Expression and Purification of Sperm Whale Myoglobin
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[1] Swarthmore College
Swarthmore College
Borough of Swarthmore, Estados Unidos
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- Localización: Journal of chemical education, ISSN 0021-9584, Vol. 87, Nº 3 (March), 2010, págs. 303-305
- Idioma: inglés
- Texto completo no disponible (Saber más ...)
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Resumen
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.
