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Sub-cellular proteomic analysis of a Medicago truncatula root microsomal fraction

机译:苜蓿根微粒体部分的亚细胞蛋白质组学分析

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摘要

Since the last decade, Medicago truncatula has emerged as one of the model plants particularly investigated in the field of plant-microbe interactions. Several genetic and molecular approaches including proteomics have been developed to increase knowledge about this plant species. To complement the proteomic data, which have mainly focused on the total root proteins from M. truncatula, we carried out a sub-cellular approach to gain access to the total membrane-associated proteins. Following the setting up of the purification process, microsomal proteins were separated on 2-DE. Ninety-six out of the 440 well-resolved proteins were identified by MALDI-TOF peptide mass fingerprinting. A high percent (83%) of successful protein identification was obtained when using M. truncatula clustered EST database for queries. During the purification process, the enrichment in membrane-associated proteins was monitored on 2-D gels. The membrane location of microsomal proteins was further confirmed using PMF identification. This study reports a fractionation process for characterizing microsomal root proteins of M. truncatula, which could be an interesting tool for investigating the molecular mechanisms involved in root symbioses.
机译:自最近十年以来,run藜苜蓿已成为模型植物之一,在植物-微生物相互作用领域进行了专门研究。已经开发了包括蛋白质组学在内的几种遗传和分子方法来增加对这种植物物种的了解。为了补充蛋白质组学数据,蛋白质组学数据主要集中在截形支原体的总根蛋白上,我们进行了亚细胞方法来获取与总膜相关蛋白的通道。在建立纯化过程后,微粒体蛋白在2-DE上分离。通过MALDI-TOF肽质谱指纹图谱鉴定了440种分辨率良好的蛋白质中的96种。当使用M. truncatula聚类EST数据库进行查询时,获得了很高百分比的蛋白质鉴定成功(83%)。在纯化过程中,在2-D凝胶上监测膜相关蛋白的富集。使用PMF鉴定进一步证实了微粒体蛋白的膜位置。这项研究报告了特征化截形支原体的微粒体根蛋白的分级分离过程,这可能是研究与根共生有关的分子机制的有趣工具。

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