Student design and characterization of visible DHFR fusions for biochemistry tools to improve learning during lab exercises.
Autor: | Park J; Department of Basic Sciences, University of Health Sciences and Pharmacy in St. Louis, St. Louis, Missouri, USA., Obeng J; Department of Basic Sciences, University of Health Sciences and Pharmacy in St. Louis, St. Louis, Missouri, USA., Spezia P; Department of Basic Sciences, University of Health Sciences and Pharmacy in St. Louis, St. Louis, Missouri, USA., Huang J; Department of Basic Sciences, University of Health Sciences and Pharmacy in St. Louis, St. Louis, Missouri, USA., Morrone DJ; Department of Basic Sciences, University of Health Sciences and Pharmacy in St. Louis, St. Louis, Missouri, USA. |
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Jazyk: | angličtina |
Zdroj: | Biochemistry and molecular biology education : a bimonthly publication of the International Union of Biochemistry and Molecular Biology [Biochem Mol Biol Educ] 2021 Jul; Vol. 49 (4), pp. 560-569. Date of Electronic Publication: 2021 Apr 08. |
DOI: | 10.1002/bmb.21508 |
Abstrakt: | Student feedback from an undergraduate biochemistry lab course suggested the use of visibly traceable proteins may assist learning. Based on this feedback, we used guided inquiry lab exercises where students developed and characterized a suite of fluorescent protein-dihydrofolate reductase (DHFR) fusions as tools for a biochemistry teaching lab. In contrast to the unfused versions, members of this suite are well-expressed, soluble, visible, highly stable, and easily characterized. The color of mCherry and EGFP fluorescent fusions with microbial DHFR allows students to visibly track their target protein from expression through purification under ambient light, while fusions with BFP are visible under UV-light. Fusions were made to both wild-type and kinetically enhanced DHFR variants. Importantly, we found that fluorescent protein fusions with DHFR did not kinetically interfere as the K (© 2021 International Union of Biochemistry and Molecular Biology.) |
Databáze: | MEDLINE |
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