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sRNA Multiplex Gene Expression Inhibition System based on Synthetic Regulatory Small RNA and Method of Preparing the Same

机译:基于合成调控小RNA的sRNA多重基因表达抑制系统及其制备方法

摘要

The present invention relates to a multiplex target gene expression inhibition system based on a synthetic regulatory sRNA and a method of preparing the same, and more specifically, to a combination of vectors comprising a variety of antibiotic-resistance genes, an origin of replication, and a synthetic regulatory sRNA; a method of preparing the same; an sRNA library consisting of the combination of vectors; and a method of screening for a bacterial strain producing a useful substance by using the same. The multiplex target gene expression inhibition system according to the present invention is a gene expression regulation system based on a synthetic regulatory sRNA that, unlike existing gene knock-out methods, can effectively inhibit the expression of a specific gene without altering the sequence of a target gene; comprises various vectors, and thus can be used compatibly with other plasmids usable in prokaryotes; and is designed to enable multiplex introduction, and thus is advantageous in a fast and efficient production of recombinant microorganisms. Furthermore, the present invention provides a technology that enables simple and fast transferring of a genome-level synthetic regulatory sRNA library to a new platform, and the technology is useful in selection of recombinant microorganisms producing useful substances in high capacity. Accordingly, the present invention is useful in the production of recombinant bacterial strains capable of dramatically increasing the production efficiency of a variety of medically useful metabolites.
机译:本发明涉及基于合成的调节性sRNA的多重靶基因表达抑制系统及其制备方法,更具体地说,涉及包含多种抗生素抗性基因,复制起点的载体的组合,以及合成的调节性sRNA;一种制备方法;由载体组合组成的sRNA文库;以及通过使用该方法筛选产生有用物质的细菌菌株的方法。根据本发明的多重靶基因表达抑制系统是基于合成调控sRNA的基因表达调控系统,与现有的基因敲除方法不同,该sRNA可以有效抑制特定基因的表达而无需改变靶序列。基因;包含各种载体,因此可以与原核生物中使用的其他质粒兼容使用;并且其被设计成能够多重引入,因此有利于快速有效地生产重组微生物。此外,本发明提供了一种技术,其使得能够将基因组水平的合成调节性sRNA文库简单且快速地转移到新平台,并且该技术可用于选择以高容量产生有用物质的重组微生物。因此,本发明可用于生产重组细菌菌株,该菌株能够显着提高多种医学上有用的代谢产物的生产效率。

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