Expression And Purification Of Sperm Whale Myoglobin
We present a multiweek laboratory exercise that exposes students to the fundamental techniques of bacterial expression and protein purification through the preparation of sperm whale myoglobin. Myoglobin, a robust oxygen-binding protein, contains a single heme that gives the protein a reddish color,...
| Published in: | Journal of Chemical Education |
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| Main Authors: | , , |
| Format: | Text |
| Language: | unknown |
| Published: |
Works
2010
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| Subjects: | |
| Online Access: | https://works.swarthmore.edu/fac-chemistry/59 https://doi.org/10.1021/ed8000842 |
| Summary: | We present a multiweek laboratory exercise that exposes students to the fundamental techniques of bacterial expression and protein purification through the preparation of sperm whale myoglobin. Myoglobin, a robust oxygen-binding protein, contains a single heme that gives the protein a reddish color, making it an ideal subject for the teaching laboratory as students can follow the progress of the protein visually in real time. The purification uses both ion-exchange and size-exclusion chromatography, thus, introducing students to two of the most common separation methods. Students evaluate the quality of the purification via sodium dodecyl sulfate−polyacrylamide gel electrophoresis. In most cases, this procedure produces sufficient quantities of pure protein for further laboratory exercises studying the properties of myoglobin. |
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