| 引用本文： |
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岑卫健,甘永亮,于凯,付强,韦宇拓,姜伯乐.基于LC-MS/MS策略建立十字花科黑腐病菌蛋白质表达谱的方法研究[J].广西科学,2014,21(2):103-107. [点击复制]
- CEN Wei-jian,GAN Yong-liang,YU Kai,FU Qiang,WEI Yu-tuo,JIANG Bo-le.Method Research of Proteomic Profile Identification on Xanthomonas campestris pv.campestris by LC-MS/MS[J].Guangxi Sciences,2014,21(2):103-107. [点击复制]
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| 摘要: |
| [目的]建立十字花科黑腐病菌(Xanthomonas campestris pv.campestris,Xcc)的蛋白质组学研究平台,用于分离、鉴定该菌的致病相关蛋白质。[方法]Xcc 8004经过液体培养,分别提取胞内蛋白质和胞外蛋白质,对所有蛋白质进行液体酶解,通过强阳离子交换色谱预分离多肽混合物,预分离后的馏分采用EASY-nLC结合LTQ-Orbitrap质谱鉴定蛋白质表达谱。[结果]建立了基于bottom-up策略的蛋白质组鉴定方法,采用第一维离子交换预分离和第二维反相色谱分离结合,实现了高通量鉴定蛋白质组表达谱的技术体系。通过SEQUEST检索,在胞内蛋白质样品中共鉴定蛋白质数目为1595个,胞外蛋白质样品共鉴定蛋白质数目为1241个。[结论]建立的LC-MS/MS方法适合用于十字花科黑腐病菌蛋白质组学的研究,为研究微生物植物相互作用奠定了方法与理论基础。 |
| 关键词: Xcc 蛋白质表达谱 LC-MS/MS LTQ-Orbitrap |
| DOI： |
| 投稿时间：2014-03-26修订日期：2014-03-31 |
| 基金项目:广西自然科学基金项目(2013GXNSFAA019097),广西大学科研基金项目(XJZ130364),广西研究生教育创新计划项目(T32401)资助。 |
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| Method Research of Proteomic Profile Identification on Xanthomonas campestris pv.campestris by LC-MS/MS |
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CEN Wei-jian1, GAN Yong-liang2, YU Kai2, FU Qiang1, WEI Yu-tuo1,2, JIANG Bo-le1,2
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| (1.State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China;2.College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China) |
| Abstract: |
| [Objective] A proteomics technical platform of Xanthomonas campestris pv.campestris (Xcc) was established for the separation and identification of pathogenic bacteria associated proteins.[Methods] The intracellular proteins and extracellular proteins of Xcc 8004 strains were extracted after liquid cultured.After in-solution digestion of total proteins, the peptides mixtures were separated by strong cation exchange (SCX) column.The proteome profiles were acquired by EASY-nLC combined LTQ-Orbitrap mass spectrometer.[Results] Based on bottom-up method, the proteins were analyzed by cation exchange column and nanoflow reversed-phase liquid chromatography.The high proteomic profile identification was realized.The mass spectrometry data were searched with SEQUEST.As a result, a total of 1595 proteins were obtained from intracellular proteins and a total of 1241 proteins were obtained from extracellular proteins.[Conclusion] LC-MS/MS workflow could be utilized successfully in the proteomics research of Xcc pathogenesis, which provides the basis for studing the interaction of microbe and plant. |
| Key words: Xcc proteome profile LC-MS/MS LTQ-Orbitrap |
