Laboratory experiment on gene subcloning for chemical engineering students

Claire F. Komives1, Sabine Rech2, and Melanie McNeil1. (1) Dept. of Chemical and Materials Engineering, San Jose State University, One Washington Square, San Jose, CA 95192-0082, (2) Department of Biological Sciences, San Jose State University, One Washington Square, San Jose, CA 95192

ABSTRACT: Biocatalysis is the use of enzymes or whole-cell organisms for enabling industrial chemical processes. More and more chemical engineers are being employed by chemical companies that use biocatalysis as a route to green processes. In our elective course, Biochemical Engineering Laboratory, we teach a five-week experiment to subclone green fluorescent protein from a cloning plasmid to a high-expression vector system. The expression levels are then compared based on SDS-PAGE and fluorescence. The high-expression system is used in the course for the fermentation experiments and the expressed protein is purified by chromatography. We have seen that giving students hands-on experience with molecular biology enables their understanding of PCR, electrophoresis and the mechanisms of gene subcloning. The protocols of the experiment will be discussed briefly along with class results and classroom logistics.