| Project/Area Number |
16550143
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Chemistry related to living body
|
|---|
| Research Institution | Kobe University |
|---|
Principal Investigator |
EBARA Yasuhito Kobe University, Fuculty of Human Development, Associate Professor, 発達科学部, 助教授 (40251657)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
UEJI Shin-ichi Kobe University, Fuculty of Human Development, Professor, 発達科学部, 教授 (40031364)
|
|---|
| Project Period (FY) |
2004 – 2005
|
| Project Status |
Completed (Fiscal Year 2005)
|
| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2005: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2004: ¥2,900,000 (Direct Cost: ¥2,900,000)
|
|---|
| Keywords | Saccharide / in vitro selection / DNA / Phenylboronic Acid / SELEX / Aptamer / Sialyl Lewis X / in vivo selection / シリアルルイスX / 糖質 / シアリルルイスx |
|---|
| Research Abstract |
Sialyl Lewis X (sLex) is one of the most important saccharide on the cell surface. It is well known that sLex play a important role on the cancer cell metastasis. A compound that recognizes sLex specifically with high affinity is expected to be efficient tumor marker or cancer cell targeting drug. But it is difficult to design such the compound because there are tremendous possibilities of sLex conformations and there are a lot of hydroxyl groups that is difficult to be recognized using conventional normal amino acids or nucleic acids. In this study, phenylboronic acid (PBA)-modified DNA library was constructed, and in vitro selection procedure was achieved to find the compounds that recognize sLex via multi point interaction. PBA has been used to recognize cis-diol like mono- and di-saccharides in alkali solution. PBA-modified novel dUTP was synthesized. The PBA-modified dUTP worked as DNA polymerase substrate and was efficiently incorporated into ssDNA containing 60 randamized nucleotides. The PBA-modified ssDNA library was incubated on a quartz crystal mocrobalance (QCM) on which biotin-sLex was immobilized via streptavidin. The QCM device is able to detect 30 pg/Hz mass change on the surface, so the PBA-modified ssDNA binding behavior to sLex surface could be monitored in situ at the each round. Specific DNAs were recoverd by adding sLex solution and the recovered PBA-modified ssDNA was amplified by asymmetric PCR and was incubated to the sLex on the QCM again. After the rounds, through cloning of the ssDNAs, it was demonstrated that the selected PBA-modified ssDNA bound to sLex with high affinity more than 10^5/ M^<-1>.
|
