海洋底泥环境DNA提取与纯化方法比较研究

为了探索有效的海洋底泥宏基因组DNA提取与纯化方法,分别采用CTAB结合SDS法、CTAB结合玻璃珠法、SDS结合蛋白酶K法和SoilMasterTM试剂盒法提取海洋底泥宏基因组DNA,用柱式腐殖酸清除试剂盒法与电洗脱法纯化宏基因组DNA,并对宏基因组DNA的产量与纯度进行评价.结果表明:SDS结合蛋白酶K法提取率最高,DNA片段完整;SoilMasterTM试剂盒法与CTAB结合SDS法次之;CTAB结合玻璃珠法有严重降解;电洗脱法可以得到高纯度、大于42kb宏基因组DNA片段.因此,SDS结合蛋白酶K法和电洗脱法提取纯化海洋近海底泥宏基因组DNA优于其他方法.     

远志属腊叶标本DNA提取方法研究

以远志属植物腊叶标本为例,研究如何从植物腊叶标本中提取到高质量的DNA。在改良CTAB法的基础上,对水浴时间、水浴温度、沉淀时间进行优化。结果表明,降低水浴温度为45℃,延长水浴时间为5-6h,延长沉淀时间为1-2h,可以从远志属腊叶标本中提取到用于PCR反应的DNA。     

槲蕨中提取柚皮苷最适宜条件研究

采用控制变量法、正交试验法探索槲蕨中提取柚皮苷最适宜的条件。控制变量法结果:提取柚皮苷最适宜条件水浴温度为60℃,水浴次数为3次,提取时间为80min,甲醇体积为40mL,甲醇浓度为60%。正交试验法结果:提取柚皮苷最适宜条件水浴温度为60℃,水浴次数为3次,提取时间为40min,甲醇体积为30mL,甲醇体积分数为60%。     

正交设计太白贝母粗多糖的不同提取工艺

对太白贝母粗多糖提取工艺进行了研究。采用水浴、超声波和煎煮提取,经蒽酮比色法进行含量的确定,通过正交实验确定最佳提取条件。水浴法:提取温度70℃,时间1 h,料液比1∶15,采用其最佳提取工艺条件提取贝母多糖提取含量为0.65 mg/g;超声波法:提取温度80℃,时间15 min,料液比1∶15,采用其最佳提取工艺条件提取贝母多糖提取含量为0.99 mg/g;煎煮法:提取温度90℃,时间3 h,料液比1∶20,采用其最佳提取工艺条件提取贝母多糖含量为0.38mg/g。     

一种直接用于PCR分析的风沙土微生物总DNA提取方法

以风沙土为材料,直接提取土壤微生物DNA.采用SDS—溶菌酶—CTAB—蛋白酶K和液氮反复冻融法裂解细胞得到DNA粗提液,以PEG对其纯化.结果表明,SDS—溶菌酶—CTAB—蛋白酶K和液氮反复冻融法裂解细胞可获得大片段的DNA,提高DNA产率.每克干土的DNA提取量为0.586～1.311μg.所提DNA片段在23Kb以上.PEG可有助于去除DNA中腐植酸,DNA不作回收纯化即可作为模板进行16SrDNAPCR扩增.SDS—溶菌酶—CTAB—蛋白酶K和液氮反复冻融裂解细胞、PEG纯化DNA的方法组合是一种简便的适合风沙土微生物总DNA的提取方法.     

活性污泥宏基因组Fosmid文库的构建

大插入片段宏基因组文库的构建是开发大片段目的基因及分析其结构与功能的基础.文中分别采用十六烷基三甲基溴化铵(CTAB)法、试剂盒及琼脂糖包埋法提取活性污泥宏基因组DNA,其中琼脂糖包埋法获得的DNA片段大于23kbp,利用此DNA成功构建了以pCC1FOS为载体的Fosmid文库,该文库含有5280个克隆,平均插入片段长度为35～40 kbp,共包含约200 Mbp的宏基因组DNA.从此文库中随机挑选200个克隆,利用活性筛选方法快速筛选到了1个含有淀粉酶的阳性克隆,表明活性污泥Fosmid文库可用于功能基因的活性筛选,具有开发新基因的潜力.     

三氯乙酸沉淀蛋白法辅助提取牛奶中环腺苷酸工艺

以牛奶为原料,通过离心去除乳中脂肪,采用不同质量分数不同用量TCA沉淀乳中蛋白质,再浓缩乳糖,分离提取环腺苷酸（cAMP）,选用最佳高效液相色谱条件测定牛奶中cAMP含量.结果表明,不同质量分数TCA处理组之间比较,质量分数为10%TCA处理组cAMP含量显著高于其他2组（P〈0.05）;不同用量TCA处理组之间比较,8 mL相同质量分数TCA处理组cAMP含量较其他2组显著降低（P〈0.05）.选用TCA沉淀乳蛋白方法提取cAMP时最佳工艺是TCA质量分数为10%,用量为4 mL,cAMP质量浓度为（29.02±1.58）μg/mL.研究通过建立牛奶中cAMP的提取工艺,可从奶酪副产物、乳清、不合格原料奶中提取cAMP,在降低cAMP提取成本、废物利用、增加牛奶附加值等方面具有重要意义.     

