费氏中华根瘤菌042BS(Rhizobium fredii 042BS)蛋白质组学初步研究

为了从蛋白质水平阐明根瘤菌的固氮机理,扩大其宿主植物范围,应用蛋白质组学技术(2-DE),分析比较了能够在大豆和苜蓿上跨宿主结瘤的费氏中华根瘤菌042BS与已经完成基因组测序的根瘤菌模式菌株苜蓿中华根瘤菌(Rhizobium meliloti 1021)的蛋白质表达图谱差异。经过图像分析检测到二者分别有436和428蛋白点。其中约有50个蛋白点初步认为二者所共有的。     

胎儿宫内发育迟缓疾病标志物的初步研究

目的筛选胎儿宫内发育迟缓大鼠血清中差异表达的蛋白质,寻找与胎儿宫内发育迟缓相关的疾病标志物。方法收集大鼠疾病正常组、IUGR组血浆各3例;TCA法沉淀血清蛋白,建立疾病组和正常组蛋白质的双向凝胶电泳图谱;ImageMaster分析软件分析差异蛋白质;飞行时间质谱（MALDI—TOF—MS）获得差异蛋白肽指纹图谱,MASCOT引擎搜索蛋白质种类。结果鉴定出个差异蛋白点3个,分别为：IGF—I、IGF-Ⅱ、IGFBP-3。结论IGF—I、IGF-II、IGFBP-3,可能是胎儿宫内发育迟缓疾病诊断的生物标志物。     

抑郁模型大鼠海马突触的差异蛋白质组学分析

目的通过抑郁模型大鼠海马突触的差异蛋白质组学分析,为抑郁症发病机制研究提供依据。方法根据糖水消耗基线值将40只健康雄性Sprague—Dawley大鼠随机分为慢性轻度不可预见性应激组和对照组（每组20只）。通过CUMS实验模式建立大鼠抑郁模型后,运用蔗糖密度梯度离心法提取两组大鼠的海马突触并用透射电镜进行检测。采用传统的双向凝胶电泳和基质辅助激光解析电离飞行时间串联质谱进行差异蛋白质组学分析。结果在行为学评价中CUMS组大鼠糖水消耗量和偏好度均降低（P〈0．01）,表明大鼠抑郁模型建立成功。通过结合双向凝胶电泳和质谱分析,共得到6个在CUMS组表达上调和10个在CUMS组表达下调的蛋白质,这些蛋白质的功能主要归为四类,即囊泡调节／递质释放／信号转导、能量代谢、细胞骨架、物质代谢。结论通过对抑郁模型大鼠海马突触的比较蛋白质组学分析及后续的蛋白质功能预测,发现了一些与突触传导功能障碍可能相关的蛋白质,从而为抑郁症发病机制的研究提供了有价值的线索。     

小菜蛾成虫蛋白质组双向电泳图谱的建立及条件优化

通过选择合适的pH范围及长度的胶条,并对蛋白样品上样量、等电聚焦伏小时数、银染时间及样品前处理等进行优化,建立了以双向电泳技术为基础的小菜蛾成虫蛋白质组学研究方法.优化结果表明使用17 cm、pH3～pH10的非线性胶条可以得到分辨率较高的胶图.该方法的建立为小菜蛾抗药性差异蛋白质组学研究奠定了基础.     

植物叶片H_2O_2胁迫应答蛋白质组学分析

植物叶片是感知外界H_2O_2胁迫信号的重要器官.整合分析了水稻(Oryza sativa)、小麦(Triticum aestivum)、二穗短柄草(Brachypodium distachyon)和柑橘(Citrus aurantium)在应对不同程度H_2O_2胁迫时蛋白质表达模式的变化特征.阐明了H_2O_2胁迫应答网络体系中的信号与代谢通路(如:光合作用、糖类与能量代谢、转录调控、蛋白质合成与命运、胁迫防御、信号转导和基础代谢等)的变化及植物叶片应答H_2O_2胁迫的分子调控机制.     

