蛋白亚基局部相互作用模拟构筑二十面体病毒衣壳

二十面体病毒衣壳结构蛋白具有不同的构象。装配过程中,正在附着的蛋白亚基构象与位置在先前蛋白亚基构象的指令下,发生相应的调整后装配到衣壳骨架上。局部相互作用指导下的装配反复进行,最终形成规整的二十面体病毒衣壳的立体结构。对T=1、3、4、7的病毒衣壳,分别建立了蛋白亚基构象局部相互作用的连接方式与增长方式,并构筑了T=4、7的立体模型。     

二十面体病毒衣壳自组装的热力学参数

球状病毒衣壳采用二十面体对称。本文就二十面体病毒蛋白质骨架的自组装提出一个统计热力学模型。统计结果表明病毒衣壳二十面体对称不是自由能最小化的结果，而是衣壳内部蛋白结构参数最优化的结果。     

斑马鱼PKR的dsRBM1和dsRBM3重组及体外功能分析

为了重组斑马鱼(Daniorerio)双链RNA依赖的蛋白激酶(DrPKR)的双链RNA结合模体1(dsRBM1)和双链RNA结合模体3(dsRBM3)基因,并对重组基因表达的蛋白进行鉴定和功能分析,设计定向删除引物,应用PCR技术扩增DrPKR的ds RBM1和dsRBM3的基因片段,通过重叠PCR技术将dsRBM1和dsRBM3基因重组在一起得到dsRBM13基因,再将dsRBM13构建到原核表达质粒pET32a上并表达和纯化蛋白;通过表达蛋白的二聚化实验和pull down实验对重组蛋白dsRBM13进行体外功能分析。结果:DrPKR的ds RBM1和dsRBM3重组的蛋白dsRBM13可以进行二聚化和多聚化,且能结合Poly I:C(人工合成的dsRNA)。因此,重组DrPKR蛋白模体dsRBM13仍然具有二聚化和dsRNA结合活性,提示串联了三个dsRBM的Dr PKR激活过程中,串联两个dsRBM对DrPKR激活是必需的,并且在DrPKR结合dsRNA的激活过程中,因dsRNA的长短和空间结构的多样性,含有三个dsRBM的DrPKR可能存在更强和更多的结合方式,因此具有更强的抗病毒免疫反应功能。     

N端延长对大肠杆菌铁蛋白结构稳定性及其自组装的影响

【目的】研究延长的N端肽链对铁蛋白结构稳定性及蛋白自组装的影响,为在N端融合生物分子构建功能性纳米材料奠定基础。【方法】在大肠杆菌铁蛋白N端添加6个组氨酸残基,所得重组蛋白采用体积排阻色谱、非变性凝胶电泳和圆二色谱等手段与野生型铁蛋白的聚合态、二级结构及热力学稳定性进行对比分析。【结果】体积排阻色谱表明野生型铁蛋白在溶液中以24聚体和二聚体混合物形式存在,但突变蛋白仅以二聚体的形式存在; 非变性凝胶电泳实验结果与体积排阻色谱结果吻合,即突变蛋白仅显示1条带,且与野生型铁蛋白的2聚体迁移位置基本相同; 圆二色谱结果表明突变蛋白的二级结构与野生型类似,呈典型的α-螺旋结构,突变蛋白的T_m值比野生型降低了1.1 ℃。【结论】在大肠杆菌铁蛋白亚基的N端增加6个组氨酸残基所得突变蛋白的二级结构没有发生明显变化,但却无法形成24聚体壳状结构,在溶液中仅以二聚体的形式存在,且热力学稳定性有所降低。N端的延长可能导致铁蛋白单体构型发生改变,因而无法进一步组装成壳状结构。     

H5N1亚型禽流感病毒M基因的克隆与分子进化分析

以一株H5N1亚型禽流感病毒RNA为模板,用RT-PCR方法,扩增M基因全长,将PCR产物克隆于pMD18-T载体,测序结果表明所克隆的982个核苷酸的片段包含了M1和M2基因的完整阅读框架,通过软件推导M1和M2基因分别编码252和97个氨基酸,将M全长序列与Genbank收录的10株H5N1亚型流感病毒M基因序列进行比较,病毒株之间M基因核苷酸序列同源性为92.3%～99.3%,编码的两个蛋白M1和M2氨基酸序列间同源性分别为为96.0%～98.8%和92.9%～99%,分子进化分析揭示了病毒株间的亲源关系.     

