Sphingolipid trafficking and purification in Chlamydia trachomatis-infected cells.
Sphingolipid trafficking and purification in Chlamydia trachomatis-infected cells.
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DOI：
10.1002/9780471729259.mc11a02s27
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发表时间：
2012-11
影响因子：
--
通讯作者：
Moore, Elizabeth R
中科院分区：
文献类型：
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作者：
Moore, Elizabeth R
沙眼衣原体是一种专性细胞内的人类病原体，它缺乏一个系统，允许遗传操作。因此，衣原体研究人员必须操纵宿主细胞，以更好地了解衣原体生物学。宿主来源的脂质获得对于衣原体在宿主内的存活至关重要。因此，跟踪和纯化鞘脂的能力/从衣原体感染的细胞已成为衣原体生物学的关键研究的一个组成部分。该单位概述了协议，提供有关标记真核细胞与外源性脂质检查高尔基体衍生的脂质贩运的衣原体列入，然后进行成像研究或脂质提取定量的细节。提供细节以允许这些方案应用于亚融合、极化或siRNA敲低细胞。此外，人们会发现重要的实验设计考虑和技术。这些方法是帮助理解C.沙眼衣原体操纵和篡夺宿主细胞运输途径。
Chlamydia trachomatis is an obligate intracellular human pathogen, which lacks a system that allows genetic manipulation. Therefore, chlamydial researchers must manipulate the host cell to better understand chlamydial biology. Host-derived lipid acquisition is critical for chlamydial survival within the host. Hence, the ability to track and purify sphingolipids in/from chlamydial infected cells has become an integral part of pivotal studies in chlamydial biology. This Unit outlines protocols that provide details about labeling eukaryotic cells with exogenous lipids to examine Golgi-derived lipid trafficking to the chlamydial inclusion and then performing imaging studies or lipid extractions for quantification. Details are provided to allow these protocols to be applied to subconfluent, polarized or siRNA knockdown cells. In addition, one will find important experimental design considerations and techniques. These methods are powerful tools to aid in the understanding of mechanisms which allow C. trachomatis to manipulate and usurp host cell trafficking pathways.