我们非常感谢中国国家自然科学基金（21205078）和中国高等教育博士研究基金（No.20123120110002）的支持。这项工作得到了中国国家重点研究发展计划（2016YFB1102303），中国国家基础研究计划（973计划，2015CB352001）和国家自然科学基金（61378060）的部分支持。

SGE芯片的基本设计<ol><li>使用任何透明塑料，如聚甲基丙烯酸甲酯（PMMA）或聚碳酸酯。 注:SGE芯片如图1B所示 。 SGE芯片由用于TBE缓冲器的圆柱形孔，用于DNA分离的通道和沿着电极孔嵌入的两条通道组成。 </li><li>使用激光雕刻机在PMMA块中制造SGE通道阵列。 注意:芯片的几何参数取决于要分离的DNA的大小。例如，如果DNA片段小于1,000bp，则SGE芯片的最佳信道参数为2.5mm×4.0mm×90mm（宽×深×长）。 </li><li>基于SGE设计，制造梳子以在芯片中创建用于加载DNA样品的孔。 </li></ol> 2.琼脂糖凝胶的制备<ol><li>通过混合10×TBE（1×TBE =89mM Tris，89mM硼酸和2mM EDTA; pH 8.4）和蒸馏水以1:19的比例。 </li><li>制备1.0％琼脂糖凝胶，分离100-bp DNA梯。 注意:0.5x TBE缓冲液中琼脂糖的浓度取决于要分离的DNA片段的大小。 </li><li>将0.1 g琼脂糖放入烧瓶中，加入10 mL 0.5x TBE缓冲液。 注意:制备溶液的体积应小于烧瓶容量的1/3。 </li><li>用防腐膜密封烧瓶，然后在微波（培养基，1.0分钟）中加热琼脂糖/缓冲液混合物。在将烧瓶放入微波炉之前，在爆炸的情况下，在防腐膜中形成一些孔。 </li><li>将2.7mL熔融的琼脂糖溶液倒入SGE芯片的4个通道。将梳子放入琼脂糖溶液中以形成孔。 注意:3分钟后，熔化的琼脂糖溶液将在室温下冷却至凝胶状态。 </li><li>从g去除梳子并加入0.9mL 0.5x TBE缓冲液以覆盖凝胶。 </li></ol> 3.在SGE芯片中运行电泳<ol><li>打开LED光源，并在光源上方放置一条425至505 nm的带通滤光片。 </li><li>使用涡旋器混合14.4μL的100bp DNA梯和1.6μL的SYBR Green。 </li><li>在SGE芯片的琼脂糖凝胶的每个孔中加入4μL混合物（ 图2 ）。 </li><li>将电极放入SGE芯片的两条通道。 </li><li>在SGE芯片上方放置550至700 nm带通滤波器。 </li><li>打开电源。将电压设置为180 V（20 V / cm）。打开智能手机记录DNA片段分离过程（ 图3 ）。 </li><li>电泳10分钟后，关掉电源和LED灯。 </li><li>清洁SGE芯片。 </li></ol>

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没有宣布利益冲突。

琼脂糖凝胶电泳广泛用于分离DNA，RNA和蛋白质。这项工作提出了一种替代传统凝胶电泳方法的新方法。结果表明，在这样一个小的组装装置中，50,100和1,000bp的DNA梯子可以很好地分离。该方法的优点在于不仅可以将化学消耗少的核酸分开，还可以记录分离过程。尽管DNA片段在图2中看起来很宽，但可以通过优化SGE芯片的参数和琼脂糖凝胶的浓度来改善结果。此外，如果在芯片?...