蚕豆萎蔫病毒中国分离物RNA2全序列及多聚蛋白切割位点

对蚕豆萎蔫病毒2(BBWV2)分离物B935 RNA2全序列进行了测定,全长由3 601个核苷酸组成(不包括3'端Poly(A)).RNA2包含一个长的开读框,该开读框起始于231位,终止于3 428位,编码一个分子量为119 ku的多聚蛋白.对外壳蛋白亚基N端氨基酸序列测定表明外壳蛋白大亚基(LCP)和小亚基(SCP)是由119 ku多聚蛋白中谷酰胺与甘氨酸及谷酰胺与丙氨酸之间的切割产生的,LCP含有402个氨基酸,SCP含有197个氨基酸.与蚕豆病毒属(Fabavirus)病毒基因组核苷酸及编码的蛋白质氨基酸序列比较表明,B935与BBWV2分离物及广藿香轻花叶病毒具有很高的同源性,而与BBWV1分离物的同源性较低.B935与豇豆花叶病毒属(Comovirus)和线虫传多面体病毒属(Nepovirus)的基因组结构相似,但序列同源性很低.B935与豇豆病毒属病毒的LCP和SCP氨基酸具有共同的保守序列.用4株单克隆抗体进行TAS-ELISA测定表明B935与BBWV1分离物(NRS和PH3)抗原上有差异.     

八肋游仆虫中心蛋白的核磁共振研究

利用液体核磁共振方法对八肋游仆虫中心蛋白全长蛋白(EoCen),LC端结构域(EoCen-LC)和N端结构域(EoCen-N)进行了分析,并研究了蛋白与钙离子的相互作用情况.研究结果表明,未加入钙离子,全长蛋白,LC端结构域和N端结构域谱图重叠严重,峰型较差,说明未折叠成良好的三级结构,存在较大的构象交换或一定程度的聚集;与钙离子结合后全长蛋白,LC端结构域三级结构有所改善,但仍然无法用液体核磁共振方法进行进一步的结构研究.N端结构域与钙离子结合后谱峰分散性好,说明N端结构域具有较好的三级结构,能够应用核磁共振的方法进行结构解析.     

室温下的小蛋白折叠研究

目前,无论在实验还是理论研究方面,研究蛋白质的从头折叠即使是小蛋白的折叠也非常困难.在室温下研究蛋白质的折叠过程可以有效的阐述蛋白质折叠机制.笔者从2I9M的线性结构出发,在室温下运行了15 ns的分子动力学模拟,发现它在6.4 ns时成功折叠到了天然态结构,此时的结构与天然态结构的均方根偏差为1.17(A),模拟结构与天然态结构符合的非常好.折叠过程是首先在C端出现了螺旋,随后N端又出现了螺旋,然后螺旋从N端鱼贯式生成,最终形成天然态结构.     

油菜中—未知BnRCH蛋白的E3连接酶活性分析

利用本实验室前期以ATP6为诱饵蛋白,通过酵母双杂交系统筛选到的一个N端含有RINGv结构(RING-variant)的蛋白,暂命名为BnRCH,构建GTK-BnRCH表达载体在原核细胞E.coli BL21(DE3)中高效表达GST-BnRCH融合蛋白,并将纯化的融合蛋白加入体外泛素反应体系,发现BnRCH在ATP、泛素、E1、E2存在的条件下,能催化多聚泛素化形成,具有E3连接酶活性.     

影响HIV-GP41N端1/2在E.coli中表达的两段氨基酸序列界定

GP41蛋白二级结构预测显示 ,前 1 /2片段的 N端 ( 4～ 2 6位氨基酸 )和 C端 ( 1 67～ 1 89位氨基酸 )各有一个富含疏水氨基酸的穿膜螺旋 (可能性 >0 .9) ,分别从 N1 (前 1 /2片段无 C端穿膜螺旋 )的 N端和 N6(前1 /2片段无 N端穿膜螺旋 )的 C端进行缺失构建一系列缺失突变体 ,只有 p ET-N2 ,p ET-N3 ,p ET-N4,p ET-N5在大肠杆菌中获得高效表达 ,其表达量占菌体总蛋白的 2 0 % ,Western blot显示表达蛋白与 HIV阳性血清有良好的反应性 .而 p ET-N1 ,p ET-N6在大肠杆菌中表达量很低或不表达 .从而证明 1～ 6位 ,1 84～ 2 0 1位氨基酸是影响 gp41基因 N端 1 /2片段表达的主要因素     

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.     

Design and analysis of generic LBS application developing platform

Location based services is promising due to its novel working style and contents.A software platform is proposed to provide application programs of typical location based services and support new applications developing efficiently. The analysis shows that this scheme is easy implemented, low cost and adapt to all kinds of mobile nework system.     

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