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Protein factories - week 50

Course code NABI009

Course description
In this course the participants will get hands-on experience of protein production and purification using bacterial and insect cell-based expression systems (baculovirus expression vector system (BEVS)). The course presents methods to express genes, which have products that are known, unknown or not well characterized, and to analyze the gene products. Theories behind the methods are discussed and experiments are performed. The methods to be presented include: optimization of recombinant gene expression in Escherichia coli and in the BEVS; affinity purification of tagged proteins; using proteases to for the removal of protein fusion tags; generation and analysis of gene fusions; molecular cloning; metabolic engineering; seleno-methionine incorporation for MAD phasing; 2H-, 13C-, 15N-metabolic labeling for NMR studies; production of perdueterated proteins for neutron-scattering experiments. Strategies for improving production of "difficult" proteins will be presented. The lectures will cover the current status of cell-based protein production systems, and theoretical aspects of the methodology. The course will be held in collaboration with the Lund Protein Production Platform, LP3 ( Study format: lectures (ca 20%), student presentations (ca 5%), and laboratory exercises (ca 75%).

Course organizer    
Claes von Wachenfeldt
Department of Biology
Faculty of Science (N)
Lund University

More information
Claes von Wachenfeldt
Tel: +46 (0)46 222 34 56