原子力显微镜观测DNA分子的表面沉积与扩展

原子力显微镜具有原子级分辨率,能够对生物样品进行观察.用原子力显微镜观察了DNA分子在Si、云母片及修饰过的云母片表面的沉积与扩展,对3种情况作了比较,用超声振荡方法可以有效地切断DNA分子链.     

使用碳纳米管AFM针尖的蛋白质高分辨率成像

原子力显微镜(AFM)是分析生物分子结构的有效手段,而目前使用的探针针尖的性质限制了高分辨率图像的获得。该文将碳纳米管安装到原子力显微镜的传统针尖上,制作出碳纳米管针尖以解决这个问题。运用碳纳米管针尖在大气常温条件下获得了由3个单元组成的小鼠抗体IgG蛋白质的Y形结构,并且分子的尺寸与X射线晶体衍射的结果非常接近,这种效果用传统针尖是无法获得的。获得的蛋白质分子超微结构的高分辨率图像为研究蛋白质分子功能提供了有价值的信息。     

Height measurement of DNA molecules with lift mode AFM

Atomic force microscopy (AFM) can be used to im-age and study the local properties of the sample surfacethrough tip/sample interactions. There are three operationmodes: contact mode, non-contact mode and tappingmode. Mostly, the measured heights of dsDNA were only0.7—0.8 nm[1—8] in air by tapping mode atomic force mi-croscopy (TMAFM), which were much less than the gen-erally accepted diameter of 2.0—2.2 nm[9]. In TMAFM, stable imaging is available when the tip is closer to sample surf…     

Local conformation transitions of linear DNA induced by cisplatin

Cisplatin is the most successful anti-tumor drug, and its pharmacological property is generally considered to derive from the modification of DNA molecules. Structural modifications of short DNA induced by cisplatin have already been investigated. However, the conformation transitions induced by cisplatin are not clear. In the present letter, we have studied the effect of low-concentration cisplatin on DNA conformation by using AFM imaging. We observed formations of micro-rod structures of linear DNA induced by cisplatin. A method is presented to quantitatively analyze the occurrence of micro-rod structures. We found that the formation of micro-rod structures depends on the DNA sequence. Based on the results, we proposed a physical mechanism to explain the local conformation transitions of DNA molecules under the influence of cisplatin.     

二氧化钛制备、表面特性及光催化的研究

以纳米二氧化钛薄膜为研究对象,用两种不同的方案制备出不同的样品,用原子力显微镜分别对样品表面微观形貌的直观的三维结构信息,及样品表面在纳米尺度上表现出来的物理、化学性质,以及光催化甲基蓝等方面进行测试比较。结果表明:AFM非常适合用于研究纳米尺度上样品表面形貌的结构,水体系制备TiO_2薄膜光催化活性比用醇体系所得薄膜强。     

Non-viral gene delivery carrier of probe type host molecule ——Interactions between DNA and β-cyclodextrin derivative complexes (I)

GENE THERAPY IS A COMMON PROCESS TO DELIVER EXTRIN- SIC GENES INTO TARGET CELLS FOR FURTHER GENE EXPRESSION, FOR THE PURPOSE OF TREATING DISEASES. A WELL DESIGNED GENE DELIVERY CARRIER CAN EFFICIENTLY PACKAGE AND PRO- TECT NUCLEIC ACID FROM BEING DIGESTED BY A VARIETY OF ENZYMES IN VIVO AND SHOULD BE ABLE TO SPECIFICALLY LO- CATE EXTRINSIC GENES INTO THE TARGET ORGANISMS. S…     

DNA bending induced by cruciform formation

Cruciform structures in DNA are of considerable interest, both as extreme examples of sequence-dependent structural heterogeneity and as models for four-way junctions such as the Holliday junction of homologous genetic recombination. Cruciforms are of lower thermodynamic stability than regular duplex DNA, and have been observed only in negatively supercoiled molecules, where the unfavourable free energy of formation is offset by the topological relaxation of the torsionally stressed molecule. From an experimental viewpoint this can be a disadvantage, as cruciform structures can be studied only in relatively large supercoiled DNA circles, and are destabilized when a break is introduced at any point. We therefore set out to construct a pseudo-cruciform junction--by generating hereroduplex formation between two inverted repeat sequences. Stereochemically, this should closely resemble a true cruciform but remain stable in a linear DNA fragment. We have now created such a junction and find that it has the expected sensitivities to endonucleases. These DNA fragments exhibit extremely anomalous gel electrophoretic mobility, the extent of which depends on the relative position of the pseudo-cruciform along the length of the molecule. Our results are very similar to those obtained by Wu and Crothers using kinetoplast DNA, and we conclude that the pseudo-cruciform junction introduces a bend in the linear DNA molecule.     

