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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >A medium-copy-number plasmid for insertional mutagenesis of Streptococcus mutans.
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A medium-copy-number plasmid for insertional mutagenesis of Streptococcus mutans.

机译：一个中等拷贝数的质粒，用于变形链球菌的插入诱变。

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摘要

We have constructed a plasmid useful for insertional mutagenesis in Streptococcus mutans. The molecule, pSU20Erm, is based on a derivative of pACYC184 known as pSU20. The plasmid described here is approximately 3.7 kb in size and has the following properties: it replicates in Escherichia coli, does not replicate in S. mutans, contains an erythromycin-resistance marker which can be selected in E. coli or the streptococci, contains a multiple cloning site with few restriction sites in the remainder of the molecule, and can be screened on X-Gal-containing medium for the presence of insertions into the multiple cloning site. We have used the plasmid to construct a library of S. mutans DNA in E. coli and show that the clones can be reintegrated into the S. mutans chromosome via homologous recombination, thereby interrupting native genes. The plasmid has been used to clone part of a homologue of the E. coli drpA gene, encoding a global regulatory element for RNA synthesis. Further, we have identified an element closely linked to drpA in S. mutans with high homology to IS861. Copyright 1998 Academic Press.
机译：我们已经构建了可用于变异链球菌中的插入诱变的质粒。分子pSU20Erm基于pACYC184的衍生物，称为pSU20。此处描述的质粒大小约为3.7 kb，具有以下特性：它在大肠杆菌中复制，在变形链球菌中不复制，包含可以在大肠杆菌或链球菌中选择的红霉素抗性标记，多克隆位点，在分子的其余部分具有很少的限制性位点，并且可以在含X-Gal的培养基上筛选是否存在插入多克隆位点的插入。我们已经使用该质粒在大肠杆菌中构建了变形链球菌DNA文库，并显示该克隆可以通过同源重组重新整合到变形链球菌染色体中，从而中断了天然基因。该质粒已用于克隆大肠杆菌drpA基因同源物的一部分，该基因编码用于RNA合成的全局调控元件。此外，我们已经确定了与IS861高度同源的变形链球菌中与drpA紧密相连的元件。版权所有1998学术出版社。

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