水稻硝酸还原酶基因5′上游序列的克隆与序列分析

以日本晴水稻幼苗叶的基因组总DNA为模板,采用PCR方法扩增获得约1 300 bp大小的DNA片段,回收该片段并与pMD18-T载体连接,转化大肠杆菌感受态,并提取质粒DNA.用凝胶电泳检测和酶切鉴定,选取阳性克隆进行测序分析.测序结果显示,该片段含1 376个核苷酸对;应用NCBI网站的BLAST软件对本试验中克隆的序列与GenBank(NC_008401)公布的水稻硝酸还原酶基因启动子序列进行比对,有3个核苷酸差异,同源性为99%,证实本试验中克隆的DNA序列为水稻硝酸诱导基因启动子.该序列含有多个与转录调控有关的保守序列(如TATA-box、CAAT-box和GAGA-box).此序列的成功克隆,为今后开展水稻等重要农作物转基因研究奠定基础.     

正常和细胞质雄性不育水稻线粒体DNA限制性内切酶的酶切图谱分析

从N,cms-WA,cms-BT和cms-RL四种不同细胞质源的水稻中制备线粒体DNA。用限制性内切酶Pst Ⅰ,Hind Ⅲ和Xho Ⅰ等酶切线粒体DNA经琼脂糖凝胶电泳,发现水稻N,cms-WA,cms-BT和cms-RL细胞质确有不同的线粒体DNA,不同的保持系之间也存着一定的差异。水稻线粒体DNA最小分子的大小在195～245kb之间。     

氧化亚铁硫杆菌Tf-55基因文库的构建

以λEMBL_3噬菌体 DNA作为载体,用BamHI/EcoRI双酶切后,与Sau3AI部分酶切的氧化亚铁硫杆菌Tf-55染色体10～23Kb DNA片段连接,体外包装成重组噬菌体,所得重组子为6×10~4Pfu(噬菌斑形成单位)达到了建库要求的理论值。并用酶切分析及分子杂交的方法对该库进行了验证。     

高质量水稻基因组文库构建的策略和方法

采用改良的CTAB法提取水稻基因组DNA,经Sau3AI部分酶切的产物经透析袋电洗脱法回收、正丁醇浓缩后,以1∶1的摩尔数比和载体臂连接.包装后测定,文库滴度达到106pfu/ml;随机酶切重组克隆显示,插入片段达到预期的15~23 kb之间,提示该方法适于植物材料高质量基因组文库的构建,有利于分离完整的基因序列及其调控区.     

K型和冈型水稻线粒体DNA的AFLP分析

用264个随机引物(E11,M24)对K型和冈型水稻(Oryza sativa L.)的不育系、保持系和F1代线粒体DNA进行AFLP分析,共筛选到了37对扩增效果较好的引物,显示出9条可能与K型水稻雄性不育相关的差异带,3条可能与冈型水稻雄性不育相关的差异带.其中引物组合E10M8(EcoR Ⅰ-ATA×Mse Ⅰ-AGC)扩增后,K型和冈型水稻不育系、F1代几乎在750 bp左右的位置都同时表现出了差异条带而在保持系相应位置表现为缺失,它与"缺陷型不育"假设一致.由此证明了K型和冈型水稻不育系线粒体DNA的特异性及其与雄性不育的相关性,为回收克隆等后续工作奠定了基础.     

提取水稻线粒体DNA的一种简易方法

介绍了一种利用常用的普通试剂提取水稻线粒体DNA的简便方法 .与其它方法相比较 ,它具有省时、省钱、高效、快捷、简单等特点 .用该方法提取的水稻线粒体DNA ,经紫外分光光度计测定其纯度、限制性内切酶进行酶切和随机引物进行RAPD反应的结果表明 ,完全可以满足作为RFLP和RAPD等分析质量的要求 .     

麦芽寡糖基海藻糖合酶基因的克隆及在大肠杆菌中的表达

应用PCR方法，从芝田硫化叶菌B12中扩增了麦芽寡糖基海藻糖合酶基因（TreY），扩增产物大小为2．2kb，将扩增片段与pUCm-T连接，得到重组质粒pUCm-T-TreY，CaCl2法转化DH5α，并筛选阳性克隆。用BamHI／Ndel分别双酶切pUCm-T-TreY和表达载体pET21a，连接后得重组pET21a-TreY，转化大肠杆菌DE3并进行了表达。表达产物经SDS-PAGE分析，得到一条分子量约为74kDa的特异条带，同核苷酸序列所推导的值相符。     

盐生盐杆菌含耐盐相关基因DNA片段的克隆

将盐生盐杆菌总DNA的BamHI不完全酶切片段与质粒pUC19连接成重组质粒,并以重组质粒转化E.coli JM101.经X-Gal-IPTG法筛选白色重组子,结合高盐平板检测,已经获得了3株比原受体菌的耐盐性提高30％－67％的转化子。重组质粒酶切电泳分析表明,被克隆的盐生盐杆菌含耐盐相关基因的DNA片段长度在1．0－3．5kb之间。     

人类扭转蛋白A的基因克隆、表达与蛋白质纯化(I)——DYT1基因克隆与原核表达

通过基因工程方法,用大肠杆菌原核表达系统表达人扭转蛋白A.从该蛋白编码序列中设计引物,以人肝cDNA文库作模板扩增到编码该蛋白的基因片段.将所得片段与pMD l8-T载体连接,转化到JM l09大肠杆菌中,从转化平板上挑出菌落,用碱裂解法提取质粒,通过PCR和酶切分析,筛选到阳性克隆,测序结果与文献报道结果一致.提取质粒,用BamHI和XhoI酶切,回收目的片段,分别克隆到原核表达载体pET28a( )和pGEX-6P-1中,转化JM l09受体菌,从JM l09受体菌中提出质粒,再转化到BL21(DE3)菌中,筛选出阳性克隆,构建了人扭转蛋白A原核表达载体.IPTG诱导该工程菌,培养并离心收集.SDS-PAGE结果表明该蛋白在两种载体中均得到了高效表达.     

Markedness and its Interpretation in Language Use

Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.     

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…     

低渗透非均质砂岩油藏启动压力梯度研究

理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。     

On Tragic Consciousness In The Works By Ernest Hemingway

As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.     

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.     

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。     

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

正~~     

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     

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