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1.
经典的蛋白质组学研究方法包括IEF/SDS-PAGE双向电泳和质谱技术的联用,但由于IEF的一些不足,限制了其应用范围。对角线电泳是蛋白质组学研究中的一项特殊分离技术,由于其原理与IEF/SDS-PAGE不同,正逐渐成为蛋白质组学中电泳分离技术的重要补充,特别是在膜蛋白和蛋白质相互关系的研究中将起到重要作用。本文综述了对角线双向电泳技术的特点、发展和在蛋白质组学研究中的最新进展,比较了双向电泳和对角线电泳的优缺点,展望了对角线电泳在蛋白质组学研究中的应用前景。  相似文献   

2.
随着拟南芥、水稻等模式植物基因组测序的完成,植物基因组学的研究重点已经转变为功能基因组学研究。蛋白质组学成为后基因组时代的重要研究手段,它有助于从分子水平上了解植物功能。主要介绍了双向电泳技术、生物质谱、蛋白质质谱数据的生物信息学分析等蛋白质组学研究的主要技术手段及植物应答病原菌胁迫的蛋白质组学研究进展,并对蛋白质组学在研究植物抗病机制方面的应用前景做出展望。  相似文献   

3.
免疫蛋白质组学及其在病原菌研究中的应用   总被引:2,自引:0,他引:2  
以双向电泳、生物质谱及生物信息学为主要技术支撑的蛋白质组学与传统免疫印迹技术相结合,产生了一门新兴的学科——免疫蛋白质组学。本主要从免疫蛋白质组学的产生、技术体系及在病原菌免疫原性蛋白质研究中的应用等3个方面对其进行综述。  相似文献   

4.
Jia LY  Wang X 《生理科学进展》2004,35(3):237-239
蛋白质组学是旨在研究蛋白质表达谱和蛋白质与蛋白质之间相互作用的新的领域。蛋白质组学的研究必须依赖高通量、自动化程度很高的技术。双向电泳、液相色谱和生物质谱技术的发展推动了蛋白质组学的研究。蛋白质组学为疾病发病机制的研究提供了新的思路和方法 ,本文重点介绍了蛋白质组学技术在心血管疾病研究中的应用  相似文献   

5.
自20世纪90年代后期.随着蛋白质组概念的提出和基因组测序工作的快速进行,尤其是双向电泳技术的改进和生物质谱的大范围应用,蛋白质组学研究进入了一个快速发展的十年。十年间.伴随着对蛋白质组分离技术分辨率和重复性的争论,以双向电泳为主的基于凝胶的分离技术和以反相色谱为主的液相色谱质谱技术都取得了迅猛而深入的发展。  相似文献   

6.
蛋白质组学中的分离检测技术   总被引:5,自引:1,他引:4  
10多年来,随着基因组学研究取得的巨大成就,蛋白质组学的研究也得到了突飞猛进的发展,并产生了许多先进的分离检测技术,包括与电泳相关的和非电泳的技术。本就蛋白质组学中的分离检测技术,如双向电泳、差异凝胶电泳、毛细管电泳、液相色谱质谱联用、蛋白质芯片等作一综述。  相似文献   

7.
随着“人类基因组计划”的完成,包括双向电泳和质谱技术的蛋白质组学作为基因组学研究的补充和终点,不断增加我们对基因功能的理解,逐渐成为医学研究的中心。原虫作为医学研究的对象和工具,研究内容十分广泛。本文将从原虫生活史、致病机理、潜在疫苗以及抗药性等方面对原虫的蛋白质组学进行综述。  相似文献   

8.
生物信息学及其在蛋白质组学中的应用   总被引:2,自引:0,他引:2  
随着基因组学和蛋白质组学的发展,生物信息学在数据处理中的应用已经越来越广泛。作为数据处理中越来越重要的分析手段,蛋白质组学数据库是蛋白质组学的主要内容之一。本文分别从生物信息学的蛋白质双向电泳数据库和基于蛋白质质谱结果的数据库两个方面,概述了发展中的蛋白质数据库的最新动态和有关信息,同时对主要的热门蛋白质组学数据库站点和资源进行了评价和分析。  相似文献   

9.
蛋白质组学是在蛋白质水平定量、动态、整体地研究生物体的一门学科。双向电泳技术、质谱技术和生物信息学是蛋白质组学的三大支撑技术。近年来,蛋白质组学技术从整体水平出发,在更贴近生命本质的层次上去发现和理解并应用于许多疾病的早期预警、诊断和治疗。我们对蛋白质组学在心血管疾病、肝病、胰腺疾病和自身免疫性疾病等研究中的应用做了简单阐述,揭示了蛋白质组学技术在许多重大疾病研究方面具有十分诱人的发展前景。  相似文献   

