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A Versatile Approach Towards Regioselective Platinated DNA Sequences
Author(s) -
Heetebrij Robert J.,
de Kort Martin,
Meeuwenoord Nico J.,
den Dulk Hans,
van der Marel Gijs A.,
van Boom Jacques H.,
Reedijk Jan
Publication year - 2003
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.200390209
Subject(s) - chemistry , guanine , chromatography , mass spectrometry , linker , nucleotide , adduct , gel electrophoresis , high performance liquid chromatography , dna , biochemistry , organic chemistry , computer science , gene , operating system
Undesired N 7 platination of 2′‐deoxyguanosine residues at predetermined sites in an oligodeoxynucleotide (ODN) sequence is prevented by applying the sterically demanding diphenylcarbamoyl (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 electrophoresis) and Maxam–Gilbert sequencing analysis.