文献类型：期刊文献

中文题名：Aerolysin蛋白纳米孔道直接检测单个DNA碱基修饰

英文题名：Direct Single-molecule Discrimination of Acetylene-linked Deoxyribonucleoside Triphosphates with an Aerolysin Nanopore

作者：路瑶[1];胡正利[1];应佚伦[1];龙亿涛[1]

机构：[1]华东理工大学化学与分子工程学院结构可控先进功能材料及其制备教育部重点实验室,上海200237

年份：2019

卷号：38

期号：2

起止页码：129

中文期刊名：分析测试学报

外文期刊名：Journal of Instrumental Analysis

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD2019_2020】；

基金：国家自然科学基金资助项目(21421004);上海市"晨光计划"项目(17CG27)

语种：中文

中文关键词：生物纳米孔道;单分子分析;DNA;电化学空间限域

外文关键词：biological nanopore;single-molecule analysis;DNA;electrochemically confined space

摘要：基于Aerolysin生物膜通道蛋白的纳米孔道电化学分析技术,因其高的电化学空间限域能力可实现超灵敏DNA单分子检测。本文利用单个Aerolysin纳米孔道在无需标记、无需扩增的条件下直接分辨3种具有单个碱基差异的单链DNA。实验结果显示,具有单个炔基侧链基团修饰的单个ss DNA在限域空间内与Aerol-ysin纳米孔道的相互作用时间较未修饰的ss DNA增长近7倍,电流阻断程度增大7%,且高斯峰半峰宽减小了44%,增强了Aerolysin纳米孔道对单个DNA分子的分辨能力。研究成果有望推动Aerolysin纳米孔在DNA直接测序及表观遗传修饰检测中的应用。
Aerolysin could self-assemble and oligomerize heptameric complex which insert into the lipid bilayer to form a nanopore. Due to the electrochemically confined effects,the aerolysin shows the high sensitivity in single-molecule analysis of DNAs. Here,aerolysin was used to directly discriminate the single acetylene-linked deoxyribonucleoside triphosphates from a single oligonucleotide without label or PCR operations. After modifying with acetylene derivative,a single oligonucleotide produced seven times longer blockade duration inside an aerolysin nanopore,increasing the blockage current amplitude by 7% and narrowing the current distribution by 44%. Therefore,the modification of the linear side-chain of acetylene derivative could be further used to improve the temporal and spatial resolution in single DNA analysis via the aerolysin nanopore.