DNA sequence at the end of IS1 required for transposition

The insertion sequence IS1 belongs to a class of bacterial transposable genetic elements that can form compound transposons in which two copies of IS1 flank an otherwise non-transposable segment of DNA. IS1 differs from other known elements of this class (such as IS10, IS50 and IS903) in several respects. It is one of the smallest known insertion elements, exhibits a relatively complex array of open reading frames, is present in the chromosomes of various Enterobacteria, in some cases in many copies, and its insertion can result in the duplication of either 8 or 9 base pairs (bp) in the target DNA. Furthermore, although, like other members of the compound class, it seems to undergo direct transposition, IS1 also promotes replicon fusion (co-integrate formation) at a relatively high frequency. Like all other elements studied to date, the integrity of the extremities of IS1 are essential for efficient transposition. We have constructed a test system to determine the minimal DNA sequences at the extremities of IS1 required for transposition. Sequential deletions of the end sequences reveal that 21-25 bp of an isolated extremity are sufficient for transposition. A specific sequence 13-23 bp from the ends, defining the edge of the minimal sequence, is implicated as an essential site. The sites, symmetrically arrayed at both ends of IS1, correspond to the apparent consensus sequence of the known binding sites for the Escherichia coli DNA-binding protein (called integration host factor or IHF) which is required for the site-specific recombination that leads to integration of bacteriophage lambda into the bacterial genome. The sites at the ends of IS1 may thus bind a host protein, such as JHF or a related protein, that is involved in regulating the transposition apparatus.     

Modified Dye for Water-Fast Ink-Jet Printing

A quaternary ammonium compound containing amino acid residue was synthesized by Converting 3-chloro-2-hydroxy-propyalkyldimethylammonium chlorides into its epoxide derivatives, then attaching an amino acid to the epoxide derivatives synthesized a quaternary ammonium compound containing amino acid residue. Modified dyes were prepared by the ionotropy of anionic dyes with the quaternary ammonium compound containing amino acid residue. It was discovered that the modified dyes exhibited an excellent pH controllable solubility. These modified dyes have good water solubility at pH> 8.0, but they were water insoluble at pH < 6.5. On the printing paper, modified dyes in water-based ink-jet print ink could convert to water insoluble form and give prints excellent water fastness.     

V型苯乙炔Bola两亲化合物的合成及其液晶、光物理性质研究

以Pd 催化的Sonogasira偶联反应为关键步骤合成V型苯乙炔Bola两亲化合物,产物结构经1H NMR、13C NMR及元素分析得到证实.采用偏光显微镜(POM),DSC等手段对化合物的液晶行为进行了研究,研究结果表明,该化合物是柱相热致性液晶.化合物的紫外光谱证实组装模型中V型芳香核间的-堆积形式为Ｈ聚集结构,由紫外光谱换算后的h-(ah)2关系图推算的能隙范围推测该化合物可作为半导体材料,荧光光谱推断它们可以作为潜在的蓝光材料,化合物较大的Stokes位移值表明该化合物在激光材料及荧光光子的放射性计量仪等方面有潜在应用.此外,高斯计算模拟量化结果显示了该化合物近乎理想的平面共轭结构.     

Structural basis for synthesis of inflammatory mediators by human leukotriene C4 synthase

Cysteinyl leukotrienes are key mediators in inflammation and have an important role in acute and chronic inflammatory diseases of the cardiovascular and respiratory systems, in particular bronchial asthma. In the biosynthesis of cysteinyl leukotrienes, conversion of arachidonic acid forms the unstable epoxide leukotriene A4 (LTA4). This intermediate is conjugated with glutathione (GSH) to produce leukotriene C4 (LTC4) in a reaction catalysed by LTC4 synthase: this reaction is the key step in cysteinyl leukotriene formation. Here we present the crystal structure of the human LTC4 synthase in its apo and GSH-complexed forms to 2.00 and 2.15 A resolution, respectively. The structure reveals a homotrimer, where each monomer is composed of four transmembrane segments. The structure of the enzyme in complex with substrate reveals that the active site enforces a horseshoe-shaped conformation on GSH, and effectively positions the thiol group for activation by a nearby arginine at the membrane-enzyme interface. In addition, the structure provides a model for how the omega-end of the lipophilic co-substrate is pinned at one end of a hydrophobic cleft, providing a molecular 'ruler' to align the reactive epoxide at the thiol of glutathione. This provides new structural insights into the mechanism of LTC4 formation, and also suggests that the observed binding and activation of GSH might be common for a family of homologous proteins important for inflammatory and detoxification responses.     

