers to express at least two genes at the same time. We used this new expression system to express one myosin heavy chain and two light chains at the same time in Dictyostelium cells. In this way, we are now under way to express large amount of the head domain of Dictyostelium myosin for crystallization.
|
