Hagemann Sylvia, Stöger Lisa, Kappelmann Melanie, Hassl Ingrid, Ellinger Adolf, Velimirov Branko
Center of Pathobiochemistry and Genetics, Medical University of Vienna, Vienna, Austria.
J Basic Microbiol. 2014 Oct;54(10):1062-72. doi: 10.1002/jobm.201300376. Epub 2013 Oct 18.
Outer membrane vesicles (OMVs) derived from the alphaproteobacterium Ahrensia kielensis and from Pseudoalteromonas marina, a gammaproteobacterium, were sampled from liquid cultures in order to extract the MV-associated DNA, establish a shotgun library, and sequence randomly chosen clones to determine the origins of their DNA. We show that OMVs from A. kielensis and from P. marina both harbour DNA larger than 20 or 30 kbp. Transmission electron microscopical inspection of OMVs of A. kielensis and P. marina showed two types of vesicles: bilayered OMVs with a diameter between 30 and 250 nm and double bilayered OMVs ranging between 80 and 200 nm. Bilayered OMVs are either characterized by the presence of a large electron-dense substance or are elctron translucent. Double bilayered OMVs contained an electron dense substance in the core region surrounded by the second bilayer. 30,094 bp of the genome from OMV of A. kielensis and 45,981 bp of that from P. marina were sequenced. The results indicated that all sequences were single copy and that all sequences, with one exception, were similar to prokaryotic sequences, inserted viral sequences were not detected.
从α-变形菌基尔恩斯阿伦斯菌（Ahrensia kielensis）和γ-变形菌海假交替单胞菌（Pseudoalteromonas marina）的液体培养物中采集外膜囊泡（OMV），以便提取与囊泡相关的DNA，建立鸟枪法文库，并对随机选择的克隆进行测序，以确定其DNA的来源。我们发现，来自基尔恩斯阿伦斯菌和海假交替单胞菌的OMV都含有大于20或30 kbp的DNA。对基尔恩斯阿伦斯菌和海假交替单胞菌的OMV进行透射电子显微镜检查，发现有两种类型的囊泡：直径在30至250 nm之间的双层OMV和直径在80至200 nm之间的双双层OMV。双层OMV的特征要么是存在大量电子致密物质，要么是电子透明的。双双层OMV在被第二双层包围的核心区域含有电子致密物质。对基尔恩斯阿伦斯菌OMV的30,094 bp基因组和海假交替单胞菌OMV的45,981 bp基因组进行了测序。结果表明，所有序列都是单拷贝的，并且除了一个例外，所有序列都与原核序列相似，未检测到插入的病毒序列。