<html>板状凝胶电泳（SGE）是DNA片段分离最有效的方法1,2,3,4,5 ，因此被认为是生物化学和生物学分析中的通用工具6,7,8。然而，许多实验表明，SGE受到以下四个问题的限制:（1）分离需要很多小时甚至几天; （2）化学品消耗量非常高; （3）需要复杂的装置（ 例如， 2D电泳池，电泳电源和凝胶成像系统）; （4）实验完成后，凝胶成像系统只能观察分离的DNA片段。此外，SGE 9中通常使用的溴化乙锭（EtBr）ass ="xref"&gt; 10，是致突变性和致癌性11,12 。因此，在装载含有EtBr的凝胶时，应始终佩戴手套。 与SGE相比，毛细管电泳（CE）具有许多优点，如自动操作，分离时间短，消耗量少等优点，13，14，15，16，17。然而，CE仪器相当昂贵。因此，为了克服这些限制，已经开发了一种用于分离DNA的系统（ 图1 ）。这样的系统不仅可以大大降低化学消耗，还可以节省SGE实验时间（&lt;8分钟），而且可以通过智能手机对琼脂糖凝胶中的DNA分离过程进行实时跟踪。按照本协议中描述的程序，学生们可以可以设计和制造SGE芯片，在芯片中制备琼脂糖凝胶，用智能手机设置简单的SGE系统，并将DNA迁移过程记录在琼脂糖凝胶中。

<table border="0" cellpadding="0" cellspacing="0" width="683">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">10&#215;TBE</td>
			<td style="width:287px;">Beijing Solarbio Science &#38; Technology Co., Ltd.</td>
			<td style="width:181px;">T1051</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">50bp DNA ladder</td>
			<td style="width:287px;">Takara Bio Inc.</td>
			<td style="width:181px;">3421A</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">100bp DNA ladder</td>
			<td style="width:287px;">Takara Bio Inc.</td>
			<td style="width:181px;">3422A</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">1kbp DNA ladder</td>
			<td style="width:287px;">Takara Bio Inc.</td>
			<td style="width:181px;">3426A</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">SYBR GREEN</td>
			<td style="width:287px;">Takara Bio Inc.</td>
			<td style="width:181px;">5760A</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Agarose</td>
			<td style="width:287px;">Sigma-Aldrich Corporate</td>
			<td style="width:181px;">V900510</td>
		</tr>
	</tbody>
</table>

real-time tracking of dna fragment separation by smartphone

板状凝胶电泳（SGE）是分离DNA片段的最常用方法;因此，它广泛应用于生物学等领域。然而，传统的SGE协议相当乏味，实验需要很长时间。此外，SGE实验中的化学消耗量非常高。这项工作提出了一种基于SGE芯片分离DNA片段的简单方法。该芯片由雕刻机制成。两个塑料片用于光信号的激发和发射波长。通过智能手机收集DNA条带的荧光信号。为了验证该方法，分离出50,100和1,000bp DNA梯。结果表明，使用该方法可以在12分钟内解析出小于5,000bp的DNA梯度，分辨率高，表明它是传统SGE方法的理想替代品。

<html> 图4 ， 图5和图6代表了在50,100和1,000bp DNA梯级的凝胶电泳之后的典型结果。实验后，DNA片段分离良好。此外，在SGE芯片的4个通道中分离相同的样品，显示出相同尺寸的DNA片段在每个实验中移动相同的距离。 DNA梯度的分离性能可以通过分析迁移距离与DNA大小对数的关系进行评估;因此，SGE可用于计算未知DNA...

Watch this Scientific Journal Video about Real-time Tracking of DNA Fragment Separation by Smartphone at JoVE.com

Real-time Tracking of DNA Fragment Separation by Smartphone

通过智能手机实时跟踪DNA片段分离

传统的平板凝胶电泳（SGE）实验需要复杂的装置和高的化学消耗。这项工作提出了一个协议，描述了一种在短时间内分离DNA片段的低成本方法。

Slab gel electrophoresis (SGE) is the most common method for the separation of DNA fragments; thus, it is broadly applied to the field of biology and ...

real-time-tracking-of-dna-fragment-separation-by-smartphone

Research

JoVE Journal

Biochemistry

14.4K 次观看.  University of Shanghai for Science and Technology. 传统的平板凝胶电泳（SGE）实验需要复杂的装置和高的化学消耗。这项工作提出了一个协议，描述了一种在短时间内分离DNA片段的低成本方法。 

