| Project/Area Number |
62870091
|
|---|
| Research Category |
Grant-in-Aid for Developmental Scientific Research
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Biological pharmacy
|
|---|
| Research Institution | Hokkaido University |
|---|
Principal Investigator |
OHTSUKA Eiko Faculty of Pharmaceutical Sciences, Hokkaido University, 薬学部, 教授 (80028836)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
IWAI Shigenori Faculty of Pharmaceutical Sciences, Hokkaido University, 薬学部, 助手 (10168544)
INOUE Hideo Faculty of Pharmaceutical Sciences, Hokkaido University, 薬学部, 助手 (80088856)
|
|---|
| Project Period (FY) |
1987 – 1988
|
| Project Status |
Completed (Fiscal Year 1988)
|
| Budget Amount *help |
¥5,200,000 (Direct Cost: ¥5,200,000)
Fiscal Year 1988: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1987: ¥3,800,000 (Direct Cost: ¥3,800,000)
|
|---|
| Keywords | Site-specific cleavage of RNA / Sequence-specific cleavage of RNA / RNase H / RNA・modified DNA hybrid / 2'-O-Methyloligoribonucleotide / Chimeric oligonucleotide / 大腸菌tRNA<met(＼)f> / 2′-O-メチルオリゴリボヌクレオチド / 大腸菌tRNA<Met( / )f> |
|---|
| Research Abstract |
The method for cleaving RNA site-specifically with RNase H was developed; RNase H is known to hydrolyze RNA only in RNA-DNA hybrids. The method invloves the use of the chimeric oligonucleotide containing a tetradeoxyribonucleotide and 2'-0-methyloligoribonucleotide(s) as a DNA splint.
Model experiments were carried out using the hybrids of oligo-ribonucleotides (9 and 18mer) and chimeric splints (9mer), and E. coli RNase H. We found that the ribo-strands were cleaved specifically at the 3'-end of the RNA DNA hybrid regions. This technique was successfully applied to the site-directed cleavage of the loop and stem regions in RNA 90mer. We also examined applicability of the method using E. coli tRNA (f Met) and found that the cleavages occurred at desired positions in the acceptor-stem region.
|
