细菌Hfq蛋白的结构、功能及作用机制

非编码小RNA(small RNA, sRNA)是细菌基因转录后调控的一个重要层次,也是近十年来原核生物研究领域的焦点之一。大多数sRNA的作用与Hfq蛋白密切相关,即Hfq可以促进sRNA与其靶标mRNA的互补配对,进而影响翻译的进行或者mRNA的稳定性。笔者对Hfq的结构、Hfq参与sRNA调节作用的机制、Hfq在多种细菌中的功能表型进行了综述。Hfq是一个保守的蛋白质,在很多细菌中广泛存在,并与真核生物中参与mRNA剪切与降解活动的Sm蛋白同源。在结构上,Hfq具有两个非等同的RNA结合面,可以结合并介导多个RNA分子的相互作用,其结构体现了和功能的高度统一性。目前,对Hfq的研究主要集中于革兰氏阴性细菌中,在革兰氏阳性细菌中,Hfq的功能尚不明晰; 此外,在许多重要的细菌中,Hfq影响功能表型的具体机制也不清楚。因此,今后有必要进一步精细研究Hfq的分子结构特征和功能特点,深入分析Hfq对细菌表型多样化的影响机制,探究Hfq影响靶标分子和功能表型的详尽机制。     

低温生物菌剂在SBR工艺中的应用及功能分析

为解决寒冷地区冬季城市污水厂低温启动周期长和运行不稳定的难题,考察了应用低温生物菌剂缩短序批式反应器(Sequencing batch reactor,SBR)冬季启动周期及提高其长期运行稳定性的可行性。构建后的低温生物菌剂和剩余污泥分别按SBR单池有效容积的1%和4%投加到生化池内,并逐步由最初接种的生化池向其他生化池导泥;系统成功启动后,分别采集冬季低温期和春季气温回升期活性污泥样品进行了Biolog分析。结果表明:在平均水温为14℃时,仅用15 d完成了8组SBR生化池的启动;成功启动后的生化处理系统在常规条件下微生物群落结构较稳定,在水质水量及环境因子发生波动时,为适应环境及营养条件的变化,微生物群落结构及代谢类型也随之呈现动态演替的态势,从而保证了工艺运行的稳定性和理想的出水水质。低温菌剂在污水厂SBR工艺低温启动及稳定运行中发挥了重要作用。     

Assembly and function of the two ABC transporter proteins encoded in the human major histocompatibility complex.

Presentation of cytoplasmic antigens to class I-restricted cytotoxic T cells implied the existence of a specialized peptide transporter. For most class I heavy chains, association with peptides of the appropriate length is required for stable assembly with beta 2-microglobulin. Mutant cells RMA-S and .174/T2 neither assemble stable class I molecules nor present intracellular antigens, and we have suggested that they have lost a function required for the transport of short peptides from the cytosol to the endoplasmic reticulum. The genetic defect in .174 has been localized to a large deletion in the class II region of the major histocompatibility complex, within which two genes (RING4 and RING11) have been identified that code for 'ABC' (ATP-binding cassette) transporters. We report here that the protein products of these two genes assemble to form a complex. Defects in either protein result in the formation of unstable class I molecules and loss of presentation of intracellular antigens. The molecular defect in a new mutant, BM36.1, is shown to be in the ATP-binding domain of the RING11/PSF2 protein. This is in contrast to the mutant .134, which lacks the RING4/PSF1 protein.     

机器人化柔性自动装配线的总体性能分析

通过实验确定了机器人化柔性自动装配线中,底板零件到达时间间隔的概率密度函数服从ｆ（ｔ）＝ａｅｘｐ（－ｂｔ）所决定的分布规律。运用随机服务系统理对该装配线进行了仿真研究,分析了底板到达时间间隔的均值、各装配工位自动装配机服务时间的分布参数和缓冲区大小对装配线性能的影响,提出了若干改进建议,使装配线运行状态更良好。     

α-N-乙酰半乳糖胺酶的表达及活性检测

为了建立新型α-N-乙酰半乳糖胺酶的筛选、检测方法，实验中提取脑膜金黄杆菌的基因组DNA，以此为模板PCR扩增出α-N-乙酰半乳糖胺酶(A4).将A4克隆至pET-24a载体，转化Bl21表达菌株进行蛋白表达.使用亲和层析方法纯化His-A4酶，选择显色底物验证酶活性.同时，改进了传统的ELISA方法，直接将红细胞膜包被于ELISA检测平板中，以红细胞膜表面抗原作为直接底物，用ELISA方法检测酶活性.此研究建立了新型ELISA实验方法，以此方法验证了A4酶的活性，证明了此酶能够有效降低红细胞表面抗原抗体反应，且具有浓度和时间依赖性.     

