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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Large-scale filter mating assay for intra- and inter-specific conjugal transfer of the promiscuous plasmid pMV158 in Gram-positive bacteria.
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Large-scale filter mating assay for intra- and inter-specific conjugal transfer of the promiscuous plasmid pMV158 in Gram-positive bacteria.

机译：革兰氏阳性细菌中混杂质粒pMV158的种内和种间结合转移的大规模滤器交配测定。

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摘要

Many conjugative or mobilizable plasmids from Gram-positive bacteria can be transferred mainly on filter-based surfaces. Such is the case of the streptococcal promiscuous plasmid pMV158 which can be mobilized by functions provided by auxiliary plasmids of the Inc18 family. We have developed a new large scale method consisting of a 96-well microtiter plate coupled with a filter so that we could perform several mating assays between strains of Streptococcus pneumoniae and between S. pneumoniae and Enterococcus faecalis cells. Transfer was monitored by employment of a derivative plasmid from pMV158 tagged with the gene encoding the Green Fluorescent Protein. Several experimental conditions, like different ratios donor to recipient cells, could be tested, thus allowing us to perform multiple mating assays in a single experiment.
机译：来自革兰氏阳性细菌的许多结合型或可动员质粒可以主要在基于过滤器的表面上转移。链球菌混杂质粒pMV158就是这种情况，其可以通过Inc18家族的辅助质粒提供的功能来动员。我们开发了一种新的大规模方法，该方法由一个96孔微量滴定板和一个过滤器组成，因此我们可以在肺炎链球菌菌株之间以及肺炎链球菌和粪肠球菌之间进行几种交配测定。通过使用来自pMV158的衍生质粒来监测转移，所述衍生质粒标记有编码绿色荧光蛋白的基因。可以测试几种实验条件，例如供体与受体细胞的比率不同，从而使我们能够在单个实验中执行多个交配测定。

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