用AFM观察小白鼠心肌核DNA片段的基因体外表达和垃圾DNA的相互作用

用AFM直接现场观察、体外表达等实验技术组合,观察到小白鼠（Balb/c）心肌核DNA片段的基因在体外表达过程中形成的n mRNA（n=9）线型链状复合体,处于垃圾DNA片段的特定的“翻译平台”上,其每种mRNA两端非共价键分别结合自己编码蛋白质（即分子开关）,中间的编码序列均非共价结合完全可解离的翻译活性因子等多种蛋白质：这些蛋白质可能均由垃圾DNA片段的极复杂的立体结构所形成的、匹配协同的、专一性蛋白质通路所调控,该通路对蛋白质按顺序分别进行特异性双向调控.核内n mRNA线型链状复合体在体外可翻译出LDH等蛋白质,并显示n mRNA翻译的“群体效应”.用AFM还观察到胞质制取的n mRNA（n=12）线型链状复合体（无垃圾DNA存在）,体外翻译出少量LDH等蛋白质,并显示n mRNA翻译的＂群体效应＂.本工作展示了未来运用AFM观察体外表达等生物学反应,研究基因表达与调控机制及其与垃圾DNA相互作用的前景.     

Analogic China map constructed by DNA

In this research,a nanoscale DNA structure of analogic China map is created. The nanostructure of roughly 150 nm in diameter with a spatial resolution of 6 nm is purely constructed by folding DNA. The picture observed by atomic force microscopy (AFM) is almost identical with the de-signed shape. The DNA origami technology invented by Rothemund in 2006 is employed in the construc-tion of this shape,which has proved the capability of constructing almost any complicated shape enabled by DNA origami,and provides new bottom-up method for constructing nanostructures.     

用构造专用接口方法实现AUTOCAD与CAPP系统联接

在ＡｕｔｏＣＡＤ１１．０版实体造型模块的基础上，开发了一个用于箱体类零件的特征造型系统ＡＦＭ，并通过ＡＦＭ生成的特征数据文件进行数据交换，实现了ＡｕｔｏＣＡＤ与ＴＪＥＸＣＡＰＰ系统的联接，本文介绍了该系统总体结构，并对系统开发中的主要问题进行了讨论。     

3D DNA origami designed with caDNAno

For about three decades, DNA-based nanotechnology has been undergoing development as an assembly method for nanostructured materials. The DNA origami method pioneered by Rothemund paved the way for the formation of 3D structures using DNA self assembly. The origami approach uses a long scaffold strand as the input for the self assembly of a few hundred staple strands into desired shapes. Herein, we present a 3D origami "roller" (75 nm in length) designed using caDNAno software. This has the potential to be used as a template to assemble nanoparticles into different pre-defined shapes. The "roller" was characterized with agarose gel electrophoresis, atomic force microscopy (AFM) and transmission electron microscopy (TEM).     

AFM观察小白鼠心肌和肝核DNA片段的基因移位

用原子力显微镜(简称AFM)直接观察、体外表达和体外转录等实验技术组合,观察到了心肌和肝的核DNA片段的基因;用磷酸缓冲液稀释,并用开关蛋白质等活性因子使其部分解离,促使核基因在核DNA片段内或核DNA片段间静态或动态移位,得到了对应核DNA片段中的相关基因,如LDH/DNA体外表达活性变化的LDH同功酶酶谱图。基因静态或动态移位均表达活性降低,且基因移位程度与其对应基因活性降低程度呈正相关性。展示了未来运用AFM和体外表达等实验技术组合研究核DNA片段的基因移位和对应基因突变机制的前景。     

Spintronic materials and devices based on antiferromagnetic metals

In this paper, we review our recent experimental developments on antiferromagnet (AFM) spintronics mainly comprising Mn-based noncollinear AFM metals. IrMn-based tunnel junctions and Hall devices have been investigated to explore the manipulation of AFM moments by magnetic fields, ferromagnetic materials and electric fields. Room-temperature tunneling anisotropic magnetoresistance based on IrMn as well as FeMn has been successfully achieved, and electrical control of the AFM exchange spring is realized by adopting ionic liquid. In addition, promising spin-orbit effects in AFM as well as spin transfer via AFM spin waves reported by different groups have also been reviewed, indicating that the AFM can serve as an efficient spin current source. To explore the crucial role of AFM acting as efficient generators, transmitters, and detectors of spin currents is an emerging topic in the field of magnetism today. AFM metals are now ready to join the rapidly developing fields of basic and applied spintronics, enriching this area of solid-state physics and microelectronics.     

