Development of a green fluorescent protein‐based laboratory curriculum

Abstract

A laboratory curriculum has been designed for an undergraduate biochemistry course that focuses on the investigation of the green fluorescent protein (GFP). The sequence of procedures extends from analysis of the DNA sequence through PCR amplification, recombinant plasmid DNA synthesis, bacterial transformation, expression, isolation, and characterization of the protein by SDS-PAGE. A survey of participants found that the majority of them were performing most of these procedures for the first time and that participants found the exercises enjoyable and considered them a significant aid to their understanding of biochemistry, cell biology, and molecular genetics.

From gene mutation to protein characterization

“…Because of its natural green color, this member of the green fluorescent protein family is an ideal protein to use in an undergraduate laboratory setting [3][4][5]. The green colored protein allows students (i) to visually monitor the expression of the protein in a recombinant host such as E. coli, (ii) to watch in real time the whereabouts of the protein during the purification process, and (iii) to easily characterize the protein with standard equipment found in a typical undergraduate laboratory [6,7].…”

Competition between student groups in the protein production challenge

“…-In order to motivate all students to engage in a laboratory project, particularly those motivated by achievement or instrumental drivers, an element of competition was integrated into the project. 14,15 Students were provided with a basic protein production protocol for each protein (Appendix A). The students were free to modify the protocol as they saw fit to enhance their performance on the project.…”

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

“…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.…”