视频: Real-time Tracking of DNA Fragment Separation by Smartphone

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis

For any enzyme, robust, quantitative methods are required for characterization of both native and engineered enzymes. For DNA polymerases, DNA synthesis can be characterized using an in vitro DNA synthesis assay followed by polyacrylamide gel electrophoresis. The goal of this assay is to quantify synthesis of both natural DNA and modified DNA (M-DNA). These approaches are particularly useful for resolving oligonucleotides with single nucleotide resolution, enabling observation of individual steps during enzymatic oligonucleotide synthesis. These methods have been applied to the evaluation of an array of biochemical and biophysical properties such as the measurement of steady-state rate constants of individual steps of DNA synthesis, the error rate of DNA synthesis, and DNA binding affinity. By using modified components including, but not limited to, modified nucleoside triphosphates (NTP), M-DNA, and/or mutant DNA polymerases, the relative utility of substrate-DNA polymerase pairs can be effectively evaluated. Here, we detail the assay itself, including the changes that must be made to accommodate nontraditional primer DNA labeling strategies such as near-infrared fluorescently labeled DNA. Additionally, we have detailed crucial technical steps for acrylamide gel pouring and running, which can often be technically challenging.

This protocol describes the characterization of DNA polymerase synthesis of modified DNA through observation of changes to near-infrared fluorescently labeled DNA using gel electrophoresis and gel imaging. Acrylamide gels are used for high resolution imaging of the separation of short nucleic acids, which migrate at different rates depending on size.

用丙烯酰胺凝胶电泳法可视化近红外荧光标记 dna 聚合酶活性

For any enzyme, robust, quantitative methods are required for characterization of both native and engineered enzymes. For DNA polymerases, DNA synthesis ...

科研

生物化学

Regeneration of Arrayed Gold Microelectrodes Equipped for a Real-Time Cell Analyzer

The label-free cell-based assay is advantageous for biochemical study because of it does not require the use of experimental animals. Due to its ability to provide more dynamic information about cells under physiological conditions than classical biochemical assays, this label-free real-time cell assay based on the electric impedance principle is attracting more attention during the past decade. However, its practical utilization may be limited due to the relatively expensive cost of measurement, in which costly consumable disposable gold microchips are used for the cell analyzer. In this protocol, we have developed a general strategy to regenerate arrayed gold microelectrodes equipped for a commercial label-free cell analyzer. The regeneration process includes trypsin digestion, rinsing with ethanol and water, and a spinning step. The proposed method has been tested and shown to be effective for the regeneration and repeated usage of commercial electronic plates at least three times, which will help researchers save on the high running cost of real-time cell assays.

This protocol describes a general strategy to regenerate commercial arrayed gold microelectrodes equipped for a label-free cell analyzer aimed at saving on the high running costs ofmicrochip-based assays. The regeneration process includes trypsin digestion, rinsing with ethanol and water, and a spinning step, which enables repeated usage of microchips.

用于实时细胞分析仪的阵列金电极再生

The label-free cell-based assay is advantageous for biochemical study because of it does not require the use of experimental animals. Due to its ability ...

Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51

The DNA strand exchange reaction mediated by Rad51 is a critical step of homologous recombination. In this reaction, Rad51 forms a nucleoprotein filament on single-stranded DNA (ssDNA) and captures double-stranded DNA (dsDNA) non-specifically to interrogate it for a homologous sequence. After encountering homology, Rad51 catalyzes DNA strand exchange to mediate pairing of the ssDNA with the complementary strand of the dsDNA. This reaction is highly regulated by numerous accessary proteins in vivo. Although conventional biochemical assays have been successfully employed to examine the role of such accessory protein in vitro, kinetic analysis of intermediate formation and its progression into a final product has proven challenging due to the unstable and transient nature of the reaction intermediates. To observe these reaction steps directly in solution, fluorescence resonance energy transfer (FRET)-based real-time observation systems of this reaction were established. Kinetic analysis of real-time observations shows that the DNA strand exchange reaction mediated by Rad51 obeys a three-step reaction model involving the formation of a three-strand DNA intermediate, maturation of this intermediate, and the release of ssDNA from the mature intermediate. The Swi5-Sfr1 complex, an accessary protein conserved in eukaryotes, strongly enhances the second and third steps of this reaction. The FRET-based assays presented here enable us to uncover the molecular mechanisms through which recombination accessary proteins stimulate the DNA strand exchange activity of Rad51. The primary goal of this protocol is to enhance the repertoire of techniques available to researchers in the field of homologous recombination, particularly those working with proteins from species other than Schizosaccharomyces pombe, so that the evolutionary conservation of the findings presented herein can be determined.