非特异性PCR产物的分离和扩增

在PCR（polymerase chain reaction)扩增产物的非特异性电泳带位置上及弥散状背景的一定间隔位置上，分别以切胶分离相应分子量的DNA分子作为模板，用产生这些非特异性DNA分子的一种引物在通用PCR条件下进行PCR扩增，仅在切胶的相应位置出现DNA分子量同样大小的电泳条带。阐述了这种模板DNA分子形成和出现非特异性电泳带与弥散状背景之间的关系。     

Enhancement of transfection efficiency for HeLa cells via incorporating arginine moiety into chitosan

Arginine-rich peptides have attracted considerable attention due to their distinct internalization mechanism. It was reported that arginine and guanidino moieties were able to translocate through cell membranes and played a critical role in the process of membrane permeation. In this work, arginine was conjugated to the backbone of chitosan to form a novel chitosan derivative, arginine modified chitosan (Arg-CS). Arg-CS/DNA complexes were prepared according to the method of coacervation process. The physicochemical properties of Arg-CS and Arg-CS/DNA complexes were characterized and the transfection activity and efficiency mediated by Arg-CS/DNA complexes were investigated taking HeLa cells as target cells. Arg-CS was characterized by FTIR and 13C NMR. Arg-CS/DNA polye- lectrolyte complexes were investigated by agarose gel retardation, dynamic light scattering (DLS) and atomic force microscopy (AFM). The results revealed that the Arg-CS/DNA complexes started to form at N/P ratio of 2:1, and the size of particles varied from 100 to 180 nm. The cytotoxicity of Arg-CS and their complexes with plasmid DNA were determined by MTT assay for HeLa cells, and the results suggested that Arg-CS/DNA complexes were slightly less toxic than Arg-CS. Moreover, the derivative alone and their complexes showed significantly lower toxicity than PEI and PEI/DNA complexes, respectively. Taking HeLa cells as target cells and using pGL3-control as reporter gene, the luciferase expression mediated by Arg-CS was greatly enhanced to about 100 folds compared with the luciferase expression mediated by chitosan at different pH media. These results suggest that Arg-CS is a promising candi- date as a safe and efficient vector for gene delivery and transfection.     

Spermidine-induced two-dimensional DNA condensations on mica surfaces: A different pathway from condensations in solution

With atomic force microscopy （AFM） we systematically studied the DNA condensations on mica surfaces induced by multivalent cation spermidine. The pattern of the DNA condensates is a flat single layer, with a core in the centre and DNA wrapping around it at high density. We assume this to be a two-dimensional condensation of free coiled DNA onto negatively charged mica surfaces by the multivalent cation. The DNA molecules condense on mica surfaces via a pathway different from the formation of toroids, rods or globules in bulk solutions. We give an explanation to why toroid structures are difficult to be observed by AFM, and further discuss the relationship between DNA condensations in solutions and on mica surfaces. The present work will be helpful for understanding the behaviors of DNA on charged surfaces, which might be significantly different from that in solutions.     

A Study on the Molecular Switch of Gene Expression of the Mouse Heart Nuclear DNA Fragments

It is observed by in situ stain that LDH(1-5)…nNAD+ can probably enter the nucleopore and can be bound bound specifically with the genes that encode them. During the in vitro expression, the dilution of heart nuclear DNA fragments could enhance the expression activity of LDH/DNA and the amount of expressed LDH(1-5) is in proportion to the amount of dissociable LDH(1-5) on the LDH/DNA. With the integration of 14CLeu to the proteins, it is also observed that the addition of LDH(1-5)…nNAD+ can suppress the in vitro expression activity of LDH/DNA. AFM bservation shows that the regulation sequence at the both ends of active genes may be bound with such active factors as proteins encoded by the genes which probably is the main molecular switch of gene expression and regulation we have been always searching for. Our work shows the prospective application of the combination of AFM and isotope labeling in the research of biological reaction.     

壳聚糖与DNA相互作用的单分子方法研究

利用原子力显微镜与磁镊技术研究不同浓度的壳聚糖与多种DNA分子的作用,对其凝聚形态及力谱曲线进行观察。在恒力情况下,壳聚糖与DNA的电荷比k分别为3和4时,从DNA的力谱曲线上可以看见0.2~0.5μm左右的跳跃阶梯及线性增长区域,原因可能是壳聚糖-DNA聚合物中出现了环状、棒状及球状结构;利用原子力显微镜观察壳聚糖与λ-DNA及bpr322-DNA作用后的凝聚形态,发现随着壳聚糖与DNA电荷比k的增加,球状与棒状聚合物的比例明显减小,而且凝聚物的尺寸也整体减小。最后根据实验结果,建立了壳聚糖导致DNA构象变化的直观模型。