Stochastic sensing of organic analytes by a pore-forming protein containing a molecular adapter

The detection of organic molecules is important in many areas, including medicine, environmental monitoring and defence. Stochastic sensing is an approach that relies on the observation of individual binding events between analyte molecules and a single receptor. Engineered transmembrane protein pores are promising sensor elements for stochastic detection, and in their simplest manifestation they produce a fluctuating binary ('on/off') response in the transmembrane electrical current. The frequency of occurrence of the fluctuations reveals the concentration of the analyte, and its identity can be deduced from the characteristic magnitude and/or duration of the fluctuations. Genetically engineered versions of the bacterial pore-forming protein alpha-haemolysin have been used to identify and quantify divalent metal ions in solution. But it is not immediately obvious how versatile binding sites for organic ligands might be obtained by engineering of the pore structure. Here we show that stochastic sensing of organic molecules can be procured from alpha-haemolysin by equipping the channel with an internal, non-covalently bound molecular 'adapter' which mediates channel blocking by the analyte. We use cyclodextrins as the adapters because these fit comfortably inside the pore and present a hydrophobic cavity suitable for binding a variety of organic analytes. Moreover, a single sensing element of this sort can be used to analyse a mixture of organic molecules with different binding characteristics. We envisage the use of other adapters, so that the pore could be 'programmed' for a range of sensing functions.     

植物基因工程的现状与展望

介绍了80年代以来植物基因工程的历史、现状和发展趋势,对利用转基因植物生产药用蛋白,用工程病毒为载体生产药用蛋白,改进植物品质及逆境适应性,提高光合作用和固氮效率,抗除草剂工程植物及工程微生物与生物医学的关系等作了可行性分析和前景预测。     

Genetically engineered organisms for bioremediation of pollutants in contaminated sites

Environmental pollution is a major problem which affects biodiversity, public health and ecosystems worldwide. This issue cannot currently be solved using conventional technology because these treatments are expensive, ineffective and time consuming. Conventional methods focus unduly on the separation, rather than the destruction of contaminants. The use of genetically engi- neered organisms for bioremediation would be an envi- ronmentally-friendly and cost-effective alternative for the management and remediation of pollutants in contaminated sites. Different types of genetically engineered microbes have been developed through recombinant DNA and RNA technologies, these have been utilized for the removal of heavy metals and toxic substances from contaminated sites. Transgenic plants can also mobilize or degrade chlorinated solvent, xenobiotic compounds, explosives and phenolic substances. A symbiotic relationship between genetic engineered microbes and transgenic plants can enhance the effectiveness of bioremediation of contaminated sites. This review examines recent developments in the use of genetically engineered microbes and transgenic plants forthe bioremediation of contaminated sites. This review will also identify the environmental factors which influence bioremediation by genetically engineered microbes and transgenic plants as well as suggesting future directions for research in these areas.     

基于改进Sobel算法的焊缝X射线图像气孔识别方法

提出一种具有自适应能力的焊缝X射线图像气孔检测方法.通过局部动态阈值分割法设计了多方向的焊缝X射线图像模板,采用Sobel算法对复杂背景下的焊缝进行边缘检测和区域标记,并在标记区域对原始图像x和y方向的灰度梯度进行分析,以增强小气孔和粘连气孔的识别能力.检测应用效果表明,所提出的方法具有良好的自适应能力且准确快速,能够有效地克服人工评片中产生的漏判与误判等缺点,并且能够存储和查询检测数据.     

用多尺度形态学方法实现成像测井电导率图像的缝洞参数表征

裂缝和溶蚀孔洞为缝洞型储层提供了主要的储集空间和渗流通道,其分布的随机性和复杂性严重影响了对缝洞储层的定量评价。基于高覆盖率和高分辨率的电成像测井数据,采用多尺度数学形态学方法提取电导率图像中的缝洞孔隙度谱。选择不同尺度和形状的结构元素,构造不同种类的形态学滤波算子,实现井壁裂缝和溶蚀孔洞的电导率异常边缘检测。针对缝洞异常的边缘检测结果,用椭圆形函数拟合溶蚀孔洞,用多项式插值函数拟合裂缝边缘,继而提取缝洞参数并获得缝洞孔隙度谱。实验结果表明,用多尺度数学形态学方法对电导率图像的边缘检测有效地实现了缝洞的自动识别,验证了该方法计算缝洞孔隙度谱的准确性。     

