Isolation of Dividing Organelles from Cyanidioschyzon merolae Cells
Cells of the red alga Cyanidioschyzon merolae contain similar types of organelles as other model eukaryotic organisms (see Chap. 2). However, C. merolae cells are distinct in that they contain fewer organelles, with only one nucleus, mitochondrion, chloroplast, peroxisome, Golgi, and endoplasmic reticulum (ER) per cell, and only a few lysosomes or vacuoles. C. merolae cells lack a rigid cell wall, and the plasma membrane is thick and elastic. These biological features facilitate the isolation of distinct types of organelles from C. merolae cells. Standardized protocols have been established to isolate organelles with bilayer membranes (e.g., chloroplasts and mitochondria) and single-layer membranes (e.g., peroxisomes and lysosomes). The isolated organelles can be used to address biological questions by performing in vitro experiments and biochemical analyses of purified target molecules. Together with imaging and genome information analyses, the resulting data will provide insights into key components involved in fundamental cellular mechanisms. In this chapter, we provide detailed protocols for the isolation of specific organelles from C. merolae cells.
KeywordsChloroplast Lysosome Mitochondria Organelle isolation Peroxisome
This work was supported by a Human Frontier Science Program Long Term Fellowship (LT000356/2011-L to Y.Y.) and grants from the Japan Society for the Promotion of Science Fellowship (no. 14 J04556 to Y.I, 11044 to F.Y.).
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