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An Investigation of Bacterial Protein Interactions as a Primary Research Project in a Sophomore-Level Molecular Biology Course

    Author: Jean A. Cardinale1
    VIEW AFFILIATIONS HIDE AFFILIATIONS
    Affiliations: 1: Division of Biology, Alfred University, Alfred, NY 14802
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • Published 01 December 2011
    • Supplemental material available at http://jmbe.asm.org
    • Author’s mailing address: Division of Biology, Alfred University, One Saxon Drive, Alfred, NY 14802. Phone: 607-871-2205. Fax: 607-871-2359. E-mail: cardinale@alfred.edu.
    • Copyright © 2011 American Society for Microbiology
    Source: J. Microbiol. Biol. Educ. December 2011 vol. 12 no. 2 194-199. doi:10.1128/jmbe.v12i2.327
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    Abstract:

    Longer term research activities that may be incorporated in undergraduate courses are a powerful tool for promoting student interest and learning, developing cognitive process skills, and allowing undergraduates to experience real research activities in which they may not otherwise have the opportunity to participate. The challenge to doing so in lower-level courses is that students may have not fully grasped the scientific concepts needed to undertake such research endeavors, and that they may be discouraged if activities are perceived to be too challenging. The paper describes how a bacterial protein:protein interaction detection system was adapted and incorporated into the laboratory component of a sophomore-level Molecular Cell Biology course. The project was designed to address multiple learning objectives connecting course content to the laboratory activities, as well as teach basic molecular biology laboratory skills and procedures in the context of a primary research activity. Pre- and posttesting and student surveys both suggest that the laboratory curriculum resulted in significant learning gains, as well as being well received and valued by the students.

Key Concept Ranking

Bacterial Proteins
0.46354717
DNA Restriction Enzymes
0.44843748
0.46354717

References & Citations

1. Cardinale JA2000Structural and functional analysis of AniA, the major anaerobically induced outer membrane protein of Neisseria gonorrhoeae[PhD Thesis]University of RochesterRochester, NY
2. Daines DA, Silver RP2000Evidence for multimerization of neu proteins involved in polysialic acid synthesis in Escherichia coli K1 using improved LexA-based vectorsJ. Bact1825267527010.1128/JB.182.18.5267-5270.200010960115 http://dx.doi.org/10.1128/JB.182.18.5267-5270.2000
3. Daines DA, Granger-Schnarr M, Dimitrova M,, Silver RP2002Use of LexA-based system to identify protein-protein interactions in vivoMethods in Enzymology35815316110.1016/S0076-6879(02)58087-312474385 http://dx.doi.org/10.1016/S0076-6879(02)58087-3
4. Dmitrova M, Younes-Cauet G, Oertel-Buchheit P,, Porte D, Schnarr M,, Granger-Schnarr M1998A new LexA-based genetic system for monitoring and analyzing protein heterodimerization in Escherichia coliMol. Gen. Genet.25720521210.1007/s0043800506409491079 http://dx.doi.org/10.1007/s004380050640
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/content/journal/jmbe/10.1128/jmbe.v12i2.327
2011-12-01
2017-06-28

Abstract:

Longer term research activities that may be incorporated in undergraduate courses are a powerful tool for promoting student interest and learning, developing cognitive process skills, and allowing undergraduates to experience real research activities in which they may not otherwise have the opportunity to participate. The challenge to doing so in lower-level courses is that students may have not fully grasped the scientific concepts needed to undertake such research endeavors, and that they may be discouraged if activities are perceived to be too challenging. The paper describes how a bacterial protein:protein interaction detection system was adapted and incorporated into the laboratory component of a sophomore-level Molecular Cell Biology course. The project was designed to address multiple learning objectives connecting course content to the laboratory activities, as well as teach basic molecular biology laboratory skills and procedures in the context of a primary research activity. Pre- and posttesting and student surveys both suggest that the laboratory curriculum resulted in significant learning gains, as well as being well received and valued by the students.

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Figures

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Fig. 1

The LexA protein interaction detection system.

General flowchart of the LexA protein interaction detection system, which allows students to identify putative proteins which interact with the gonococcal protein, AniA. A: Schematic of constructed plasmids. pANI contains the gene for the target protein in a LexDBD low copy plasmid and is made and provided by the instructor. Students use a shotgun technique to generate a library of plasmids, each containing a section of the gonococcal genome in frame with the wild type LexDBD. B: Expression of chimeric proteins. Chimeric proteins containing LexDBD represent (in theory) all possible gonococcal proteins. C: If there is binding between AniA and a putative protein, the heterodimer binds to a chimeric operator (op/op) and prevents transcription of a reporter gene from the promoter (P). Colonies whose cloned proteins do not interact with AniA will express LacZ and appear purple on a MacConkey plate.

Source: J. Microbiol. Biol. Educ. December 2011 vol. 12 no. 2 194-199. doi:10.1128/jmbe.v12i2.327
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Fig. 2

Normalized learning gains.

Pre- and posttests were graded anonymously. The rubric (provided in Appendix 4 reflected a maximum score of 4 for each question, with a score of 3 representing the expected understanding. Normalized learning gains ((Score - Score)/(4-Score)) were calculated for each question for each individual student, and then averaged for each question (listed in Table 3 ). Class averages are indicated by semester (Fall ‘07, Spring ‘09, Fall ‘09).

Source: J. Microbiol. Biol. Educ. December 2011 vol. 12 no. 2 194-199. doi:10.1128/jmbe.v12i2.327
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Image of Fig. 3

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Fig. 3

Raw pretest vs. raw posttest scores.

Raw scores for pre- and posttests were plotted for individual students. Both tests had a maximum raw score of 40 points; however, a score of 30 represented expected understanding was achieved. The dashed line indicates where equivalent pre-and posttest scores would lie on the graph. The placement of points above this dashed line indicates that all students did improve their understanding of the concepts presented with the activity. Students in the fall ‘09 class tended to score lower on the pretest; however, posttest scores are evenly distributed over the three years.

Source: J. Microbiol. Biol. Educ. December 2011 vol. 12 no. 2 194-199. doi:10.1128/jmbe.v12i2.327
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