人乳铁蛋白基因的克隆及序列分析与重组逆转录病毒质粒载体PLNCX2-hLF的构建

从正常人外周血中性粒白细胞中提取总RNA，用RT-PCR的方法扩增人乳铁蛋白（hLF）cDNA，将其克隆到pMD18-T Simple载体上并测序．用限制性内切酶Xho Ⅰ和Hind Ⅲ将目的片段与逆转录病毒载体PLNCX2进行双酶切、并用T4连接酶进行连接；然后转化到top10菌中，再次进行DNA序列测定．前后两次序列测定结果一致，没有突变，全长为2，136bp，说明所克隆的为hLF基因的cDNA序列；与GeneBank中登录的序列相比，同源性达99％以上．     

全长人lrp基因及其截短和定点突变体真核表达载体的构建

为了对脂多糖应答基因（lrp）的功能进行深入的研究,实验用PCR及双PCR法扩增出lrp基因的截短体（lrp△C）和突变体（lrpm）序列,测序正确后将正常lrp基因及其截短体和突变体序列均连入真核表达载体pcDNA3．1（＋）,构建重组质粒pcDNA3．1（＋）-lrp、pcDNA3．1（＋）-lrp△C和pcDNA3．1（＋）-lrpm。DNA测序结果显示,PCR反应成功得到了lrp基因的截短体和突变体序列;重组质粒酶切鉴定结果显示：人lrp基因及其截短体、突变体成功连入了真核表达载体pcDNA3．1（＋）。     

可溶性人干细胞因子的原核表达和纯化

用Trizol提取人骨髓总RNA，通过RT-PCR扩增人干细胞因子DNA，克隆到pMD18-T载体中并测序．将其定向连入原核表达载体pET32a( )，获得了重组表达载体pET32a( )／hSCF，其cDNA长度为504bp，测序结果与已知序列吻合．然后在大肠杆菌BL21中经IPTG诱导表达，获得37kD的融合蛋白，约占菌体总蛋白量的30％．经Ni^2 -NTA树脂亲和层析柱纯化获得纯度大约90％的重组蛋白．     

海水养殖真鲷肝CYP1A基因的克隆与表达

根据国外已发表野生真鲷（Pagrus major）肝CYP1A cDNA同源序列，利用反转录-聚合酶链式反应（RT-PCR）从我国海水养殖真鲷（Pagrus major）的肝脏中扩增出P4501A基因全长cDNA序列（1548bp）．用基因重组技术将P4501A cDNA克隆于pTrc-CKS载体，在大肠杆菌TOP10F＇诱导表达，将细菌总蛋白进行SDS聚丙烯酰胺凝胶电泳分析；其表达产物为89ku，获得了表达融合蛋白CKS-CYP1A．该项研究将为利用免疫学技术研究有机污染物的毒性效应机制以及探讨P450酶作为毒性效应的生物标志物奠定基础．     

绵羊防御素sBD-1 cDNA的克隆及序列分析

从成年绵羊舌表皮组织分离提取总RNA，经反转录PCR（RT-PCR)扩增出绵羊防御素sBD-lcDNA，将扩增片段回收，重组插入克隆载体T—vector，经限制性酶切鉴定和DNA序列测定，结果扩增出cDNA215bp，与发表的绵羊sBD-lcDNA序列完全相同。该cDNA编码以个氨基酸，其中信号肽与前片段相同，成熟肽位于前片段的羧基端，由38个氨基酸残基组成。     

百合花发育相关基因LLGLO1的RNAi载体构建

百合花发育相关基因的研究是开展花卉分子育种的重要基础。以麝香百合(Lilium longi-florumThumb.)未开放的花蕾为材料提取总RNA。根据LLGLO1序列的同源比对选取特异区段,设计引物扩增区段cDNA并引入酶切位点。将PCR扩增片断连接到T载体上,经酶切及PCR验证后进行测序。测序结果表明:用于RNAi的cDNA片断与LLGLO1序列完全相同。通过中间载体pSK+intron引入一段内含子间隔区,RNA干扰片断经过多次的酶切和连接过程最终连入表达载体pCAMBIA1301中形成ihpRNA结构。PCR及双酶切鉴定结果显示,构建的RNA干扰载体的ihpRNA结构完整。     

