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首页> 外文期刊>Cell motility and the cytoskeleton >Recovery of flagellar dynein function in a Chlamydomonas actin/dynein-deficient mutant upon introduction of muscle actin by electroporation.
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Recovery of flagellar dynein function in a Chlamydomonas actin/dynein-deficient mutant upon introduction of muscle actin by electroporation.

机译：通过电穿孔引入肌动蛋白后，衣藻肌动蛋白/动力蛋白缺陷型突变体中鞭毛动力蛋白的功能恢复。

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摘要

Flagellar and ciliary inner-arm dyneins contain actin as a subunit; however, the function of this actin subunit remains unknown. As a first step toward experimental manipulation of actin in dynein, we developed a method for introducing exogenous actin into Chlamydomonas cells by electroporation. A non-motile mutant, ida5oda1, lacking inner-arm dyneins due to the absence of conventional actin, was electroporated in the presence of rabbit skeletal muscle actin. About 20% of the electroporated cells recovered motility under optimal conditions. In addition, by taking advantage of their phototactic behavior, the rescued cells could be concentrated. Motility was also recovered with fluorescently labeled actin; in this case, axonemes became fluorescent after electroporation, suggesting that actin was in fact incorporated as a dynein subunit. The feasibility of incorporating a substantial amount of macromolecules by electroporation will be useful not only for studying actin function, but also for a variety of studies using Chlamydomonas in which no efficient methods have been developed for expressing or introducing foreign proteins and other macromolecules. Copyright 2001 Wiley-Liss, Inc.
机译：鞭毛和睫状内臂动力蛋白含有肌动蛋白作为亚基。然而，该肌动蛋白亚基的功能仍然未知。作为在肌动蛋白中进行肌动蛋白实验操作的第一步，我们开发了一种通过电穿孔将外源肌动蛋白引入衣藻的方法。在兔骨骼肌肌动蛋白存在的情况下，电穿孔了由于缺乏常规肌动蛋白而缺乏手臂动力蛋白的非运动型突变体ida5oda1。在最佳条件下，约20％的电穿孔细胞恢复了运动能力。另外，通过利用它们的光战术行为，可以浓缩所拯救的细胞。还用荧光标记的肌动蛋白恢复了运动能力。在这种情况下，轴蛋白在电穿孔后发荧光，表明肌动蛋白实际上是作为动力蛋白亚基结合的。通过电穿孔掺入大量大分子的可行性不仅对于研究肌动蛋白的功能是有用的，而且对于使用衣藻的各种研究也将是有用的，其中尚未开发出表达或引入外来蛋白质和其他大分子的有效方法的研究。版权所有2001 Wiley-Liss，Inc.

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