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A Versatile Approach Towards Regioselective Platinated DNA Sequences

机译:一种多功能的区域选择性镀铂DNA序列。

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摘要

Undesired N~7 platination of 2'-deoxyguanosine residues at predetermined sites in an oligodeoxynucleotide (DPC) as an O~6-protecting group. The presence of a base-labile oxalyl linker between the immobilized 3'-nucleotide and controlled pore glass (CPG) allows cleavage of the protected ODN from the support and leaves DPC protection unaffected. This method provides an ODN with specifically blocked guanine-N~7 sites for platination. In the hexanucleotides prepared in this study, 5'-GGBGGT-3'(for B=T, C and A), a platinum GG adduct is introduced at G4,G5. These site-specific platinated hexamers were isolated in a yield of 65%, and were fully characterized by using reversed-phase HPLC (high performance liquid chromotography), LCMS (liquid chromatography-mass spectrometry), MALDI-TOF MS (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry), PAGE (polyacrylamide gel electropho-resis) and Maxam-Gilbert sequencing analysis.
机译:在寡聚脱氧核苷酸(DPC)中作为O-6保护基的预定位点的2'-脱氧鸟苷残基进行了不希望的N〜7电镀。固定的3'-核苷酸和可控孔玻璃(CPG)之间存在碱不稳定的草酰连接基,可从载体上裂解保护的ODN,而DPC保护不会受到影响。该方法为ODN提供了特别封闭的鸟嘌呤-N〜7位用于电镀。在本研究中制备的六核苷酸5'-GGBGGT-3'(对于B = T,C和A)中,在G4,G5处引入了铂金GG加合物。分离出这些位点特异性的铂化六聚体,产率为65%,并通过使用反相HPLC(高效液相色谱),LCMS(液相色谱-质谱),MALDI-TOF MS(矩阵辅助激光)进行了全面表征解吸/电离飞行时间质谱),PAGE(聚丙烯酰胺凝胶电泳)和Maxam-Gilbert测序分析。

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