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

生物相容性的人工 DNA 接头,可被 DNA 聚合酶读取,且在大肠杆菌中发挥作用

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作者:Afaf H El-Sagheer, A Pia Sanzone, Rachel Gao, Ali Tavassoli, Tom Brown

Abstract

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.

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