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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Low copy expression vectors for use in Yersinia sp. and related organisms
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Low copy expression vectors for use in Yersinia sp. and related organisms

机译：用于耶尔森氏菌的低拷贝表达载体。和相关生物

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摘要

In Yersinia, the most commonly used expression vectors for genetic studies such as gene complementation do not effectively allow for both induction and repression of gene expression. Additionally, there is no expression system available that can be induced in bacteria growing in vitro as well as in vivo, e.g. in eukaryotic cell lines or in living animal models. Here, we present a series of novel inducible low copy expression vectors that are well suited for use in the Yersinia species. Their tet operator/promoter/repressor system makes them distinct from other vectors, and gene transcription in bacteria can easily be induced by addition of anhydrotetracyline (ATc) either to the growth medium, to tissue culture medium during bacterial infections of cell lines or by injection into animals infected with bacteria. Researchers can choose between two different antibiotic resistances (kanamycin or spectinomycin), between two copy numbers (5 or 12-22) as well as between two different versions for expression from either the native RBS and ATG or RBS and ATG encoded in the plasmid. The whole vector series contains the same multi-cloning site from pBluescript II KS+ that allows for easy subcloning. Moreover, these vectors are built in a modular fashion that makes it simple to adapt them for other purposes. Finally, in addition to their use in Yersinia they are suitable for use in many other Enterobacteriaceae.
机译：在耶尔森氏菌中，用于基因研究的最常用表达载体（例如基因互补）不能有效地诱导和抑制基因表达。另外，没有可用的表达系统可在体外以及体内生长的细菌中诱导。在真核细胞系或活体动物模型中。在这里，我们介绍了一系列非常适合用于耶尔森氏菌物种的新型诱导型低拷贝表达载体。它们的tet操纵子/启动子/阻遏物系统使其与其他载体截然不同，并且可以通过在细胞系细菌感染过程中向生长培养基，组织培养基中添加无水四氢大麻酚（ATc）或通过注射轻松地诱导细菌中的基因转录进入被细菌感染的动物。研究人员可以选择两种不同的抗生素抗性（卡那霉素或壮观霉素），两种拷贝数（5或12-22）以及两种不同的版本，以从质粒中编码的天然RBS和ATG或RBS和ATG表达。整个载体系列包含来自pBluescript II KS +的相同多克隆位点，可轻松进行亚克隆。而且，这些向量以模块化的方式构建，这使得将其轻松用于其他目的变得很容易。最后，除了在耶尔森菌中使用外，它们还适用于许多其他肠杆菌科。

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