Chutkan Halima, Macdonald Ian, Manning Andrew, Kuehn Meta J
Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC, USA.
Methods Mol Biol. 2013;966:259-72. doi: 10.1007/978-1-62703-245-2_16.
Gram-negative bacterial outer membrane vesicle production and function have been studied using a variety of quantitative and qualitative methods. These types of analyses can be hampered by the use of impure vesicle preparations. Here we describe a set of techniques that are useful for the quantitative analysis of vesicle production and for preparative yields of highly purified vesicles for studies of vesicle function or composition. Procedures and advice are also included for the purification of vesicles from encapsulated and low-yield strains.
革兰氏阴性菌外膜囊泡的产生和功能已通过多种定量和定性方法进行了研究。使用不纯的囊泡制剂可能会妨碍这类分析。在此,我们描述了一套技术,这些技术对于囊泡产生的定量分析以及用于囊泡功能或组成研究的高纯度囊泡的制备产量很有用。还包括从包膜菌和低产菌株中纯化囊泡的程序和建议。