CLN8P相互作用蛋白靶基因的获取与初步鉴定

为了获得CLN8P相互作用蛋白靶基因并对其进行初步鉴定,实验经抽提酵母质粒DNA,电转化大肠杆菌,选择性培养基分离质粒,酶切鉴定、测序、生物信息学分析,结果获得了CLN8P相互作用蛋白阳性克隆的一个靶基因BAG5;利用酵母双杂交共转化法初步验证,表明BAG5蛋白与CLN8P具有相互作用,并可能因此影响到神经细胞的正常生长.这将有利于进一步深入研究CLN8的功能及阐明NCLs发病机制.     

邻苯二酚与脱氧核糖核酸相互作用的研究

用循环伏安法、荧光光谱法、紫外可见光谱法、量子化学方法研究了pH为7．42的磷酸盐缓冲溶液中邻苯二酚与脱氧核糖核酸（DNA）的相互作用,在DNA开链过程中,邻苯二酚与脱氧核糖核酸作用后的荧光光谱曲线、紫外光谱曲线的吸收峰改变较大,循环伏安法峰氧化峰正向移动,电流增大,量化计算表明邻苯二酚位于磷酸基团近侧,邻苯二酚与脱氧核糖核酸主要以静电方式结合。     

人NOD8基因启动子绿色荧光蛋白表达载体的构建与鉴定

目的:构建人NOD8基因启动子的绿色荧光蛋白表达载体.方法:用特定的限制性内切酶位点,以人基因组DNA为模板,PCR扩增含有人NOD8基因启动子不同长度2段序列,并进行酶切以切除启动子的pEGFP-C2作框架结构,插入表达载体pEGFP-C2中,构建含有人NOD8基因启动子驱动的绿色荧光蛋白载体pEGFP-C2-NOD8(520 bp)、pEGFP-C2-NOD8(760 bp),用Vsp Ⅰ和Nhe Ⅰ双酶切和PCR鉴定重组质粒,再将重组质粒进行DNA序列分析.构建的重组质粒经脂质体(lipofectamine)TM2000介导转染HEK293、K562和HeLa细胞,转染48 h后在倒置荧光显微镜下观察.结果:pEGFP-C2-NOD8(520 bp)、pEGFP-C2-NOD8(760 bp)分别经酶切鉴定和序列测定证实目的基因已插入重组质粒;细胞转染结果表明,构建的2段重组质粒转染HEK293、K562及HeLa细胞均能表达绿色荧光,其中构建的pEGFP-C2-NOD8(760 bp)重组质粒绿色荧光表达强于pEGFP-C2-NOD8(520 bp).结论:成功构建2段不同长度的人NOD8基因启动子绿色荧光蛋白表达载体.     

SO(8)模型中对一对与四极一四极相互作用

使用对一对与四极一四极相互作用,描述振动到γ—不稳定区域的原子核。这个相互作用有三种极限:su(2) so(5)、so(7)和so(6)极限。同位素~(98-104)Ru位于so(7)—so(6)区域。     

紫外线吸收剂PBSA与DNA相互作用的光谱研究

采用荧光光谱法和紫外光谱法, 在模拟生理条件下(pH＝7.23)对紫外线吸收剂2-苯基苯并咪唑-5-磺酸(PBSA)与脱氧核糖核酸(DNA)的相互作用进行了研究.结果表明PBSA能够与DNA相结合形成复合物,引起了PBSA紫外吸收的降低和荧光的猝灭;PBSA与DNA的相互作用为静态猝灭过程,两者间的结合位点数为1.11.     

光谱法研究7-[(8-喹啉)偶氮]-8-羟基喹啉-5-磺酸与蛋白质相互作用

应用紫外吸收光谱法和荧光光谱法研究了7-[(8-喹啉)偶氮]-8-羟基喹啉-5-磺酸(简称主体)与蛋白质的相互作用.在p H=7.40的Tris-HCl缓冲溶液中,主体分子在400 nm处发射弱荧光,其荧光强度随白蛋白的加入明显增强.据此建立测定微量蛋白质的新方法.在相同的实验条件下,测定牛血清白蛋白和人血清白蛋白,线性范围分别为1.0×10-6~2.7×10-5mol/L和2.5×10-7~9.5×10-6mol/L,检测限分别为8.09×10-7mol/L、1.05×10-7mol/L.同时讨论7-[(8-喹啉)偶氮]-8-羟基喹啉-5-磺酸对白蛋白内源荧光的猝灭机理,测定牛血清白蛋白和人血清白蛋白结合常数分别为3.38×105和7.28×106,结合位点数分别为1.218和1.412.依据Forster非辐射能量转移理论,确定了主体-受体间的结合距离rBSA=3.98 nm,rHSA=3.04 nm及能量转移效率EBSA=0.038,EHSA=0.161,用同步荧光技术考察主体对蛋白质构象的影响.1     

