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采用二维荧光差异凝胶电泳结合质谱技术对不同弓形虫基因型进行比较蛋白质组学分析。

Comparative proteomic analysis of different Toxoplasma gondii genotypes by two-dimensional fluorescence difference gel electrophoresis combined with mass spectrometry.

机构信息

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, P. R. China.

出版信息

Electrophoresis. 2014 Feb;35(4):533-45. doi: 10.1002/elps.201300044. Epub 2013 Dec 2.

Abstract

Toxoplasma gondii is a protozoan parasite infecting almost all warm-blooded animals and humans. There are three infective stages of T. gondii: the tachyzoites, the bradyzoites, and the oocysts. The tachyzoite is a rapidly multiplying stage and the main pathogenic factor. In North America and Europe, T. gondii is consisted of four major clonal lineages (namely Types I, II, III, and Type 12). In this study, we explored the proteomic profiles of different genotypes (Type I-RH strain, Type II-PRU strain, Type II-TgQHO strain, and ToxoDB 9-TgC7 strain) of T. gondii tachyzoites by using 2D DIGE combined with MALDI-TOF MS. Totally, 110 differentially abundant protein spots were selected. Of these, 98 spots corresponding to 56 proteins from T. gondii were successfully identified. These included surface antigen (SAG1), heat shock protein 70 (Hsp 70), disulfide isomerase, coronin, heat shock protein 60 (Hsp 60), pyruvate kinase, receptor for activated C kinase 1, and peroxiredoxin. Gene ontology enrichment analysis revealed that most of the differentially abundant proteins were involved in biological regulation, metabolic process, response to stress, binding, antioxidant activity, and transporter activity. According to the KEGG metabolic pathway maps of T. gondii, some identified proteins were involved in the glycolytic/gluconeogenesis pathway. The present study identified differentially abundant proteins among different genotypes of T. gondii and these findings have implications for the better understanding of the phenotypic differences among the examined T. gondii genotypes, which in turn may contribute to the better control of toxoplasmosis.

摘要

刚地弓形虫是一种寄生在几乎所有温血动物和人类中的原生动物寄生虫。刚地弓形虫有三个感染阶段:速殖子、缓殖子和卵囊。速殖子是一种快速繁殖的阶段,也是主要的致病因素。在北美和欧洲,刚地弓形虫由四个主要的克隆谱系(即 I 型、II 型、III 型和 12 型)组成。在这项研究中,我们通过 2D DIGE 结合 MALDI-TOF MS 技术探索了不同基因型(I 型 RH 株、II 型 PRU 株、II 型 TgQHO 株和 ToxoDB 9-TgC7 株)刚地弓形虫速殖子的蛋白质组谱。总共选择了 110 个差异丰度蛋白点。其中,98 个对应于 56 个刚地弓形虫蛋白的斑点成功被鉴定。这些蛋白包括表面抗原(SAG1)、热休克蛋白 70(Hsp 70)、二硫键异构酶、冠蛋白、热休克蛋白 60(Hsp 60)、丙酮酸激酶、激活蛋白激酶 C 受体 1 和过氧化物还原酶。基因本体富集分析显示,大多数差异丰度蛋白参与生物调节、代谢过程、应激反应、结合、抗氧化活性和转运活性。根据刚地弓形虫的 KEGG 代谢途径图谱,一些鉴定出的蛋白参与了糖酵解/糖异生途径。本研究鉴定了不同基因型刚地弓形虫之间的差异丰度蛋白,这些发现有助于更好地理解所研究的刚地弓形虫基因型之间的表型差异,从而有助于更好地控制弓形虫病。

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