Implementing a Hybrid Expression Method That Allows Upper-Division Biochemistry Lab Students To Engage in a Full Protein Production Experience While Allowing Ample Time for Characterization Experiments

Protein structure, function, and signaling are a large portion of biochemistry. Because of this, proteins are often used as model systems in biochemistry laboratory courses, where a course-long project might comprise protein expression, purification, and characterization. Two common protein expressi...

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Veröffentlicht in:Journal of chemical education 2019-11, Vol.96 (11), p.2606-2610
Hauptverfasser: Johnson, Josiah W, Mitchell, Christian D, Deloach, Anna M, Simpson, Hannah E, Dunlap, Tori B
Format: Artikel
Sprache:eng
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Zusammenfassung:Protein structure, function, and signaling are a large portion of biochemistry. Because of this, proteins are often used as model systems in biochemistry laboratory courses, where a course-long project might comprise protein expression, purification, and characterization. Two common protein expression methods are isopropyl β-d-1-thiogalactopyranoside (IPTG) induction, which utilizes easy-to-make media but requires extensive cell-growth monitoring that is time-intensive, and autoinduction, which employs multicomponent media that are time-consuming to make but require no cell-growth monitoring. A protein expression method that is a hybrid of IPTG induction and autoinduction is presented. The hybrid method utilizes the medium of IPTG induction and the no-cell-growth-monitoring induction process of autoinduction, saving hands-on time in the protein expression phase to allow more time for protein characterization while still having students execute each step.
ISSN:0021-9584
1938-1328
DOI:10.1021/acs.jchemed.8b00674