白芷细胞外21kDCaMBP抗体制备

植物细胞外存在钙调素（CaM），并且胞外CaM具有促进白茫悬浮培养细胞增殖“’及原生质体第一次分裂、壁再生功能“‘．唐军”’首次在白花悬浮培养细胞外检测并纯化了一种与CaM结合依赖于Ca’“的分子量约为21kD的钙调素结合蛋白（CaMBP），那么，对此蛋白深入研究将有助于揭示胞外Ca’”·CaM信号分子的作用机制．由于ZIkDCaMBP主要存在于细胞壁区，含量较低，提纯困难，因此，对此蛋白的大量纯化及其抗体制备就成为对ZIkDCaMBP定量、定位及其生理功能深入研究的关键．本实验利用唐军”’建立的SephadexG—100凝胶过滤及CM…     

花椰菜天然钙调素抗体的特性及其ELISA定量法的建立

钙调素是一种具有多种调节功能的钙结合蛋白质，其免疫原性很弱，本文采用两次基础免疫和多次加强免疫的方法，获得了免疫花椰菜天然钙调素抗血清，用免疫双扩散法鉴定，其效价为１：３２钙调节不易与固相载体结合，我们先用０．２％戊二醛处理聚苯乙烯板载体，再用于包被钙调素，在上述基础上建立了定量测定植物钙调素的竞争性酶联免疫吸附测定（ＥＬＩＳＡ）技术及检测动物血清中抗钙调素抗体的间接ＥＬＩＳＡ技术。     

马铃薯钙调素表位的微机多参数预测

据文献报道［１－３］,分析蛋白氨基酸序列的某些参数,如 Ｈｏｐｐ　＆ Ｗｏｏｄｓ　亲水性和Ｗｅｌｌｉｎｇ抗原性,可达到预测蛋白抗原分子表位的目的。本研究借助微机,对马铃薯钙调素分子表位进行了预测和分析,结果表明,该种钙调素分子存在７个表位,其中至少有３个表位可能严生相应抗体。鉴于钙调素分子一级结构具有高度保守性,以上预测结果对研制各种植物钙调素单克隆抗体均有较大的参考价值。     

小麦高分子量谷蛋白亚基多克隆抗体的制备、纯化及鉴定

利用丙酮沉淀法对小麦高分子量谷蛋白亚基（HMW—GS）进行快速高效纯化，以纯化后的蛋白做为抗原，免疫白兔，制备了相应的多克隆抗体．与大肠杆菌裂解液共温育去除交叉反应性抗体，并用饱和硫酸铵沉淀法对抗体进行了初步纯化．电泳及免疫印迹结果表明，所制备的多克隆抗体能与小麦的高分子量谷蛋白亚基发生强烈反应，而与小麦中的水溶性蛋白、醇溶蛋白及低分子量谷蛋白亚基不反应．该抗体的制备为进一步研究小麦高分子量谷蛋白亚基的功能及其品质改良提供了基础材料和有用工具．     

人表皮生长因子(hEGF)基因的合成、鉴定及植物表达质粒的构建

用PCR方法合成了人表皮生长因子（hEGF）基因，构建了原核表达质粒p 20T-hEGF，研究了原核表达的重组蛋白hEGF的生物学活性，并构建了植物表达质粒，研究表明：无论是融合蛋白GST－hEGF或纯化的hEGF蛋白，都有相当高的生物活性，hEGF蛋白对Hela细胞的增殖有很好的促进作用，免疫小鼠亦能产生很高的免疫应答反应。     

BIG-3的原核表达、纯化及其多克隆抗体的制备与鉴定

原核表达、纯化骨形态发生蛋白诱导基因（BIG-3,BMP-2-induced gene 3kb）所编码的融合蛋白并制备其多克隆抗体。将BIG-3基因插入原核表达载体PGEX-4T-2的多克隆位点获得pGEX-4T-2-BIG-3重组表达载体,转化大肠杆菌BL21（DE3）菌株,诱导表达,SDS-PAGE鉴定融合蛋白表达情况及Sepharose4B层析柱亲和层析法纯化的融合蛋白;用纯化的融合蛋白免疫新西兰大白兔制备其多克隆抗体,并以Western blot鉴定其特异性。结果在大肠杆菌中获得BIG-3-GST融合蛋白高水平的诱导表达,经Sepharose4B层析柱亲和层析,GST-BIG-3融合蛋白在电泳图片上显示较为清晰的单一条带,所制备的多克隆抗体具有较高的特异性。说明在大肠杆菌中成功表达且以亲和层析法纯化得到了BIG-3-GST融合蛋白,并制备了特异性较高的多克隆抗体。     

恶性疟原虫复合多表位基因的原核表达和多克隆抗体制备

原核表达恶性疟原虫多期多表位基因并制备其多克隆抗体。在前期构建恶性疟原虫复合多期多表位重组真核表达载体的基础上,将测序正确的AMAMEG基因克隆入原核表达载体pGEX4T—1并诱导表达。采用包涵体洗涤的方式纯化目的蛋白,以AMA—1抗体对纯化蛋白进行Western—blot分析。将纯化的融合蛋白免疫小鼠制备多克隆抗体。通过PCR成功获得长度为1 000 bp的恶性疟原虫AMA—1胞外域基因片段,获得了与多表位基因连接后的AMAMEG基因。通过诱导表达,显示在相对分子量约95 000处有预计大小的特异性条带,表明成功表达出融合蛋白。经此纯化的融合蛋白免疫的小鼠能产生特异性抗体,其滴度达到1∶105。纯化的融合蛋白和多克隆抗体为研究新型疟疾疫苗奠定一定的理论和实践基础。     