鄂尔多斯盆地东南部上古生界储集砂岩的成岩作用

研究鄂尔多斯盆地东南部上古生界砂岩储层特征,以及不同的成岩作用类型与储集性能的关系。通过分析取芯井的岩石薄片、铸体薄片、扫描电镜、能谱分析数据,发现储集砂岩中发育有粒内溶孔、粒间孔、粒间溶孔、晶间孔、溶缝等多种孔缝类型,其中最主要的是粒内溶孔和粒间溶孔;主要成岩作用类型有压实作用、溶蚀作用、碳酸盐矿物、硅质矿物和黏土矿物的形成作用。结合不同砂岩组分与孔隙面孔率的相关性,得到如下结论:(1)压实作用是导致砂岩孔隙度较差的重要原因;(2)增加砂岩整体孔隙度最重要的是溶蚀作用;(3)碳酸盐矿物的形成会导致砂岩孔隙度减小;(4)硅质矿物会一定程度减少砂岩粒间孔;(5)高岭石的形成对增加岩石整体孔隙作用有限,水云母和绿泥石对溶孔和晶间孔的形成不利。     

获得四环素抗性的温敏同源重组载体质粒pRNT15的构建

 苏云金芽孢杆菌工程菌株TnX对蚊幼虫具有高毒力,但红霉素抗性基因的存在限制了其商品化。根据转座子Tn917载体的基因序列,利用重叠延伸PCR法克隆同源重组序列到pRN5101上得到pRN15,根据pBU4载体上的四环素抗性基因序列,对pRN15进行改造,使其失去红霉素抗性,获得四环素抗性,载体命名为pRNT15。重组载体四环素抗性的获得,使之扩大了使用范围,也为工程菌株染色体中红霉素抗性基因的敲除奠定了基础。     

富有机质页岩有机质孔发育差异性探讨——以四川盆地五峰组-龙马溪组笔石页岩为例

探讨富有机质页岩有机质孔发育的差异性,为页岩气勘探开发提供理论依据。在1 500倍和25 000倍电镜下,对四川盆地南部上奥陶统五峰组-下志留统龙马溪组富有机质笔石页岩进行观察。结果表明,页岩中笔石、干酪根、固体沥青等有机显微组分丰富,有机质孔总体发育;但不同有机显微组分、同一有机显微组分的有机质孔发育均存在差异,个体较大的固体沥青有机质孔最为发育,干酪根有机质孔较为发育,笔石碎片有机质孔不发育。有机质孔是随着有机碳向烃类和碳质残渣转化而形成的,笔石富碳贫氢的特征导致其有机质孔不发育;不同母质来源的干酪根生烃潜力存在差异决定了其有机质孔发育存在差异;固体沥青有机质孔是在原油裂解过程中形成的,其发育程度可能主要与原油赋存的粒间孔隙空间大小相关。     

储层孔隙结构特征及其对开发的影响-以辽河盆地西部凹陷某试验区于楼油层为例

以辽河盆地西部凹陷某试验区为例,综合7口取心井岩心、镜下薄片、分析测试以及400口井测井精细解释储层物性等多种资料,从微观和宏观角度对稠油热采储层孔隙结构进行研究。研究区储层为高孔高渗,平均孔隙度31.25%,平均渗透率1 829.3 md。从微观成因角度,将研究区目的层孔隙结构划分为原生孔隙和次生孔隙等2大类,同时进一步细分为粒间孔隙、粒内孔隙、基质内微孔、解理缝、粒间溶孔、粒内溶孔、铸模孔、特大溶蚀粒间孔、构造缝和溶蚀缝等10种亚类,其中以粒间孔隙和粒间溶孔为主。宏观上,基于精细测井解释资料,绘制了不同单层孔隙结构分布特征图。高孔隙度的区域主要呈北西-南东向条带状分布与水下分流河道主流线部位,指示孔隙度主要受沉积微相控制。在蒸汽吞吐转蒸汽驱开发方式转换时,应该充分考虑储层孔隙度的影响作用,在保证注汽井和采油井位于孔隙度高值区与的同时,应该尽量使得同一井组位于高孔隙度连片区域,已取得最好的注采对应效果。同时应该适当控制注入压力,防止蒸汽沿着高孔隙度发育条带突进,从而降低波及体积和提高石油采收率。     

