Solid-phase synthesis of oligonucleotides containing site-specific N-(2'-deoxyguanosin-8-yl)-2-(acetylamino)fluorene adducts using 9-fluorenylmethoxycarbonyl as the base-protecting group.
Eight oligodeoxyribonucleotides containing a site-specific N-(2'-deoxyguanosin-8-yl)-2-(acetyl-amino)fluorene (dG-C8-AAF) adduct were prepared successfully by solid-phase DNA synthesis using the 2-cyanoethyl N,N-diisopropylphosphoramidites of dA, dC, dG, dT, and dG-C8-AAF, with 9-fluorenyl-methoxycarbonyl (Fmoc) as the base-protecting group. The oligonucleotides were deprotected and released from the support by 1:9 piperidine/MeOH at room temperature for 22-36 h or by 1:1 diisopropylamine in MeOH at 55 degrees C for 15 h, purified by HPLC, and fully characterized. About 6 mg of HPLC-purified d[GTGGCG(C8-AAF)CCAAGT] and 7 mg of d[GTGATG(C8-AAF)ATAAGT] were obtained from the 10-mumol-scale synthesis, and their 1D 1H NMR spectra were consistent with the presence of a dG-C8-AAF adduct. The dG-C8-AAF oligonucleotides were also deacetylated to afford the corresponding dG-C8-AF oligonucleotides. d[GTGGCG(C8-AAF)CCAAGT] formed stable 1:1 duplexes with both the fully complementary 12-mer and a GC-deleted (across the adduct) 10-mer complement, and identical melting temperatures were observed for both duplexes. The multidimensional NMR study of these duplexes is presently under investigation.
Zhou
Y
Y
Department of Chemistry, Wayne State University, Detroit, Michigan 48202.
Romano
L J
LJ
D015073:2-Acetylaminofluorene / Q000031:analogs & derivatives* / Q000032:analysis; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005449:Fluorenes / Q000737:chemistry*; D006147:Guanine / Q000032:analysis; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis* / Q000737:chemistry
D005449:Fluorenes; D009838:Oligodeoxyribonucleotides; C018776:N-(deoxyguanosin-8-yl)acetylaminofluorene; D006147:Guanine; D015073:2-Acetylaminofluorene; D003849:Deoxyguanosine