3 D DISTRIBUTION OF CYTOSKELETAL ANTIGENS

我们正在继续努力对重要的蛋白质进行免疫定位 在哺乳动物细胞和酵母中的细胞骨架功能 将抗体应用于已被 通过冷冻保存和冷冻替代良好固定，并嵌入 洛维克利或LR-怀特。最近从以下来源获得的四种抗体 合作伙伴，其中两个已经通过以下方式实现了良好的本地化 经过严格的统计测试，高于背景的信号。 因此，Ahn提供的其他优秀抗体对活性 MAP激酶的不同形式及芽孢杆菌对抗端粒结合蛋白的作用 RAP1未能给出良好的定位，而来自 事实证明是成功的，发表了一篇论文和一篇论文 手稿正在准备中。每一篇论文都描述了一些新的 肌动蛋白与肌动蛋白相关蛋白关系的研究进展 (Arps)在分裂酵母中。我们在以下方面也取得了一些成功 MYC标记的蛋白质版本，只要有足够的标记 现在时。一个和三个Mycs之前都没有成功，但 有了麦科勒姆，我们现在已经成功地在一个基因上使用了13个真菌 称为sid2+，它在指定 分裂酵母中的分裂沟，导致了另一篇论文 现在正在做好提交的准备。 C10

We are continuing our efforts to immunolocalize proteins important for cytoskeletal function in both mammalian cells and in yeast by applying antibodies to thin sections of samples that have been well-fixed by cryopreservation and freeze-substitution and embedded in the Lowicryls or LR-White. Of four antibodies recently obtained from collaborators, two have yielded well-controlled localizations with signals that are above background by statistically rigorous tests. Thus, otherwise excellent antibodies provided by Ahn against active forms of MAP kinase and by Pillus against the telomere-binding protein RAP1 have failed to give good localizations, while two antibodies from Gould have proven successful, yielding one published paper and one manuscript in preparation. Each of these papers describes some new aspect of the relationship between actin and actin-related proteins (ARPs) in fission yeast. We have also had some success with myc-tagged versions of protein, so long as there are enough tags present. One and three mycs have previously been unsuccessful, but with McCollum, we have now been successful using 13 mycs on a gene called sid2+, which is important in specifying the position of the cleavage furrow in fission yeasts, leading to another paper that is now being readied for submission. C10