检测汞的全细胞生物传感器构建

汞（Hg）是环境中主要的重金属污染物,暴露于环境中的汞对人体健康有严重危害。汞的精确检测对控制环境污染和减少对健康的危害有着重要意义。该实验是基于MerR蛋白调节的启动子对应答汞毒性基因的调控机理,首先通过试验确定红色荧光蛋白mCherry为报告基因。将MerR基因片段链接到质粒pUC57得到质粒pUC57-MerR,以其为模版得到质粒pUCRR,转入到E. coli DH5α中,得到工程菌株E. coli UCRR。为提高菌株的性能和灵敏度,通过易错PCR技术获得MerR基因突变体库,筛选出菌株V109E在培养4 h时,荧光信号强度远远高于其他菌株荧光信号,对Hg2＋有高度专一性,检测Hg2＋的最低浓度为0.5nM。     

现值法与年值法的等价条件

本文从理论上阐明：经济评价准则中现值法和年值法只有在一定的条件下才是等价的，通常在多个水平年规划中，年值法不可随意使用。本文导出了有关现值法的计算公式。论文所作结论和导出的计算公式已被作者用于长江三峡电源发展规划的优化数学模型中，并可以推广到其他工业部门的可行性研究中去。     

Preparation and properties of microencapsulated genetically engineered bacteria cells for oral therapy of uremia

~~Preparation and properties of microencapsulated genetically engineered bacteria cells for oral therapy of uremia~~     

辽河西部凹陷曙103块潜山油藏储层特征

辽河西部凹陷曙 10 3块潜山由中元古界长城系高于庄组泥粉晶白云岩、灰质白云岩和白云质灰岩组成 ,其主要的储集空间包括溶蚀孔洞、构造裂缝及微孔微缝等。该区溶蚀孔洞非常发育 ,根据其规模可分为大、中、小型溶洞及溶孔 ,其中大、中型溶洞常因坍塌和充填而失去储集能力 ,而小型溶洞和溶孔等则成为该潜山的主要储集空间。高角度构造缝对该储层储集空间的贡献最大 ,其优势发育方位为北东—南西向 ,与区域构造线方位一致。压力测试结果表明 ,该区储层中显微级的微孔和微裂缝经过一段时间的开采后 ,替代宏观溶孔和裂缝而成为主要的供油空间 ,这表明它也是很重要的储集空间。对各井开采现状及采油层段的对比研究表明 ,曙 10 3块潜山储层的发育受古地貌位置、构造位置及沉积旋回的控制 ,好的储层段主要分布在潜山顶面及地貌高点     

Supramolecular self-assembled molecules as organic directing agent for synthesis of zeolites

Solid materials with uniform micropores, such as zeolites, can act as selective catalysts and adsorbents for molecular mixtures by separating those molecules small enough to enter their pores while leaving the larger molecules behind. Zeolite A is a microporous material with a high void volume. Despite its widespread industrial use in, for example, molecular separations and in detergency, its capability as a petroleum-refining material is limited owing to its poor acid-catalytic activity and hydrothermal stability, and its low hydrophobicity. These characteristics are ultimately a consequence of the low framework Si/Al ratio (normally around one) and the resulting high cationic fraction within the pores and cavities. Researchers have modified the properties of type-A zeolites by increasing the Si/Al compositions up to a ratio of three. Here we describe the synthesis of zeolite A structures exhibiting high Si/Al ratios up to infinity (pure silica). We synthesize these materials, named ITQ-29, using a supramolecular organic structure-directing agent obtained by the self-assembly, through pi-pi type interactions, of two identical organic cationic moieties. The highly hydrophobic pure-silica zeolite A can be used for hydrocarbon separations that avoid oligomerization reactions, whereas materials with high Si/Al ratios give excellent shape-selective cracking additives for increasing propylene yield in fluid catalytic cracking operations. We have also extended the use of our supramolecular structure-directing agents to the synthesis of a range of other zeolites.     