浮萍植物中一个衰老下调基因的初步鉴定

本报道对一个衰老下调基因的鉴定．从绿色和正在衰老的紫萍叶状体组织中分别提取总RNA，然后使用一个oligo—dT引物进行逆转录合成cDNA.cDNA用oligo—dT和简并引物进行PCR扩增，然后进行琼脂糖凝胶电泳．电泳后发现，一条cDNA带存在于由绿色组织而不存在于由正在衰老的组织所得到的样品中．将这一cDNA回收，克隆进T—vector并且进行了测序．Northern blot分析表明，该cDNA对应基因的表达受衰老调节而减少．根据第一个cDNA序列设计了一个特异性引物，进行RT-PCR扩增，并且克隆到另外一个部分cDNA．测序结果表明，第二个克隆具有与第一个cDNA相重叠的40bp的片段．将第一个cDNA序列与删去40bp重叠区的第二个cDNA序列连接，我们得到一个322个核苷酸的cDNA3'端部分序列．用BLASTN程序将该序列翻译后与NCBI non—redundant database中已知的蛋白进行比较，结果表明所克隆的cDNA代表的基因与光呼吸途径的丝氨酸-乙醛酸氨基转移酶编码基因有很高的相似性．     

一种快捷有效的构建cDNA文库的方法

构建cDNA文库是研究基因组功能基因一种有效的手段，传统的建库方法通常需要在双链cDNA两端分别加上带限制性酶切位点的接头序列，通过酶切使之产生与质粒匹配的粘性末端，然后与质粒连接构成文库。本文介绍一种简单而且不需要核酸内切酶的构建cDNA文库的方法。该方法的主要步骤包括：（1）通过反转录合成cDNA第一条链，通过置换底物的方式在cDNA3’末端合成接头序列。（2）利用cDNA两端的接头引物，通过TaqDNA合成酶PER扩增合成cDNA第二条链。（3）将PCR产生的双链cDNA直接与T-载体连接。（4）将连接物转化大肠杆菌，得到cDNA文库。利用本方法，我们成功构建红树植物红海榄的cDNA文库。PCR结果显示插入的片段分布在O．5～3．0kb之间，大部分cDNA的长度在500—1000bp之间，表明cDNA具有较好的完整性。本方法具有操作简单、高效率、低成本、RNA模板需要量少的特点，而且适用于任何高等生物RNA样品。     

猪肝脏和肌肉肉碱脂酰转移酶I基因片段克隆及序列分析

分别从金皮F2代猪的肝脏、第十肋背最长肌中提取总RNA,并根据报道的猪L—CPT I cDNA和M—CPT I cDNA序列分别设计引物1和引物2,用反转录一聚合酶链式反应（RT—PCR）进行cDNA扩增,获得两条长度分别为733bp和510bp的片段,克隆于pUCm-TVector后进行序列分析。结果表明,长度为733bp的片段为L—CPTI基因cDNA的部分序列,编码243个氨基酸,而长度为510bp的片段则为M—CPTI基因cDNA的部分序列,编码169个氨基酸。得到的两条基因片段与报道的猪L—CPT IcDNA和M—CPT IcDNA部分序列同源性分别为99.59％和99．61％。     