面向虚拟装配的复杂产品装配建模技术

针对复杂产品特点,提出并分析了虚拟环境下复杂产品的装配建模技术:虚拟环境下的实体模型表达和装配模型.实体模型表达采用精确模型和面片模型结合的混合方式进行,在单元划分基础上,构造了复杂产品的基于单元的4级多层次关联装配模型.基于装配建模,建立了虚拟装配操作的基本流程,以一个船舶产品的装配建模实例验证了其有效性.     

X-ray structure of a bacterial oligosaccharyltransferase

Asparagine-linked glycosylation is a post-translational modification of proteins containing the conserved sequence motif Asn-X-Ser/Thr. The attachment of oligosaccharides is implicated in diverse processes such as protein folding and quality control, organism development or host-pathogen interactions. The reaction is catalysed by oligosaccharyltransferase (OST), a membrane protein complex located in the endoplasmic reticulum. The central, catalytic enzyme of OST is the STT3 subunit, which has homologues in bacteria and archaea. Here we report the X-ray structure of a bacterial OST, the PglB protein of Campylobacter lari, in complex with an acceptor peptide. The structure defines the fold of STT3 proteins and provides insight into glycosylation sequon recognition and amide nitrogen activation, both of which are prerequisites for the formation of the N-glycosidic linkage. We also identified and validated catalytically important, acidic amino acid residues. Our results provide the molecular basis for understanding the mechanism of N-linked glycosylation.     

基于装配特征的虚拟自动装配的研究和表达方法的描述

装配特征的概念是实际自动装配的基本问题,首先给出了装配特征的定义．为了实现虚拟自动装配,在CAD系统中引入了装配特征的概念。最后在UGⅡ环境下,结合VC＋＋对虚拟自动装配进行了描述。     

Identification and characterization of a bacterial hydrosulphide ion channel

The hydrosulphide ion (HS(-)) and its undissociated form, hydrogen sulphide (H(2)S), which are believed to have been critical to the origin of life on Earth, remain important in physiology and cellular signalling. As a major metabolite in anaerobic bacterial growth, hydrogen sulphide is a product of both assimilatory and dissimilatory sulphate reduction. These pathways can reduce various oxidized sulphur compounds including sulphate, sulphite and thiosulphate. The dissimilatory sulphate reduction pathway uses this molecule as the terminal electron acceptor for anaerobic respiration, in which process it produces excess amounts of H(2)S (ref. 4). The reduction of sulphite is a key intermediate step in all sulphate reduction pathways. In Clostridium and Salmonella, an inducible sulphite reductase is directly linked to the regeneration of NAD(+), which has been suggested to have a role in energy production and growth, as well as in the detoxification of sulphite. Above a certain concentration threshold, both H(2)S and HS(-) inhibit cell growth by binding the metal centres of enzymes and cytochrome oxidase, necessitating a release mechanism for the export of this toxic metabolite from the cell. Here we report the identification of a hydrosulphide ion channel in the pathogen Clostridium difficile through a combination of genetic, biochemical and functional approaches. The HS(-) channel is a member of the formate/nitrite transport family, in which about 50 hydrosulphide ion channels form a third subfamily alongside those for formate (FocA) and for nitrite (NirC). The hydrosulphide ion channel is permeable to formate and nitrite as well as to HS(-) ions. Such polyspecificity can be explained by the conserved ion selectivity filter observed in the channel's crystal structure. The channel has a low open probability and is tightly regulated, to avoid decoupling of the membrane proton gradient.     

Crystal structure and mechanism of a bacterial fluorinating enzyme

Fluorine is the thirteenth most abundant element in the earth's crust, but fluoride concentrations in surface water are low and fluorinated metabolites are extremely rare. The fluoride ion is a potent nucleophile in its desolvated state, but is tightly hydrated in water and effectively inert. Low availability and a lack of chemical reactivity have largely excluded fluoride from biochemistry: in particular, fluorine's high redox potential precludes the haloperoxidase-type mechanism used in the metabolic incorporation of chloride and bromide ions. But fluorinated chemicals are growing in industrial importance, with applications in pharmaceuticals, agrochemicals and materials products. Reactive fluorination reagents requiring specialist process technologies are needed in industry and, although biological catalysts for these processes are highly sought after, only one enzyme that can convert fluoride to organic fluorine has been described. Streptomyces cattleya can form carbon-fluorine bonds and must therefore have evolved an enzyme able to overcome the chemical challenges of using aqueous fluoride. Here we report the sequence and three-dimensional structure of the first native fluorination enzyme, 5'-fluoro-5'-deoxyadenosine synthase, from this organism. Both substrate and products have been observed bound to the enzyme, enabling us to propose a nucleophilic substitution mechanism for this biological fluorination reaction.     

