DNA光学逻辑门研究进展

DNA逻辑门是一种重要的新型计算单元,为下一代计算机的研发提供了新的思路。光学检测器拥有快速简便的优势,因此在DNA逻辑门的研究中显得十分重要。本文综述了近些年来DNA光学逻辑门的研究进展,对荧光、比色和表面增强拉曼光谱为基础的逻辑门的研究进行了阐述和评价,并对DNA光学逻辑门的发展前景进行了展望。     

以分子信标为报告分子的凝血酶检测新方法

以分子信标为报告分子,核酸适体为识别分子,发展了一种新的凝血酶检测方法.含有分子信标互补序列的核酸适体探针与凝血酶结合后,分子信标的荧光信号下降,从而得到凝血酶的浓度信息.该方法快速、灵敏,核酸适体探针无需荧光标记、设计简单,检测限达到0.83nmol/L.     

我国研制出新型DNA逻辑门奠定发展DNA计算机基础

中科院上海应用物理研究所的樊春海研究员与上海交通大学Bio-X中心的贺林院士、张治洲教授通过学科交叉研究与合作,应用DNA核酶研制成功一类新型的“DNA逻辑门”,为发展DNA计算机奠定了基础。DNA核酶是一种通过体外进化筛选出来的具有特定酶活性的核酸结构,在该项研究中采用的是具有DNA水解酶活性的DNA核酶。这种具有锤头状结构的核酶可以在铜离子辅助下催化氧化并切割底物DNA。DNA逻辑门即是在这种DNA核酶结构基础上通过模块设计（modular design）研制出来的。输入信号通过特定的生物分子传感可以产生输出信号,从而实现“YES”、“NOT”等逻辑判断,并可以组合成复杂的三输入逻辑门“AND（A,NOT（B）,NOT（C））”。该逻辑门系统的新特色在于排除以往DNA逻辑门设计中RNA核苷的参与,仅单纯应用DNA分子,从而避免了RNA核苷带来的系统不稳定性。相关研究结果已发表在2006年3月出版的著名化学杂志（（Angew．Chem．Int．Ed．》上。     

适体酶的筛选及应用

适体酶也称为别构效应核酶或别构效应脱氧核酶,是一种新的人工合成酶,它兼具适体的靶物质特异性结合与核酶或脱氧核酶的催化活性优点。这种酶是从大量随机序列库中针对各种效应分子筛选获得的目的酶。适体酶为效应分子的定量分析提供了新的思路。适体酶不仅在基因组学和蛋白质组学研究中得到应用,而且在生物传感器和DNA分子逻辑研究中也具有潜在的应用前景。     

基于CiteSpace分析核酸适体（Aptamer）技术研究现状

利用可视化分析软件,客观、全面地展示和分析适体的研究现状、研究热点和发展趋势,为适体的研究提供指导和参考。以SCI数据库Web of Science平台为数据源,采用CiteSpace 5.5.R2软件对1900年1月1日至2020年12月31日的适体相关文献进行分析。共纳入文献14 939篇。在该领域发表论文最多的作者是WEIHONG TAN、KHALIL ABNOUS等;近年来突现的主题词是“wide linear range”“pg ml”“clinical diagnosis”“early diagnosis”等,是目前该领域的研究热点;作者共被引分析和文献共被引分析表明,该领域的研究主要集中在适体的体外筛选、适体构象的变化、适体在药物靶向治疗中的应用及适体生物传感器。结果表明,利用CiteSpace软件对Web of Science数据库中已发表的适体文献进行分析,揭示了该领域的研究热点从体外筛选和适体构象变化（1992—2002）到适体在药物靶向治疗和适体生物传感器中的应用（2002—2020）。结合近年来突现的主题词预测适体在药物靶向治疗和适体生物传感器中的应用仍将是未来的研究热点。     

适体酶Aptazymes的研究进展

适体酶是从寡核苷酸随机序列库中筛选获得的针对各种效应分子的一种新的人工合成酶.它具有适体的特异性和核酶的催化活性.适体酶作为催化分子信标,为效应分子的定量分析提供了新的思路.适体酶不仅在基因组学和蛋白质组学中应用,而且在生物传感器和DNA分子逻辑研究中具有潜在的应用前景.     