Fluorescence resonance energy transfer-based real-time observation systems of the DNA strand exchange reaction mediated by Rad51 were developed. Using the protocols presented here, we are able to detect the formation of reaction intermediates and their conversion into products, while also analyzing the enzymatic kinetics of the reaction.

rad51 介导的 dna 链交换反应的实时观测

The DNA strand exchange reaction mediated by Rad51 is a critical step of homologous recombination. In this reaction, Rad51 forms a nucleoprotein filament ...

DNA Sequence Recognition by DNA Primase Using High-Throughput Primase Profiling

DNA primase synthesizes short RNA primers that initiate DNA synthesis of Okazaki fragments on the lagging strand by DNA polymerase during DNA replication. The binding of prokaryotic DnaG-like primases to DNA occurs at a specific trinucleotide recognition sequence. It is a pivotal step in the formation of Okazaki fragments. Conventional biochemical tools that are used to determine the DNA recognition sequence of DNA primase provide only limited information. Using a high-throughput microarray-based binding assay and consecutive biochemical analyses, it has been shown that 1) the specific binding context (flanking sequences of the recognition site) influences the binding strength of the DNA primase to its template DNA, and 2) stronger binding of primase to the DNA yields longer RNA primers, indicating higher processivity of the enzyme. This method combines PBM and primase activity assay and is designated as high-throughput primase profiling (HTPP), and it allows characterization of specific sequence recognition by DNA primase in unprecedented time and scalability.&#160;

Protein binding microarray (PBM) experiments combined with biochemical assays link the binding and catalytic properties of DNA primase, an enzyme that synthesizes RNA primers on template DNA. This method, designated as high-throughput primase profiling (HTPP), can be used to reveal DNA-binding patterns of a variety of enzymes.

DNA序列识别利用高通量原酶分析的DNAPrimase

DNA primase synthesizes short RNA primers that initiate DNA synthesis of Okazaki fragments on the lagging strand by DNA polymerase during DNA replication. ...

Real-Time, Semi-Automated Fluorescent Measurement of the Airway Surface Liquid pH of Primary Human Airway Epithelial Cells

In recent years, the importance of mucosal surface pH in the airways has been highlighted by its ability to regulate airway surface liquid (ASL) hydration, mucus viscosity and activity of antimicrobial peptides, key parameters involved in innate defense of the lungs. This is of primary relevance in the field of chronic respiratory diseases such as cystic fibrosis (CF) where these parameters are dysregulated. While different groups have studied ASL pH both in vivo and in vitro, their methods report a relatively wide range of ASL pH values and even contradictory findings regarding any pH differences between non-CF and CF cells. Furthermore, their protocols do not always provide enough details in order to ensure reproducibility, most are low throughput and require expensive equipment or specialized knowledge to implement, making them difficult to establish in most labs. Here we describe a semi-automated fluorescent plate reader assay that enables the real-time measurement of ASL pH under thin film conditions that more closely resemble the in vivo situation. This technique allows for stable measurements for many hours from multiple airway cultures simultaneously and, importantly, dynamic changes in ASL pH in response to agonists and inhibitors can be monitored. To achieve this, the ASL of fully differentiated primary human airway epithelial cells (hAECs) are stained overnight with a pH-sensitive dye in order to allow for the reabsorption of the excess fluid to ensure thin film conditions. After fluorescence is monitored in the presence or absence of agonists, pH calibration is performed in situ to correct for volume and dye concentration. The method described provides the required controls to make stable and reproducible ASL pH measurements, which ultimately could be used as a drug discovery platform for personalized medicine, as well as adapted to other epithelial tissues and experimental conditions, such as inflammatory and/or host-pathogen models.

We present a protocol to make dynamic measurements of the airway surface liquid pH under thin film conditions using a plate-reader.

一次人气道上皮细胞气道表面液体 pH 值的实时、半自动荧光测量

In recent years, the importance of mucosal surface pH in the airways has been highlighted by its ability to regulate airway surface liquid (ASL) ...

