1997 Fiscal Year Final Research Report Summary
Development of a chemiluminescent reagent that recognizes guanine in nucleic acid, and its use for chemiluminescence detection of DNA
| Project/Area Number |
08457640
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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 |
Laboratory medicine
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
KAI Masaaki Kyushu University Pharmaceutical Sciences TITLE OF POSITION (Associate Professor), 薬学部, 助教授 (00160953)
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| Project Period (FY) |
1996 – 1997
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| Keywords | Nucleic acid / DNA / Chemiluminescence / Imaging detection / Arylglyoxalreagents / CCD camera / Selective detection / Hybridization |
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| Research Abstract |
A novel reagent, 3', 4', 5'-trimethoxyphenylglyoxal has been found to specifically convert guanine bases of a nucleic acid in a neutral or alkaline phosphate solution to the corresponding chemiluminescent derivatives. The chemiluminogenic derivatization technique offers a facile method of the sensitive imaging detection for a DNA probe tailing with deoxyriboguanylic acid oligomers. A cooled charge-coupled device camera is usable to detect visually the chemiluminescence emitted from the derivatized probe that was hybridized with a target DNA fragment on a Nylon membrane. The proposed method permits the quantitative detection of the probe covalently attached with a 5'- (G) _<15>TT (G) _<15>TT deoxyoligomer at 3' terminus, indicating a detection limit of 0.1 pmol on the membrane. The detection sensitivity can be increased with larger contents of deoxyriboguanylic acid in the probe molecule.
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