Abstract：The present research was to develop the exciplex-based fluorescence detection of DNA. A SNP-containing region of cytochrome P450 2C9 DNA systems was evaluated to define some of the structural and associated requirement of this new class of exciplex-formed probe, and a 24-base target was selected which contains single-nucleotide polymorphisms (SNP) in genes coding for cytochrome P450. The two probes were all 12-base to give coverage of a 24-base target region to ensure specificity within the human genome. Exciplex partners used in this study were prepared using analogous phosphoramide attachment to the 3’- or 5’-phosphate group of the appropriate oligonucleotide probes. The target effectively assembled its own detector by hybridization from components which were non-fluorescent at the detection wavelength, leading to the huge improvement in terms of decreased background. This research provides details of the effects of different partner, position of partners and different excitation wavelengths for the split-oligonucleotide probe system for exciplex-based fluorescence detection of DNA. This study demonstrates that the emission intensity of the excimer formed by new pyrene derivative is the highest in these excimer and exciplex, and the excimer is easy to be formed and not sensitive to the position of partners. However the exciplex formed by the new pyrene derivative and naphthalene emitted strongly at ～505 nm with large Stokes shifts (120-130 nm), and the monomer emission at 390 and 410 nm is nearly zero. Excitation wavelength of 400 nm is the best for Ie505/Im410 (exciplex emission at 505 nm/monomer emission at 410 nm) of the exciplex. This method features lower background and high sensitivity. Moreover the exciplex is sensitive to the steric factor, different position of partners and microenvironment, so this exciplex system is promising and could be tried to identify the SNP genes.
Key words：New fluorescent probes;Exciplex;Detection;Specific sequence DNA
李庆勇，祖元刚，吕宏艳，王丽敏 . 新基激复合物探针检测指定序列DNA[J]. 光谱学与光谱分析, 2009, 29(07): 1962-1966.
LI Qing-yong, ZU Yuan-gang, Lü Hong-yan, WANG Li-min . A New Class of Exciplex-Formed Probe Detect of Specific Sequence DNA . SPECTROSCOPY AND SPECTRAL ANALYSIS, 2009, 29(07): 1962-1966.
[1] GUO Liang-qia, YE Fang-gui, LIN Xu-cong, et al(郭良洽, 叶芳贵, 林旭聪，等). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2006, 26(1): 121. [2] WANG Dong-dong, MENG Qing-xiang, CHEN Dan-dan, et al(王东冬, 孟庆翔, 陈丹丹，等). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2007, 27(3): 510. [3] Birks B, Photophysics of Aromatic Molecules. London: Wiley, 1970. [4] Birks J B. Rep. Prog. Phys., 1975, 38: 903. [5] Ebata K, Masuko M, Ohtani H, et al. Nucleic Acid Symp. Ser., 1995, 34: 187. [6] Ebata K, Masuko M, Ohtani H, et al. Photochem. Photobiol., 1995, 62: 836. [7] Masuko M, Ebata K, Ohtani H. Nucleic Acids Symp. Ser., 1996, 35: 165. [8] Masuko M, Ohtani H, Ebata K, et al. Nucleic Acids Res., 1998, 26: 5409. [9] Shimadzu A, Ohtani H, Ohuchi S, et al. Nucleic Acids Symp Ser., 1998, 39: 45. [10] Paris P L, Langenhan J M, Kool E T. Nucleic Acids Res., 1998, 26: 3789. [11] Bichenkova E V, Gbaj A, Walsh L, et al. Org. Biomol. Chem., 2007, 5: 1039. [12] Bichenkova E V, Savage H E, Sardarian A R, et al. Biochem. Biophys. Res. Commun., 2005, 332: 956. [13] Bichenkova E V, Sardarian A R, Wilton A N, et al. Org. Biomol. Chem., 2006, 4: 367. [14] Masaki T. Journal of the Physical Society of Japan, 1976, 41: 1394. [15] Swinnen A M, Ruttens F, Van Der Auweraer, et al. Chemical Physics Letters, 1985, 116: 217. [16] Winnik F M. Chem. Rev., 1993, 93: 587.
