Easy labeling of proliferative phase and sporogonic phase of microsporidia Nosema bombycis in host cells.
Easy labeling of proliferative phase and sporogonic phase of microsporidia Nosema bombycis in host cells.
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轻松标记宿主细胞中家蚕微孢子虫的增殖期和孢子发生期
DOI：
10.1371/journal.pone.0179618
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Pan G
中科院分区：
文献类型：
--
作者：
Chen J;Guo W;Dang X;Huang Y;Liu F;Meng X;An Y;Long M;Bao J;Zhou Z;Xiang Z;Pan G
微孢子虫是一种真核、单细胞寄生虫，已被研究了150多年。这些生物具有非同寻常的能力，能够入侵包括脊椎动物和无脊椎动物在内的各种宿主，如人类和具有重要商业价值的动物。由于缺乏合适的标记方法，限制了对细胞周期和细胞内蛋白质定位的研究。在本报告中，发展了一种简便的荧光标记方法来标记微孢子虫在宿主细胞中的增殖期和产孢期。在甲壳素存在的基础上，用硫代荧光白M2R标记孢子相，用β-微管蛋白抗体与荧光二抗结合免疫荧光标记增殖期。该方法简单、高效，既可用于感染细胞，也可用于组织切片，在微孢子虫研究中具有很大的应用潜力。
Microsporidia are eukaryotic, unicellular parasites that have been studied for more than 150 years. These organisms are extraordinary in their ability to invade a wide range of hosts including vertebrates and invertebrates, such as human and commercially important animals. A lack of appropriate labeling methods has limited the research of the cell cycle and protein locations in intracellular stages. In this report, an easy fluorescent labeling method has been developed to mark the proliferative and sporogonic phases of microsporidia Nosema bombycis in host cells. Based on the presence of chitin, Calcofluor White M2R was used to label the sporogonic phase, while β-tubulin antibody coupled with fluorescence secondary antibody were used to label the proliferative phase by immunofluorescence. This method is simple, efficient and can be used on both infected cells and tissue slices, providing a great potential application in microsporidia research.