Methane oxidation by an extremely acidophilic bacterium of the phylum Verrucomicrobia

Aerobic methanotrophic bacteria consume methane as it diffuses away from methanogenic zones of soil and sediment. They act as a biofilter to reduce methane emissions to the atmosphere, and they are therefore targets in strategies to combat global climate change. No cultured methanotroph grows optimally below pH 5, but some environments with active methane cycles are very acidic. Here we describe an extremely acidophilic methanotroph that grows optimally at pH 2.0-2.5. Unlike the known methanotrophs, it does not belong to the phylum Proteobacteria but rather to the Verrucomicrobia, a widespread and diverse bacterial phylum that primarily comprises uncultivated species with unknown genotypes. Analysis of its draft genome detected genes encoding particulate methane monooxygenase that were homologous to genes found in methanotrophic proteobacteria. However, known genetic modules for methanol and formaldehyde oxidation were incomplete or missing, suggesting that the bacterium uses some novel methylotrophic pathways. Phylogenetic analysis of its three pmoA genes (encoding a subunit of particulate methane monooxygenase) placed them into a distinct cluster from proteobacterial homologues. This indicates an ancient divergence of Verrucomicrobia and Proteobacteria methanotrophs rather than a recent horizontal gene transfer of methanotrophic ability. The findings show that methanotrophy in the Bacteria is more taxonomically, ecologically and genetically diverse than previously thought, and that previous studies have failed to assess the full diversity of methanotrophs in acidic environments.     

西非L-2000区块巨厚盐膏层钻井液技术

西非L-2000区块是中国石化在海外的重点勘探区块,埃詹加组是一套巨厚盐膏层,最大厚度达到1 226 m。前期施工井在埃詹加组均出现过缩径、卡钻、划眼困难、测井和下套管遇阻等井下复杂情况,其中2口井造成工程报废。对巨厚盐膏层钻井液技术难点进行了分析,制定了针对性技术对策。通过主处理剂优选,在原聚合物饱和盐水钻井液的基础上,开发了复合离子欠饱和盐水钻井液配方。室内实验表明,复合离子欠饱和盐水钻井液具有良好的沉降稳定性、抗钙侵、抗劣质土污染能力和防塌抑制性,根据其特点,制定了钻井液现场维护措施。LT-1井和LT-2井的应用表明,复合离子欠饱和盐水钻井液现场维护处理简单、性能稳定,盐膏层段井径规则,无缩径、卡钻复杂情况发生,平均井径扩大率分别为17.8%和12.5%。复合离子欠饱和盐水钻井液体系是适合该区块巨厚盐膏层钻井施工的成功钻井液体系。     

Biological degradation of TCDD in rats

Apart from its high toxicity, the chemical stability and persistence of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in the environment bring about additional hazards. However, until now there has been no evidence for degradation of this dioxin in biological systems, and it is considered unusual that a compound which is insusceptible to biological attack should be so extremely toxic. In in vitro studies with microsomal preparation from mouse, rat and rabbit liver, as well as in vivo, no metabolite of TCDD has been detected by several workers. Recent data, showing a lower toxicity of TCDD in rats after stimulation of the hepatic mixed-function oxidase, suggested possible metabolic transformation of this compound. In the present study, we used tritiated TCDD to search for metabolites in the bile of rats and found marked changes in lipophilicity and mobility in TLC of the excreted radioactivity. Treatment with diazomethane yielded at least two new compounds. These observations hinted at the formation of phenolic hydroxyl groups in the dioxin molecule.     

Structure and function of the AAA+ protein CbbX, a red-type Rubisco activase

Ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyses the fixation of atmospheric CO(2) in photosynthesis, but tends to form inactive complexes with its substrate ribulose 1,5-bisphosphate (RuBP). In plants, Rubisco is reactivated by the AAA(+) (ATPases associated with various cellular activities) protein Rubisco activase (Rca), but no such protein is known for the Rubisco of red algae. Here we identify the protein CbbX as an activase of red-type Rubisco. The 3.0-? crystal structure of unassembled CbbX from Rhodobacter sphaeroides revealed an AAA(+) protein architecture. Electron microscopy and biochemical analysis showed that ATP and RuBP must bind to convert CbbX into functionally active, hexameric rings. The CbbX ATPase is strongly stimulated by RuBP and Rubisco. Mutational analysis suggests that CbbX functions by transiently pulling the carboxy-terminal peptide of the Rubisco large subunit into the hexamer pore, resulting in the release of the inhibitory RuBP. Understanding Rubisco activation may facilitate efforts to improve CO(2) uptake and biomass production by photosynthetic organisms.     

