Preparation of Outer Membrane Vesicle from Escherichia coli
从大肠杆菌分离外膜囊泡
发布: 2013年12月05日第3卷第23期 DOI: 10.21769/BioProtoc.995 浏览次数: 22014
评审: Anonymous reviewer(s)
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Apr 2013
Abstract
Outer membrane vesicles (OMVs) are spherical bilayered phospholipids of 20-200 nm in size produced from all Gram-negative bacteria and Gram-positive bacteria investigated to date. Previous biochemical and proteomic studies have revealed that the Gram-negative bacteria-derived OMVs are composed of various components like outer membrane proteins, lipopolysaccharides, outer membrane lipids, periplasmic proteins, DNA, and RNA. Here, in this protocol, we describe the method to isolate the OMVs from the culture supernatant of Escherichia coli (E. coli).
Keywords: Outer membrane vesicles (外膜囊泡)
Bacterial extracellular vesicles (细菌胞外囊泡)
Vesicle isolation (囊泡分离)
Materials and Reagents
- Phosphate buffered saline (PBS) (Gibco®, catalog number: 70013-032 )
- E. coli (Isolated from the peritoneal lavage fluid of cecal ligation and puncture-operated mice)
- Luria-Bertani broth (LB) medium (Merck KGaA, catalog number: 1.10285.0500 ) (see Recipes)
Equipment
- 2 L Flasks
- Shaking incubator
- Centrifuge
- 500 ml Bottle top filter 43 mm neck (0.45 μm and 0.22 μm) (Corning, catalog number: 430514 , 430513 )
- QuixStand Benchtop System (Amersham Biosciences, catalog number: 56-4107-44 )
- 100-kDa hollow-fiber membrane (Amersham Biosciences, catalog number: 56-4101-33 )
- Vacuum pump
Note: All centrifuge tubes and flasks should be autoclaved before use to avoid contamination
Procedure
文章信息
版权信息
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Kim, O. Y., Hong, B. S., Park, K., Yoon, Y. J., Choi, S. J., Lee, W. H., Roh, T., Kim, Y. and Gho, Y. S. (2013). Preparation of Outer Membrane Vesicle from Escherichia coli. Bio-protocol 3(23): e995. DOI: 10.21769/BioProtoc.995.
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分类
微生物学 > 微生物细胞生物学 > 细胞器分离
细胞生物学 > 细胞器分离 > 外膜囊泡
生物化学 > 脂质 > 胞外脂质
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