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Author:
Date:
2015-03-05
[Abstract] Recently, membrane vesicle (MV) production was described in Gram-positive bacteria, which harbor a variety of components such as toxins, antibiotic resistance proteins, proteases, DNA, and immune modulators. Free lipids have the ability to form micelles, thus it is important to rule out spontaneous association of lipids into vesicle-like structures and rather, that MVs are produced naturally by a metabolically active cell. Here, we describe a protocol utilizing the polysaccharide, glucuronoxylomannan (GXM) from Cryptococcus neoformans (C. neoformans) as a marker to differentiate naturally produced MVs from vesicles that form spontaneously in the Gram-positive model organism, Bacillus subtilis (B. subtilis). MVs are purified from bacterial cultures grown ...
[摘要] 最近，在革兰氏阳性细菌中描述了膜囊泡（MV）产生，其含有多种组分，例如毒素，抗生素抗性蛋白，蛋白酶，DNA和免疫调节剂。游离脂质具有形成胶束的能力，因此排除脂质自发缔合成囊泡样结构是重要的，并且更确切地说，MV由代谢活性细胞天然产生。在这里，我们描述了利用来自新隐孢子虫的多糖葡萄糖醛酸甘露聚糖（GXM）（ C。新型隐球菌）作为标记物来区分天然产生的MV与自发形成的囊泡革兰氏阳性模式生物，枯草芽孢杆菌（<枯草芽孢杆菌）。从在GXM存在下生长的细菌培养物中纯化MV;由细胞天然产生的MV在管腔中不含有GXM，而在培养基中形成的囊泡结构可包封GXM，并且这可通过免疫金属透射电子显微镜可视化。
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