G-四链体序列筛选及G-四链体等温信号扩增体系的构建  

G-quadruplex Sequences Screening and Construction of G-quadruplex Isothermal Signal Amplification System

作　　者：何杉 盛磊 高进[1] 苗朝华[1] 刘卫晓 宛煜嵩[1] 金芜军[1] He Shan;Sheng Lei;Gao Jin;Miao Chaohua;Liu Weixiao;Wan Yusong;Jin Wujun(Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing,100081)

机构地区：[1]中国农业科学院生物技术研究所,北京100081

出　　处：《分子植物育种》2022年第23期7812-7818,共7页Molecular Plant Breeding

基　　金：转基因新品种培育重大专项(2018ZX08012-001)资助。

摘　　要：G-四链体序列可与Hemin结合形成具有过氧化物酶活性的DNAzyme,被广泛应用于生物传感器的构建中。G-四链体的过氧化物酶活性高低对生物传感器构建至关重要。本研究利用圆二色谱比较分析了生物传感器中常用G-四链体序列的空间构型,筛选出过氧化物酶活性高的G-四链体序列,并比较了G-四链体中鸟苷酸聚集单元数、K+及Hemin对其过氧化物酶活性的影响。本研究应用高过氧化物酶活性的G-四链体作为可视检测信号核酸分子设计了一种可用于核酸检测的G-四链体等温信号扩增体系,不同于通常核酸检测方法直接对目标序列进行扩增,该体系在等温条件下,通过剪切与置换进一步大量扩增作为信号分子的G-四链体序列,致使整个核酸检测过程更为简便,为生物传感器构建、新型核酸检测技术研发提供了新的思路和技术支持。G-quadruplex sequences can bind Hemin to form DNAzyme with peroxidase activity, which is widely used in the construction of biosensors. The peroxidase activity of G-quadruplex is critical for the construction of biosensor.In this study, the spatial configurations of G-quadruplex sequences commonly used in biosensors were analyzed by circular dichroism, and the G-quadruplex sequences with high peroxidase activity were screened. out The effects of the number of guanosine aggregation units, K+and Hemin on the peroxidase activity of G-quadruplex sequences were compared. In this study, a G-quadruplex isothermal signal amplification system for nucleic acid detection was designed by using G-quadruplex with high peroxidase activity as a visual detection signal nucleic acid molecule. Different from the usual nucleic acid detection methods to directly amplify the target sequence, the system further amplified the G-quadruplex sequence as the signal molecule in large quantities under isothermal conditions by cutting and replacement, which made the whole nucleic acid detection process more convenient. It will provide new ideas and technical supports for biosensor construction and new nucleic acid detection technology development.

关 键 词：G-四链体 DNAZYME 等温信号扩增 

分 类 号：Q503[生物学—生物化学]