高等学校化学学报Issue(11)：2353-2361,9.
四色荧光标记二硫键可逆终端的合成及在DNA合成测序中的应用
Synthesis and Application of Four Fluorescence Labeled Nucleotides Through Disulfide as Reversible Terminators in DNA Sequencing by Synthesis
摘要
Abstract
DNA sequencing is one of the important means of biological research. The development of accurate and high-throughput DNA-sequencing methods has a strong impact on biomedical research. Sequencing by synthesis( SBS) approaches is one of the most important techniques of DNA sequencing. A series of SBS methods have been investigated and implemented. In SBS methods, the principal thing is to synthesize four fluorescently labelled cyclic reversible terminators as the sequencing reagents. These cyclic reversible termina-tors need to connect the fluorescein through a cleavable linker. In chemical biology, disulfide bridges, which are known to be efficiently and rapidly cleaved by mild reducing agents, have been used in a wide range of ap-plications including drug delivery, functional and structural proteomics, DNA and tumor imaging. The disul-fide-based cleavable linker is commonly used due to its rapid cleavage and straightforward synthesis. In this work, starting from 5-iodo-2'-deoxyuridine, 5-iodo-2'-deoxycyridine, 7-deazo-7-iodo-2'-deoxyadenosine and 7-deazo-7-iodo-2'-deoxyguanosine, four kinds of fluorescence labelled necleotides with four different bases through disulfide were synthesized and characterized. The synthesized fluorescence labelled nucleotides can be recognized and accepted by an identified DNA polymerase, incorporated faithfully into DNA strand, followed by cleaved completely by DTT within 1 min. These results demonstrate that the fluorescence labelled nucleo-tides have potential value as reversible terminators in DNA sequencing by synthesis.关键词
DNA合成测序/可逆终端/可裂解连接单元/四色荧光标记Key words
DNA sequencing by synthesis/Reversible terminator/Cleavable linker/Four fluorescence labelled nucleotide分类
化学化工引用本文复制引用
汤道年,江敏,李小卫,康亚妮,伍新燕,龚兵,邵志峰,赵小东,沈玉梅..四色荧光标记二硫键可逆终端的合成及在DNA合成测序中的应用[J].高等学校化学学报,2014,(11):2353-2361,9.基金项目
国家自然科学基金(批准号:81071250,91227109)和科技重大专项“重大新药创制”项目(批准号:2011ZX09501-001-05)资助 （批准号:81071250,91227109）