Monitoring GPCR-&#946;-arrestin1/2 Interactions in Real Time Living Systems to Accelerate Drug Discovery

Interactions between G-protein coupled receptors (GPCRs) and &#946;-arrestins are vital processes with physiological implications of great importance. Currently, the characterization of novel drugs towards their interactions with &#946;-arrestins and other cytosolic proteins is extremely valuable in the field of GPCR drug discovery particularly during the study of GPCR biased agonism. Here, we show the application of a novel structural complementation assay to accurately monitor receptor-&#946;-arrestin interactions in real time living systems. This method is simple, accurate and can be easily extended to any GPCR of interest and also it has the advantage that it overcomes unspecific interactions due to the presence of a low expression promoter present in each vector system. This structural complementation assay provides key features that allow an accurate and precise monitoring of receptor-&#946;-arrestin interactions, making it suitable in the study of biased agonism of any GPCR system as well as GPCR c-terminus &#8216;phosphorylation codes&#8217; written by different GPCR-kinases (GRKs) and post-translational modifications of arrestins that stabilize or destabilize the receptor-&#946;-arrestin complex.

GPCR-&#946;-arrestin interactions are an emerging field in GPCR drug discovery. Accurate, precise and easy to set up methods are necessary to monitor such interactions in living systems. We show a structural complementation assay to monitor GPCR-&#946;-arrestin interactions in real time living cells, and it can be extended to any GPCR.

监测GPCR-β-阻丁1/2实时生命系统中的相互作用,加速药物发现

Interactions between G-protein coupled receptors (GPCRs) and &#946;-arrestins are vital processes with physiological implications of great importance. ...

Measuring Nucleotide Binding to Intact, Functional Membrane Proteins in Real Time

We have developed a method to measure binding of adenine nucleotides to intact, functional transmembrane receptors in a cellular or membrane environment. This method combines expression of proteins tagged with the fluorescent non-canonical amino acid ANAP, and FRET between ANAP and fluorescent (trinitrophenyl) nucleotide derivatives. We present examples of nucleotide binding to ANAP-tagged KATP ion channels measured in unroofed plasma membranes and excised, inside-out membrane patches under voltage clamp. The latter allows for simultaneous measurements of ligand binding and channel current, a direct readout of protein function. Data treatment and analysis are discussed extensively, along with potential pitfalls and artefacts. This method provides rich mechanistic insights into the ligand-dependent gating of KATP channels and can readily be adapted to the study of other nucleotide-regulated proteins or any receptor for which a suitable fluorescent ligand can be identified.

This protocol presents a method for measuring adenine nucleotide binding to receptors in real time in a cellular environment. Binding is measured as F&#246;rster resonance energy transfer (FRET) between trinitrophenyl nucleotide derivatives and protein labeled with a non-canonical, fluorescent amino acid.

实时测量核苷酸与完整功能性膜蛋白的结合

We have developed a method to measure binding of adenine nucleotides to intact, functional transmembrane receptors in a cellular or membrane environment. ...

Enabling Real-Time Compensation in Fast Photochemical Oxidations of Proteins for the Determination of Protein Topography Changes

Fast photochemical oxidation of proteins (FPOP) is a mass spectrometry-based structural biology technique that probes the solvent-accessible surface area of proteins. This technique relies on the reaction of amino acid side chains with hydroxyl radicals freely diffusing in solution. FPOP generates these radicals in situ by laser photolysis of hydrogen peroxide, creating a burst of hydroxyl radicals that is depleted on the order of a microsecond. When these hydroxyl radicals react with a solvent-accessible amino acid side chain, the reaction products exhibit a mass shift that can be measured and quantified by mass spectrometry. Since the rate of reaction of an amino acid depends in part on the average solvent accessible surface of that amino acid, measured changes in the amount of oxidation of a given region of a protein can be directly correlated to changes in the solvent accessibility of that region between different conformations (e.g., ligand-bound versus ligand-free, monomer vs. aggregate, etc.) FPOP has been applied in a number of problems in biology, including protein-protein interactions, protein conformational changes, and protein-ligand binding. As the available concentration of hydroxyl radicals varies based on many experimental conditions in the FPOP experiment, it is important to monitor the effective radical dose to which the protein analyte is exposed. This monitoring is efficiently achieved by incorporating an inline dosimeter to measure the signal from the FPOP reaction, with laser fluence adjusted in real-time to achieve the desired amount of oxidation. With this compensation, changes in protein topography reflecting conformational changes, ligand-binding surfaces, and/or protein-protein interaction interfaces can be determined in heterogeneous samples using relatively low sample amounts.

