棉花根总蛋白双向电泳方法的优化

以陆地棉(Gossypium hirsutumL.)根为材料,比较分析了三种总蛋白提取方法及电泳参数和上样量等因素对双向电泳结果的影响.结果表明:选择pH 4⁷IPG胶条,采用丙酮沉淀/2-D Clean-up Kit法提取总蛋白,电泳时设置合适的电压,上样量为250μg蛋白进行双向电泳能够检测到1 000多个清晰的蛋白点,适合做质谱分析.     

蛋白质组学在药物及疾病标志物研究中的应用进展

蛋白质组学提供了整体水平研究蛋白质的表达、翻译后修饰,以及蛋白质的功能的思路,由此获得蛋白质水平上的关于疾病发生、细胞代谢等过程的全面认识,逐渐被广泛应用于药物及生物标志物研究中.本文介绍了蛋白质组学的经典技术路线和新策略,并综述了其在药物及疾病标志物研究中应用进展.     

Proteomic analysis of a toxic dinoflagellate Alexandrium catenella under different growth phases and conditions

Alexandrium is a widely spread dinoflagellate genus throughout many regions of the world,which not only causes the harmful algal blooms(HABs) but also results in the paralytic shellfish poisoning(PSP) throughout the world.This study compared protein profiles of A.catenella grown under different growth phases and conditions using a proteomic approach,and identified the differentially expressed proteins.The results showed that the expressions of proteins identified in three different regions of the gels,the groups 1,2 and 3 proteins,varied significantly with the growth phases and conditions.Group 1 proteins and six Group 2 proteins were highly expressed at the initial,exponential and stationary growth phases,eight Group 2 proteins were highly expressed only at the initial phase,and Group 3 proteins were highly expressed at the exponential and/or stationary phases.However,all these proteins were expressed at low levels or were barely visible at the dissipation phase.The expressions of groups 1 and 2 proteins were low or barely visible in various growth conditions except in continuous darkness they were highly expressed.Group 3 proteins,on the other hand,were overexpressed in continuous illumination and expressed at low levels or barely visible in continuous darkness or under nitrate-starvation.The data from MALDI-TOF-TOF mass spectrometry demonstrated that these differentially expressed proteins were associated with macromolecular biosynthesis,photosynthesis,tRNA synthesis and DNA stability,stress response and cell division regulation.Synthetase was the major component of the altered proteins.This is one of the first comprehensive proteomic study of a dinoflagellate,A.catenella,that provides a fundamental understanding of the proteins involved in A.catenella growth and response to environmental stresses,and potential physiological indicator proteins related to growth and environmental stress have been identified.     

新生物技术在现代疫苗研究中的应用

生物信息学、基因芯片技术、蛋白质组学等新生物技术的出现使得现代疫苗学的研究取得了质的飞跃,以前无法获得的疫苗现在成为可能.     

A complex of Shc and Ran-GTPase localises to the cell nucleus

The three isoforms of the adaptor protein Shc play diverse roles in cell signalling. For example, the observation of p46 Shc in the nuclei of hepatocellular carcinoma cells suggests a function quite distinct from the better characterised cytoplasmic role. Ligands responsible for the transport of various Shc isoforms into organelles such as the nucleus have yet to be reported. To identify such ligands a far western approach was used to determine the p52 Shc interactome. The Ran-GTPase nuclear transport protein was identified and found to bind to p52 Shc in vitro with low micromolar affinity. Co-immunoprecipitation, pull down and fluorescence lifetime imaging microscopy experiments in stable cells confirmed cellular interaction and nuclear localisation. The nuclear transport factor protein NTF2, which functions in cohort with Ran, was shown to form a complex with both RAN and Shc, suggesting a mechanism for Shc entry into the nucleus as part of a tertiary complex. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Received 20 October 2008; received after revision 04 December 2008; accepted 15 December 2008     

Serum amyloid A: An acute-phase protein involved in tumour pathogenesis

The synthesis of acute-phase protein serum amyloid A (SAA) is largely regulated by inflammation- associated cytokines and a high concentration of circulating SAA may represent an ideal marker for acute and chronic inflammatory diseases. However, SAA is also synthesized in extrahepatic tissues, e.g. human carcinoma metastases and cancer cell lines. An increasing body of in vitro data supports the concept of involvement of SAA in carcinogenesis and neoplastic diseases. Accumulating evidence suggests that SAA might be included in a group of biomarkers to detect a pattern of physiological events that reflect the growth of malignancy and host response. This review is meant to provide a broad overview of the many ways that SAA could contribute to tumour development, and accelerate tumour progression and metastasis, and to gain a better understanding of this acute-phase reactant as a possible link between chronic inflammation and neoplasia. Received 11 June 2008; received after revision 10 July 2008; accepted 28 July 2008     

缓冲液成分对固相pH梯度等电聚焦的影响

分别从水质、DTT(二硫苏糖醇)含量及含盐缓冲液三个方面对等电聚焦过程进行了对比。通过电学参数变化,研究了三者对聚焦进程的影响。结果表明:高质量的水、低含量的DTT和盐可以缩短聚焦时间,从而提高聚焦质量。可以把电学参数的变化作为优化和控制等电聚焦过程的指标,为正确设计合理的双向电泳实验流程提供依据。     

SARS冠状病毒核蛋白的质谱鉴定

利用质谱技术,对纯化的SARS冠状病毒的核蛋白进行初步鉴定与分析.所获得的质谱分析数据中,分子量为2881.19Da的酶切肽段的氨基酸序列与其它冠状病毒的同源性比较分析表明,SARS冠状病毒和其它冠状病毒均含有一个高度保守的基序“FYYLGTGP”.     

Des(rhamnosyl) verbascoside抗HBV作用的定量蛋白质组学研究

为探究des(rhamnosyl) verbascoside体外抗乙型肝炎病毒(HBV)的活性作用和机制,本研究以des(rhamnosyl) verbascoside为实验药物对HepG2.2.15细胞进行干预,实验分为药物干预组和对照组,采用串联质谱标签(Tandem Mass Tag, TMT)蛋白质组学方法对提取的总蛋白进行分析。结果表明,共筛选得到300个差异表达蛋白,其中有109个上调蛋白,191个下调蛋白。基因本体论(Gene Ontology, GO)分析结果显示,差异蛋白主要参与DNA复制(DNA replication)、鞘糖脂代谢(Glycosphingolipid metabolic process)、细胞增殖(Cell proliferation)、寡糖分解代谢(Oligosaccharide catabolic process)等生物学过程,以及DNA聚合酶活性(DNA polymerase activity)、丝氨酸型羧肽酶活性(Serine-type carboxypeptidase activity)、DNA引物酶活性(DNA primase acti...     

锯缘青蟹对病原菌感染的急性反应功能蛋白质组

为研究锯缘青蟹对常见重要病原菌的急性反应蛋白质组．将锯缘青蟹随机分为四组．分别注射生理盐水、副溶血弧菌、鳗弧菌和嗜水气单胞菌．继而提取肌肉蛋白进行双向电泳．通过比较各组肌肉蛋白的双向电泳图谱获得三个差异表达的蛋白．对这三个差异蛋白进行肽质量指纹图谱及其生物信息分析．鉴定为Calexeitin、无翅蛋白片断、速激肽相关肽．这些结果对研究锯缘青蟹的抗病蛋白及其机理具有重要意义．