双向凝胶电泳中三种蛋白沉淀方法的比较
omparison of three protein precipitation procedures in two-dimensional gel electrophoresis
本文比较了不同蛋白沉淀方法对鸡毒支原体总蛋白双向凝胶电泳结果的影响。应用超声辅助的尿素裂解法提取总蛋白，采用2 D Clean-up Kit沉淀、丙酮沉淀法及三氯乙酸(TCA) /丙酮沉淀法分别沉淀鸡毒支原体的蛋白质组,采用固相pH梯度等电聚焦为第一向和垂直SDS-PAGE为第二向对沉淀蛋白进行双向电泳,观察双向电泳的蛋白质点数及质量。采用2D Clean-up Kit法沉淀后，鸡毒支原体双向电泳图谱的蛋白质点数较丙酮沉淀法、三氯乙酸(TCA) /丙酮沉淀法多，图谱背景更清晰，分辨率也更高，提高了鸡毒支原体双向凝胶电泳的蛋白检测灵敏度，为下一步的蛋白组学分析奠定了基础。
hree different protein precipitation procedures were compared in the two-dimensional (2 D) gel electrophoresis of Mycoplasma gallisepticum proteome. After ultrasound-assisted urea lysis of M. gallisepticum, 2 D clean-up kit, acetone, and trichloroacetic acid / acetone were used for protein precipitation. The quantity and quality of protein spots in 2 D map of M.Gallisepticum were observed and analyzed. Compared to acetone, and trichloroacetic acid / acetone precipitation, 2 D clean-up kit increased the protein spot number and improved background and resolution in the 2 D map. The 2 D clean-up kit was able to enhance the detection sensitivity of 2 D gel electrophoresis and preferred for further proteomic analysis.
夏曦、崔耀文、沈建忠
生物科学研究方法、生物科学研究技术生物化学微生物学
蛋白沉淀双向凝胶电泳蛋白组学
Protein precipitationtwo-dimensional gel electrophoresisproteomics
夏曦,崔耀文,沈建忠.双向凝胶电泳中三种蛋白沉淀方法的比较[EB/OL].(2012-06-08)[2025-08-06].http://www.paper.edu.cn/releasepaper/content/201206-139.点此复制
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