Powers Matthew J, Herrera Carmen M, Tucker Ashley T, Davies Bryan W, Trent M Stephen
Department of Microbiology, University of Georgia, Athens, GA, USA.
Department of Infectious Diseases, University of Georgia, Athens, GA, USA.
Methods Mol Biol. 2019;1946:233-252. doi: 10.1007/978-1-4939-9118-1_22.
With the increasing occurrence of antibiotic resistance among Acinetobacter sp., the race is on for researchers to not only isolate resistant isolates but also utilize basic and applied microbiological techniques to study mechanisms of resistance. For many antibiotics, the limit of efficacy against Gram-negative bacteria is dependent on its ability to permeate the outer membrane and access its target. As such, it is critical that researchers be able to isolate and analyze the lipid components of the cell envelope from any number of Acinetobacter sp. that are either resistant or sensitive to antibiotics of interest. The following chapter provides in-depth protocols to confirm the presence or absence of lipooligosaccharide (LOS) in Acinetobacter sp., isolate lipid A, and glycerophospholipids and analyze them using qualitative (mass spectrometry) and semiquantitative (thin-layer chromatography) methods.
随着不动杆菌属中抗生素耐药性的日益增加，研究人员不仅要分离出耐药菌株，还要利用基础和应用微生物技术来研究耐药机制。对于许多抗生素来说，其对革兰氏阴性菌的疗效极限取决于其穿透外膜并作用于靶点的能力。因此，研究人员能够从对感兴趣的抗生素耐药或敏感的任何不动杆菌属中分离和分析细胞膜的脂质成分至关重要。以下章节提供了深入的实验方案，以确认不动杆菌属中脂寡糖（LOS）的存在与否，分离脂质A和甘油磷脂，并使用定性（质谱）和半定量（薄层色谱）方法对它们进行分析。
