Interaction between PⅡ and NifA in Azospirillum brasilense Sp7

The interaction between PⅡ and NifA in A.brasilense Sp7 was investigated by using the yeast two-hybrid system.Our experimental results showed that PⅡ directly interacted with the entire NifA protein and its N-terminal domain,but did not interact with the central domin and the C-terminal domain of NifA.No interaction happened if glnB coding for PⅡ was frame-shift mutated.Pz,a homolog of PⅡ,had no interation with NifA.     

PAS domain of the deduced Org35 protein mediates the interaction with NifA

PAS domains are sensory input domains and pro- tein-protein interaction sites that have been identified recently in a family of sensory proteins from all king-doms of life[1,2]. A variety of environmental stimuli such as light, oxygen, redox potential, an…     

Upstream CRP-binding site is not essential for CRP-cAMP-mediated inhibition on the nifU promoter

When the NifA-mediated activation of Klebsiella pneumoniae nifU promoter is recreated in Escherichia coli, it has been observed that CRP-cAMP has an inhibitory effect on the nifU promoter. Sequence analysis indicates that there is a strong CRP-binding site located upstream of the nifU promoter, overlapping completely with a previously identified NifA-binding site. In vitro gel retardation analysis indicates that this putative CRP-binding site has similar affinity for CRP, when compared with that at the lac promoter, suggesting that CRP could effectively compete with NifA for such a binding site under physiological conditions. When this putative CRP-binding site on nifU was mutated, in vitro gel retardation analysis indicates that CRP can no longer bind to the mutant promoter. However, when constitutively expressed NifA is used as the activator, CRP-cAMP-mediated inhibitory effect on this mutant nifU promoter has no significant difference when compared with that obtained from its wild-type promoter. These results suggest that direct interaction between CRP and Eσ54, other than the DNA binding site(s) competition between CRP and NifA, plays the principal role in the CRP-cAMP-mediated inhibitory effect on nifU.     

Screening of the interacting proteins with NifA in <Emphasis Type="Italic">Azospirillum brasilense</Emphasis> Sp7 by the yeast two-hybrid system

NifA in AzospiriUum brasilense plays a key role in regulating the synthesis and activity of nitrogenase in response to ammonia and oxygen available. In this work we used the yeast two-hybrid system to identify the proteins that interact with NifA. The nifA gene was fused to the yeast two-hybrid vector pGBD-C2, and three A. brasilense Sp7 genomic libraries for use in yeast two-hybrid studies were constructed. Screening of the libraries identified four clones encoding proteins that interact with NifA. The confirmation of the interactions of each gene product of the four clones and NifA were carried out by exchanging the vectors for nifA and the four clones and by mutageneses of the four clones with shift reading frame experiments in yeast two-hybrid studies. DNA sequence analyses showed that two clones encode proteins containing PAS domains that play an important role in signal transduction. One clone has high similarity with the fhuE gene of Escherichia coli, whose gene product is involved in iron uptake and transportation, and the other clone encodes an unknown protein.     

捕食线虫真菌丝氨酸蛋白酶PII结构的同源模建

通过同源模建方法构建捕食线虫真菌丝氨酸蛋白酶PII的三维结构模型,并结合以往研究对其结构进行分析。结果表明,蛋白酶PII的结构具有枯草杆菌丝氨酸蛋白酶特有的a／B脚手架折叠模式,具有保守的催化三聚体和氧负离子孔结构组织,整体表现为坚固的球状折叠构型,不具备二硫键。研究结果将为进一步深入研究PII的结构和功能之间的关系奠定结构基础。     

Transcriptome analysis of Sinorhizobium meliloti nodule bacteria in nifA mutant background

Sinorhizobium meliloti is one genus of gram-nega- tive soil bacteria that can fix atmospheric nitrogen in root nodules of its symbiotic leguminous host plants[1]. Specific recognition and progressive differentiation ofboth bacteria and host cells are requ…     

用酵母双杂交检测BAR与PDZ结构域的相互作用

用酵母双杂交系统探索P ICK 1分子中BAR结构域与PDZ结构域的相互作用.分别构建了pGAD-PDZ和pGBK-BAR重组子,共转入酵母细胞后,涂布于SD(L eu-,T rp-,H is-)固体平板培养基,经滤纸复印和显色反应,在8 h内观察到只有pGAD-PDZ/pGBK-BAR的表达产物呈阳性,而其它对照pGADT 7/pGBKT 7、pGAD-PDZ/pGBKT 7和pGADT 7/pGBK-BAR均呈阴性反应,说明BAR与PDZ结构域具有相互作用.     

日本赭纤虫第一类肽链释放因子C端结构域分析

eRF1的C结构域与eRF3的C结构域相互作用对于蛋白质翻译终止过程中的快速反应至关重要.通过计算机同源建模对日本赭纤虫第一类肽链释放因子C结构域Bj-eRF1C进行结构模拟,发现在Bj-eRF1C结构域中有些肽段直接参与了eRF1-eRF3相互作用,特别是Bj-eRF1C结构域中V294和D297位点高度保守.通过定点突变与pull-down分析,表明在Bj-eRF1C结构域中V294和D297是eRF1-eRF3相互作用的关键位点.     

