头孢西丁诱导产ACT-1型AmpC酶大肠埃希菌的比较蛋白质组学研究

Comparative proteomics analysis of Escherichia coli producing ACT-1 type class C beta-lactamase induced with Cefoxitin

目的研究产AmpC酶大肠埃希菌在头孢西丁诱导前后的蛋白质谱差异，筛选有价值的靶位。方法提取产ACT-1AmpC酶大肠埃希菌在头孢西丁诱导前后的全菌蛋白，应用双向凝胶电泳技术分析和Bluesilver法染色。凝胶图像分析后，对差异蛋白质进行MALDI-TOF质谱分析。结果在所鉴定的8个差异蛋白当中，AmpC酶、外膜蛋白Ⅱ和转座酶表达显著上调，而磷酸转移酶系统的葡萄糖特异性组分ⅡA蛋白、抗碲酸盐蛋白、丙三醇磷酰基二酯酶、硫醇过氧化物酶及细菌分化蛋白FtsZ表达下调。结论所鉴定的这些差异蛋白将为阐明产AmpC酶大肠埃希菌耐药产生的机制提供线索，也为筛选具有潜在价值的靶位提供理论依据。

Objective To study protein alteration profile between before and after the AmpC production of E. coli induced by cefoxitin. Methods The extracted proteins were separated by 2-Di mensional electrophoresis（DE） and stained with blue silver staining. After gel image analysis, the differential staining spot were identified by matrix assisted laser desorption ionization time of flight mass spectrometry （ MALDITOF MS） technology. Results Of the identified eight differential proteins, AmpC beta-lactamase, outer membrane protein II,transposase were up-regulated after induction. While glucose-specific component IIA of the PTS system, tellurite resistance, glycerophosphoryl diester phosphordiesterase, thiol peroxidase, cell division protein FtsZ were down-regulated after induction. Conclusion These differentially expressed proteins give some clues to elucidate the the resistant mechanism of E. coli producing AmpC, and provide the basis of searching for potential target.