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Production of selenomethionyl-derivatized proteins in baculovirus-infected insect cells

机译:在杆状病毒感染的昆虫细胞中产生硒代蛋氨酸衍生的蛋白质

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摘要

A protocol is described for the production of both intracellularly expressed and secreted selenomethionyl-derivatized recombinant proteins in baculovirus-infected insect cells. The method results in the production of recombinant soluble proteins with an SeMet occupancy of ∼75% and with a recovery of ∼20% that of native protein expression. The method is independent of the percentage methionine content of the protein and is reliable and consistent. Similar results are obtained using either Spodoptera frugiperda Sf9 or Trichoplusia ni High Five insect cells as the expression host, and when cultures are grown in either shake flasks or in Wave BioReactors.
机译:描述了在杆状病毒感染的昆虫细胞中产生细胞内表达的和分泌的硒代甲硫酰基衍生的重组蛋白的方案。该方法可产生重组可溶性蛋白质,SeMet占75%,天然蛋白质表达回收率约20%。该方法与蛋白质中蛋氨酸含量的百分比无关,并且可靠且一致。当在摇瓶中或Wave BioReactors中培养培养物时,使用节食夜蛾Sf9或Trichoplusia ni High Five昆虫细胞作为表达宿主,可获得相似的结果。

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