10.
植物盐胁迫应答蛋白质组学研究的技术策略   总被引:2,自引:1,他引:1  
Zhang H  Dai SJ 《应用生态学报》2011,22(8):2201-2210
土壤盐渍化是限制植物生长和分布的关键因素之一.揭示植物响应盐胁迫的分子机理是借助分子生物学手段提高植物耐盐性的基础,也是当前植物生理与分子生态学研究的热点问题.高通量的蛋白质组学技术体系包括双向电泳技术、蓝色自然胶电泳技术、双向荧光差异凝胶电泳技术、液相色谱技术,以及各种生物质谱技术,已经被广泛应用于植物应答盐胁迫研究,为解析植物耐盐分子机制提供了重要信息.本文综述了应用于植物盐胁迫响应蛋白质组学研究的技术策略.  相似文献   

11.
蛋白质组研究是近年兴起的生命科学的前沿领域,是生命科学进入后基因组时代的标志之一。蛋白质组研究中的主要技术是双向凝胶电泳技术和质谱技术。本文简要综述了双向凝胶电泳和质谱技术的现状及存在的问题。  相似文献   

12.
The recent upsurge in proteomics research has been facilitated largely by streamlining of two-dimensional (2-D) gel technology and the parallel development of facile mass spectrometry for analysis of peptides and proteins. However, application of these technologies to the mitochondrial proteome has been limited due to the considerable complement of hydrophobic membrane proteins in mitochondria, which precipitate during first dimension isoelectric focusing of standard 2-D gels. In addition, functional information regarding protein:protein interactions is lost during 2-D gel separation due to denaturing conditions in both gel dimensions. To resolve these issues, 2-D blue-native gel electrophoresis was applied to the mitochondrial proteome. In this technique, membrane protein complexes such as those of the respiratory chain are solubilized and resolved in native form in the first dimension. A second dimension sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel then denatures the complexes and resolves them into their component subunits. Refinements to this technique have yielded the levels of throughput and reproducibility required for proteomics. By coupling to tryptic peptide fingerprinting using matrix-assisted laser desorption/ionization-time of flight mass spectrometry, a partial mitochondrial proteome map has been assembled. Applications of this functional mitochondrial proteomics method are discussed.  相似文献   

13.
Comparative analysis has long been utilized in biological research to interpret protein interactions in both drug na?ve versus drug challenged and normal versus diseased tissues. The technology of proteomics today allows researchers to provide insight into old and still open questions related to biological mechanisms while offering the opportunity to discover novel details in cellular lifecycles. Perhaps the most powerful way to execute these differential displays is in the combination of two-dimensional (2-D) gel electrophoresis and mass spectrometry. While these two techniques together are well suited for abundant and soluble proteins found in cells, rare proteins and integral membrane proteins are still problematic. Recently, a series of novel zwitterionic detergents has been reported in the literature that shows a substantial improvement in solubilizing integral membrane proteins. We show that the amidosulfobetaine, 4-octylbenzol amidosulfobetaine, is better than 3-[(3-cholamidopropyl)dimethylamino]-1-propanesulfonate (CHAPS) at solubilizing both an ion channel and a G-protein coupled receptor (GPCR), while another amidosulfobetaine, myristic amidosulfobetaine (ASB-14), was better than CHAPS at solubilizing a GPCR. Neither membrane protein was visible after staining with colloidal Coomassie blue, silver nor Sypro Ruby. However, a comparison against a duplicate immunoblot allowed for the localization and identification of the ion channel from a 2-D gel by liquid chromatography-tandem mass spectrometry.  相似文献   

14.
Proteomic analysis of skeletal muscle presents particular challenges when trying to identify valid biomarkers of phenotypic change in small biopsies from genetically diverse human subjects. Currently, two-dimensional (2-D) gel electrophoresis and mass spectrometry are the chosen analytical strategies but 2-D gels are not appropriate for analyzing proteins less than 11 kDa, they can suffer from problems of reproducibility and in routine use are not a viable high-throughput technique. We have evaluated an integrated proteomic strategy employing Ciphergen ProteinChip arrays, one-dimensional polyacrylamide gel electrophoresis and mass spectrometry. Protein fingerprints characteristic of fast and slow contracting muscles from normal and kyphoscoliosis (ky) mutant mice were obtained from Ciphergen protein arrays. Eight statistically validated protein biomarkers have so far been identified capable of discriminating fast from slow muscle. Five of these showed further differential expression in ky versus normal BDL soleus muscles. Several biomarkers have been formally identified, and were myosin light chain isoforms shown previously to be expressed differentially by fast versus slow skeletal muscles. This integrated experimental approach using a model mouse muscle system shows the potential of Ciphergen protein array technology for proteomic analysis of small proteins in small muscle samples and its applicability for phenotypic characterization of skeletal muscle in general.  相似文献   