Fast photochemical oxidation of proteins is an emerging technique for the structural characterization of proteins. Different solvent additives and ligands have varied hydroxyl radical scavenging properties. To compare the protein structure in different conditions, real-time compensation of hydroxyl radicals generated in the reaction is required to normalize reaction conditions.

实现蛋白质快速光化学氧化的实时补偿，用于确定蛋白质地形变化

Fast photochemical oxidation of proteins (FPOP) is a mass spectrometry-based structural biology technique that probes the solvent-accessible surface area ...

Workflow and Tools for Crystallographic Fragment Screening at the Helmholtz-Zentrum Berlin

Fragment screening is a technique that helps to identify promising starting points for ligand design. Given that crystals of the target protein are available and display reproducibly high-resolution X-ray diffraction properties, crystallography is among the most preferred methods for fragment screening because of its sensitivity. Additionally, it is the only method providing detailed 3D information of the binding mode of the fragment, which is vital for subsequent rational compound evolution. The routine use of the method depends on the availability of suitable fragment libraries, dedicated means to handle large numbers of samples, state-of-the-art synchrotron beamlines for fast diffraction measurements and largely automated solutions for the analysis of the results.
Here, the complete practical workflow and the included tools on how to conduct crystallographic fragment screening (CFS) at the Helmholtz-Zentrum Berlin (HZB) are presented. Preceding this workflow, crystal soaking conditions as well as data collection strategies are optimized for reproducible crystallographic experiments. Then, typically in a one to two-day procedure, a 96-membered CFS-focused library provided as dried ready-to-use plates is employed to soak 192 crystals, which are then flash-cooled individually. The final diffraction experiments can be performed within one day at the robot-mounting supported beamlines BL14.1 and BL14.2 at the BESSY&#160; II electron storage ring operated by the HZB in Berlin-Adlershof (Germany). Processing of the crystallographic data, refinement of the protein structures, and hit identification is fast and largely automated using specialized software pipelines on dedicated servers, requiring little user input.
Using the CFS workflow at the HZB enables routine screening experiments. It increases the chances for successful identification of fragment hits as starting points to develop more potent binders, useful for pharmacological or biochemical applications.

Crystallographic fragment screening at the Helmholtz-Zentrum Berlin is performed using a workflow with dedicated compound libraries, crystal handling tools, fast data collection facilities&#160;and largely automated data analysis. The presented protocol intends to maximize the output of such experiments to provide promising starting points for downstream structure-based ligand design.

赫尔姆霍尔茨-曾特鲁姆柏林水晶碎片筛选工作流程和工具

Fragment screening is a technique that helps to identify promising starting points for ligand design. Given that crystals of the target protein are ...

Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique

Chromatin is a higher-order structure that packages eukaryotic DNA. Chromatin undergoes dynamic alterations according to the cell cycle phase and in response to environmental stimuli. These changes are essential for genomic integrity, epigenetic regulation, and DNA metabolic reactions such as replication, transcription, and repair. Chromatin assembly is crucial for chromatin dynamics and is catalyzed by histone chaperones. Despite extensive studies, the mechanisms by which histone chaperones enable chromatin assembly remains elusive. Moreover, the global features of nucleosomes organized by histone chaperones are poorly understood. To address these problems, this work describes a unique single-molecule imaging technique named DNA curtain, which facilitates the investigation of the molecular details of nucleosome assembly by histone chaperones. DNA curtain is a hybrid technique that combines lipid fluidity, microfluidics, and total internal reflection fluorescence microscopy (TIRFM) to provide a universal platform for real-time imaging of diverse protein-DNA interactions.Using DNA curtain, the histone chaperone function of Abo1, the Schizosaccharomyces pombe bromodomain-containing AAA+ ATPase, is investigated, and the molecular mechanism underlying histone assembly of Abo1 is revealed. DNA curtain provides a unique approach for studying chromatin dynamics.

DNA curtain, a high-throughput single-molecule imaging technique, provides a platform for real-time visualization of diverse protein-DNA interactions. The present protocol utilizes the DNA curtain technique to investigate the biological role and molecular mechanism of Abo1, a Schizosaccharomyces pombe bromodomain-containing AAA+ ATPase.

DNA帘法破译组蛋白组装的分子机制

Chromatin is a higher-order structure that packages eukaryotic DNA. Chromatin undergoes dynamic alterations according to the cell cycle phase and in ...