HIV-1复合多表位基因的原核表达和多克隆抗体制备

为表达HIV-1复合多表位基因并制备其多克隆抗体.设计引物,以HIV-1标准株全长cDNA序列为模板,PCR扩增获得p24基因片段;合成改造后的多个表位基因MEG并且与p24片段相连接,克隆入大肠杆菌.将测序正确的p24MEG基因克隆入原核表达载体pET32a并诱导表达.采用Ni2 -NTA亲和柱纯化目的蛋白,以p24抗体对纯化蛋白进行Western-blot分析.将纯化的融合蛋白免疫家兔制备多克隆抗体.通过PCR成功获得长度为651 bp的HIV-1的p24片段,获得了与多表位基因连接后的p24MEG基因,通过诱导表达,显示在相对分子量约45 000处有预计大小的特异性条带,表明成功表达出融合蛋白.经此纯化的融合蛋白免疫的家兔能产生特异性抗体,其滴度达到1:3 200.纯化的融合蛋白和多克隆抗体为研究新型HIV疫苗奠定一定的理论和实践基础.     

绿色荧光蛋白GFP多克隆抗体的制备及在多种细胞中的特异性鉴定

绿色荧光蛋白是当前在生物及生物医学研究当中应用最为广泛的一种荧光标记蛋白.为开发一种高灵敏度、高特异性以及简单经济的GFP多克隆抗体,并使之适用于在跨物种细胞里表达的GFP蛋白进行检测,本文利用gfp基因cDNA全序列分别构建了带有6×His和GST标签的原核表达载体pET28A-gfp和p GEX-2T-gfp,并将其转入大肠杆菌(Escherichia coli)BL21(DE3)细胞进行诱导表达,而后进行聚丙烯酰胺凝胶电泳(SDS-PAGE)表达鉴定,分别获得了相对分子质量为2.6×10~4和5.2×10~4的重组6×His/GST-GFP融合蛋白.将亲和纯化的6×His-GFP融合蛋白免疫新西兰大白兔,采集第5次免疫后血清用ELISA测定效价为1﹕32,000.抗血清依次经Protein A亲和纯化和GST-GFP抗原抗体纯化后,用免疫印迹实验(Western blot)方法检测抗体特异性,结果表明制备的多克隆抗体能很好地识别在莱茵衣藻(Chlamydomonas reinhardtii)、HEK293细胞和大肠杆菌中表达的GFP,对继续开展GFP相关研究具有重要意义.     

重组人源抗血管内皮生长因子单克隆抗体的制备及鉴定

通过优化密码子获得带有信号肽的人源抗血管内皮生长因子（VEGF）抗体的重链和轻链DNA序列, 分别构建表达载体pcDNA VEGFH和pcDNA VEGFV, 将表达载体共转染HEK 293细胞后, 与C6胶质瘤细胞共培养, 并将所得蛋白进行分离纯化. 实验结果表明: pcDNA VEGFH和pcDNA VEGFV共转染HEK 293F细胞, 可显著抑制C6细胞增殖（P<0.01）; 转染24~144 h后的细胞培养上清液中均有蛋白表达; 所得纯化蛋白和标准品一致; 纯化所得抗体蛋白能明显抑制C6细胞增殖（P<005~001）, 与标准品抑制效果相似.     

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.     

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.     

An Analysis of Catherine＇s Character——Wuthering Heights

Wuthering Heights, Emily Bronte＇s only novel, was published in December of 1847 under the pseudonym Ellis Bell. The book did not gain immediate success, but it is now thought one of the finest novels in the English language. Catherine is the key character of this masterpiece, because everybody and everything center on her though she had a short life. We can understand this masterpiece better if we know Catherine well.     

Lower Paleozoic oil relationships within Williston Basin, Canada

The Williston Basin is a significant petroleum province, containing oil production zones that include the Middle Cambrian to Lower Ordovician, Upper Ordovician, Middle Devonian, Upper Devonian and Mississippian and within the Jurassic and Cretaceous. The oils of the Williston Basin exhibit a wide range of geochemical characteristics defined as "oil families", although the geochemical signature of the Cambrian Deadwood Formation and Lower Ordovician Winnipeg reservoired oils does not match any "oil family". Despite their close stratigraphic proximity, it is evident that the oils of the Lower Palaeozoic within the Williston Basin are distinct. This suggests the presence of a new "oil family" within the Williston Basin. Diagnostic geochemical signatures occur in the gasoline range chromatograms, within saturate fraction gas chromatograms and biomarker fingerprints. However, some of the established criteria and cross-plots that are currently used to segregate oils into distinct genetic families within the basin do not always meet with success, particularly when applied to the Lower Palaeozoic oils of the Deadwood and Winnipeg Formation.     

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     

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