基于局部搜索策略的车间作业调度问题的算法研究

为车间作业调度问题提供了一个快速、易于实现的近似算法．该算法基于局部搜索策略,采用特殊的邻域构造方法,即邻域的构造仅与关键路径上的工序相关．该算法找到了所测试的14个标准算例中12算例的最优解,而且在PⅡ233的计算机上每个算例的计算时间不超过1s。     

The heteromeric cyclic nucleotide-gated channel adopts a 3A:1B stoichiometry

Cyclic nucleotide-gated (CNG) channels are crucial for visual and olfactory transductions. These channels are tetramers and in their native forms are composed of A and B subunits, with a stoichiometry thought to be 2A:2B (refs 6, 7). Here we report the identification of a leucine-zipper-homology domain named CLZ (for carboxy-terminal leucine zipper). This domain is present in the distal C terminus of CNG channel A subunits but is absent from B subunits, and mediates an inter-subunit interaction. With cross-linking, non-denaturing gel electrophoresis and analytical centrifugation, this CLZ domain was found to mediate a trimeric interaction. In addition, a mutant cone CNG channel A subunit with its CLZ domain replaced by a generic trimeric leucine zipper produced channels that behaved much like the wild type, but less so if replaced by a dimeric or tetrameric leucine zipper. This A-subunit-only, trimeric interaction suggests that heteromeric CNG channels actually adopt a 3A:1B stoichiometry. Biochemical analysis of the purified bovine rod CNG channel confirmed this conclusion. This revised stoichiometry provides a new foundation for understanding the structure and function of the CNG channel family.     

近邻和四自旋相互作用对铁电体畴结构影响的模拟

在二维Ising模型基础上,采用蒙特卡罗方法研究了二自旋和四自旋相互作用下,铁电体畴结构的动态演化和极化反转过程。数值模拟结果表明二自旋相互作用下极化反转过程中显示不出明显的畴纵向生长趋势。而四自旋相互作用下,方型的四自旋相互作用对于畴的纵向生长起主要作用,T型的四自旋相互作用可以导致反铁电相的出现。     

TD-PSOLA技术在汉语语音波形编码合成中的应用

以时域基音同步叠加（ＰＳＯＬＡ）技术和一个全汉语单音节库为合成单元进行汉语语音波形编码合成,针对汉语语音的音高、时长、音强以及音节之间的协同发音效应等影响合成语音质量的主要因素,建立相应音节的声调曲线、时长规则和音节之间协同发音规则等韵律规则,并利用时域基间同步叠加法原理调整合成语音的音高和时长,从而使合成的语音比较清晰自然。     

地基动力阻抗的双自由度集总参数模型

在土－结构动力相互作用分析中，下覆地基对基础与结构的阻抗函数通常强烈地依赖于外加激振频率。当结构为线性时，耦联体系的动力分析可采用子结构法在频域上有效地实施。但对于非线性结构，在时域上要用与频率有关的动力阻抗进行相互作用分析就变得十分困难与复杂。而单靠采用某一频率下动力阻抗函数固定值的单自由度集总参数模型不能充分地包含所有频率成分而难以反映实际性态。     

与畴界有关的介电损耗、介电常数和内耗研究Ⅱ.理论部分

提出了畴界与外场、晶格及缺陷间的互作用模型以及畴界间的互作用模型,并在该模型基础上,系统地解释了与畴界运动有关的介电损耗、介电常数的和内耗(f=1KHz～10KHz)。本文还对介电损耗的理论和实验结果进行了比较     

土-结构动力相互作用的振型分解法

采用振型分解法对土-结构相互作用体系进行分析．基于上部结构具有经典正交振型的假设,对控制方程进行解耦,使之成为单自由度体系,直接运用振型叠加法求解．本文方法在求解土-结构相互作用问题时与基础固定时的动力响应频域分析方法一致,只需要调整地震动输入,就可得到问题的解答．数值计算表明,本文方法的计算值与试验值吻合较好．     

Crystal structure of a streptococcal protein G domain bound to an Fab fragment.

Protein G is a cell-surface protein from Streptococcus which binds to IgG molecules from a wide range of species with an affinity comparable to that of antigen. The high affinity of protein G for the Fab portion of IgG poses a particular challenge in molecular recognition, given the variability of heavy chain subclass, light chain type and complementarity-determining regions. Here we report the crystal structure of a complex between a protein G domain and an immunoglobulin Fab fragment. An outer beta-strand in the protein G domain forms an antiparallel interaction with the last beta-strand in the constant heavy chain domain of the immunoglobulin, thus extending the beta-sheet into the protein G. The interaction between secondary structural elements in Fab and protein G provides an ingenious solution to the problem of maintaining a high affinity for many different IgG molecules. The structure also contrasts with Fab-antigen complexes, in which all contacts with antigen are mediated by the variable regions of the antibody, and to our knowledge provides the first details of interaction of the constant regions of Fab with another protein.     

列车载荷下隧道联络通道动态响应的并行计算

以上海某大型双线隧道为例,建立了列车 隧道 联络通道 土体的大型三维有限元模型.基于上海超算中心曙光5000A,设计了列车隧道动态耦合均衡的分区方法,解决了因模型规模庞大而造成计算量大的问题.分析了列车载荷下联络通道结构的动态响应,比较了不同分区方案的计算效率.结果表明：列车载荷在联络通道与隧道连接部位引起的附加动应力较小,但该位置处于较高的应力水平,应尽量避免在联络通道位置汇车;列车隧道耦合均衡的分区方法比常用的递归坐标二分法具有更高的并行效率.     

Membrane protein association by potential intramembrane charge pairs.

The transmembrane domain of the alpha chain of the T-cell receptor is responsible both for its assembly with the CD3 delta chain and for rapid degradation of the unassembled chain within the endoplasmic reticulum. The determinant for both assembly and degradation is located in a segment of eight residues containing two basic amino acids. We show here that placement of a single basic residue in the transmembrane domain of the Tac antigen can induce interaction with the CD3 chain, through its transmembrane acidic residue. This interaction is most favoured when the interacting residues are located at the same level in the membrane. The ability to induce protein-protein interaction by placing charge pairs within transmembrane domains suggests an approach to producing artificial dimers.