The objective of this course is to provide undergraduate students the opportunity to conduct research in the areas of Cell Biology, Biochemistry and Molecular Biology. The research course in this semester will be focused on producing recombinant enzymes in E.coli, some of which may hydrolyze N-acylethanolmines (NAEs) as substrates. NAEs are a class of fatty acid derivatives that play a role in plant growth and development. Related lipids in humans function as part of the endocannabinoid signaling pathway that regulates many physiological and behavioral processes. Here we will be examining members of the fatty acid amide hydrolase (FAAH) in various plant species which hydrolyze and inactivate NAEs and other acylamide signaling molecules. In the class, we will clone the coding sequences for novel FAAH proteins (with unknown substrate profiles and physiological functions) into expression vectors designed to produce recombinant proteins in E.coli under inductive conditions. Proteins will be extracted from the host, purified and tested for enzyme activity toward potential acylamide lipid substrates in an effort to understand more about the nature of these enzymes. We will compare these biochemical properties to the recombinant human FAAH protein produced and purified in a similar manner. During the course, students will read the scientific literature, design experiments and evaluate/interpret their own results. Emphasis will be placed on critical thinking, data collection and analysis, and presentation of their scientific findings.
This course is based on real-world experimentation and will provide first-hand knowledge of the process of scientific discovery with its triumphs and frustrations. Students will be part of research teams and responsible for their own experimental results. It is expected that findings from student research will be of the highest quality and suitable for research publication.