基于三链核酸的DNA计算

一种研究DNA计算的新模型——三链DNA计算模型在本文中提出。此模型是在近年三链核酸的研究成果的基础上建立的。并应用于求解可满足性问题（SAT）,这是一个困难的NP-完全问题。不同于以住的DNA计算方法,基于三链核酸的分子算法通过寡聚脱氧核苷酸（ODN）在RecA蛋白的介导下与同源的双链DNA匹配成三链DNA进行基本的运算,这样可以有效减少计算中的错误。依据分子生物学的实验方法,该算法切实可行并且有效。     

一种新的基于适体和酶切保护分析的超灵敏蛋白质检测方法

以凝血酶适体(aptamer)为例,利用适体和核酸外切酶特性,通过定量PCR扩增建立一种高灵敏的蛋白质检测方法.首先合成3段寡核苷酸序列即凝血酶适体探针,上游连接子和下游连接子.将适体探针与凝血酶温育结合后,再加入核酸外切酶I降解未能结合的探针.接着将保护下来的探针与连接子杂交、连接和对连接产物进行定量PCR .分别建立连接产物标准品浓度与Ct 值的标准曲线和凝血酶浓度与连接产物浓度的标准曲线,通过定量PCR对凝血酶进行定量.结果显示,基于适体的外切酶保护凝血酶检测方法灵敏度较高,连接产物标准品浓度的对数值和Ct 值之间的方程为y =- 2 95x + 33 6 5 (R2 =0. 990 ,P <0 .0 1) ;凝血酶浓度和连接产物浓度对数值之间的方程为y =0 94x - 0 . 2 9(R2 =0 . 998,P <0 . 0 1) ,还对可能影响检测的有关参数举行了探讨.     

基于适体技术的桔青毒素检测方法的建立

桔青毒素（citrinin,CTN）是以玉米、谷物、奶酪等为主要成分的食品和动物饲料中常见的、由桔青霉菌产生的酮类真菌毒素,可引起人和动物的慢性中毒或癌症,一直缺少灵敏的快速检测方法。本文通过指数富集适体系统进化技术（简称SELEX）对可能与CTN结合的特异性适体进行了筛选,经过15轮循环,得到17条适体。通过二级结构分析、亲和力检测发现适体13（Apt-13）对CTN有较好的亲和度,解离常数Kd为0.06μmol/L。进一步利用非荧光标记染料PicoGreen,利用其与双链DNA结合的原理,建立了桔青毒素非标记荧光检测方法,30min完成检测,最低检测限达到国家标准（50ppb）且与其他毒素无交叉反应。本研究建立的基于适体的桔青毒素检测技术成本低,可以替代传统的基于抗体的检测方法,为霉菌毒素的精准检测技术的开发提供了实验证据。     

Gold Nanoparticle Based FRET for DNA Detection

The nanoscience revolution that sprouted throughout the 1990s is having great impact in current and future DNA detection technology around the world. In this review, we report our recent progress on gold nanoparticle based fluorescence resonance energy transfer assay to monitor DNA hybridization as well as the cleavage of DNA by nucleases. We tried to discuss a reasonable account of the science and the important fundamental work carried out in this area. We also report the development of a compact, highly specific, inexpensive and user-friendly optical fiber laser-induced fluorescence sensor based on fluorescence quenching by nanoparticles to detect single-strand DNA hybridization at femtomolar level.     

