The goal of this project is to incorporate the critical aspects of basic scientific research while training the students in the fundamentals of molecular biology. Although students must learn the required techniques of molecular biology in order to be successful in professional school or in the work force, too often these techniques are learned in isolated experiments where the connections between the experiments are unclear. Such experiments also tend to focus on a particular technique without considering the experiment in the larger context of scientific inquiry. Integrating the learning of laboratory techniques, experimental design and experimental analysis is critical for the training of future researchers that can adapt to the necessary changes that occur in molecular biology. This project will teach students molecular biology techniques in the framework of a research project. The students will be presented with a scientific problem and they will attempt to solve the problem. Specifically, the students will be introduced to the RET receptor, a newly characterized signaling molecule and a protein involved in the early stages of kidney development. They will be told about the ligand for this receptor and the three co-receptors that may be involved in the receptors activation. Finally, the students will learn that the three co-receptors have similar amino acid sequences but vary in their ability to activate the receptor in the presence of the ligand. The students will then proceed to spend the rest of the semester devising experiments to determine the important regions of the co-receptor for RET activation. Although a seemingly daunting task for untrained molecular biologists, the students will learn how to approach this question in a step-by-step fashion. The students will first learn how to align protein sequences and compare similarities between the co-receptors. The aligned co-receptors will then be analyzed to allow construction of receptor hybrids using standard cloning and amplification techniques. Finally, with the help of a technician, the students will study the expression of their designed protein looking at both the levels of expression and the ability of their co-receptor to activate the RET receptor in the presence of the appropriate ligand. In the process of performing these experiments, the students will learn how to analyze data in preparation for later experiments, understand the connections between experiments and gain insight into the methods of experimental design by studying a timely research problem. Finally, the data obtained in these experiments will help answer the question of co-receptor function as well as provide reagents and hypotheses to be studied by future student laboratories.
