DMMC Course: TECHNIQUES & STRATEGIES IN MOLECULAR MEDICINE

0930-1015 Wednesday 12 December 2007, Panoz institute, LTEE2, TCD

Protein expression and purification

Dr Henry Windle (Institute of Molecular Medicine, TCD)

This lecture will cover the basics of protein expression and purification. Emphasis will be placed on alternative strategies and issues that should be considered prior to selection of specific expression systems and purification strategies. As protein purification methodologies are generally well described and accessible, only a brief overview of these will be given but with emphasis on common problems that can arise, particularly for those about to attempt purification for the first time. The following books from The Practical Approach series by IRL Press are an invaluable aid with detailed and reliable protocols: Protein Purification Applications; Protein Purification Methods (2001, Editor Simon Roe).
Gallus (chicken) and Mus Musculus (mouse) as model organisms will be described. The life-cycle, generation time, embryo development and amenability of these organisms to genetic manipulation will be discussed. An emphasis will be placed on describing mutagenesis screens. This technique, in which the genes in the genome are randomly inactivated, has been extensively applied to the fly/fish models and has accelerated our understanding of gene function (functional genomics).