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鹌鹑输卵管分离纤毛中肌动蛋白的免疫检测

Immunological detection of actin in isolated cilia from quail oviduct.

作者信息

Chailley B, Bork K, Gounon P, Sandoz D

出版信息

Biol Cell. 1986;58(1):43-52. doi: 10.1111/j.1768-322x.1986.tb00487.x.

Abstract

Cilia from quail oviduct were isolated with their membrane. The ultrastructural study revealed a good preservation of cilia in the purified fraction. Electrophoresis on SDS-PAGE showed a reproducible pattern of ciliary proteins, the major bands being those of tubulins 57 kDa and dyneins above 250 kDa. Among the minor bands, an immunological study was focused on a 43 kDa molecular mass protein, using monospecific antibodies against actin. Presence of actin was then detected by immunoblotting of isolated cilia fractions as well as of demembranated cilia, suggesting that actin is associated with the axoneme. The presence of actin in the cilia was confirmed by immunofluorescence. The cilia were found stained only on the proximal part, suggesting an heterogeneous distribution of actin within the axonemal length.

摘要

鹌鹑输卵管的纤毛与其细胞膜一起被分离出来。超微结构研究表明,纯化后的纤毛部分保存良好。SDS - PAGE电泳显示纤毛蛋白呈现出可重复的模式,主要条带为57 kDa的微管蛋白和250 kDa以上的动力蛋白条带。在次要条带中,使用抗肌动蛋白的单特异性抗体对一种分子量为43 kDa的蛋白质进行了免疫学研究。随后通过对分离的纤毛部分以及去膜纤毛进行免疫印迹检测到了肌动蛋白的存在,这表明肌动蛋白与轴丝相关。通过免疫荧光证实了纤毛中存在肌动蛋白。发现纤毛仅在近端部分被染色,这表明轴丝长度范围内肌动蛋白分布不均。

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