Teaching the Principles of DNA cloning, Recombinant Protein Expression, and Purification using a Purple Chromoprotein

Abstract

The Freshman Research Initiative (FRI) program at the University of Texas at Austin offers first-year students access to cutting edge research in science. The Biobricks for Molecular Machines FRI stream focuses on training first-year undergraduates (with little to no lab experience) in basic molecular and biochemical skills in their first 9-weeks in the stream. We use the purple chromogenic protein (Meffcp_GBR22), derived from Montipora efflorescens, to allow visual confirmation of various molecular cloning and recombinant protein expression and purification steps, which highly enhances the learning experience of undergraduates. By conducting this lab curriculum in a research-based setting, students learn to troubleshoot and examine sources of error by experiencing the repetitive nature of research where results are not always as expected and each lab module is linked the next. In addition to hands-on experiences, students also learn the important skills of analyzing and presenting their experimental results in bi-weekly laboratory reports. The robust laboratory modules described here have been tested and tried for the past decade and can be easily modified to incorporate any gene of interest. Further students apply the lab techniques learned on research projects that are in collaboration with other faculty labs at the university. With the use of common molecular and biochemical equipment found in basic teaching wet labs, this curriculum can be easily adapted to any university/college or advanced high school biotechnology courses.