School of Chemistry, University of Southampton, Highfield, Southampton SO17 1BJ, United Kingdom.
Proc Natl Acad Sci U S A. 2011 Jul 12;108(28):11338-43. doi: 10.1073/pnas.1101519108. Epub 2011 Jun 27.
A triazole mimic of a DNA phosphodiester linkage has been produced by templated chemical ligation of oligonucleotides functionalized with 5'-azide and 3'-alkyne. The individual azide and alkyne oligonucleotides were synthesized by standard phosphoramidite methods and assembled using a straightforward ligation procedure. This highly efficient chemical equivalent of enzymatic DNA ligation has been used to assemble a 300-mer from three 100-mer oligonucleotides, demonstrating the total chemical synthesis of very long oligonucleotides. The base sequences of the DNA strands containing this artificial linkage were copied during PCR with high fidelity and a gene containing the triazole linker was functional in Escherichia coli.
通过 5'-叠氮化物和 3'-炔烃官能化的寡核苷酸的模板化学连接，合成了 DNA 磷酸二酯键的三唑类似物。单个叠氮化物和炔烃寡核苷酸通过标准的亚磷酰胺方法合成，并使用简单的连接程序组装。这种酶促 DNA 连接的高效化学等价物已用于从三个 100 -mer 寡核苷酸组装 300-mer，证明了非常长的寡核苷酸的全化学合成。含有这种人工键的 DNA 链的碱基序列在 PCR 中具有高保真度进行复制，并且含有三唑接头的基因在大肠杆菌中具有功能。
