穿梭质粒pNBC11的构建及性质研究

质粒 pUB110与 pUC18分别册 EcoRI 酶切,T_4DNA 连接酶进行连接,得到重组质粒 pBC11。琼脂糖凝胶电泳分析,该质粒分子量为4.8×10~6道尔顿。pBC11是一种穿梭质粒,能转化大肠杆菌和枯草杆菌,它对大肠杆菌 C600、JM101的转化率分别为:4×10~5、1.5×10~5转化体/μgDNA.对枯草杆菌 BR151、QB1130的转化率分别为1.5×10~3、2.15×10~3转化体/μgDNA。在基因表达方面,pBC11上的 K_m~r 基因能在大肠杆菌和枯草杆菌中复制和表达,但 pBC11的 Ap~r 基因不能在枯草杆菌中表达。     

厦门霉素生物合成基因簇在白色链霉菌及吸水链霉菌FR-008中的异源表达

将携带有厦门霉素生物合成的整合质粒p LMO09404,通过供体菌大肠杆菌ET12567(p UZ8002)与受体菌白色链霉菌及吸水链霉菌FR-008进行接合转移,将得到的接合转移菌株经PCR验证后进行发酵,发酵液经HPLC和质谱检测.结果显示,厦门链霉菌来源厦门霉素基因簇可以在白色链霉菌及吸水链霉菌FR-008中异源表达.     

氧化硫硫杆菌重组质粒pSDT125的构建及其在大肠杆菌之间的转移

对氧化硫硫杆菌(Thiobacillus thiooxidans)隐蔽性质粒pTt12进行了限制性酶切分析,并绘出了限制性酶切图谱,其大小为13.7Kb.将pTt12质粒与pBR325质粒重组,得到了重组质粒pSDT125(Cm',Ap',Tc~s).氧化硫硫杆菌重组质粒pSDT125本身不具有接合转移能力,但它在广泛寄主范围的转移性质粒RP4的带动下以较高的频率在大肠杆菌(Escherichia coli)之间转移,表明在氧化硫硫杆菌质粒pTt12上存在着与接合转移有关的DNA序列.     

从猪粪中分离抗铜大肠杆菌及其抗铜基因的初步定位

 从云南昆明周边的4个养猪场采集不同年龄段的猪粪样品,分离猪粪中的大肠杆菌.从分离到的大肠杆菌中发现了具有金属铜抗性的菌株并统计出抗性菌株的发生率.摸索了测定硫酸铜对抗铜性大肠杆菌的最低抑菌浓度(MIC)的方法,接合实验确定抗铜基因是否位于可移动的接合质粒上.实验结果初步表明抗铜大肠杆菌的发生率与饲料中添加硫酸铜的量成正相关性,且72%的抗性菌株中的抗铜基因位于可移动的接合质粒上.     

家蚕(Bombyx mori)基因文库的建立和鉴定

组建家蚕Bombyx mori品种754和东肥的基因文库.用cosmid 质粒pHC 79在BamH1和Pst1位点进行休外重组和包装,转导至E.coli 受体菌HB101中,获得10~7以上的重组体克隆,统计分析表明库内已包括了将近100%的家蚕基因组的遗传信息.试验分析了包装混合物BHB2688/BHB2690体外连接的家蚕/载体DNA 的比值以及家蚕总DNA 酶解程度对重组建库的影响,使建库效率达到10~6—10~7CFU/μg 载体DNA,双抗值低至5%以下.在对基因文库进行鉴定时发现了克隆的家蚕DNA 片段对HB101的leu pro 基因突变的互补(校正)效应,频率为10~(-5).     

La(Ⅲ)的抑菌作用研究

进行了La()抑菌实验,采用比浊法、平板计数法、滤纸片法对大肠杆菌(Escherichiacoli)、金黄色葡萄球菌(Staphulococcusaureus)和枯草芽孢杆菌(Bacillussubtilis)进行了测定.结果表明:所用抑菌物对供试菌均有不同程度的抑制作用,当La()的浓度为1.0×10-6~1.0×10-4mol/L时抑制作用最明显,且随着La()离子浓度增大,抑菌作用趋势增强.     

