温度对重组人凝血酶-2在大肠杆菌BL21(DE3) ArcticExpress中表达的影响

S. Silaban, M. Simorangkir, S. Gaffar, I. Maksum, T. Subroto
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引用次数: 2

摘要

大肠杆菌产生的许多蛋白质在包涵体中积累。本研究旨在检测温度在e.c oli BL21 (DE3) Arctic Express宿主表达重组人凝血酶前2过程中减少包涵体形成的作用。在本研究中,我们创建了一个宿主生长曲线,以找出添加诱导剂的合适时间。本实验使用的诱导剂为IPTG 0.1 mM,发酵过程温度分别为12℃和22℃。结果表明,重组人凝血酶-2在大肠杆菌BL21 (DE3) Arctic Express中以蛋白可溶性的形式成功表达,最佳表达温度为12℃。在SDS-PAGE上从可溶性部分得到63kDa蛋白带。发酵温度越高,不溶性部分蛋白质含量越高。结果表明,发酵温度影响凝血酶-2的表达率。关键词:大肠杆菌BL21(DE3) ArcticExpress,凝血酶前2,可溶性,温度
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Temperature effect on expression of recombinant human prethrombin-2 in Escherichia coli BL21(DE3) ArcticExpress
Many proteins produced in E. coli accumulate in inclusion bodies. This study aims to detect the role of temperature in reducing the formation of inclusion bodies during recombinant human prethrombin-2expressed in E. coli BL21 (DE3) Arctic Express host. In this study, we created a host growth curve to find out the right time to add an inducer. The inducer used in our experiment was IPTG 0.1 mM. The fermentation process use a temperature of 12°C and 22°C. The results showed that recombinant human prethrombin-2 was successfully expressed as protein soluble using an optimum temperature of 12°C in E. coli BL21 (DE3) Arctic Express. It was indicated from the 63kDa protein band obtained from the soluble fraction on SDS-PAGE. The higher temperature of fermentation increased the amount of protein in the insoluble fraction due. It concluded that the fermentation temperature affect the rate of prethrombin-2 expression. Keywords: E. coli BL21(DE3) ArcticExpress, prethrombin-2, soluble, temperature
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