小鼠脂多糖应答蛋白mlrpS的载体构建及融合蛋白表达

为构建小鼠mlrpS-cDNA基因原核、真核表达载体,大肠杆菌表达其融合蛋白。采用反转录-聚合酶链反应从经脂多糖刺激的鼠NIH3T3细胞cDNA中,扩增出编码mlrpS的cDNA。用限制性内切酶KpnⅠand XhoⅠ消化后,插入原核表达载体pTAT中,经酶切鉴定与测序证实后,转化大肠杆菌BL21（DE3）菌株。异丙基β—D硫代半乳糖苷（IPTG）诱导产生融合蛋白。经KpnⅠ和XhoⅠ酶切回收mlrpS-cDNA,插入pcDNA3．1载体中;pcDNA3．1-mlrpS再经KpnⅠ和ApaⅠ酶切后,插入到pEGFP—c1载体上,构建pEGFP-mlrpS融合的真核表达载体。构建mlrpS表达载体经测序证实,与GenBank登录的序列完全一致;双酶切鉴定证实,克隆基因正确插入载体pEGFP及pTAT;SDS—PAGE证实融合蛋白表达成功。说明：成功地构建了mlrpS原核、真核表达载体,成功正确表达了6His／mlrpS融合蛋白。     

Characterization of Ordovician carbonate reservoirs, southeastern Saskatchewan, Canada

The discovery of the prolific Ordovician Red River reservoirs in 1995 in southeastern Saskatchewan was the catalyst for extensive exploration activity which resulted in the discovery of more than 15 new Red River pools. The best yields of Red River production to date have been from dolomite reservoirs. Understanding the processes of dolomitization is, therefore, crucial for the prediction of the connectivity, spatial distribution and heterogeneity of dolomite reservoirs.The Red River reservoirs in the Midale area consist of 3～4 thin dolomitized zones, with a total thickness of about 20 m, which occur at the top of the Yeoman Formation. Two types of replacement dolomite were recognized in the Red River reservoir: dolomitized burrow infills and dolomitized host matrix. The spatial distribution of dolomite suggests that burrowing organisms played an important role in facilitating the fluid flow in the backfilled sediments. This resulted in penecontemporaneous dolomitization of burrow infills by normal seawater. The dolomite in the host matrix is interpreted as having occurred at shallow burial by evaporitic seawater during precipitation of Lake Almar anhydrite that immediately overlies the Yeoman Formation. However, the low δ18O values of dolomited burrow infills (-5.9‰～ -7.8‰, PDB) and matrix dolomites (-6.6‰～ -8.1‰, avg. -7.4‰ PDB) compared to the estimated values for the late Ordovician marine dolomite could be attributed to modification and alteration of dolomite at higher temperatures during deeper burial, which could also be responsible for its 87Sr/86Sr ratios (0.7084～0.7088) that are higher than suggested for the late Ordovician seawaters (0.7078～0.7080). The trace amounts of saddle dolomite cement in the Red River carbonates are probably related to "cannibalization" of earlier replacement dolomite during the chemical compaction.     

A computer generator for randomly layered structures

AcomputergeneratorforrandomlylayeredstructuresYUJia shun1,2,HEZhen hua2(1.TheInstituteofGeologicalandNuclearSciences,NewZealand;2.StateKeyLaboratoryofOilandGasReservoirGeologyandExploitation,ChengduUniversityofTechnology,China)Abstract:Analgorithmisintrod…     

海南岛地体及其毗邻陆缘晚中生代—新生代古地磁研究和构造演化

本文叙述了对海南岛及其毗邻大陆边缘白垩纪到第四纪地层岩石进行古地磁研究的全部工作过程。通过分析岩石中剩余磁矢量的磁偏角及磁倾角的变化,提出海南岛白垩纪以来经历的构造演化模式如下:早期伴随顺时针旋转而向南迁移,后期伴随逆时针转动并向北运移。联系该地区及邻区的地质、地球物理资料,对海南岛上述的构造地体运动提出以下认识:北部湾内早期有一拉张作用,主要是该作用使湾内地壳显著伸长减薄,形成北部湾盆地。从而导致了海南岛的早期构造运动,而海南岛后期的构造运动则主要是受南海海底扩张的影响。海南地体运动规律的阐明对于了解北部湾油气盆地的形成演化有重要的理论和实际意义。     

Exciton Seebeck in Molecular Systems

Various applications relevant to the exciton dynamics,such as the organic solar cell,the large-area organic light-emitting diodes and the thermoelectricity,are operating under temperature gradient.The potential abnormal behavior of the exicton dynamics driven by the temperature difference may affect the efficiency and performance of the corresponding devices.In the above situations,the exciton dynamics under temperature difference is mixed with     

