应用二维凝胶电泳和质谱技术分析空肠弯曲菌 NCTC11168 的体外蛋白质表达谱

In vitro protein expression profile of Campylobacter jejuni strain NCTC11168 by two-dimensional gel electrophoresis and mass spectrometry.

机构信息

Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.

出版信息

Biomed Environ Sci. 2013 Jan;26(1):48-53. doi: 10.3967/0895-3988.2013.01.006.

DOI:10.3967/0895-3988.2013.01.006

PMID:23294615

Abstract

OBJECTIVE

To investigate the protein expression profiles of the major food-borne pathogen Campylobacter jejuni NCTC11168.

METHODS

Membrane and soluble cellular proteins were extracted from the genome-sequenced C. jejuni strain NCTC11168. Protein expression profiles were determined using two-dimensional gel electrophoresis (2-DE). All the detected spots on the 2-DE map were subjected to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/TOF) analysis.

RESULTS

A total of 537 and 333 spots were detected from the whole cell and membrane-associated proteins of C. jejuni NCTC11168 cultured on Columbia agar medium at 42 °C by 2-DE and Coomassie Brilliant Blue staining, respectively. Analyses of whole cell and membrane-associated proteins included 399 and 133 spots, respectively, which included 182 and 53 functional proteins identified by MALDI-TOF/TOF analysis.

CONCLUSION

The comprehensive expression protein profiles of C. jeuni NCTC11168 obtained in this study will be useful for elucidating the roles of these proteins in further pathogenesis investigation.

摘要

目的

研究主要食源性病原体空肠弯曲菌 NCTC11168 的蛋白质表达谱。

方法

从基因组测序的空肠弯曲菌 NCTC11168 株中提取膜和可溶性细胞蛋白。使用二维凝胶电泳（2-DE）测定蛋白质表达谱。对 2-DE 图谱上所有检测到的斑点进行基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF/TOF）分析。

结果

通过 2-DE 和考马斯亮蓝染色，从 42°C 哥伦比亚琼脂培养基中培养的空肠弯曲菌 NCTC11168 的全细胞和膜相关蛋白中分别检测到 537 和 333 个斑点。全细胞和膜相关蛋白分析分别包括 399 和 133 个斑点，其中通过 MALDI-TOF/TOF 分析鉴定了 182 个和 53 个功能蛋白。