Synthesis of the H-cluster framework of iron-only hydrogenase

The metal-sulphur active sites of hydrogenases catalyse hydrogen evolution or uptake at rapid rates. Understanding the structure and function of these active sites--through mechanistic studies of hydrogenases, synthetic assemblies and in silico models--will help guide the design of new materials for hydrogen production or uptake. Here we report the assembly of the iron-sulphur framework of the active site of iron-only hydrogenase (the H-cluster), and show that it functions as an electrocatalyst for proton reduction. Through linking of a di-iron subsite to a {4Fe4S} cluster, we achieve the first synthesis of a metallosulphur cluster core involved in small-molecule catalysis. In addition to advancing our understanding of the natural biological system, the availability of an active, free-standing analogue of the H-cluster may enable us to develop useful electrocatalytic materials for application in, for example, reversible hydrogen fuel cells. (Platinum is currently the preferred electrocatalyst for such applications, but is expensive, limited in availability and, in the long term, unsustainable.).     

Aminoglycoside antibiotics induce bacterial biofilm formation

Biofilms are adherent aggregates of bacterial cells that form on biotic and abiotic surfaces, including human tissues. Biofilms resist antibiotic treatment and contribute to bacterial persistence in chronic infections. Hence, the elucidation of the mechanisms by which biofilms are formed may assist in the treatment of chronic infections, such as Pseudomonas aeruginosa in the airways of patients with cystic fibrosis. Here we show that subinhibitory concentrations of aminoglycoside antibiotics induce biofilm formation in P. aeruginosa and Escherichia coli. In P. aeruginosa, a gene, which we designated aminoglycoside response regulator (arr), was essential for this induction and contributed to biofilm-specific aminoglycoside resistance. The arr gene is predicted to encode an inner-membrane phosphodiesterase whose substrate is cyclic di-guanosine monophosphate (c-di-GMP)-a bacterial second messenger that regulates cell surface adhesiveness. We found that membranes from arr mutants had diminished c-di-GMP phosphodiesterase activity, and P. aeruginosa cells with a mutation changing a predicted catalytic residue of Arr were defective in their biofilm response to tobramycin. Furthermore, tobramycin-inducible biofilm formation was inhibited by exogenous GTP, which is known to inhibit c-di-GMP phosphodiesterase activity. Our results demonstrate that biofilm formation can be a specific, defensive reaction to the presence of antibiotics, and indicate that the molecular basis of this response includes alterations in the level of c-di-GMP.     

含偶氮介晶基元的环氧化合物的合成

为了制得一种新型液晶环氧树脂,合成了一种含偶氧介晶基元的环氧化合物,研究了反应物配比对产物结构的影响,利用红外光谱、紫外光谱、核磁共振氢谱、示差扫描热分析和元素分析等手段确认了其结构,并给出了适合偶氮环氧体系的平均相对分子量测定方法。正交偏光场观察显示４,４’－二缩水甘油醚偶氮苯与４,４’－二氨基二苯甲烷的固化过程出现了从各向同性到各向异性的转变,产物为液晶有序的不溶不熔的环氧树脂。     

气田水平井防砂筛管类型优选与精度优化试验

机械防砂筛管是疏松砂岩气藏防砂完井管柱的关键组成部分,其总体结构和挡砂介质性能决定了挡砂效果和气井防砂后产能以及总体服务期限。南海X气田为疏松砂岩易出砂气藏,为选择合适的水平井二次防砂机械筛管类型并优化挡砂精度,使用自行研制开发的挡砂介质性能评价径向流驱替试验装置,模拟该气田地层砂和生产条件,对单层网布复合筛管、双层网布复合筛管、复合绕丝筛管、多层精密筛管、孔网复合筛管等5种类型的筛管样品进行了综合性能评价试验;根据动态试验数据,提出筛管的流通性能、挡砂性能及其评价指标的计算方法,系统地评价5种筛管的各单项性能和综合性能,并得到具体的量化评价指标。为优化挡砂精度,使用从0.08~0.18mm的5种不同精度的双层网布复合筛管分别阻挡粒度中值0.08mm的模拟地层,测试与评价各精度筛管的综合防砂性能。结果表明,对于南海X气田不产水粉细砂气藏水平井,双层网布绕丝筛管和精密筛管的流通性能、挡砂性能等综合指标较高,依次推荐为气田二次防砂最佳筛管类型;最终推荐该气田二次防砂挡砂精度为0.1~0.12mm。     

凹凸棒负载型TiO_2光催化剂的制备及改性研究

大部分化工废水成分复杂,毒性高,难以生物降解,文章以降解对苯二酚为例研究光催化氧化法的处理效果.实验以凹凸棒为载体,通过溶胶-凝胶法,以钛酸丁酯为主要原料,加入所制备的凹凸棒载体制成包覆粒子凹凸棒-TiO2光催化剂;以催化剂对对苯二酚的降解效果作为评价凹凸棒-TiO2样品光催化活性的标准,对其制备条件进行了优化,得到了光催化效果很好的复合光催化剂.采用水解沉淀法,以SnCl4·nH2O为主要原料制备了SnO2·nH2O,对凹凸棒-TiO2光催化剂改性,制成了凹凸棒-SnO2-TiO2光催化剂;通过对对苯二酚的降解效果,得到了催化剂各组分的最佳比例,并进行了模拟太阳光源的研究;最后用XRD和TEM等手段对样品进行了表征.     

