Real time monitoring of nucleic acids ligation based on molecular beacon

The replication, ligation and repair of nucleic acids are essential life processes that based the survival, pro-creation and evolution of creatures. These interactions involving proteins (enzymes) and nucleic acids have fas-cinated great interest of scientists[1]. The nucleic acids ligation has been revealed that it is catalyzed by ligase and co-enzyme (ATP or NAD)[2—4]. Along with in-depth re-search, the nucleic acids ligation becomes a significant and common tool employed in bioengineerin…     

Synthesis and photo-activity of peptide nucleic acids containing an azobenzene unit

We prepared the azobenzene PNA unit and synthesized various PNAs containing an azobenzene unit by a typical tBoc method. Both the azobenzene PNA unit and PNA containing an azobenzene unit （N-PNA） showed reversible photoisomerization with UV and visible light irradiation. The hybridization of N-PNA with complementary DNA resulted in a considerable decrease of the photoisomerization rate constant of the azobenzene.     

Macromolecules Mimicking Backbones of Nucleic and Teichoic Acids.Synthesis,Some Properties and Applications

1 Results Several methods have been elaborated in this laboratory allowing preparation of macromolecules with phosphodiester bonds,and having sequence of atoms similar as in the chains of biomacromolecules - nucleic or teichoic acids (TA),namely:-(C)n-O-PO-,where n=2 (for teichoic acids) or 3.These methods,to be discussed in the lecture,are based on the ring-opening polymerization,transesterification,and recently elaborated direct addition of phosphoric acid to diepoxides.For the first time an attempt has been made to directly reacting phosphoric acid with diols in the presence and absence of catalysts.The major side reaction,preventing high molar mass polymers formation,results from dealkylation,i.e.nucleophilic attack of the OH groups on the C atom,instead attack on the P atom.The other two methods allowed reaching high molar mass macromolecules,with Mn up to 105.Following these studies phosphorylation of polymers with side groups containing -OH groups were studied and a series of diblock copolymers with ionic and nonionic blocks were prepared.     

Study on Marine Application Potentiality of CMODIS／SZ-3

The third spaceship SZ-3 of China was launched in March 2002, in which the payload is Chinese moderate imaging spectra radiometer (CMODIS) . In this paper, first, the properties and characteristics of CMODIS are briefly introduced; second, the quality and availahility are evaluated by means of the Complex signal noise ratio (CSNR) which is simulated theoretically； third, the received CMODIS data are compared with the Sea wide field-of-view sensor (SeaWiFS) / SeaSTAR data to understand the accuracy of radiance measurement by CMODIS;finally, the remote sensing products of ocean color and temperature are mapped by CMODIS to study its marine application potentiality. The results show that CMODIS has its latent capability for the application of marine environment detection, the management and protection of marine resources, and the national rights and interests. Meanwhile some suggestions are proposed to modify the next generation sensor on the Chinese spaceship.     

Editorial note for the special topic on ＂Nanopore Analysis＂

The concept of nanopore analysis, using the pore-forming protein a-hemolysin to detect individual nucleic acids at a single-molecule level, was first proposed in 1996. Over the past two decades, tremendous progress has been made in the nanopore field, and nanopore analysis has become a label-free and high-throughput method for probing bio- molecules and other analytes with single-molecule sensi- tivity, especially holds the promising for ＂third generation＂ DNA sequencing. However, challenges still remain in the experimental strategies and the design of whole nanopore-based instruments. Here, we proudly present a special topic dedicated to the topic of ＂Nanopore Analysis＂, with 8 reviews/articles providing up to date coverage of the experimental strategies, theoretical calcu- lations and simulations, and instrument design. Reviews and articles on the experimental strategies cover control of DNA partitioning into a nanopore, detection of target DNA, and the advantages of nanopore-based DNA sequencing. The theoretical calculations and simulations discuss the translocation behavior of DNA, and an inte- grated measurement system and data analysis software are presented for instrument design.     

Field experiment on a robust hierarchical metropolitan quantum cryptography network

A hierarchical metropolitan quantum cryptography network upon the inner-city commercial telecom fiber cables is reported in this paper. The seven-user network contains a four-node backbone net with one node acting as the subnet gateway, a two-user subnet and a single-fiber access link, which is realized by the Faraday-Michelson interferometer set-ups. The techniques of the quantum router, optical switch and trusted relay are assembled here to guarantee the feasibility and expandability of the quantum cryptography network. Five nodes of the network are located in the government departments and the secure keys generated by the quantum key distribution network are utilized to encrypt the instant video, sound, text messages and confidential files transmitting between these bureaus. The whole implementation including the hierarchical quantum cryptographic communication network links and the corresponding application software shows a big step toward the practical user-oriented network with a high security level.     

