Difference between revisions of "Team:CU-Boulder/Isolation"
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| + | <p><i>Figure 1: EutS compartment containing fusion red tagged with Eut C | ||
| + | 2. Electron Microscopy with various radiological stains on a copper grid.</i></p> | ||
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| − | <p>Figure 2: EutS compartment with negative stain</p> | + | <p><i>Figure 2: EutS compartment with negative stain</i></p> |
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Revision as of 23:16, 29 October 2017
Bacterial microcompartments are protein-based organelles that can enclose enzymes and other proteins, however without successful isolation of the compartments they do not have much use. We used methods previously described [1] to isolate the microcompartments through lysis and ultracentrifugation then confirmed isolation of the micro compartments in two ways: 1. Fluorescent foci (Fusion Red-LVA and Fusion Red Luciferase LVA) to identify the compartments in fluorescent imaging. With Eut-C tags, the fluorescent markers accumulate inside the compartment, allowing them to be clearly visible inside the compartment. The LVA protein degradation tag targets any excess fluorescent proteins for degradation to allow a sufficient background for viewing the compartments in vivo.
Figure 1: EutS compartment containing fusion red tagged with Eut C 2. Electron Microscopy with various radiological stains on a copper grid.
Figure 2: EutS compartment with negative stain
