Suppr超能文献

利用杆状病毒表达系统表达人白细胞介素-2受体α链和β链。

Expression of human IL-2 receptor alpha- and beta-chains using the baculovirus expression system.

作者信息

Lindqvist C, Engberg C, Ojala P, Karp M, Akerman K, Oker-Blom C

机构信息

Abo Akademi University, Department of Biochemistry and Pharmacy, Turku, Finland.

出版信息

Scand J Immunol. 1993 Sep;38(3):267-72. doi: 10.1111/j.1365-3083.1993.tb01724.x.

Abstract

The genes encoding the alpha- and beta-chains of the human interleukin-2 receptor were expressed in lepidopteran insect cells using the baculovirus expression vector system. The corresponding genes were inserted under the polyhedrin promoter of the Autographa california nuclear polyhedrosis virus and expressed in the Spodoptera frugiperda insect cell line during viral infection. The recombinant receptor proteins were identified in the insect cell lysates by using protein dot blot and ELISA techniques. At 36 h post infection the corresponding proteins were clearly detected using anti-IL-2 alpha- and beta-receptor-specific antibodies. A large amount of the alpha-chain was also found in the supernatant culture media at 72 h post infection and metabolic labelling with [35S]-methionine indicated that it was proteolytically cleaved into a 32 kDa soluble form. A similar soluble or secreted form of the beta-chain was, however, not observed. Both receptor proteins were expressed on the surface of the insect cells as determined by flow cytometry analysis. Studies performed with the different IL-2 receptor forms (alpha- and beta-chains alone or in combination) in the presence or absence of rIL-2 suggest that the receptor proteins when expressed in infected insect cells are non-functional with respect to tyrosine phosphorylation.

摘要

利用杆状病毒表达载体系统,将编码人白细胞介素-2受体α链和β链的基因在鳞翅目昆虫细胞中进行表达。将相应基因插入苜蓿银纹夜蛾核型多角体病毒的多角体蛋白启动子下游,并在病毒感染期间在草地贪夜蛾昆虫细胞系中表达。通过蛋白质斑点印迹和酶联免疫吸附测定技术在昆虫细胞裂解物中鉴定重组受体蛋白。感染后36小时,使用抗IL-2α和β受体特异性抗体可清晰检测到相应蛋白质。感染后72小时,在上清培养液中也发现大量α链,用[35S]-甲硫氨酸进行代谢标记表明它被蛋白水解切割成32 kDa的可溶性形式。然而,未观察到类似的β链可溶性或分泌形式。通过流式细胞术分析确定,两种受体蛋白均在昆虫细胞表面表达。在存在或不存在重组白细胞介素-2的情况下,对不同白细胞介素-2受体形式(单独的α链和β链或两者组合)进行的研究表明,在感染昆虫细胞中表达时,受体蛋白在酪氨酸磷酸化方面无功能。

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验