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ERIC Number: EJ877819
Record Type: Journal
Publication Date: 2010
Pages: 6
Abstractor: As Provided
Reference Count: 10
ISSN: ISSN-1470-8175
Expression, Purification, and Analysis of Unknown Translation Factors from "Escherichia Coli": A Synthesis Approach
Walter, Justin D.; Littlefield, Peter; Delbecq, Scott; Prody, Gerry; Spiegel, P. Clint
Biochemistry and Molecular Biology Education, v38 n1 p17-22 Jan-Feb 2010
New approaches are currently being developed to expose biochemistry and molecular biology undergraduates to a more interactive learning environment. Here, we propose a unique project-based laboratory module, which incorporates exposure to biophysical chemistry approaches to address problems in protein chemistry. Each of the experiments described herein contributes to the stepwise process of isolating, identifying, and analyzing a protein involved in a central biological process, prokaryotic translation. Students are provided with expression plasmids that harbor an unknown translation factor, and it is their charge to complete a series of experiments that will allow them to develop hypotheses for discovering the identity of their unknown (from a list of potential candidates). Subsequent to the identification of their unknown translation factor, a series of protein unfolding exercises are performed employing circular dichroism and fluorescence spectroscopies, allowing students to directly calculate thermodynamic parameters centered around determining the equilibrium constant for unfolding as a function of denaturant (temperature or chemical). The conclusion of this multi-part laboratory exercise consists of both oral and written presentations, emphasizing synthesis of the roles of each translation factor during the stepwise process of translation. (Contains 1 table and 3 figures.)
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Publication Type: Journal Articles; Reports - Research
Education Level: Higher Education
Audience: N/A
Language: English
Sponsor: N/A
Authoring Institution: N/A