TRAC 16: Expression of Recombinant Proteins: Overcoming Challenges & Optimization for Downstream Applications
This 3 day lecture laboratory course will give the student a good theoretical background and practical experience in expression and purification of recombinant proteins from a variety of expression systems. Furthermore, the course will address new methods to overcome traditional challenges in recombinant protein expression and purification. Finally, students will be introduced to methods to prepare their samples for downstream applications such as immunofluorescence studies.
Topics: Introduction to Protein Purification Techniques; Expression of Proteins in Prokaryotic and Eukaryotic Cells: Requirements, Advantages and Disadvantages; Prokaryotic Expression Systems: Strains, Tags and Purification Systems; Expression in Insect Cells: Baculovirus and Drosophila Systems; Expression in Mammalian Cells: Detection and Purification (Immunohistochemistry, Reporter Gene Assays, GFP, Tags for Expression/Purification Systems); Generation of Antibodies to Recombinant Proteins: System considerations and Antibody Purification; Expression in Yeast Cells (Pichia).