Biocompatible artificial DNA linker that is read through by DNA polymerases and is functional in Escherichia coli

摘要

A triazole mimic of a DNA phosphodiester linkage has been produced by templated chemical ligation of oligonucleotides func tionalized with 5 -azide and 3 -alkyne. The individual azide and alkyne oligonucleotides were synthesized by standard phosphora midite 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 synth esis 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.