©CLONTECH	BIOL 340 Biochemical Methods Course Information 1/22/98-2/24/98 Studying Protein-Protein Interactions Using a GST Fusion Protein and Studying mRNA levels Using RT-PCR
Instructor:Marie Pizzorno Department of Biology Bucknell University Office: BB205 Phone: (570) 577-3084 pizzorno@bucknell.edu

Course Description:

In this section of BIOL340 - Biochemical Methods you will investigate two separate experimental procedures. In the first you will study a a specific protein-protein interaction using a bacterially expressed protein. The protein we will examine is Grb2, a mammalian protein involved in several signaling pathways. The open reading frame for Grb2 has been cloned into a plasmid as an in-frame fusion with the enzyme glutathione-S-transferase. We will express the GST-Grb2 protein in bacteria, purify it using affinity chromatography and study its interaction with dynamin obtained from rat brain using a Western blot assay. At the same time you will design your own experiment to test the effect of a particular condition on the ability of Grb2 and dynamin to associate with each other. The second experimental procedure will involve the use of reverst transcriptase - polymerase chain reaction to determine the level of a particular mRNA in various cell types.

Schedule of Activities