Analysis of components of conserved “backbone sequences”among genomes of Shigella spp. strains

Difference in the genomic compositions of prokaryotes is the basis of the diversity in their biological characters. However, besides their flora-or strain-specific genes,those floras with closer relationship in the evolution also have conserved “backbone sequences”, which reveal the marks of their origin and evolution, and these “backbone sequences”are just the basis of their elementary living abilities and common biological properties. Shigella is very closely related to E. coli in the origin and evolution, and may turn out to belong to the same genus. In this study, a microarray containing E. coli K-12 whole genome and Sf301 specific ORFs is used to investigate the genomic components of four Shigella strains. The results indicate that 16%-22% K-12 ORFs sequences are not detected in the genome of Shigella strains while the genome of Shigella contains at least 2800 conserved ORFs, which compose the common “backbone sequences”.Advanced analysis indicated that the “backbone sequences”are the essential components in maintaining the normal physiological activities of intestinal bacteria. Furthermore,only 20% Sf301-specific ORFs exist in other strains simultaneously, which demonstrate the great genome heterogeneity and the genetic diversity among the strains.     

Study and Application of Internal Coating Technique to Drag Reduction of the Trunk Pipeline for the West-East Gas Pipeline

Coverage layer coated in the internal wall of pipeline enables the friction drag to be reduced, the throughput and the gas transmission efficiency to be increased, the frequency of pigging and the number of the intermediate compressor station to be reduced, and the power consumption of the compressor to be decreased etc. The drag reduction is a high advanced scientific technique with outstanding economical benefit. The study and application of internal coating technique for drag reduction of 4000km trunk pipeline in West-East gas transmission pipeline (WEGTP) project are described, in which the main points are the drag reduction principle, coating process and the indoor study of this technique with own-decided knowledge property right at home.     

Coordination chemistry mimics of nuclease-activity in the hydrolytic cleavage of phosphodiester bond

In vivo, there occurs the synthesis and degradation of the nucleic acids, involving the formation and cleavage of phosphodiester bond. The cleavage modes of phosphodiester bond can be divided into two types, oxidative and hydrolytic, only the hydrolytic products are sticky and connectable, allowing to be religated by ligases. In recent years, great progresses have been made in chemical mimics in the hydrolytic cleavage of phosphodiester bond. Among the found metal complexes with the activity of nucleic acid cleavage, there are mononuclear and dinuclear metal complexes, the adopted metal ions including transitional and lanthanide metal ions. However, the reaction rates are still several orders less than those of the natural enzymes. It should be a prosperous and expedient direction to mimic the hydrolytic cleavage of phosphodiester bond by taking the advantages of dinuclear metal complexes.     

Ways to produce and measure atto- and femtosecond soft X-ray pulses

The ways to produce and measure atto-and femtosecond soft X-ray pulses are reported. The laser phase relation of high-order harmonic generation(HHG) shows two different radiation energy distribu-tions in time(or laser phase) domain. These energy-phase relations are helpful for realizing the dy-namic processes of HHG. Two presented parameterized formulas can be used to calculate the durations of the energy distributions with a bandwidth of the pulse. These formulas are useful in calculating and simulating pulses transports and interactions with mediums. The time structures of atto-and femto-second soft X-ray pulses can be directly measured with photoelectron spectrum transfer equations and the related laser phase determination methods without any previous pulse shape and the instantane-ous frequency assumptions. These equations and methods can be used to evaluate and improve the technical parameters of the ultra-short X-ray sources. They have wide measurement ranges and high time resolutions,which may enable ultra-fast measurements to reach metrological precisions,and lead to a new tide of scientific researches in physics,chemistry,biochemistry,etc. The application of atto-and femtosecond X-rays as well as the theoretical and technical problems in measurements are briefly discussed.     

肽核酸研究的现状与未来

肽核酸 (PNA)是DNA的类似物 ,具有由氨乙基甘氨酸单位组成的假肽骨架 ,能以高亲和性和高生物稳定性与DNA和RNA特异性杂交 .PNA以链侵占方式与DNA结合 ,抑制转录或提供人工强启动子促进转录 ;与mRNA结合 ,阻止蛋白质的合成 ;此外 ,PNA还能导向核糖核蛋白的RNA部分 ,抑制其酶活性 .肽核酸的其它特点如抗核酸酶和蛋白酶作用、方便而灵活的固相合成法 (SPS)等 ,使其有望成为一个基因靶向药物 .该文综述了近年有关肽核酸研究的生物化学特性及其应用前景     

