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大肠杆菌无细胞裂解物中的膜蛋白生产

Membrane protein production in Escherichia coli cell-free lysates.

作者信息

Henrich Erik, Hein Christopher, Dötsch Volker, Bernhard Frank

机构信息

Institute of Biophysical Chemistry, Centre for Biomolecular Magnetic Resonance, J.W. Goethe-University, Frankfurt-am-Main, Germany.

Institute of Biophysical Chemistry, Centre for Biomolecular Magnetic Resonance, J.W. Goethe-University, Frankfurt-am-Main, Germany.

出版信息

FEBS Lett. 2015 Jul 8;589(15):1713-22. doi: 10.1016/j.febslet.2015.04.045. Epub 2015 May 1.

Abstract

Cell-free protein production has become a core technology in the rapidly spreading field of synthetic biology. In particular the synthesis of membrane proteins, highly problematic proteins in conventional cellular production systems, is an ideal application for cell-free expression. A large variety of artificial as well as natural environments for the optimal co-translational folding and stabilization of membrane proteins can rationally be designed. The high success rate of cell-free membrane protein production allows to focus on individually selected targets and to modulate their functional and structural properties with appropriate supplements. The efficiency and robustness of lysates from Escherichia coli strains allow a wide diversity of applications and we summarize current strategies for the successful production of high quality membrane protein samples.

摘要

无细胞蛋白质合成已成为迅速发展的合成生物学领域的一项核心技术。特别是膜蛋白的合成,这在传统细胞生产系统中是极具挑战性的蛋白质,是无细胞表达的理想应用。可以合理设计出各种各样的人工和自然环境,以实现膜蛋白的最佳共翻译折叠和稳定。无细胞膜蛋白生产的高成功率使得能够专注于个别选定的目标,并通过适当的补充剂来调节其功能和结构特性。大肠杆菌菌株裂解物的效率和稳健性允许进行广泛多样的应用,我们总结了目前成功生产高质量膜蛋白样品的策略。

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