DNA Logic Gate Based on Metallo-Toehold Strand Displacement

DNA is increasingly being used as an ideal material for the construction of nanoscale structures, circuits, and machines. Toehold-mediated DNA strand displacement reactions play a very important role in these enzyme-free constructions. In this study, the concept of metallo-toehold was utilized to further develop a mechanism for strand displacement driven by Ag⁺ ions, in which the intercalation of cytosine–cytosine mismatched base pairs on the toeholds provides additional control by varying of the concentration of Ag⁺ ions. The characteristics of displacement reaction in response to different concentration of Ag⁺ ions are investigated by fluorescence spectral and non-denaturing polyacrylamide gel electrophoresis. The reaction can successfully occur when the concentration of Ag⁺ ions is suitabe; excess Ag⁺ ions block the reaction. Furthermore, the displacement reaction can be tuned and controlled most efficiently under the condition of two C:C mismatched base pairs placed on the six-nt toehold. Based on our research, a mechanism was developed to construct Boolean logic gate AND and OR by employing strand displacement reaction as a tool, Ag⁺ and Hg²⁺ as input.     

Aptamer DNA: A New Type of Thrombin Inhibitors

The formation of complexes between various thrombin preparations and 30-mer aptamer DNA was comparatively studied, and a correlation between the complex formation and the fibrinogen-hydrolyzing activity of thrombin was found. The aptamer DNA was shown to inhibit the formation of fibrin from fibrinogen.     

Logic Gate Operation by DNA Translocation through Biological Nanopores

Logical operations using biological molecules, such as DNA computing or programmable diagnosis using DNA, have recently received attention. Challenges remain with respect to the development of such systems, including label-free output detection and the rapidity of operation. Here, we propose integration of biological nanopores with DNA molecules for development of a logical operating system. We configured outputs “1” and “0” as single-stranded DNA (ssDNA) that is or is not translocated through a nanopore; unlabeled DNA was detected electrically. A negative-AND (NAND) operation was successfully conducted within approximately 10 min, which is rapid compared with previous studies using unlabeled DNA. In addition, this operation was executed in a four-droplet network. DNA molecules and associated information were transferred among droplets via biological nanopores. This system would facilitate linking of molecules and electronic interfaces. Thus, it could be applied to molecular robotics, genetic engineering, and even medical diagnosis and treatment.     

基于信号放大技术的适体生物传感器研究进展

信号放大技术因其能实现低浓度分子检测,灵敏度高而在多个研究领域发展非常迅速。而适体作为识别分子已成功应用于多种生物传感器平台,在医疗诊断、环境检测、生化分析中显示出良好的应用前景。近年来,以适体为识别元件的生物传感器越来越受到人们的关注。综述了近3年来基于信号放大技术的适体生物传感器研究新发展。     

SELEX技术与适体

SELEX技术是从随机寡核苷酸评议库中筛选与靶分子特异结合序列的组合化学技术。随机寡核苷酸的多种空间结构是筛选基础。该技术对靶分子无特殊要求,筛选出的寡核苷酸被称为适体,适体最突出的特征是与靶分子的特异性高亲和力。     

与非门(NAND)的DNA计算模型

近年来,随着DNA计算研究的深入,基于DNA的布尔电路的模拟成为其中一个热门的研究方向.分子信标是一种特殊的探针分子,广泛应用于各种生物技术的检测方面,具有结果稳定,特异性强的优点,本文提出了一种基于诱导"发夹"形式的DNA与非门模型,和已有的模型相比,该模型具有简单,可靠性更高且可以重复使用等优点.     

A DNA Aptamer Based Method for Detection of SARS-CoV-2 Nucleocapsid Protein

Altogether, we have identified a novel method for the detection of SRAS-CoV-2 N protein using DNA based aptamers. Although the aptamers used in this study were designed based on an aptamer previously selected for SARS-CoV N protein, they bind to SRAS-CoV-2 N protein with a high affinity. Most importantly, SARS-CoV-2 N protein shares very low similarity (16%–38%) with N protein from other five known human coronaviruses except for SARS-CoV. Because there are no SARS-CoV cases reported since 2004, our aptamer-based method can be used as a supplementation to the current diagnosis of SARS-CoV-2.