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Effects of a recombinant gene expression on ColE1-like plasmid segregation in Escherichia coli

机译:重组基因表达对大肠杆菌中类似于ColE1的质粒分离的影响

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摘要

BackgroundSegregation of expression plasmids leads to loss of recombinant DNA from transformed bacterial cells due to the irregular distribution of plasmids between the daughter cells during cell division. Under non-selective conditions this segregational instability results in a heterogeneous population of cells, where the non-productive plasmid-free cells overgrow the plasmid-bearing cells thus decreasing the yield of recombinant protein. Amongst the factors affecting segregational plasmid instability are: the plasmid design, plasmid copy-number, host cell genotype, fermentation conditions etc. This study aims to investigate the influence of transcription and translation on the segregation of recombinant plasmids designed for constitutive gene expression in Escherichia coli LE392 at glucose-limited continuous cultivation. To this end a series of pBR322-based plasmids carrying a synthetic human interferon-gamma (hIFNγ) gene placed under the control of different regulatory elements (promoter and ribosome-binding sites) were used as a model.
机译:背景表达质粒的分离由于在细胞分裂过程中子细胞之间质粒的不规则分布而导致转化DNA从重组细菌细胞中丢失。在非选择性条件下,这种分离的不稳定性导致细胞的异质群体,其中非生产性的无质粒细胞使携带质粒的细胞过度生长,从而降低了重组蛋白的产量。影响分离质粒不稳定性的因素包括:质粒设计,质粒拷贝数,宿主细胞基因型,发酵条件等。本研究旨在研究转录和翻译对重组质粒分离的影响,所述重组质粒用于大肠杆菌中组成型基因表达限制葡萄糖连续培养的大肠杆菌LE392。为此目的,将一系列携带合成人干扰素-γ(hIFNγ)基因的基于pBR322的质粒置于不同调控元件(启动子和核糖体结合位点)的控制下。

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