一种从小麦干种子中快速提取DNA的优化方法

对植物组织DNA抽提是在分子水平上对植物进行系统分析与研究的基础性工作．本文提供了一种从小麦干种子中快速提取总DNA的优化方法．该方法不需液氮冷冻或是冻干组织脱水和水浴，用氯仿对材料进行预处理。使用1．5％的SDS抽提缓冲液和氯仿／异戊醇处理，并用预冷的无水乙醇沉淀DNA，即可在短时间内得到小麦基因组总DNA．实验证明，用该方法每300mg干种子可得到约30-40μg的总DNA，其产量和质量均适合进行SSR分析．     

半仿生及超声波辅助半仿生法提取杜仲叶黄酮

探讨半仿生法及超声波辅助半仿生法提取杜仲黄酮的实验条件.半仿生法考察提取温度、提取时间、料液比对黄酮提取率的影响.以半仿生法提取在超声波协同作用的基础上,采用正交试验、单因素考察超声时间、超声温度、料液比对黄酮提取率的影响.结果表明,半仿生法在提取时间1.5 h,提取温度60 ℃,料液比1∶20（g∶mL）时,黄酮提取率高达325.0 mg/g;超声波辅助法提取的最佳条件为时间50 min,温度65 ℃,料液比1∶40（g∶mL）,黄酮提取率为264.4 mg/g.     

荧光原位杂交法检测环境硝化细菌实验条件优化及应用

通过正交试验筛检荧光原位杂交(FISH)技术在检测环境中硝化细菌时的最佳及适合的实验条件.结果显示,样品预处理较优的条件为:热固定2h,乙醇脱水3min;FISH技术检测环境样本中亚硝化细菌的最佳实验条件为:杂交温度46℃,杂交时间3h,洗脱液中NaCl浓度20mmol/L;检测硝酸菌最佳实验条件为:杂交温度46℃,杂交时间5 h,洗脱液中NaCl浓度60 mmol/L.应用该方案对太湖、玄武湖、南京锁金村污水处理厂活性污泥中硝化细菌分布进行分析,结果显示,环境样本中亚硝化细菌和硝酸菌含量分别是:太湖1.68×103, 0.54×103cells/mL;玄武湖1.34×103, 0.38×103cells/mL;锁金村污水处理厂活性污泥8.30×106, 0.52×106cells/g.FISH技术检测环境样品中硝化细菌与传统检测方法相比具有快速、简便、准确的优点,在研究环境微生物方面有较好的应用前景.     

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.     

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.     

Achieving Unusual States of Matter Under High Pressure

正The periodicity of the elements and the non-reactivity of the inner-shell electrons are two related principles of chemistry,rooted in the atomic shell structure.Within compounds,Group I elements,for example,invariably assume the+1 oxidation state,and their chemical properties differ completely from those of the p-block elements.These general rules govern our understanding of chemical structures and reactions.Using first principles calcula-     

Exchange-Correlation and Electronic Excitation Energies from Pairing Matrix Fluctuations

We have developed an adiabatic connection to formulate the ground-state exchange-correlation energy in terms of pairing matrix linear fluctuations.This formulation of the exchange-correlation energy opens a new channel for density functional approximations based on the many-body perturbation theory.We illustrate the potential of such approaches with an approximation based on the particle-particle Random Phase Approximation(pp-RPA).This re-     

Density Functional Theory for the Response of Periodic Systems to Electric Fields Based on the Vector Potential Approach

正The electronic and nuclear(structural/vibrational)response of 1D-3D nanoscale systems to electric fields gives rise to a host of optical,mechanical,spectral,etc.properties that are of high theoretical and applied interest.Due to the computational difficulty of treating such large systems it is convenient to model them as infinite and periodic(at least,in first approximation).The fundamental theoretical/computational problem in doing so is that     

On the Thermodynamic Limit for Responses to Static Electromagnetic Fields

For molecular systems,the quantum-mechanical treatment of their responses to static electromagnetic fields usually employs a scalar-potential treatment of the electric field and a vector-potential treatment of the magnetic field.Although the potential for each field separately is associated with the choice of an(unphysical)origin,the precise choice of the origin for the electrostatic field has little consequences for the results.This is different for the     

An affordable accurate evaluation of the binding conformations predicted by molecular docking studies in bio-system

Franck-Condon factors bridge the gap between theoretical modeling and experimental observations for molecular electronic spectroscopy and electron transfer.Under the displaced harmonic oscillator approximation,multidimensional Franck-Condon factors are decomposed into a product of many one-dimensional(1D)Franck-Condon(FC)factors,and each 1D-FC factor is associated with one Huang-Rhys factor that determines the leading contribution of     

Call for Papers

<正>"The Journal of Shanghai Normal University:Mathematics"is published by Shanghai Normal University as regular issues of The Journal of Shanghai Normal University each year from 2014 in English.The editors-in-chief of the issues are professors Yuhao Cong and Maoan Han.The Journal of Shanghai Normal University was started in 1958 with     

Journal of Donghua University ( English Edition) Contents of Volume 31

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