Developing a new interdisciplinary lab course for undergraduate and graduate students: Plant cells and proteins

Abstract

Studies of protein function increasingly use multifaceted approaches that span disciplines including recombinant DNA technology, cell biology, and analytical biochemistry. These studies rely on sophisticated equipment and methodologies including confocal fluorescence microscopy, mass spectrometry, and X-ray crystallography that are beyond the scope of traditional laboratory courses. To equip the advanced undergraduate and beginning graduate students with an enabling base of knowledge and initial experience with advanced protein research methodologies, a laboratory course entitled Plant Cells and Proteins was developed in a partnership between Washington University and the Donald Danforth Plant Science Center in St. Louis. In this one semester course, 10-12 students obtain hands-on experience with plant tissue culture, gene transformation, subcellular localization of fluorescent recombinant proteins using confocal microscopy, purification of affinity-tagged recombinant proteins, isolation of total protein extracts, enzymatic assays, one-and two-dimensional gel electrophoresis, MALDI-TOF and ESI-Q-TOF mass spectrometry, protein crystallization, and X-ray diffraction. The course is taught as a series of modules, each led by an expert researcher. Students are evaluated based on a series of graded written reports and tests of their mastery of key concepts, interpretations, and the limitations of the experimental methods.Keywords: Undergraduate, graduate, teaching laboratory, interdisciplinary, fundamental skills.The conceptual and technical skills of upper-level undergraduate students and beginning graduate students are partly the product of traditional laboratory courses dedicated to specific topics and methods. Although teaching laboratories provide solid foundations in the principles and techniques of their respective fields, traditional classes are often limited by available facilities or faculty expertise. Classic biochemistry classes often do not reflect contemporary research practices that blend multiple disciplines, modern instrumentation, and a variety of experimental approaches. Likewise, traditional plant biology laboratories tend to focus on whole plant physiology or genetics rather than molecular-based problems. In many curricula, implementation of research-oriented teaching leads to major revisions in existing courses [1][2][3]. With the goal of addressing this issue, a new laboratory class, which integrates current biochemistry, proteomics, microscopy, and plant biology approaches, was developed to provide students with the breadth of understanding and scientific skills to successfully apply such methods to advanced topics in plant biochemistry and molecular biology. Course DevelopmentAs part of a curriculum update, a new combined lecture and laboratory class was designed to effectively introduce a range of methodologies that are defining current experimental practice to research-oriented senior undergraduates and first or second year graduate students. The new course serves students interested in bioc...

Beyond the Teaching Assistantship: CURE Leadership as a Training Platform for Future Faculty

“…Development of course materials is a major component of the trainee's teaching experience in the cotaught CURE model. At our institution, trainees modify an existing protein biochemistry CURE framework 36 by selecting a protein relevant to their research efforts and designing a research question for the CURE (for examples of course materials from our CURE, see the Supporting Information). During the course design process, which begins in the semester before the CURE is offered, the professor guides the trainee on how to adapt the research project into the course flow and how to anticipate logistical issues, such as specialized equipment and lab materials, and the timing of planned experiments.…”

Peptide Mass Fingerprinting of Egg White Proteins

“…Use of advanced mass spectrometry techniques in the undergraduate setting has burgeoned in the past decade, in particular using electrospray ionization time-of-flight mass spectrometry (ESI-TOF-MS) or matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) to determine the accurate mass of a pure protein. − A few experiments in this Journal − and another biochemistry education journal − have used advanced MS techniques to determine the identity of a pure protein using peptide digest data and database searching, but none have started with a mixture of proteins as is done here. Peptide mass fingerprinting is an analytical technique for protein identification.…”

Using Crystallographic Data To Facilitate Students’ Discovery of How Protein Models Are Produced—An Activity Illustrating the Effect of Resolution on Model Quality

“…The illustration of techniques through authentic activities is important in deepening student learning and can give students a stronger sense of fulfillment and satisfaction . Other authentic activities that illustrate protein crystallography include crystallization , and collecting diffraction data . The activity described here complements these by giving students an opportunity to work with real experimental data to explore how molecular models are built.…”

“…4 Other authentic activities that illustrate protein crystallography include crystallization 5,6 and collecting diffraction data. 7 The activity described here complements these by giving students an opportunity to work with real experimental data to explore how molecular models are built. The activity has been completed by three cohorts of 65−80 biochemistry students at a UK university.…”