Escherichia coli is one of the most widely used hosts for the production of heterologous proteins. Within this host, the choice of cloning vector constitutes a key factor for a satisfactory amplified expression of a target gene. We aimed to develop novel, unpatented expression vectors that enable the stable maintenance and efficient overproduction of proteins in E. coli. A series of expression vectors based on the ColE1-like pIGDM1 plasmid were constructed. The vectors named pIGDMCT7RS, pIGDM4RS and pIGDMKAN carry various antibiotic resistance genes: chloramphenicol, ampicillin or kanamycin, respectively. Two derivatives contain the inducible T7 promoter while the third one bears the constitutive pms promoter from a clinical strain of Klebsiella pneumoniae. The pIGDM1-derivatives are compatible with other ColE1-like plasmids commonly used in molecular cloning. The pIGDMCT7RS and pIGDM4RS vectors contain genes encoding AGA and AGG tRNAs, which supplement the shortage of these tRNAs, increasing the efficiency of synthesis of heterologous proteins. In conclusion, pIGDMCT7RS, pIGDM4RS and pIGDMKAN vectors, with significantly improved features, including compatibility with vast majority of other plasmids, were designed and constructed. They enable a high-level expression of a desired recombinant gene and therefore constitute a potential, valuable tool for pharmaceutical companies and research laboratories for their own research or for the production of recombinant biopharmaceuticals.

中文翻译：

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基于pIGDM1质粒的新型表达载体。

大肠杆菌是产生异源蛋白质的最广泛使用的宿主之一。在该宿主中，克隆载体的选择是令人满意地扩增靶基因表达的关键因素。我们旨在开发新颖，未专利的表达载体，该载体能够稳定地维持大肠杆菌中的蛋白质并有效地过量生产。构建了基于类似ColE1的pIGDM1质粒的表达载体。名为pIGDMCT7RS，pIGDM4RS和pIGDMKAN的载体分别带有多种抗生素抗性基因：氯霉素，氨苄青霉素或卡那霉素。两种衍生物包含可诱导的T7启动子，而第三种则带有来自临床肺炎克雷伯菌的组成型pms启动子。pIGDM1衍生物与分子克隆中常用的其他ColE1样质粒兼容。pIGDMCT7RS和pIGDM4RS载体包含编码AGA和AGG tRNA的基因，这些基因弥补了这些tRNA的不足，从而提高了异源蛋白质合成的效率。总之，设计并构建了pIGDMCT7RS，pIGDM4RS和pIGDMKAN载体，这些载体具有显着改善的功能，包括与绝大多数其他质粒的相容性。它们能够高水平表达所需的重组基因，因此对于制药公司和研究实验室进行自己的研究或生产重组生物药物而言，是一种潜在的有价值的工具。这补充了这些tRNA的短缺，提高了异源蛋白质的合成效率。总之，设计并构建了pIGDMCT7RS，pIGDM4RS和pIGDMKAN载体，这些载体具有显着改善的功能，包括与绝大多数其他质粒的相容性。它们能够高水平表达所需的重组基因，因此对于制药公司和研究实验室进行自己的研究或生产重组生物药物而言，是一种潜在的有价值的工具。这补充了这些tRNA的短缺，提高了异源蛋白质的合成效率。总之，设计并构建了pIGDMCT7RS，pIGDM4RS和pIGDMKAN载体，这些载体具有显着改善的功能，包括与绝大多数其他质粒的相容性。它们能够高水平表达所需的重组基因，因此对于制药公司和研究实验室进行自己的研究或生产重组生物药物而言，是一种潜在的有价值的工具。