Gustafsson E, Thorén K, Larsson T, Davidsson P, Karlsson K A, Nilsson C L
Institute of Medical Biochemistry, Göteborg University, Box 440, SE-405 30 Göteborg, Sweden.
Rapid Commun Mass Spectrom. 2001;15(6):428-32. doi: 10.1002/rcm.243.
Escherichia coli is a gram-negative bacterium that causes sepsis and infections of the nervous system, and the digestive and urinary tracts. The availability of the complete nucleotide sequence encoding the E. coli K-12 genome has made this organism an excellent model for proteomic studies. Semi-preparative two-dimensional electrophoresis, including liquid phase isoelectric focusing (IEF), one-dimensional sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) and gel elution, have for the first time been used in combination with matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS), electrospray tandem mass spectrometry and database searching for rapid separation of proteins from a uropathogenic strain of E. coli. The identity of 30 proteins, including the membrane protein nmpC, was obtained using this approach.
大肠杆菌是一种革兰氏阴性菌，可导致败血症以及神经系统、消化系统和泌尿系统感染。编码大肠杆菌K-12基因组的完整核苷酸序列的可得性，使这种生物体成为蛋白质组学研究的极佳模型。半制备二维电泳，包括液相等电聚焦（IEF）、一维十二烷基硫酸钠（SDS）聚丙烯酰胺凝胶电泳（PAGE）和凝胶洗脱，首次与基质辅助激光解吸/电离飞行时间质谱（MALDI-TOFMS）、电喷雾串联质谱以及数据库搜索相结合，用于从一株致病性大肠杆菌中快速分离蛋白质。使用这种方法确定了30种蛋白质的身份，包括膜蛋白nmpC。
