TY - JOUR
T1 - Fluorescence detection of cytosine/guanine transversion based on a hydrogen bond forming ligand
AU - Nishizawa, Seiichi
AU - Yoshimoto, Keitaro
AU - Seino, Takehiro
AU - Xu, Chun Yan
AU - Minagawa, Masakazu
AU - Satake, Hiroyuki
AU - Tong, Aijun
AU - Teramae, Norio
N1 - Funding Information:
This work was partially supported by Grants-in-Aid for Scientific Research (A), No. 14204074, and for the COE Project, Giant Molecules and Complex Systems, 2003, from the Ministry of Education, Culture, Sports, Science and Technology, Japan. Partial support by the Japan–China Cooperative Research Program and RFTF from the Japan Society for Promotion of Science (JSPS) are also acknowledged.
PY - 2004/5/10
Y1 - 2004/5/10
N2 - In combination with abasic site (AP site)-containing oligodeoxynucleotides (ODNs), we demonstrate potential use of a hydrogen bond forming ligand, 2-amino-7-methyl-1,8-naphthyridine (AMND), for the fluorescence detection of the cytosine (C)/guanine (G) mutation sequence of the cancer repression gene p53. Our method is based on construction of the AP site in ODN duplexes, which allows small synthetic ligands to bind to target nucleobases accompanied by fluorescence signaling: an AP site-containing ODN is hybridized with a target ODN so as to place the AP site toward a target nucleobase, by which hydrophobic microenvironments are provided for ligands to recognize target nucleobases through hydrogen-bonding. In 10mM sodium cacodylate buffer solutions (pH, 7.0) containing 100mM NaCl and 1.0mM EDTA, AMND is found to strongly bind to C (Kd=1.5×10-6M) in the target ODN while the binding affinity for G is relatively moderate (Kd=50×10-6M). Significant fluorescence quenching of AMND is observed only when binding to C, making it possible to judge the C/G transversion with the naked eye.
AB - In combination with abasic site (AP site)-containing oligodeoxynucleotides (ODNs), we demonstrate potential use of a hydrogen bond forming ligand, 2-amino-7-methyl-1,8-naphthyridine (AMND), for the fluorescence detection of the cytosine (C)/guanine (G) mutation sequence of the cancer repression gene p53. Our method is based on construction of the AP site in ODN duplexes, which allows small synthetic ligands to bind to target nucleobases accompanied by fluorescence signaling: an AP site-containing ODN is hybridized with a target ODN so as to place the AP site toward a target nucleobase, by which hydrophobic microenvironments are provided for ligands to recognize target nucleobases through hydrogen-bonding. In 10mM sodium cacodylate buffer solutions (pH, 7.0) containing 100mM NaCl and 1.0mM EDTA, AMND is found to strongly bind to C (Kd=1.5×10-6M) in the target ODN while the binding affinity for G is relatively moderate (Kd=50×10-6M). Significant fluorescence quenching of AMND is observed only when binding to C, making it possible to judge the C/G transversion with the naked eye.
KW - Abasic site
KW - Fluorescence detection
KW - Hydrogen bond
KW - Single nucleotide polymorphism
KW - Synthetic ligand
KW - Transversion
UR - http://www.scopus.com/inward/record.url?scp=1842734660&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=1842734660&partnerID=8YFLogxK
U2 - 10.1016/j.talanta.2003.09.027
DO - 10.1016/j.talanta.2003.09.027
M3 - Article
C2 - 18969416
AN - SCOPUS:1842734660
VL - 63
SP - 175
EP - 179
JO - Talanta
JF - Talanta
SN - 0039-9140
IS - 1
ER -
