Probing Changes in the Conformation of tRNAPhe: An Integrated Biochemistry Laboratory Course

摘要

We have designed a new guided-inquiry laboratory for an advanced biochemistry course. This integrated laboratory focuses on the biomolecule tRNAPhe and combines elements of bioorganic and bioinorganic chemistry with biochemistry. Throughout the semester students work together to study tRNAPhe structure and ligand binding by (i) assaying metal-catalyzed hydrolysis using gel electrophoresis; (ii) probing thermal denaturation with UV–vis spectrophotometry; and (iii) characterizing ligand associated changes in conformation via fluorescence spectroscopy. Students begin the course with a common laboratory experience and then diverge into parallel projects, ending with a novel self-designed project. In their final report, students interpret experimental results from the entire class and draw conclusions regarding the effects of various ligands on the structure of tRNAPhe.