Cell-free Protein Synthesis
Cell-free Protein Synthesis
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DOI：
10.1002/0471684228.egp01977
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发表时间：
2013-05
期刊：
影响因子：
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通讯作者：
N. Sohrabi;M. Taheri;Fatemeh Sadat Tabatabaei Mirakabad;Mohammad Rahmati
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文献类型：
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作者：
N. Sohrabi;M. Taheri;Fatemeh Sadat Tabatabaei Mirakabad;Mohammad Rahmati
蛋白质组学和生物技术领域需要简单快速的方法将基因数据转化为蛋白质。虽然活细胞中的重组蛋白表达是一个耗时的过程，但体外翻译在数小时内从添加的线性PCR DNA指导蛋白合成，而不需要载体和克隆过程。此外，无细胞蛋白质合成（CFPS）的开放性和各种原核和真核细胞裂解物的可及性为体内系统不可能产生的“困难”蛋白质提供了灵活的条件选择。无细胞系统的几个优点包括易于修改反应条件，以使蛋白质折叠，降低对产物毒性的敏感性和适用于高通量策略。无细胞蛋白质表达的最新发展在生物技术、蛋白质组学和基础生物学研究中产生了新的应用。最近CFPS产量超过每升反应体积产生的蛋白质克数，成本已经降低，反应规模已经达到100升的里程碑。在该领域中仍然存在的一些挑战包括在通用平台中可靠地合成任何生物活性蛋白质，缺乏具有成本效益和可扩展的真核CFPS平台，并且不能产生含有类似于人谱的糖基化模式的蛋白质。但我们预计这一趋势将继续下去，CFPS将是一个真正的替代细胞为基础的蛋白质表达系统。
Introduction: Proteomics and biotechnology fields need simple and rapid methods to convert the genetic data into proteins. While recombinant protein expression in living cells is a time-consuming procedure, in vitro translation directs protein synthesis in hours from added linear PCR DNA without the requirement for a vector and cloning process. In addition, the open nature of cell-free protein synthesis (CFPS) and accessibility various prokaryotic and eukaryotic cellular lysates offer a flexible choice of conditions for production of "difficult" proteins that are not possible by in vivo systems. Several advantages of cell-free systems over common cell-based expression methods include the easy modification of reaction conditions to desirableprotein folding, decreased sensitivity to product toxicity and suitability for high-throughput strategies. Recent developments in cell-free protein expression causenovel applications in biotechnology, proteomics and fundamental biological studies.Conclusion: Recently CFPSyields exceed grams of protein produced per liter reaction volume, costs have been decreased and reaction scale has reached the 100-liter milestone.some challenges that remain in this field consist ofsynthesizing any biologically active protein reliably in a universal platform , lacking a cost-effective and scalable eukaryotic CFPS platform and inability to creating proteins containing glycosylation patterns resembling human profile. But we expected that this trend will continue and that CFPS will be a true alternative to cell-based protein expression systems.