Question

"5.

Briefly describe how you would use specimen images to explore the change of the Golgi during cell cycle and what you find in less than one page. Plots or charts are expected. Hint: 1) You can expand the nucleus ROI so that it also includes the Golgi ROI. 2) You can plot the amount of the Golgi associated GM130 (the brightness of the Golgi) vs the nuclear DNA content (cell cycle). 3) Simple Excel plotting techniques are required."

Answer 1

Golgi apparatus of cells was fluorescently tagged with a green fluorescent protein (GFP), localised by attachment to the NH2 terminal retention signal of N-acetylglucosaminyltransferase I (NAGT I). The location was confirmed by immunogold and immunofluorescence microscopy using a variety of Golgi markers. By metaphase, cells contain a constant number of Golgi frangments dispersed throughout yeh cytoplasm. Conventional and cryoimmunoelctron microscopy showed that the NAGT I-GFP chimera (NAGFP) positive fragments were tubulo-vesicular mitotic Golgi clusters.