Proto-cell Model Exhibiting Synchronized Self-replication Dynamics of Giant Vesicles and DNA
| Project/Area Number |
17350066
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Functional materials chemistry
|
|---|
| Research Institution | The University of Tokyo |
|---|
Principal Investigator |
SUGAWARA Tadashi The University of Tokyo, Graduate School of Arts and Sciences, Professor (50124219)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
TOYOTA Taro Chiba University, Graduate School and faculty of Engineering, An assistant (80422377)
|
|---|
| Project Period (FY) |
2005 – 2007
|
| Project Status |
Completed (Fiscal Year 2007)
|
| Budget Amount *help |
¥13,930,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥330,000)
Fiscal Year 2007: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2006: ¥4,700,000 (Direct Cost: ¥4,700,000)
Fiscal Year 2005: ¥7,800,000 (Direct Cost: ¥7,800,000)
|
|---|
| Keywords | Proto-cell Model / Self-reproducing vesicle / PCR in giant vesicle / Anionic membrance molecule / Lipid-DNA conjugate molecule / 原子細胞モデル / 自己複製ベシクル / ベシクル内膜上PCR / 脂質-DNA合分子 / ジャイアントベシクル / PCR / DNA複製 / ベシクル内酵素反応 / PAGE / 人工細胞モデル / ジャイアント・ベシクル / 自己生産 / 自己複製 / セルソーター |
|---|
| Research Abstract |
1. Self-reproducing giant vesicles composed of anionic membrane molecules A novel membrane molecule with an anionic polar head comprised of hetero-phosphate diester was prepared from 10-(p-formylphenoxy) decanol and dodecanol through an amidite method in a 62 % yield. This anionic membrane molecule constructed nearly uni-lamella giant vesicles in a phosphate buffer solution containing 15% methanol. Moreover the anionic vesicles exhibited self-reproduction dynamics when a membrane-precursor was added (The 88th Annual Meeting of Chemical Society of Japan, 3L3-17).
2. Polymerase chain reaction (PCR) in giant vesicles composed of POPC Replication of template DNA consisting of 1229 by that can express GFP was performed inside giant vesicles (GV) of POPC (palmitoyl-oleoyl-phosphatidyl choline) using a PCR technique. After 20 thermal cycles, the fluorescence emitted from SYBR Green I absorbed on the DNA-double strands became clearly visible (two orders increase in average) by fluorescence micr
… More
oscope. The exact replication of the template DNA was confirmed by PAGE (polyacrylamide gel electrophoresis). Flow-cytometric analysis revealed that ratio of the effective replication of DNA inside GVs was ca 10 % (The 88th Annual Meeting of Chemical Society of Japan, 3C6-25).
3. Coupling of dynamics of self-replication of DNA and self-reproduction of giant vesicles For the purpose titled as above, a conjugate molecule consisting of a DNA oligomer (15mer), a spacer, and cholesterol was prepared. The DNA oligomer serves as a primer hybridizing with a template DNA (100mer) inside GV, and cholesterol works as, an anchor to fix the conjugate molecule with the vesicular membrane. The progress of the complimentary replication of the template DNA on the inner surface of vesicular membrane was directly observed by a fluorescent microscope using a primer tagged with a fluorescence-probe, and the replication of the template DNA was confirmed by PAGE analysis. This approach guarantees the possible coupling between the two dynamics, which is crucial for giving rise to an artificial cell model (Soft Matter, 2005). Less
|
Report
(4 results)
Research Products
(114 results)
-
-
-
-
-
-
[Journal Article] Population Study of Sizes and Components of Self-Reproducing Giant Multilamellar Vesicles2008
Author(s)
Toyota, T., Takakura, K, Kageyama, Y, Kurihara, K., Maru, N., Ohnuma, K., Kaneko, K., Sugawara, T.
-
Journal Title
Langmuir 24
Pages: 3037-3044
Related Report
Peer Reviewed
-
-
[Journal Article] Population Study of Sizes and Components of Self-Reproducing Giant Multilamellar Vesicles2008
Author(s)
Toyota, T., Takakura, K., Kageyama, Y., Kurihara, K., Maru, N., Ohnuma, K., Kaneko, K., Sugawara, T.
-
Journal Title
Langmuir 24
Pages: 3037-3044
Related Report
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
[Journal Article] An autonomous molecular computing system in cell-sized liposome. NanoBio-Tokyo2006
Author(s)
Shohda, K., Kiga, D., Takinoue, M., Kageyama, Y., Sugawara, T., Suyama, A
-
Journal Title
NanoBio-Tokyo 2006
Pages: 357-358
Related Report
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
[Presentation] PCR in giant phospholipid vesicles2008
Author(s)
Mieko Tamura, Yoshiyuki Kageyama, Koh-ichiroh Shohda, Kentaro Suzuki, Tadashi Sugawara
Organizer
The Chemical Society of Japan 88th Spring Meeting
Place of Presentation
Tokyo
Description
「研究成果報告書概要(欧文)」より
Related Report
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
