Fares Hanna, van der Bliek Alexander M
Department of Molecular and Cellular Biology, University of Arizona, Tucson, AZ, USA.
Methods Cell Biol. 2012;107:239-63. doi: 10.1016/B978-0-12-394620-1.00008-4.
This chapter describes methods for studying membrane traffic and organelle biogenesis in Caenorhabditis elegans. These processes have traditionally been studied with yeast or mammalian cells, but C. elegans is emerging as an attractive alternative model system for cell biologists. C. elegans is well known for the ease of manipulation through classic and molecular genetic techniques. In addition, C. elegans is transparent, so fluorescent proteins can be observed in live animals. These properties have aided the development of functional assays for tracking cell biological processes in situ. Localization results obtained with fluorescent proteins can be validated with immunofluorescence and with biochemical methods, such as subcellular fractionation, adapted from methods developed for other organisms. C. elegans thus combines powerful genetics with a range of cell biological techniques to study subcellular processes in a tractable multicellular organism.
本章描述了研究秀丽隐杆线虫中膜运输和细胞器生物发生的方法。传统上，这些过程是通过酵母或哺乳动物细胞进行研究的，但秀丽隐杆线虫正逐渐成为细胞生物学家颇具吸引力的替代模型系统。秀丽隐杆线虫以其通过经典和分子遗传学技术易于操作而闻名。此外，秀丽隐杆线虫是透明的，因此可以在活体动物中观察荧光蛋白。这些特性有助于开发原位追踪细胞生物学过程的功能测定法。用荧光蛋白获得的定位结果可以通过免疫荧光以及生化方法（如亚细胞分级分离）进行验证，这些生化方法是根据为其他生物体开发的方法改编而来的。因此，秀丽隐杆线虫将强大的遗传学与一系列细胞生物学技术相结合，以研究一种易于处理的多细胞生物体中的亚细胞过程。