逆境胁迫下水稻蛋白质组学研究进展

水稻在生长发育过程中要面对各种非生物胁迫如干旱、高盐、温度、重金属和生物胁迫如病虫害等,通过观察水稻在各种胁迫条件下的蛋白质组表达情况,动态分析水稻的蛋白质组变化,对新逆境相关蛋白质进行分离与功能鉴定,这不仅能揭示参与胁迫耐受的蛋白质翻译后调控机制,而且可以增进对耐受胁迫分子机理的认识.综述了水稻应答逆境胁迫蛋白质组学研究的最新研究进展,探讨了存在的主要问题.     

罕见病诊断的相关技术及发展

 罕见病大多数是遗传性疾病,其发病率低,种类繁多且表型复杂多样,导致临床上难以进行及时和准确的诊断。随着分子遗传学、分子诊断技术、基因测序技术及组学技术的进步,罕见病诊断取得了重大发展。在传统基因检测技术基础上,二代测序技术迅速发展并广泛应用于罕见病的诊断和研究中,三代测序技术也展示出潜在应用价值。虽然传统的酶学检测技术仍占有重要地位,但已不能满足罕见病诊断的需求;蛋白质组学、代谢组学的崛起,使多种罕见病的准确诊断成为可能。同时结合分子影像技术和生物信息技术,计算机辅助诊断也展现出了广泛的应用前景。     

遗传算法-偏最小二乘法用于卵巢癌血清蛋白质组数据的特征挑选

统计学t检验结合引入的变量筛选方法——遗传算法-偏最小二乘法（GAPLS）对卵巢癌SELDI-TOF MS数据进行特征筛选,从15154个原始变量中筛选得到4个特征质荷比值,采用支持向量机（SVM）模型的留一法交叉验证结果为95.26%.结果表明这4个质荷比值具有重要的生物学意义,它们或许可以作为卵巢癌的生物标记物,同时GAPLS可以作为一种有效的蛋白质组数据的特征筛选方法.     

Proteomic analysis of long-term salinity stress-responsive proteins in Thellungiella halophila leaves

Salinity is one of the most severe environmental factors that may impair crop productivity. A proteomic study based on two-dimensional gel electrophoresis is performed in order to analyze the long-term salinity stress response of Thellungiella halophila, an Arabidopsis-related halophyte. Four-week-old seedlings are exposed to long-term salinity treatment. The total crude proteins are extracted from leaf blades, separated by 2-DE, stained with Coomassie Brilliant Blue, and differentially displayed spots are identified by MALDI-TOF MS or QTOF MS/MS. Among 900 protein spots reproducibly detected on each gel, 30 spots exhibit significant change and some of them are identified. The identified proteins include not only some previously characterized stress-responsive proteins such as TIR-NBS-LRR class disease resistance protein, ferritin-1, and pathogenesis-related protein 5, but also some proteins related to energy pathway, metabolism, RNA processing and protein degradation, as well as proteins with unknown functions. The possible functions of these proteins in salinity tolerance of T. halophila are discussed and it is suggested that the long-term salinity tolerance of T. halophila is achieved, at least partly, by enhancing defense system, adjusting energy and metabolic pathway and maintaining RNA structure.     

Mass spectrometry-based proteomics in the life sciences

Over the last 20 years, mass spectrometrybased proteomics has become an indispensable tool in the cellular and molecular life sciences. This has been enabled by the soft ionisation techniques of electrospray and matrix-assisted laser desorption-ionisation, which allow the gentle ionisation and vaporisation of large, thermally labile biomolecules. Innovative instrumentation designs and biochemical strategies have brought success in the large-scale identification and quantification of proteins, as well as the characterisation of their complexes and post-translational modifications. This review describes the instrumentation used for proteomics research. It presents an overview of the current applications of mass spectrometry-based proteomics to the cellular and molecular life sciences, and discusses challenges that exist for research in the future.Received 7 January 2005; accepted 27 January 2005