Structure of a bacterial multidrug ABC transporter

Multidrug transporters of the ABC family facilitate the export of diverse cytotoxic drugs across cell membranes. This is clinically relevant, as tumour cells may become resistant to agents used in chemotherapy. To understand the molecular basis of this process, we have determined the 3.0 A crystal structure of a bacterial ABC transporter (Sav1866) from Staphylococcus aureus. The homodimeric protein consists of 12 transmembrane helices in an arrangement that is consistent with cross-linking studies and electron microscopic imaging of the human multidrug resistance protein MDR1, but critically different from that reported for the bacterial lipid flippase MsbA. The observed, outward-facing conformation reflects the ATP-bound state, with the two nucleotide-binding domains in close contact and the two transmembrane domains forming a central cavity--presumably the drug translocation pathway--that is shielded from the inner leaflet of the lipid bilayer and from the cytoplasm, but exposed to the outer leaflet and the extracellular space.     

Design principles of a bacterial signalling network

Cellular biochemical networks have to function in a noisy environment using imperfect components. In particular, networks involved in gene regulation or signal transduction allow only for small output tolerances, and the underlying network structures can be expected to have undergone evolution for inherent robustness against perturbations. Here we combine theoretical and experimental analyses to investigate an optimal design for the signalling network of bacterial chemotaxis, one of the most thoroughly studied signalling networks in biology. We experimentally determine the extent of intercellular variations in the expression levels of chemotaxis proteins and use computer simulations to quantify the robustness of several hypothetical chemotaxis pathway topologies to such gene expression noise. We demonstrate that among these topologies the experimentally established chemotaxis network of Escherichia coli has the smallest sufficiently robust network structure, allowing accurate chemotactic response for almost all individuals within a population. Our results suggest that this pathway has evolved to show an optimal chemotactic performance while minimizing the cost of resources associated with high levels of protein expression. Moreover, the underlying topological design principles compensating for intercellular variations seem to be highly conserved among bacterial chemosensory systems.     

串行装配环境下最优装配序列的求解

为了使得基于装配与或图的装配序列寻优方法支持串行装配环境下的最优装配序列的求解,设计了从装配树列举装配序列的递归算法,并讨论了装配序列的评价问题.首先给出了串行装配环境下最优装配序列的求解思路,接着将装配树转换成表示装配任务优先关系的有向图,进而从有向图列举出所有可行的装配序列,最后讨论串行装配环境下装配序列的评价问题.文中给出的实例说明本文方法的可行性,本文的实验结果说明本文方法的可接受性.     

装配操作过程中装配对象的几何特性分析

以凸多面体为装配对象，分析装配操作过程中装配对象的几何特性，提出了二元素的几何表示法，进行装配对象的空间描述，并分析了装配对象间的允许运动方向、几何约束特征及几何不确定性，用二元素几何表示法分析不确定性条件下装配对象的几何特性，简化了误差情况下装配对象各几何元素的数学表达式，本文内容是机器人装配操作过程分析与建模的基体。     

基于混合循环算法的复杂装配体装配序列智能规划

针对单一算法无法实现复杂装配体装配序列智能规划以及缺乏深度人机交互而导致的改进效果不佳等问题,提出一种混合循环算法.该算法以遗传算法为主体,利用干涉矩阵和接触矩阵调整随机生成的装配序列,以装配方向及工具的统一性构建适应度函数;其次结合模拟退火算法,在迭代前加入退火操作,利用Metropolis准则接受交叉和变异后的个体序列;引入粒子群算法的跟踪极值思想,直接选择个体最优和群体最优序列与后代交叉;最后结合虚拟现实技术建立装配模拟平台,从装配稳定性及工具操作空间两个维度进一步优化序列.基于该方法以汽车后桥总成装配序列规划为例进行验证,表明所得装配序列符合实际生产,该方法切实有效.