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首页> 外文期刊>Biochemical Engineering Journal >Comparative production of human interleukin-2 fused with green fluorescent protein in several recombinant expression systems
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Comparative production of human interleukin-2 fused with green fluorescent protein in several recombinant expression systems

机译：在几种重组表达系统中与绿色荧光蛋白融合的人白介素2的比较生产

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摘要

The selection of an optimal recombinant expression system is important for successful protein production.Here,we compared production of human interleukin-2 (hTL-2)-green fluorescent protein (GFP) fusion proteins in several expression systems such as bacteria Escherichia coli,yeast Pichia pastoris,insect Spodoptera frugiperda Sf-9 cells,insect Tricoplusia ni larvae,and insect Drosophila melanogaster S2 cells.Due to the highly hydrophobic nature of hIL-2,the GFP/hIL-2 fusion protein was expressed as an inclusion body in the E.coli system,resulting in minimal green fluorescence;however,Western blot analysis revealed the proper fusion band.In all other cases,the fusion proteins were expressed intracellularly or secreted as a functional form;green fluorescence was observed in each of these expression systems.We determined the linear relationships between GFP fluorescence and hIL-2 concentration in each case and used these correlations for comparison of the various expression systems in terms of production yield,productivity,product solubility (for intracellular expression systems),secretion efficiency (for secretion systems),and even functionality by simple measurement of GFP fluorescence.Even though the culture conditions were not optimized for each expression system,this comparison can be used as preliminary criteria for the selection of a proper expression system for recombinant protein production.
机译：最佳重组表达系统的选择对于成功生产蛋白质非常重要。在此，我们比较了人白介素2（hTL-2）-绿色荧光蛋白（GFP）融合蛋白在几种表达系统中的生产，例如大肠杆菌，酵母巴斯德毕赤酵母，小菜蛾（Spodoptera frugiperda）Sf-9细胞，粉虱（Tricoplusia ni larvae）昆虫和果蝇（Drosophila melanogaster）S2细胞。由于hIL-2具有高度疏水性，因此GFP / hIL-2融合蛋白被表达为包涵体。 E.coli系统，产生的绿色荧光极少;但是，Western印迹分析显示了适当的融合带。在所有其他情况下，融合蛋白均在细胞内表达或以功能形式分泌;在每个表达系统中均观察到绿色荧光我们确定了每种情况下GFP荧光与hIL-2浓度之间的线性关系，并使用这些相关性来比较术语中各种表达系统产量，生产率，产品溶解度（对于细胞内表达系统），分泌效率（对于分泌系统）甚至功能，只需简单地测量GFP荧光即可。即使未针对每个表达系统优化培养条件，该比较也可以作为选择重组蛋白生产的合适表达系统的初步标准。

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