Zhu T, Tung C H, Breslauer K J, Dickerhof W A, Stein S
Center for Advanced Biotechnology and Medicine, Piscataway, New Jersey 08854.
Antisense Res Dev. 1993 Winter;3(4):349-56. doi: 10.1089/ard.1993.3.349.
Oligodeoxynucleotides (12-mers) having either a 3' aminolinker or both 3' and 5' aminolinker groups were synthesized and then reacted with N-iodoacetoxysuccinimide. Separately, a series of peptides, consisting of cysteine carboxyamide and a varying number of residues of ornithine coupled through their side-chain amino groups, was prepared, leaving on the final Fmoc protecting group. Reaction of each Fmoc-peptide with an activated oligodeoxynucleotide yielded the desired conjugates, which were purified by an Fmoc-on/Fmoc-off two-step reversed-phase HPLC procedure. On hybridization with a complementary unmodified 12-mer oligodeoxynucleotide, it was found that there is a gradual increase in melting temperature with the number of ornithine residues, whereas the appended peptide did not perturb the B form of the duplex.
合成了带有3'氨基连接基团或同时带有3'和5'氨基连接基团的12聚体寡脱氧核苷酸，然后使其与N-碘代乙酰氧基琥珀酰亚胺反应。另外，制备了一系列由半胱氨酸羧酰胺和通过其侧链氨基连接的不同数量鸟氨酸残基组成的肽，保留最终的芴甲氧羰基（Fmoc）保护基团。每个Fmoc肽与活化的寡脱氧核苷酸反应产生所需的缀合物，通过芴甲氧羰基保留/芴甲氧羰基去除两步反相高效液相色谱法进行纯化。在与互补的未修饰12聚体寡脱氧核苷酸杂交时，发现随着鸟氨酸残基数量的增加，解链温度逐渐升高，而附加的肽并未扰乱双链体的B型结构。