氧化亚铁硫杆菌质粒pTf—52限制图谱和嵌合质粒pSDF—1的构建

从一株氧化亚铁硫杆菌(Thiobacillus ferrooxidans)中分离到一个分子量为4.5×10~6Md 的质粒 pTf—52,用 Pst Ⅰ、BglⅡ、Kpn Ⅰ酶切此质粒,进行限制性酶切分析,作出了 pTf—52的限制图谱;并将 pTf—52插入到 pBR325的 Pst Ⅰ位点,体外重组,转化 E.coli HB101,构建成一个能够在 E.coli 中进行复制的嵌合质柱 pSDF—1(Tc~rCm~rAp~s),分子量为8.2×10~6Md。经限制性酶切分析,确证 pSDF—1是由 pBR325和 pTf—52重组成的,并确定了这两种质粒在 pSDF—1中的连接方向。     

Sanglifehrin A产生菌Streptomyces flaveolus DSM 9954遗传转化系统的建立

Sanglifehrin A的产生菌淡黄色链霉菌Streptomyces flaveolus DSM 9954,通过筛选,得到其菌丝培养基为TSB, 生孢培养基为ISP-4,抗性实验显示淡黄色链霉菌Streptomyces flaveolus DSM 9954对所选抗生素都较敏感. 利用原生质体转化和接合转移都成功地构建了两种遗传转移系统,并且从接合转移成功的链霉菌转化子中回收质粒pKC 1139,经EcoR Ⅰ单酶切电泳,验证了质粒确实通过接合转移从大肠杆菌转移至宿主链霉菌, 这为研究Sangl     

氧化硫硫杆菌质粒pTt12复制原点片段分离及其重组质粒pSDT125转移性能

氧化硫硫杆菌重组质粒ｐＳＤＴ１２５在ＲＰ４质粒的带动下以高出ｐＢＲ３２５两个数量级的频率在大肠杆菌之间转移．转移后,ＲＰ４和ｐＳＤＴ１２５以分离的形式共存于接合转移子中．该质粒不能被ＳＭ１０菌株染色体上的ｔｒａ基因带动转移．以ｐＳＤＴ１２５质粒为基础,通过亚克隆构建了两个质粒ｐＳＤＴ１２５０,ｐＳＤＴ１２５１．二者可以被ＲＰ４质粒带动在大肠杆菌之间转移,转移频率与ｐＳＤＴ１２５相近,没有明显降低．还构建了含有氧化硫硫杆菌质粒ｐＴｔ１２片段的穿梭质粒ｐＳＤＭ２１１,可以被ＲＰ４带动在大肠杆菌与氧化硫硫杆菌之间转移．通过限制性内切酶分析,初步将ｐＴｔ１２的复制原点确定在３．０ｋｂ的片段内．     

氧化硫硫杆菌质粒pTt12复制原点片段分离及其重组质粒pSDT …

氧化硫硫杆菌重组质粒ｐＳＤＴ１２５在ＲＰ４质粒的带动下以高出ｐＢＲ３２５两个数量级的频率在大肠杆菌之间转移。转移后,ＲＰ４和ｐＳＤＴ１２５以分离的形式共存于接合转移子中。该质粒不能被ＳＭ１０菌株染色体上的ｔｒａ基因带动转移。以ｐＳＤＴ１２５质粒为基础,通过亚克隆构建了两个质粒ｐＳＤＴ１２５０,ｐＳＤＴ１２５１。二者可以被ＲＰ４质粒带动在大肠杆菌之间转移,转移频率与ｐＳＤＴ１２５相近,没有明显降低。     

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.     

Features of Women's language

Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women＇s language from these perspectives： pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women＇s language, more attention should be paid to language use in social context. What＇s more, the linguistic phenomena in a speaking community can be understood more thoroughly.     

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.     

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。     

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.     

Jane Eyre——A Feminist In The 19th Century

In the 19th century the society was controlled by men, and women were just appendants of them, they had not any rights and freedom. But Jane was an exception, she showed some characteristics of early feminist. Jane showed her characteristics of feminism in three aspects: rebellion, equality, and independence. These characteristics were helpful to her success, and feminism is the only way out for women of that time.     

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

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