Application of a dsDNA imprinted polymer to detection for bacterial toxin
| Project/Area Number |
18350080
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Environmental chemistry
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| Research Institution | Tokyo University of Technology |
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Principal Investigator |
MINOURA Norihiko Tokyo University of Technology, School of Bionics, 教授 (10358111)
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| Co-Investigator(Kenkyū-buntansha) |
SHINBO Toshio National Institute of Advanced Industrial Science and Technology, Research Center of Advanced Bionics, 副センター長 (70357250)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥17,420,000 (Direct Cost: ¥15,200,000、Indirect Cost: ¥2,220,000)
Fiscal Year 2007: ¥9,620,000 (Direct Cost: ¥7,400,000、Indirect Cost: ¥2,220,000)
Fiscal Year 2006: ¥7,800,000 (Direct Cost: ¥7,800,000)
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| Keywords | Molecular imprinting / Polymer gel / Electrophoresis / dsDNA / DNA sequence / Molecular recognition / Template / Verotoxin / 分子認織 / ペロ毒素 |
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| Research Abstract |
Molecularly imprinted polymers(MIPs) are materials that are attractive for research applications because they have an artificial recognition cavity for the target molecule. We prepared a MIP for recognizing a specific dsDNA sequence by using 2-vinyl-4, 6-diamino-1, 3, 5-triazine(VDAT) as a functional monomer and used it in an electrophoretic gel matrix.
Firstly, interaction between VDAT analog as a new functional monomer and dsDNA was investigated to improve the specificity of target dsDNA to MIP gel. From the result, some candidate compounds were selected as new functional monomer and application of these compounds to MIP is now in progress.
To achieve a miniaturization of detection device, a label-free sample, and a real time detection, we demonstrate a novel dsDNA detection system with dsDNA-imprinted polymer gel by using VDAT as a functional monomer. The miniature-sized gel matrix was used in this system. The dsDNA eluted through the MIP gel during electrophoresis was stained with fluorescence molecule at an edge of the gel and was carried to fluorescence detector by carrier solution. In this system, elution time of dsDNA with the target sequence should be elongated in comparison with the non-imprinted polymer gel by the capture effect of the binding sites in the MIP gel. Under optimum measurement condition, the elongation of elution time on λDNA 564bp fragment was confirmed in a λDNA 564bp fragment-imprinted polymer gel.
Application of this system to a bacterial dsDNA detection was also investigated. However, the target bacterial dsDNA was not detected. The DNA size of bacterial dsDNA(34bp) is smaller in comparison with the λDNA 564bp fragment. It suggests that there is an influence of size of DNA on detection of target dsDNA, since the number of functional groups surrounding the binding sites in MIP gel increases with increasing the size of target dsDNA. We are planning to study the relationship between size of target dsDNA and detection limit in this system.
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Report
(3 results)
Research Products
(26 results)
