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黏细菌总蛋白质组双向电泳条件的建立

Establishment of two-dimensional electrophoresis conditions for proteome analysis of Myxococus xanthus

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【作者】 丁蕾谢维毛晓华

【Author】 DING Lei,XIE Wei,MAO Xiao-hua (Department of Genetics and Development Biology,School o f Bas ic Medical Science,Southeast University,Nanjing 210009,China)

【机构】 东南大学基础医学院遗传及发育生物学系东南大学基础医学院遗传及发育生物学系 江苏南京210009江苏南京210009江苏南京210009

【摘要】 目的 :建立可以对黏细菌蛋白质组进行阵列分离的双向电泳条件。方法 :以黏细菌模式菌株DK162 2为材料 ,不同方法制备DK162 2在营养性生长阶段的蛋白质组样品 ,双向电泳阵列菌体总蛋白。通过不同条件下电泳图谱的比较建立蛋白质组样品的制备方法和双向电泳的技术条件。结果 :两性离子去污剂CHAPS及还原剂的浓度是影响双向电泳图谱分辨力的重要因素 ;蛋白质组样品的处理必须在低温下快速完成 ,以避免蛋白降解。结论 :初步建立了黏细菌总蛋白质组的双向电泳条件 ,为在细胞水平分析该模式生物发育时期蛋白表达谱奠定了基础

【Abstract】 Objective To develop appropriate condit ions for profiling cell proteome of Myxococcus xanthus. Methods Total cellular proteins from Myxococcus xanthus DK1622 negetative cells were extracted and subjected t o two-dimensional gel electrophoresis under different conditions. Optimal cond i tions for sample preparation and 2-DE was established based on the comparison o f a series of protein patterns. Results The concentration of z witterionic deter gent CHAPS and reductant was decisive for obtaining good resolution of 2-DE pat t ern. Protein sample should also be processed as quickly as possible under low t emperature to prevent proteins from degradation. Conclusion Op timal conditions of 2-DE for proteomic analysis of Myxococcus xanthus has been established, which paves t he way for global profiling of protein expression patterns during the developmen t of this model organism.

【基金】 国家自然科学基金资助项目 (30 370 775)
  • 【文献出处】 东南大学学报(医学版) ,Journal of Southeast University , 编辑部邮箱 ,2004年06期
  • 【分类号】Q936
  • 【被引频次】1
  • 【下载频次】316
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