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).