Beckers C J, Keller D S, Balch W E
Cell. 1987 Aug 14;50(4):523-34. doi: 10.1016/0092-8674(87)90025-0.
We introduce a new method that removes portions of the plasma membrane of eukaryotic cells to form semi-intact cells. During preparation, these cells lose their soluble cytoplasmic contents, but retain secretory organelles such as the ER and Golgi complex in an intact form. Transport of protein between the ER and Golgi can be functionally reconstituted in vitro using these semi-intact cells by incubation in the presence of cytosol and ATP. Export of the vesicular stomatitis virus strain tsO45 G protein from the ER in vitro is temperature-sensitive, similar to the result observed in vivo. These cells allow direct access of chemicals and antibodies to the cytoplasmic domain of the cell and may be a widely applicable model system for study of a broad range of problems in cell biology.
我们介绍了一种新方法，该方法可去除真核细胞的部分质膜以形成半完整细胞。在制备过程中，这些细胞会失去其可溶性细胞质内容物，但会完整保留诸如内质网和高尔基体等分泌细胞器。通过在胞质溶胶和ATP存在的条件下孵育，利用这些半完整细胞可在体外功能上重建内质网和高尔基体之间的蛋白质转运。体外水泡性口炎病毒株tsO45 G蛋白从内质网的输出对温度敏感，这与体内观察到的结果相似。这些细胞使化学物质和抗体能够直接接触细胞的细胞质结构域，并且可能是用于研究细胞生物学中广泛问题的广泛适用的模型系统。
