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Using yeast to determine the functional consequences of mutations in the human p53 tumor suppressor gene: An introductory course‐based undergraduate research experience in molecular and cell biology

The opportunity to engage in scientific research is an important, but often neglected, component of undergraduate training in biology. We describe the curriculum for an innovative, course‐based undergraduate research experience (CURE) appropriate for a large, introductory cell and molecular biology...

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Autores principales: Hekmat‐Scafe, Daria S., Brownell, Sara E., Seawell, Patricia Chandler, Malladi, Shyamala, Imam, Jamie F. Conklin, Singla, Veena, Bradon, Nicole, Cyert, Martha S., Stearns, Tim
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5516179/
https://www.ncbi.nlm.nih.gov/pubmed/27873457
http://dx.doi.org/10.1002/bmb.21024
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author Hekmat‐Scafe, Daria S.
Brownell, Sara E.
Seawell, Patricia Chandler
Malladi, Shyamala
Imam, Jamie F. Conklin
Singla, Veena
Bradon, Nicole
Cyert, Martha S.
Stearns, Tim
author_facet Hekmat‐Scafe, Daria S.
Brownell, Sara E.
Seawell, Patricia Chandler
Malladi, Shyamala
Imam, Jamie F. Conklin
Singla, Veena
Bradon, Nicole
Cyert, Martha S.
Stearns, Tim
author_sort Hekmat‐Scafe, Daria S.
collection PubMed
description The opportunity to engage in scientific research is an important, but often neglected, component of undergraduate training in biology. We describe the curriculum for an innovative, course‐based undergraduate research experience (CURE) appropriate for a large, introductory cell and molecular biology laboratory class that leverages students′ high level of interest in cancer. The course is highly collaborative and emphasizes the analysis and interpretation of original scientific data. During the course, students work in teams to characterize a collection of mutations in the human p53 tumor suppressor gene via expression and analysis in yeast. Initially, student pairs use both qualitative and quantitative assays to assess the ability of their p53 mutant to activate expression of reporter genes, and they localize their mutation within the p53 structure. Through facilitated discussion, students suggest possible molecular explanations for the transactivation defects displayed by their p53 mutants and propose experiments to test these hypotheses that they execute during the second part of the course. They use a western blot to determine whether mutant p53 levels are reduced, a DNA‐binding assay to test whether recognition of any of three p53 target sequences is compromised, and fluorescence microscopy to assay nuclear localization. Students studying the same p53 mutant periodically convene to discuss and interpret their combined data. The course culminates in a poster session during which students present their findings to peers, instructors, and the greater biosciences community. Based on our experience, we provide recommendations for the development of similar large introductory lab courses. © 2016 by The International Union of Biochemistry and Molecular Biology, 45(2):161–178, 2017.
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spelling pubmed-55161792017-08-02 Using yeast to determine the functional consequences of mutations in the human p53 tumor suppressor gene: An introductory course‐based undergraduate research experience in molecular and cell biology Hekmat‐Scafe, Daria S. Brownell, Sara E. Seawell, Patricia Chandler Malladi, Shyamala Imam, Jamie F. Conklin Singla, Veena Bradon, Nicole Cyert, Martha S. Stearns, Tim Biochem Mol Biol Educ Laboratory Exercise The opportunity to engage in scientific research is an important, but often neglected, component of undergraduate training in biology. We describe the curriculum for an innovative, course‐based undergraduate research experience (CURE) appropriate for a large, introductory cell and molecular biology laboratory class that leverages students′ high level of interest in cancer. The course is highly collaborative and emphasizes the analysis and interpretation of original scientific data. During the course, students work in teams to characterize a collection of mutations in the human p53 tumor suppressor gene via expression and analysis in yeast. Initially, student pairs use both qualitative and quantitative assays to assess the ability of their p53 mutant to activate expression of reporter genes, and they localize their mutation within the p53 structure. Through facilitated discussion, students suggest possible molecular explanations for the transactivation defects displayed by their p53 mutants and propose experiments to test these hypotheses that they execute during the second part of the course. They use a western blot to determine whether mutant p53 levels are reduced, a DNA‐binding assay to test whether recognition of any of three p53 target sequences is compromised, and fluorescence microscopy to assay nuclear localization. Students studying the same p53 mutant periodically convene to discuss and interpret their combined data. The course culminates in a poster session during which students present their findings to peers, instructors, and the greater biosciences community. Based on our experience, we provide recommendations for the development of similar large introductory lab courses. © 2016 by The International Union of Biochemistry and Molecular Biology, 45(2):161–178, 2017. John Wiley and Sons Inc. 2016-11-22 2017 /pmc/articles/PMC5516179/ /pubmed/27873457 http://dx.doi.org/10.1002/bmb.21024 Text en © 2016 The International Union of Biochemistry and Molecular Biology This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial (http://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Laboratory Exercise
Hekmat‐Scafe, Daria S.
Brownell, Sara E.
Seawell, Patricia Chandler
Malladi, Shyamala
Imam, Jamie F. Conklin
Singla, Veena
Bradon, Nicole
Cyert, Martha S.
Stearns, Tim
Using yeast to determine the functional consequences of mutations in the human p53 tumor suppressor gene: An introductory course‐based undergraduate research experience in molecular and cell biology
title Using yeast to determine the functional consequences of mutations in the human p53 tumor suppressor gene: An introductory course‐based undergraduate research experience in molecular and cell biology
title_full Using yeast to determine the functional consequences of mutations in the human p53 tumor suppressor gene: An introductory course‐based undergraduate research experience in molecular and cell biology
title_fullStr Using yeast to determine the functional consequences of mutations in the human p53 tumor suppressor gene: An introductory course‐based undergraduate research experience in molecular and cell biology
title_full_unstemmed Using yeast to determine the functional consequences of mutations in the human p53 tumor suppressor gene: An introductory course‐based undergraduate research experience in molecular and cell biology
title_short Using yeast to determine the functional consequences of mutations in the human p53 tumor suppressor gene: An introductory course‐based undergraduate research experience in molecular and cell biology
title_sort using yeast to determine the functional consequences of mutations in the human p53 tumor suppressor gene: an introductory course‐based undergraduate research experience in molecular and cell biology
topic Laboratory Exercise
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5516179/
https://www.ncbi.nlm.nih.gov/pubmed/27873457
http://dx.doi.org/10.1002/bmb.21024
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