| Project/Area Number |
11358012
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (A)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 展開研究 |
|---|
| Research Field |
Biophysics
|
|---|
| Research Institution | Yokohama City University |
|---|
Principal Investigator |
NISHIMURA Yoshifumi Graduate School of Integrated Science, Professor, 大学院・総合理学研究科, 教授 (70107390)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
SUZUKI Ichirou AJINOMOTO, Co, LTd, Analytical Chemistry Laboratories Central Research Laborstories, Senior Researcher, 中央研究所・分析研究部, 主席研究員
|
|---|
| Project Period (FY) |
1999 – 2001
|
| Project Status |
Completed (Fiscal Year 2001)
|
| Budget Amount *help |
¥33,500,000 (Direct Cost: ¥31,400,000、Indirect Cost: ¥2,100,000)
Fiscal Year 2001: ¥9,100,000 (Direct Cost: ¥7,000,000、Indirect Cost: ¥2,100,000)
Fiscal Year 2000: ¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1999: ¥17,400,000 (Direct Cost: ¥17,400,000)
|
|---|
| Keywords | transcription tactors / DNA-binding proteins / drug / flow-type automated NMR / double-strand DNA chip / telomeres / TRF / hRapl / テロメア結合タンパク質 / DNA結合ドメイン / 表面構造 / hTRF1 / Myb / hRap1 / NMR / 基本転写因子 / 転写活性化ドメイン / 薬物の探索と設計 / TFIIE / PhoB |
|---|
| Research Abstract |
We have developed new technologies to analyze DNA-binding protein function and the interaction between proteins and other small molecules. New approach under development include a double-stranded DNA chip to detect DNA-binding specificities of proteins and a Protein Recycling Flow NMR system that will allow for faster screening of compounds that interact with proteins, while using much smaller amounts of the protein being screened.
By using the newly developed double-stranded DNA chip we have detected rapidly the sequence specificity of human TRF1 that binds double stranded telomeric DNA. Many double-stranded DNA with different sequences were chipped on a glass slide and then hTRF1 was spotted on the glass slide. The intensities of the fluorescence labeled DNA indicate the sequence specificities of hTRF1. The each intensity of a specific DNA sequence bound hTRF1 is well correlated to the binding dissociation constant of hTRF1 for each DNA sequence examined by using BIACORE. By using BIACORE it takes seven days to analyze the binding constants of hTRF1 for seven different sequences, while by using our new double-stranded DNA chip it takes half an hour.
|
