Journal: Genome Biology and Evolution

Article Title: Proteomics Reveals Plastid- and Periplastid-Targeted Proteins in the Chlorarachniophyte Alga Bigelowiella natans

doi: 10.1093/gbe/evs115

Figure Lengend Snippet: Bigelowiella natans subcellular fractionation and protein isolation. ( A ) Three-step sucrose density gradient of French press-disrupted B. natans cells. Three distinct bands are apparent. ( B ) Fluorescence microscopy of material present in “fraction 3.” The image is an overlay of two different channels showing DAPI-stained nuclei (blue) and chlorophyll autofluorescence from the plastids under a rhodamine filter (red). ( C ) Fluorescence microscopy showing material in fraction 1, which was enriched in plastid–nucleomorph complexes and largely devoid of host cell nuclei. Faint blue spots corresponding to nucleomorphs (NM) can be seen in close association with plastids (PL). A large blue spot corresponding to a nucleus (N) can also be seen. ( D ) Transmission electron micrograph of isolated B. natans plastids obtained from fraction 1 of the sucrose gradients after subcellular fractionation. ( E ) TEM of an isolated plastid–nucleomorph complex from B. natans . ( F ) One-dimensional SDS-PAGE of B. natans proteins isolated from fraction 1. Fourteen discrete bands were excised from the gel, as shown, for proteomic analysis (refer to text for further details).

Article Snippet: The sample was fractionated on a 2.4 cm 1D sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) (7–15% acrylamide gradient) using a Hoefer SE 600 Rubys system (GE Health Care).

Techniques: Fractionation, Isolation, Fluorescence, Microscopy, Staining, Transmission Assay, SDS Page