基于油水分流原理的密闭取心饱和度校正方法

为力求准确求取密闭取心井油水饱和度,在分析流体饱和度损失变化的主要影响因素的基础上,以覆压岩石孔渗压缩校正为基础,以油水相渗分流率计算原理为核心,建立了密闭取心井由于孔隙及油水体积变化和降压脱气排液造成流体饱和度损失变化的数学校正模型。为油层损失前真实油水饱和度的校正提高了可靠的依据,具有重要的理论和实用意义。     

铜导线短路熔痕组织特征与烟粒子浓度的关系

针对火灾中铜导线短路形成时的不同环境条件,利用光学显微镜研究了环境条件中烟粒子质量浓度对铜导线短路熔痕组织中晶界、孔洞等特征的影响.结果表明:随着烟粒子质量浓度的增大,短路熔痕组织中晶界由连续逐渐向不连续转变,孔洞数量逐渐增多,孔洞的面积分数逐渐增大,孔洞面积分数与烟粒子质量浓度存在一定的对应关系;相同烟粒子质量浓度条件下,黏滞性大的烟粒子对熔痕内部孔洞的生成影响较黏滞性小的烟粒子大.     

Biological pacemaker created by gene transfer

The pacemaker cells of the heart initiate the heartbeat, sustain the circulation, and dictate the rate and rhythm of cardiac contraction. Circulatory collapse ensues when these specialized cells are damaged by disease, a situation that currently necessitates the implantation of an electronic pacemaker. Here we report the use of viral gene transfer to convert quiescent heart-muscle cells into pacemaker cells, and the successful generation of spontaneous, rhythmic electrical activity in the ventricle in vivo. Our results indicate that genetically engineered pacemakers could be developed as a possible alternative to implantable electronic devices.     

A bacteriolytic agent that detects and kills Bacillus anthracis

The dormant and durable spore form of Bacillus anthracis is an ideal biological weapon of mass destruction. Once inhaled, spores are transported by alveolar macrophages to lymph nodes surrounding the lungs, where they germinate; subsequent vegetative expansion causes an overwhelming flood of bacteria and toxins into the blood, killing up to 99% of untreated victims. Natural and genetically engineered antibiotic-resistant bacilli amplify the threat of spores being used as weapons, and heighten the need for improved treatments and spore-detection methods after an intentional release. We exploited the inherent binding specificity and lytic action of bacteriophage enzymes called lysins for the rapid detection and killing of B. anthracis. Here we show that the PlyG lysin, isolated from the gamma phage of B. anthracis, specifically kills B. anthracis isolates and other members of the B. anthracis 'cluster' of bacilli in vitro and in vivo. Both vegetative cells and germinating spores are susceptible. The lytic specificity of PlyG was also exploited as part of a rapid method for the identification of B. anthracis. We conclude that PlyG is a tool for the treatment and detection of B. anthracis.     

Au nanochannels technique and its application in immunoassay

Au nanotubules (channels) can be electrolessly plated within the pores of polycarbonate microporous filtration membranes. When an electric field was applied on the cell consisting of the membrane and Pt electrodes, the response current decreased due to a baffle effect from big molecules when the electrolyte ion and big molecules passed through the channels modified with chemical groups. Based on this principle, a nanotubules-based sensing technique has been developed. This method can be applied to the determination of human IgG with a detection limit of 0.34 ng/mL.     

Capacitance steps and fusion pores of small and large-dense-core vesicles in nerve terminals

The vesicles that package neurotransmitters fall into two distinct classes, large dense-core vesicles (LDCVs) and small synaptic vesicles, the coexistence of which is widespread in nerve terminals. High resolution capacitance recording reveals unitary steps proportional to vesicle size. Measurements of capacitance steps during LDCV and secretory granule fusion in endocrine and immune cells have provided important insights into exocytosis; however, extending these measurements to small synaptic vesicles has proven difficult. Here we report single vesicle capacitance steps in posterior pituitary nerve terminals. These nerve terminals contain neuropeptide-laden LDCVs, as well as microvesicles. Microvesicles are similar to synaptic vesicles in size, morphology and molecular composition, but their contents are unknown. Capacitance steps of two characteristic sizes, corresponding with microvesicles and LDCVs, were detected in patches of nerve terminal membrane. Both types of vesicles fuse in response to depolarization-induced Ca(2+) entry. Both undergo a reversible fusion process commonly referred to as 'kiss-and-run', but only rarely. Fusion pores seen during microvesicle kiss-and-run have a conductance of 19 pS, 11 times smaller than LDCV fusion pores. Thus, LDCVs and microvesicles use structurally different intermediates during exocytosis.