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首页> 外文期刊>Protein Expression and Purification >A less laborious approach to the high-throughput production of recombinant proteins in Escherichia coli using 2-liter plastic bottles
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A less laborious approach to the high-throughput production of recombinant proteins in Escherichia coli using 2-liter plastic bottles

机译：使用2升塑料瓶在大肠杆菌中高通量生产重组蛋白的省力方法

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摘要

Contemporary approaches to biology often call for the high-throughput production of large amounts of numerous proteins for structural or functional studies. Even with the highly efficient protein expression systems developed in Escherichia coli, production of these proteins is laborious and time-consuming. We have simplified established protocols by the use of disposable culture vessels: common 2-liter polyethylene terephthalate beverage bottles. The bottles are inexpensive, fit conveniently in commonly available flask holders, and, because they are notched, provide sufficient aeration to support the growth of high-density cultures. The use of antibiotics and freshly prepared media alleviates the need for sterilization of media and significantly reduces the labor involved. Uninoculated controls exhibited no growth during the time required for protein expression in experimental cultures. The yield, solubility, activity, and pattern of crystallization of proteins expressed in bottles were comparable to those obtained under conventional culture conditions. After use, the bottles are discarded, reducing the risk of cross-contamination of subsequent cultures. The approach appears to be suitable for high-throughput production of proteins for structural or functional studies. (C) 2003 Elsevier Science (USA). All rights reserved. [References: 21]
机译：当代生物学方法常常要求高通量产生大量用于结构或功能研究的大量蛋白质。即使利用在大肠杆菌中开发的高效蛋白质表达系统，这些蛋白质的生产也是费力且费时的。我们通过使用一次性培养容器简化了已建立的协议：普通的2升聚对苯二甲酸乙二醇酯饮料瓶。瓶子价格便宜，可方便地装入常用的烧瓶架中，并且由于它们带有缺口，因此可提供足够的通气量来支持高密度培养物的生长。抗生素和新鲜制备的培养基的使用减轻了培养基灭菌的需要，并显着减少了所涉及的劳动。未接种的对照在实验培养中蛋白质表达所需的时间内没有生长。在瓶中表达的蛋白质的收率，溶解度，活性和结晶模式与常规培养条件下的蛋白质相当。使用后，将瓶子丢弃，降低了后续培养物交叉污染的风险。该方法似乎适用于高通量蛋白质的结构或功能研究。 （C）2003 Elsevier Science（美国）。版权所有。 [参考：21]

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