Freiwald Anja, Sauer Sascha
Max Planck Institute for Molecular Genetics, Otto-Warburg-Laboratory, Berlin, Germany.
Nat Protoc. 2009;4(5):732-42. doi: 10.1038/nprot.2009.37.
Bacteria are a convenient source of intrinsic marker proteins, which can be detected efficiently by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The patterns of protein masses observed can be used for accurate classification and identification of bacteria. Key to the reliability of the method is a robust and standardized procedure for sample preparations, including bacterial culturing, chemical treatment for bacterial cell wall disruption and for protein extraction, and mass spectrometry analysis. The protocol is an excellent alternative to classical microbiological classification and identification procedures, requiring minimal sample preparation efforts and costs. Without cell culturing, the protocol takes in general <1 h.
细菌是内在标记蛋白的便捷来源,可通过基质辅助激光解吸/电离飞行时间质谱法高效检测。所观察到的蛋白质质量模式可用于细菌的准确分类和鉴定。该方法可靠性的关键在于稳健且标准化的样品制备程序,包括细菌培养、用于破坏细菌细胞壁和提取蛋白质的化学处理以及质谱分析。该方案是经典微生物分类和鉴定程序的极佳替代方案,所需的样品制备工作和成本极少。无需细胞培养,该方案通常耗时不到1小时。