抗－DNA免疫复合物性质的研究

用聚乙二醇（ＰＥＧ）和硫酸铵从系统性红斑狼疮（ＳＬＥ）患者血清中分离出免疫复合物（ＩＣ）和血浆核酸（ＰＮＡ）。分析结果表明，ＰＮＡ的主要成分是ＲＮＡ，仅有少量ＤＮＡ存在，并且ＰＮＡ中ＤＮＡ的ｄＧ＋ｄＣ含量（５５％）明显高于正常值（４２％）。一旦经过ＲＮａｓｅ处理，ＰＮＡ抑制抗－ＤＮＡ抗体结合ＤＮＡ的能力降低，说明抗－ＤＮＡ抗体和ＲＮＡ有交叉反应。动力学研究观察到，抗－ＤＮＡ免疫复合物中抗原抗体的分子比是１：１，结合能为３７ｋＪ／ｍｏｌ。这提示，除ＤＮＡ／抗－ＤＮＡ外，其他抗－ＤＮＡ免疫复合物也可能同ＳＬＥ患者的组织损伤有关。     

中氮茚衍生物-花菁离子缔合平衡体系在核酸测定中的应用

应用中氮茚衍生物-花菁离子缔合平衡体系建立了一种测定痕量核酸的方法.中氮茚衍生物的最大发射波长在385nm,在一定量花菁存在下,其荧光强度显著猝灭,核酸的加入又可使其荧光回复.在最佳条件下,荧光回升与核酸浓度成正比.标准曲线线性范围:小牛胸腺DNA(CTDNA)为2-500ng/mL;鱼精子DNA(FSDNA)为2-400ng/mL.相应的检测限:CTDNA为0.92ng/mL;FSDNA为0.85ng/mL.应用本法对3个实际样品进行了测定,结果令人满意.     

近红外分光光度法测定核酸

以阳离子型近红外花菁染料做为生色探针,研究了该染料与核酸的结合反应,在ＰＨ６．０的条件下,该染料在７７１ｎｍ处有最大吸收峰,在核酸存在下,其吸收峰显著下降,而在６４０ｎｍ处出现新的吸收峰。据此,以该近红外花菁染料为生色探针,在７７１ｎｍ处依其吸光度的下降可定量测定核酸。     

MIL-101-NH2介导的核酸分子递送与基因沉默研究

核酸是DNA和RNA的总称,能够参与生物体内基因表达的调控.然而,核酸分子极易被降解,如果直接通过口服或静脉注射给药,生物利用率极低,因此需要合适的载体进行核酸的递送.选取了生物兼容性较好的铁基金属-有机框架（MOF）MIL-101-NH₂,分别负载单链DNA和siRNA来探究其细胞学行为.MIL-101-NH₂能够有效递送单链DNA和siRNA进入细胞,且siRNA在细胞内能够发生溶酶体逃逸,并发挥基因沉默的效应.结果表明：基于MIL-101-NH₂的纳米复合物是一种具有潜力的核酸递送与基因调控策略.     

Formation of a stable triplex from a single DNA strand

Homopurine.homopyrimidine DNA sequences have been shown to form triple-stranded structures readily under appropriate conditions. Interest in DNA triplexes arises from potential applications of intermolecular triplexes as antisense inhibitors of gene expression and from the possibility that intramolecular triplexes may have a role in gene expression and recombination. We recently presented NMR evidence for triplex formation from the DNA oligonucleotides d(GA)4 and d(TC)4, which showed unambiguously that the second pyrimidine strand is Hoogsteen base paired and the cytosines are protonated at N3 as required. To obtain a more well defined triplex, and to provide a model for in vivo triplex structures, we have designed and synthesized a 28-base DNA oligomer with a sequence that could potentially fold to form a triplex containing both T.A.T and C+.G.C triplets. Our NMR results indicate that the conformation at pH 5.5 is an intramolecular triplex and that a significant amount of triplex remains even at pH 8.0.     

The hybridization between peptide nucleic acid containing azobenzene and DNA labeled nanoparticle on chip surfaces studied by atomic force microscopy

Peptide nucleic acid containing azobenzene （N-PNA） was covalently bound on chip surfaces. Complementary DNA probes labeled by gold nanoparticles hybridizated with PNA on chip surfaces, The hybridization was successfully detected with AFM by the microscopy mothed. The influences of the position and the cis-trans isomerization of azobenzene unit on hybridization were investigated with AFM.     

吡啰红Y共振光散射法测定纳克级核酸

研究了二苯并吡喃类染料吡口罗红Y(PY)与核酸作用的共振光散射光谱,在pH11.5～12.0Tris-NaOH缓冲溶液中,1.75×10-5mol/L的PY溶液加入核酸后,体系的共振光散射增强,在364.0nm处,存在一共振光散射增强峰,其增强强度(△IRLS=I-I0)与核酸浓度呈线性关系,据此建立了一种测定纳克级核酸的简便灵敏的方法.对于fsDNA,方法的线性范围为27.0～625ng/mL,检出限为8.1ng/mL;对于ctD NA,方法的线性范围为39.0～500ng/mL,检出限为11.7ng/mL;对于yRNA,方法的线性范围为59.0～375ng/mL,检出限为17.7ng/mL.建立的方法已用于六种合成样品的测定,结果令人满意.