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在杆状病毒感染的昆虫细胞中生产具有生物活性的人RelA(p65)

Production of biologically active human RelA (p65) in baculovirus-infected insect cells.

作者信息

Chabot-Fletcher M, Ho M L, Breton J J, Hanning C R, Amegadzie B Y

机构信息

Department of Immunopharmacology, SmithKline Beecham Pharmaceuticals, King of Prussia, PA 19406-0939, USA.

出版信息

Cell Biol Int. 1996 Nov;20(11):777-9. doi: 10.1006/cbir.1996.0100.

Abstract

A recombinant baculovirus was constructed to express a cDNA encoding RelA (p65), a member of the NF-kappa B/Rel family of proteins. Infection of Spodoptera frugiderda insect cells with the recombinant baculovirus resulted in the production of the biologically active protein as measured by immunoblotting using RelA-specific antisera and by electrophoretic mobility shift assays. The recombinant protein bound specifically to an oligonucleotide containing the NF-kappa B consensus motif but not to that containing the unrelated Oct-1 consensus motif. Thus insect cell-derived RelA possess properties similar to the native protein and may be used in physical, biochemical, and pharmacological studies.

摘要

构建了一种重组杆状病毒,用于表达编码RelA(p65)的cDNA,RelA是NF-κB/Rel蛋白家族的成员。用重组杆状病毒感染草地贪夜蛾昆虫细胞,通过使用RelA特异性抗血清的免疫印迹和电泳迁移率变动分析测定,结果产生了具有生物活性的蛋白质。重组蛋白特异性结合含有NF-κB共有基序的寡核苷酸,但不结合含有无关的Oct-1共有基序的寡核苷酸。因此,昆虫细胞来源的RelA具有与天然蛋白相似的特性,可用于物理、生化和药理学研究。

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