灯盏花CHI基因的再克隆及其绿色荧光蛋白表达载体构建

根据灯盏花转录组所注释的CHI unigene片段,设计了RACE相关引物,克隆到灯盏花CHI的cDNA全长序列,序列全长717bp,编码238个氨基酸.该氨基酸序列与我们前期所克隆的灯盏花CHI cDNA序列相比,在345、351位点发生了突变,碱基均由C突变为T,两个位点的突变均属于同义突变,编码的氨基酸分别为115、117位异亮氨酸、酪氨酸.同时将CHI基因片段插入到pDM 18-T,构建pDM 18-T-eBCHI中间载体.经测序验证后,根据pBI121-EGFP图谱,设计带有SalI、SpeI酶切位点的引物,以pDM 18-T-eBCHI中间载体为模板,扩增带酶切位点的CHI序列,扩增产物经回收、纯化后与双酶切的pBI121-EGFP链接,构建了重组表达载体pBI121-EGFP-CHI,CHI基因插入植物表达载体pBI121-EGFP的6xHis组氨酸标签与GFP基因间.经SalI、SpeI双酶切和PCR验证重组质粒,均能得到约750bp的目的片段,通过进一步测序证明,CHI基因已成功地连接到pBI121-EGFP-eBCHI中.采用电击转化法将重组质粒导入根癌农杆菌GV3101,用叶盘法转化灯盏花叶片,筛选出具有卡那霉素抗性的愈伤组织,经PCR扩增和叶绿素荧光成像检测证明,重组基因已成功地转化到灯盏花愈伤组织中.为利用转基因技术研究CHI基因的表达及其亚细胞定位打下基础.     

烟草反义Mlo基因表达载体的构建

获得烟草反义Mlo基因的植物表达载体pBI 121-Mlo,为进行烟草遗传转化,获得该基因表达的缺陷型植株打下基础.从烟草叶片中提取总RNA,利用RT-PCR技术扩增得到Mlo基因的c DNA,以此为模板设计反义引物,通过PCR扩增出反义Mlo基因,将此反义Mlo基因与T载体连接,测序正确后再将此反义片段与植物表达载体pBI 121连接,构建烟草反义Mlo基因的植物表达载体pBI 121-Mlo.经Kan选择筛选出反义重组菌落,碱裂解法小量提取质粒后,用Xba I和Bam HI双酶切后再进行电泳鉴定.结果表明,目的基因已与植物表达载体pBI 121连接成功.成功构建了烟草反义Mlo基因表达载体pBI 121-Mlo.     

农杆菌介导牧草蔗42遗传转化体系的建立

将拟南芥叶片衰老特异启动子sag12与异戊烯基转移酶基因ipt构成的嵌合基因经农杆菌介导转入牧草蔗42中，建立了农杆菌转化牧草蔗42的遗传转化系统，已经获得转基因再生植株，PCR及Southern杂交检测证实ipt基因确实转移到牧草蔗42中，报告基因GUS在再生植株中也得到表达。     

抗汉滩病毒单抗3G1 scFv植物表达载体的构建

利用PCR方法，从含有3G1 scFv基因的重组质粒中扩增出抗体基因，并使基因两端携带合适的限制性酶切位点．将其克隆人植物表达载体pBI121，构建获得3G1 scFv-pBI121重组质粒．酶切鉴定及测序结果均证明重组质粒构建成功，将重组植物表达载体转入农杆菌LBA4404，为进一步构建转基因植物的研究奠定了基础．     

农杆菌介导的AtPCS1基因转化陇东苜蓿的初步研究

利用植物表达载体pBI121-AtPCS1转化农杆菌LBA4404,经菌落PCR鉴定获得阳性菌,利用叶盘法以重组的农杆菌侵染陇东苜蓿的子叶.采用GUS组织化学染色和对部分转基因再生植株进行PCR鉴定,初步结果表明AtPCS1基因已成功整合到苜蓿基因组.     

甜味蛋白thaumatin表达载体的构建

将克隆在不同质粒上的thaumatin基因的两个片段连接起来，连接产物克隆到质粒pUC18上．测序结果表明，连接产物是一个完整的thaumatin cDNA基因．将此基因通过基因重组技术，克隆到植物表达载体pBI121中，构建了一个新的转化植物的表达载体-pBIl21-tha.     

抗汉坦病毒单链抗体双元载体的构建

利用PCR方法，从含有1A8 scFv基因的重组质粒中扩增出抗体基因，并使基因两端携带合适的限制性酶切位点。经多步连接将其克隆入植物表达载体pBI121或pCAMBIA1305．2。酶切结果证明1A8 scFv基因被成功克隆入植物表达载体pBI121及pCAMBIA1305．2，构建获得1A8 scFv-p BI121及1A8 scFv pCAMBI A1305．2重组质粒，并将其转入农杆菌GV3101。抗汉坦病毒mAb 1A 8scFv植物双元表达载体的获得，为进一步在植物中表达该抗体片段奠定了基础。     

角化细胞生长因子的获得及植物表达载体的构建

从鼠肺中提取RNA,设计相应的引物,经RT PCR扩增出角化细胞生长因子cDNA序列;插入pMD18 T载体,经DNA测序证实为KGF基因.将KGF基因经酶切后连接于植物CaMV35S启动子和NOS终止子之间,单酶切插入pBI1301植物双元表达载体,转化大肠杆菌,阳性克隆提取质粒后转化农杆菌EHA105,实现KGF植物表达载体的构建,为KGF的进一步研究奠定基础.     

拟南芥抗旱相关基因的表达载体构建及转基因植株鉴定

文章以拟南芥Col-0植株的基因组DNA和cDNA为模板扩增出LWT2基因全长和CDS片段,用pEASY-Blunt Zero Cloning Kit试剂盒连接至中间载体,转化到Trans1-T1感受态细胞内,通过单克隆挑选、菌落聚合酶链式反应(polymerase chain reaction,PCR)鉴定及质粒测序验证获得DNA阳性克隆。利用限制性内切酶双酶切获得含有黏性末端的目的片段和载体片段,进行DNA连接反应可得最终的互补和过表达构建。提取质粒DNA测序确认后将2个构建分别转化至农杆菌菌株GV3101,菌落PCR鉴定后通过浸花转化法转化拟南芥lwt2-1突变体植株,最后通过转基因筛选与遗传鉴定获得相应构建的转基因阳性植株。     

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).     

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.     

Tracking data management using topology of moving object database system for moving object constrained trajectory in LBS application

Advances in wireless technologies and positioning technologies and spread of wireless devices, an interest in LBS (Location Based Service) is arising. To provide location based service, tracking data should have been stored in moving object database management system (called MODBMS) with proper policies and managed efficiently. So the methods which acquire the location information at regular time intervals then, store and manage have been studied. In this paper, we suggest tracking data management techniques using topology that is corresponding to the moving path of moving object. In our techniques, we update the MODBMS when moving object arrived at a street intersection or a curved road which is represented as the node in topology and predict the location at past and future with attribute of topology and linear function. In this technique, location data that are corresponding to the node in topology are stored, thus reduce the number of update and amount of data. Also in case predicting the location,because topology are used as well as existing location information, accuracy for prediction is increased than applying linear function or spline function.