| 摘要 |
Modified nucleotides, and methods to modify nucleotides with a moiety or label, such as biotin, that permits their detection and results in a modified nucleotide, and methods of use of the modified nucleotide in quantitative and qualitative assays are outlined. In particular the modified nucleotide is a nucleoside bis-phosphate of structure (I):P1-P2-Nus-Alk-Lnk-Obs (I)or a salt, conjugate base, tautomer or ionised form thereof, wherein P1 and P2 are phosphate groups; Nus is a nucleoside moiety comprising a sugar bound to a purine or pyrimidine base; Alk is a connecting group having the structure—(CH2)m-Y- where Y is a bond or NH, NR, O, C(O),S(O)2, S or Si, where m is 3-6; Lnk is a linking group of formula—A1-CH2-(CH2OCH2)n-CH2-A2- or—A1-X-A3-CH2-(CH2OCH2)n-CH2-A2- where n is 2-48; Al is CH2. C(O), C(O)NH, C(O)O, S(O)2, C(S)NH, C(NR)NH, NH, NR or O; A2and A3are NH, NR, NHC(O), NHS(O)2, NHC(S)NH, NHC(NR)NH, NHC(O)NH, OC(O), SC(O), C(O), C(O)NH or C(O)); X is a cleavable group that can undergo silicon-carbon, redox, photochemical, enzymatic or exchange-based cleavage; and Obs is an observable label such as biotin, a fluorophore or an azide. A preferred compound of formula (I) is:
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