Achieving Unusual States of Matter Under High Pressure

The elongation method,originally proposed by Imamura was further developed for many years in our group.As a method towards O(N)with high efficiency and high accuracy for any dimensional systems.This treatment designed for one-dimensional(ID)polymers is now available for three-dimensional(3D)systems,but geometry optimization is now possible only for 1D-systems.As an approach toward post-Hartree-Fock,it was also extended to     

《Chinese Science Bulletin》SUBJECT INDEX TO VOLUME 59(2014)

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An on-line scaling method for improving scalability of a database cluster

The explosive growth of the Internet and database applications has driven database to be more scalable and available, and able to support on-line scaling without interrupting service. To support more client's queries without downtime and degrading the response time, more nodes have to be scaled up while the database is running. This paper presents the overview of scalable and available database that satisfies the above characteristics. And we propose a novel on-line scaling method. Our method improves the existing on-line scaling method for fast response time and higher throughputs. Our proposed method reduces unnecessary network use, i.e. , we decrease the number of data copy by reusing the backup data. Also, our on-line scaling operation can be processed parallel by selecting adequate nodes as new node. Our performance study shows that our method results in significant reduction in data copy time.     

An improved R-tree based on childnode''s probability

R-Tree is a good structure for spatial searching. But in this indexing structure,either the sequence of nodes in the same level or sequence of traveling these nodes when queries are made is random. Since the possibility that the object appears in different MBR which have the same parents node is different, if we make the subnode who has the most possibility be traveled first, the time cost will be decreased in most of the cases. In some case, the possibility of a point belong to a rectangle will shows direct proportion with the size of the rectangle. But this conclusion is based on an assumption that the objects are symmetrically distributing in the area and this assumption is not always coming into existence. Now we found a more direct parameter to scale the possibility and made a little change on the structure of R-tree, to increase the possibility of founding the satisfying answer in the front sub trees. We names this structure probability based arranged R-tree (PBAR-tree).     

An Index mechanism for multi-scale view in spatial databases

There are numerous geometric objects stored in the spatial databases. An importance function in a spatial database is that users can browse the geometric objects as a map efficiently. Thus the spatial database should display the geometric objects users concern about swiftly onto the display window. This process includes two operations:retrieve data from database and then draw them onto screen. Accordingly, to improve the efficiency, we should try to reduce time of both retrieving object and displaying them. The former can be achieved with the aid of spatial index such as R-tree, the latter require to simplify the objects. Simplification means that objects are shown with sufficient but not with unnecessary detail which depend on the scale of browse. So the major problem is how to retrieve data at different detail level efficiently. This paper introduces the implementation of a multi-scale index in the spatial database SISP (Spatial Information Shared Platform) which is generalized from R-tree. The difference between the generalization and the R-tree lies on two facets: One is that every node and geometric object in the generalization is assigned with a importance value which denote the importance of them, and every vertex in the objects are assigned with a importance value,too. The importance value can be use to decide which data should be retrieve from disk in a query. The other difference is that geometric objects in the generalization are divided into one or more sub-blocks, and vertexes are total ordered by their importance value. With the help of the generalized R-tree, one can easily retrieve data at different detail levels.Some experiments are performed on real-life data to evaluate the performance of solutions that separately use normal spatial index and multi-scale spatial index. The results show that the solution using multi-scale index in SISP is satisfying.     

Integrating GIS Web services based on mediating architecture

The geographic information service is enabled by the advancements in general Web service technology and the focused efforts of the OGC in defining XML-based Web GIS service. Based on these models, this paper addresses the issue of services chaining,the process of combining or pipelining results from several interoperable GIS Web Services to create a customized solution. This paper presents a mediated chaining architecture in which a specific service takes responsibility for performing the process that describes a service chain. We designed the Spatial Information Process Language (SIPL) for dynamic modeling and describing the service chain, also a prototype of the Spatial Information Process Execution Engine (SIPEE) is implemented for executing processes written in SIPL. Discussion of measures to improve the functionality and performance of such system will be included.