Strategies for the expression of SUMO-modified target proteins in Escherichia coli.
Strategies for the expression of SUMO-modified target proteins in Escherichia coli.
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DOI：
10.1007/978-1-59745-566-4_14
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发表时间：
2009-02
影响因子：
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通讯作者：
H. Saitoh;Junsuke Uwada;Kawasaki Azusa
中科院分区：
文献类型：
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作者：
H. Saitoh;Junsuke Uwada;Kawasaki Azusa
我们之前描述了一个双元载体系统的建立，该系统允许相扑结合酶和目的蛋白的共表达，从而导致高效的相扑修饰，并在大肠杆菌中产生大量的重组相扑修饰蛋白。这样做的好处。大肠杆菌表达/修饰方法包括实验的可扩展性、低成本、快速生长以及缺乏切割相扑与目标蛋白之间的异肽链的蛋白酶。因此，这就是。Colimethod提供了一种有用的替代真实相扑修饰分析的方法，如体外相扑结合和使用杆状病毒或哺乳动物细胞培养的体内相扑修饰，这些通常复杂、耗时和昂贵。
We previously described the establishment of a binary vector system that allows co-expression of SUMO conjugation enzymes and a target protein of interest, leading to efficient SUMO modification and the production of a large amount of recombinant SUMO-modified proteins inEscherichia coli. The advantages of thisE. coliexpression/modification approach include scalability of experiments, low cost, fast growth, and a lack of proteases that cleave the isopeptide linkage between SUMO and the target protein. Thus, thisE. colimethod provides a useful alternative to authentic SUMO modification assays, such as in vitro SUMO conjugation and in vivo SUMO modification using baculovirus or mammalian cell culture, that are usually complicated, time-consuming and expensive.