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引用次数: 0
摘要
我们设计并合成了一种带有可裂解三氮烯连接体的荧光标记核苷酸,作为 DNA 合成测序(SBS)的潜在可逆终止子。通过对氨基修饰的核苷酸进行三嗪化,然后进行荧光标记,成功合成了关键的中间体--三嗪修饰的核苷酸,并得到了所需的产物。合成的三嗪修饰核苷酸可以有效地作为克伦诺片段(外)DNA 聚合酶的底物,并被整合到 DNA 链中。一旦第一个修饰的核苷酸被整合进去,就不可能再进一步延伸,即使是未被阻断的 3'-OH 基团也是如此。在酸性条件下,荧光标签被完全去除后,三嗪可逆终止剂可再次与 DNA 链结合,从而实现 DNA 测序循环。
Synthesis and Evaluations of Cleavable Triazene‐Modified Nucleotide for DNA Sequencing
A fluorescence‐labeled nucleotide with a cleavable triazene linker was designed and synthesized as a potential reversible terminator for DNA sequencing by synthesis (SBS). The key intermediate, a triazene‐modified nucleotide, was successfully synthesized through the triazenylation of an amino‐modified nucleotide, followed by fluorescence labeling to yield the desired product. The synthesized triazene‐modified nucleotide can effectively serve as a substrate for Klenow Fragment (exo‐) DNA polymerase and be incorporated into DNA strands. Once the first modified nucleotide is incorporated, no further extension is possible, even with an unblocked 3'‐OH group. After the fluorescent label is completely removed under acidic conditions, the triazene reversible terminator can be incorporated into DNA strands again, thereby enabling the DNA sequencing cycles.
期刊介绍:
Organic chemistry is the fundamental science that stands at the heart of chemistry, biology, and materials science. Research in these areas is vigorous and truly international, with three major regions making almost equal contributions: America, Europe and Asia. Asia now has its own top international organic chemistry journal—the Asian Journal of Organic Chemistry (AsianJOC)
The AsianJOC is designed to be a top-ranked international research journal and publishes primary research as well as critical secondary information from authors across the world. The journal covers organic chemistry in its entirety. Authors and readers come from academia, the chemical industry, and government laboratories.