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Overcoming challenges for amplified expression of recombinant proteins using Escherichia coli

机译:使用大肠杆菌克服重组蛋白的扩增表达的挑战

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A thorough characterisation of the genetics, physiology and metabolism of Escherichia coli has led to the availability of a large number of strains and vectors suitable for recombinant protein expression. Despite the relative ease in using E. coli for achieving amplified expression of many recombinant proteins, for some proteins this can be a frustrating and time-consuming process leading to very low expression or no expression at all. This is especially true for membrane proteins, which introduce additional challenges. A number of factors can be considered and optimised for achieving required levels of amplified expression of recombinant proteins in E. coli that are broadly classified as host strain, expression vector and growth conditions. In this paper we summarise these factors and consolidate the common challenges encountered and approaches to overcome them, focusing in particular on cases where there is low amplified expression or no expression at all of the desired recombinant protein, due to various reasons.
机译:彻底表征大肠杆菌的遗传学,生理学和代谢的表征导致了适用于重组蛋白表达的大量菌株和载体的可用性。尽管使用大肠杆菌相对容易实现许多重组蛋白的扩增表达,但对于一些蛋白质来说,这可能是导致非常低的表达或根本没有表达的令人沮丧和耗时的过程。对于膜蛋白尤其如此,引入额外的挑战。可以考虑许多因素,并优化以实现大肠杆菌中重组蛋白的扩增表达的所需水平,其被广泛地归类为宿主菌株,表达载体和生长条件。在本文中,我们总结了这些因素,并巩固遇到的共同挑战和克服它们的方法,特别是由于各种原因在所有所需重组蛋白中存在低扩增表达或表达的情况。

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