Biochemistry Laboratory

Experimental biochemistry emphasizing the isolation, purification and characterization of enzymes and the cloning of genes and expression of recombinant protein. Traditional biochemical techniques such as UV-VIS spectroscopy, gel filtration, ion exchange and affinity chromatography, electrophoresis, and immunoblotting will be used in the investigation of several enzymes. Specific experiments will emphasize enzyme purification, enzyme kinetics, and enzyme characterization by biochemical and immunochemical methods. An experiment utilizing intrinsic radiolabeling and radioimmunoprecipitation is used to ask specific questions about bacterial metabolism and protein synthesis. Major techniques in molecular biology will be introduced through an extended experiment that will include DNA purification, polymerase chain reaction, bacterial transformation, DNA sequencing, and the expression, purification, and characterization of the recombinant protein. Class discussions emphasize the underlying principles of the biochemical and molecular techniques employed in the course, and how these experimental tools are improved for particular applications. Laboratory reports stress experimental design, data presentation, and interpretation of results. (CHEM 0322) 2 hr. lect., 6 hrs. lab.