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周海涛, 曾华书, 李学云, 陈润莉, 侯红斌, 梁伟, 刘奋, 金利泰. 离子对染色体系细菌蛋白染色效果分析[J]. 中国公共卫生, 2011, 27(5): 657-659. DOI: 10.11847/zgggws-2011-27-05-64
引用本文: 周海涛, 曾华书, 李学云, 陈润莉, 侯红斌, 梁伟, 刘奋, 金利泰. 离子对染色体系细菌蛋白染色效果分析[J]. 中国公共卫生, 2011, 27(5): 657-659. DOI: 10.11847/zgggws-2011-27-05-64
ZHOU Hai-tao, ZENG Hua-shu, LI Xue-yun, . Application of ion pair staining system in process of bacterial protein staining[J]. Chinese Journal of Public Health, 2011, 27(5): 657-659. DOI: 10.11847/zgggws-2011-27-05-64
Citation: ZHOU Hai-tao, ZENG Hua-shu, LI Xue-yun, . Application of ion pair staining system in process of bacterial protein staining[J]. Chinese Journal of Public Health, 2011, 27(5): 657-659. DOI: 10.11847/zgggws-2011-27-05-64

离子对染色体系细菌蛋白染色效果分析

Application of ion pair staining system in process of bacterial protein staining

  • 摘要: 目的 评价蛋白离子对染色体系(使用2种带反向电荷物质)对细菌蛋白染色效果,并比较其与同类试剂的异同。方法 纯化扩增金黄色葡萄球菌、副溶血弧菌等菌株,提取菌体蛋白,定量,4倍梯度稀释,十二烷基磺酸钠(SDS)聚丙烯酰胺凝胶电泳。分别用离子对染色体系与商品化进口试剂盒对凝胶进行染料染色和银染,对比分析染色效果。结果 离子对染料最低可使0.16μg细菌蛋白显出条带,与进口染色试剂盒接近,但背景更干净,条带更清晰;离子对银染与国外进口试剂银染比较,操作更加简便,蛋白条带着色迅速而清晰,且可耐受较长时间显色(10~20 min);离子对染色体系中染料是银染的敏化剂,染料染色后再行银染与直接银染效果一致,且可省去敏化过程。结论 离子对染色体系的染料染色与银染有很好的兼容性,染色结果重复性明显提高,更贴近蛋白分离染色检测的需要。

     

    Abstract: Objective To evaluate a new proteinion pair staining system(adopting two kinds of mate rial with reverse charge)for bacterial prote in staining,and to compareits difference with smiilar foreign commercial staining kit.Methods Bacterial proteome of strains of Staphy lococcus aureus and Vibrio parahaemolyticus were extracted with thermo solution and quantified with the RCDC kit and diluted with 4-folds gradient,and then followed by sodium dodecyl sulfate polyacry lamide gelelectrophoresis.The gels were treated with dye staining and resilver staining by contrastion pair system and miported commercial staining kits.Results The new type of dyes resulted in a more clear and obviouslane than that of foreign commercial staining kit with a bacterial prote in lmiitation as low as 0.16μg.Compared to foreign commercial staining kit,the operation for ion pairs silver staining system was more smiple and the backg round was clearer;the display of bands was quick and clear.With ion pairs silver staining,the gels could withstand along erperiod staining.In the process of ion pairs staining,the dye is a sensitizing agent for silver staining.The results of resilver staining after dyeing were similar to the direct silver staining,and the process of sensitizing could beomited.Conclusion The dye used ion pair staining system has good compatibility with silver staining and is more suitable for the requirement of bacte rial proteomics.

     

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