A new atmospherically relevant oxidant of sulphur dioxide

Atmospheric oxidation is a key phenomenon that connects atmospheric chemistry with globally challenging environmental issues, such as climate change, stratospheric ozone loss, acidification of soils and water, and health effects of air quality. Ozone, the hydroxyl radical and the nitrate radical are generally considered to be the dominant oxidants that initiate the removal of trace gases, including pollutants, from the atmosphere. Here we present atmospheric observations from a boreal forest region in Finland, supported by laboratory experiments and theoretical considerations, that allow us to identify another compound, probably a stabilized Criegee intermediate (a carbonyl oxide with two free-radical sites) or its derivative, which has a significant capacity to oxidize sulphur dioxide and potentially other trace gases. This compound probably enhances the reactivity of the atmosphere, particularly with regard to the production of sulphuric acid, and consequently atmospheric aerosol formation. Our findings suggest that this new atmospherically relevant oxidation route is important relative to oxidation by the hydroxyl radical, at least at moderate concentrations of that radical. We also find that the oxidation chemistry of this compound seems to be tightly linked to the presence of alkenes of biogenic origin.     

环二鸟苷酸对细菌运动调控的研究进展

细菌中的第二信使环二鸟苷酸(c-di-GMP)对细菌的运动性有调节作用.c-di-GMP调控鞭毛的生物合成、菌毛形成和菌毛蛋白的组成,以及其他一些与运动相关的蛋白的合成.细菌的运动性与其毒力、致病性、粘附性、趋化性、生物膜组成等密切相关.在革兰氏阴性细菌中,关于c-di-GMP的信号通路的研究较为清晰,而在革兰氏阳性细菌中,关于该信号转导通路的研究较少.此外,有关c-di-GMP的信号通路的研究主要集中在病原菌.该文主要综述了一些常见病原菌中c-di-GMP对其运动性的调控机制,为研究其他细菌c-di-GMP信号通路提供思路.     

ConformationsofCytochromeP─450Modelsby1Dand2DNMR

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The Srs2 helicase prevents recombination by disrupting Rad51 nucleoprotein filaments

Homologous recombination is a ubiquitous process with key functions in meiotic and vegetative cells for the repair of DNA breaks. It is initiated by the formation of single-stranded DNA on which recombination proteins bind to form a nucleoprotein filament that is active in searching for homology, in the formation of joint molecules and in the exchange of DNA strands. This process contributes to genome stability but it is also potentially dangerous to cells if intermediates are formed that cannot be processed normally and thus are toxic or generate genomic rearrangements. Cells must therefore have developed strategies to survey recombination and to prevent the occurrence of such deleterious events. In Saccharomyces cerevisiae, genetic data have shown that the Srs2 helicase negatively modulates recombination, and later experiments suggested that it reverses intermediate recombination structures. Here we show that DNA strand exchange mediated in vitro by Rad51 is inhibited by Srs2, and that Srs2 disrupts Rad51 filaments formed on single-stranded DNA. These data provide an explanation for the anti-recombinogenic role of Srs2 in vivo and highlight a previously unknown mechanism for recombination control.     

基于Logistic-Sine-Cosine映射的图像加密算法

为了提高数字图像加密的速度和算法的安全性,提出了基于Fridrich框架的加密算法。通过Fisher-Yates变换置乱原始图像打破像素间的强相关性,再利用滤波器在RGB平面上进行滤波得到密文。Logistic-Sine-Cosine复合混沌系统具有良好的混沌特性,且时间复杂度低。密钥由输入参数和明文的SHA-512值共同决定,对明文高度敏感。二维滤波器扩散效果良好,其滤波器模板由明文和密钥决定并引入了伪随机像素值,在提升扩散效果的同时增强了系统抗差分攻击的能力。仿真结果表明,密文图像像素分布近似均匀、像素关联性弱、密钥空间足够大、密钥敏感性高,能够有效抵抗暴力、裁剪、差分等常见攻击,具有较高的安全性,且算法时间复杂度较低。