肌酸激酶MM亚型不连续琼脂糖凝胶电泳法的建立

肌酸激酶同工酶(CK)主要分为CK—MM、CK—MB、CK—BB,而CK—MM可以进一步分离为MM_1、MM_2、MM_3等亚型。急性心肌梗塞(AMI)时,CK—MM_3大量入血而升高,特别是MM_3/MM_1的比值升高,对AMI具有极好的特异性和早期诊断的价值。此外,对患者是否使用溶栓疗法具有临床应用意义。我们采用不连续缓冲系统电泳分离CK—MM亚型,方法简便,分辨率好,便于普及推广。     

粘度法研究SDS与PEO之间的相互作用

采用粘度法测定了不同温度下 ( 2 98 1 5K ,30 8 1 5K ,31 8 1 5K)阴离子表面活性剂十二烷基硫酸钠 (SDS)—水溶性聚合物聚氧乙烯 (PEO)体系在有无电解质 (NaOH)存在时溶液的宏观粘度 ,研究了SDS—PEO体系分子间相互作用对PEO构型的影响 ,得到了不同SDS—PEO体系的临界聚集浓度 (CAC)和聚合物饱和浓度 (PSP) ,并根据表面活性剂—聚合物体系相互作用模型计算了SDS胶束被吸附到聚合物PEO过程的吉布斯函数变 ,讨论了SDS—PEO体系分子间相互作用强度 结果表明 ,在所研究范围内 ,SDS—PEO体系有无电解质溶液的比浓粘度均随聚合物PEO浓度的减小而急剧升高 ,呈现典型的聚合电解质行为 ,而且SDS的浓度越大 ,电粘效应愈明显 由此可以推断 ,聚合物PEO与表面活性剂SDS之间主要是通过疏水键相互结合 ,PEO分子链处于更伸展的状态 NaOH的存在使聚合物PEO分子与胶束结合位置减少 ,聚集体的体积减小 ,或者介电常数增大 ,因此表现出电粘效应弱于不含碱的表面活性剂—聚合物体系 ,而且使其分子间相互作用强度降低     

λ噬菌体调节蛋白与操纵基因相互作用的研究

根据λ噬菌体调节蛋白和特异的ＤＮＡ部位的相互作用提出一种线笥三位点的结合模型，从理论上解释了调节蛋白结合左右操纵基因的协同性，得出耦合自由能的分配。     

The structure of ribosomal protein S5 reveals sites of interaction with 16S rRNA.

Understanding the process whereby the ribosome translates the genetic code into protein molecules will ultimately require high-resolution structural information, and we report here the first crystal structure of a protein from the small ribosomal subunit. This protein, S5, has a molecular mass of 17,500 and is highly conserved in all lifeforms. The molecule contains two distinct alpha/beta domains that have structural similarities to several other proteins that are components of ribonucleoprotein complexes. Mutations in S5 result in several phenotypes which suggest that S5 may have a role in translational fidelity and translocation. These include ribosome ambiguity or ram, reversion from streptomycin dependence and resistance to spectinomycin. Also, a cold-sensitive, spectinomycin-resistant mutant of S5 has been identified which is defective in initiation. Here we show that these mutations map to two distinct regions of the molecule which seem to be sites of interaction with ribosomal RNA. A structure/function analysis of the molecule reveals discrepancies with current models of the 30S subunit.     

The interaction between cognition and emotion

Cognition and emotion have long been thought of as independent systems. However, recent research in the cognitive and neurobiological sciences has shown that the relationship between cognition and emotion is more interdependent than separate. Based on evidence from behavioral and neuroscientific research, researchers have realized that it is necessary to propose a new conceptual framework to describe the relationship between cognition and emotion. In this article, recent research from behavioral, neuroscientific and developmental research on the interaction between cognition and emotion is summarized, and how the interaction of cognition and emotion might affect computer science and artificial intelligence is discussed. It especially focuses on the implications for affective computing.     