15.
Hancock WS  Wu SL  Shieh P 《Proteomics》2002,2(4):352-359
This paper will review the challenges of developing a proteomics strategy. A key issue is the integration of the two-dimensional (2-D) gel platform with mass spectrometry measurements. The use of both matrix-assisted laser/desorption ionization (on off-line coupling) and electrospray (on-line) ionization are complementary. While the use of one-dimensional and 2-D gels are essential to many aspects of proteomics research (sample preparation, preliminary fractionation and quantitation, storage of protein components), the emergence of shotgun sequencing based on high performance liquid chromatography and tandem mass spectrometry offers a powerful new approach. The latter has particular utility in the characterization of low level samples and complex post-translational modifications. The development of capillary columns, such as 75 to 150 micron, that can be packed in a reproducible manner has been a key step in the development of high sensitivity liquid chromatography/mass spectrometry analysis.  相似文献   

16.
Sun N  Jang J  Lee S  Kim S  Lee S  Hoe KL  Chung KS  Kim DU  Yoo HS  Won M  Song KB 《Proteomics》2005,5(6):1574-1579
Cytosolic proteins of Schizosaccharomyces pombe were separated by two-dimensional (2-D) gel electrophoresis, to construct the first 2-D reference map. In the pI range 4-7, more than 500 spots were detected by silver staining, and 70 different proteins corresponding to 111 spots were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry and tandem mass spectrometry, where necessary. In the pI range 6-9, approximately 330 spots were detected, and 31 proteins corresponding to 38 spots were identified by mass spectrometry. More than 50% of the identified proteins were involved in amino acid, carbohydrate or nucleotide metabolism, and energy production. A second large group of identified proteins comprises heat shock and other stress related proteins and chaperones.  相似文献   

17.
We have combined high-resolution two-dimensional (2-D) gel electrophoresis with mass spectrometry to identifying proteins represented in a 2-D gel database of Drosophila melanogaster ribosomes. First, we purified ribosomes from third instar Drosophila larvae and constructed a high-resolution 2-D gel database containing 58 Coomassie blue stained polypeptides. Next, we carried out preparative 2-D PAGE to isolate some of the polypeptides and characterize them by MALDI-TOF. Using this strategy we identified 52 ribosomal spots in the database, and in each case confirmed their identity by MALDI-TOF/TOF. The database can be used to analyze Minute mutants of Drosophila.  相似文献   

18.
A Proteomics Approach to Characterizing Tick Salivary Secretions   总被引:1,自引:0,他引:1  
The saliva of ticks contains a complex mixture of bioactive molecules including proteins that modulate host responses ensuring successful feeding. The limited amount of saliva that can be obtained from ticks has hampered characterization of salivary proteins using traditional protein chemistry. Recent improvements in two-dimensional gel electrophoresis, mass spectrometry, and bioinformatics provide new tools to characterize small amounts of protein. These methods were employed to characterize salivary proteins from Amblyomma americanum and Amblyvomma maculatum. Salivation was induced by injection of dopamine and theophylline. It was necessary to desalt and concentrate saliva before analysis by 2-D electrophoresis. Comparison of 1-D and 2-D gel patterns revealed that the major protein component of saliva did not appear on 2-D gels. Characterization of this protein showed that it was identical to the major protein present in the hemolymph of both tick species. Protein profiles obtained by 1-D and 2-D gel electrophoresis were similar for both tick species, however, higher concentrations of lower molecular weight proteins were present in A. maculatum. Protein analysis by MALDI-TOF mass spectrometry and western blot analysis showed that except for the most abundant protein with a molecular weight of 95 kDa, all of the proteins detected were of host origin. It is not known if this is an artifact of the collection method or has physiological significance. In either case, in these species of ticks, host proteins will have to be removed from saliva samples prior to 2-D analysis in order to characterize lower abundance proteins of tick origin.  相似文献   

19.
基于Make2D—DBII软件包构建了一个水稻二维电泳-质谱联动数据库Rice2DDB,介绍了该数据库的构建方法和步骤,为快速发展的水稻蛋白质组学研究提供了数据管理和交流的平台,对其他物种的蛋白质组学研究也具有参考价值。  相似文献   

20.
在后基因组时代,蛋白质组学成为新的研究热点。蛋白质组学的研究目标是为复杂蛋白质样品建立一个高通量、大规模、自动化的分离分析技术平台,从而实现准确、快速地筛选功能蛋白质。蛋白质的分离分析在蛋白组学研究中起着非常重要的作用。本文主要综述在蛋白质组学研究中二维凝胶电泳、毛细管电泳及其与质谱联用、多维液相分离技术及其与质谱联用和蛋白质芯片等高效分离分析技术的应用研究进展。  相似文献   

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