The Eps8 protein coordinates EGF receptor signalling through Rac and trafficking through Rab5

How epidermal growth factor receptor (EGFR) signalling is linked to EGFR trafficking is largely unknown. Signalling and trafficking involve small GTPases of the Rho and Rab families, respectively. But it remains unknown whether the signalling relying on these two classes of GTPases is integrated, and, if it is, what molecular machinery is involved. Here we report that the protein Eps8 connects these signalling pathways. Eps8 is a substrate of the EGFR, which is held in a complex with Sos1 by the adaptor protein E3bl (ref. 2), thereby mediating activation of Rac. Through its src homology-3 domain, Eps8 interacts with RN-tre. We show that RN-tre is a Rab5 GTPase-activating protein, whose activity is regulated by the EGFR. By entering in a complex with Eps8, RN-tre acts on Rab5 and inhibits internalization of the EGFR. Furthermore, RN-tre diverts Eps8 from its Rac-activating function, resulting in the attenuation of Rac signalling. Thus, depending on its state of association with E3b1 or RN-tre, Eps8 participates in both EGFR signalling through Rac, and trafficking through Rab5.     

光谱法研究杯[8]芳烃与蛋白质的相互作用

用荧光光度法研究了杯[8]芳烃磺酸钠与蛋白质的相互作用.实验结果表明,牛血清蛋白可产生λex,max=280 nm,λem,max=349 nm的自身荧光.杯[8]芳烃磺酸钠在一定条件下能静态猝灭蛋白质的荧光.求出了杯[8]芳烃磺酸钠与蛋白质相互作用的形成常数,络合比和结合位点数,确定了反应的最佳条件为pH 7.5,0.1 mol.L-1NaCl用量1 mL,初步探讨了体系的主要作用力为范德华力.     

Analysis of correlations between protein complex and protein-protein interaction and mRNA expression

Protein-protein interaction is a physical interaction of two proteins in living cells. In budding yeast Saccharomyces cerevisiae, large-seale protein-protein interaction data have been obtained through high-throughput yeast two-hybrid systems (Y2H) and protein complex purification techniques based on mass-spectrometry. Here, we collect 11855 interactions between total 2617 proteins. Through seriate genome-wide mRNA expression data, similarity between two genes could be measured. Protein complex data can also be obtained publicly and can be translated to pair relationship that any two proteins can only exist in the same complex or not. Analysis of protein complex data, protein-protein interaction data and mRNA expression data can elucidate correlations between them. The results show that proteins that have interactions or similar expression patterns have a higher possibility to be in the same protein complex than randomized selected proteins, and proteins which have interactions and similar expression patterns are even more possible to exist in the same protein complex. The work indirates that comprehensive integration and analysis of public large-seale bioinformatical data, such as protein complex data, protein-protein interaction data and mRNA expression data, may help to uncover their relationships and common biological information underlying these data. The strategies described here may help to integrate and analyze other functional genomic and proteomic data, such as gene expression profiling, protein-localization mapping and large-scale phenotypic data, both in yeast and in other organisms.     

The lambda phage att site: functional limits and interaction with Int protein

Site specific integrative recombination of bacteriophage lambda involves unequal partners. The minimal phage att site is composed of approximately 240-base pairs and four distinct binding sites for Int protein, at least three of which are crucial for function. This 'donor site' recombines efficiently with a smaller 'recipient site' that lacks the extensive interactions with Int protein.     

Identification of in vivo interaction between rabbit hemorrhagic disease virus capsid protein and minor structural protein

We investigated the ability of the rabbit hemorrhagic disease virus(RHDV) capsid protein(VP60) to interact specifically with the minor structural protein VP10,using an in vivo cell-based CheckMate Mammalian Two-Hybrid System.RHDV VP60 protein interacted specifically with VP10.Immunofluorescence analysis and co-immunoprecipitation with specific antibodies revealed the existence of biologically important VP60/VP10 complexes.However,when VP60 was divided into two fragments,the interaction between VP60 and VP10 was impaired dramatically.These results will be helpful for further investigating the mechanism of RHDV particle assembly.     

高校体育与城市社区体育发展的互动性

高等院校具有人才、场地和教育功能等优势,应在促进社区体育发展、完善社区功能、促进全民健身等方面起到积极作用.因此,高校体育与城市社区体育要相互促进、协调发展,并就如何互动性发展提出了一些新思路.