Instruct-FRISBI course: Preparation & characterization of macromolecular complexes
Monday, November 19th - Friday, November 23rd 2018
- Conference room E1031, CBI
Hosted by Catherine BIRCK & Arnaud POTERSZMAN
Co-organizer: Jean-Paul Concordet, MNHN, Paris FR
No registration fees
Place limited to 16 participants
This Instruct-FRISBI practical course will provide participants with an overview of strategies for preparation and biophysical characterization of macromolecular complexes in view of biochemical and structural studies. The emphasis of the workshop will be placed on purification of macromolecular assemblies from endogenous sources using engineered cell lines and in reconstitution of multiprotein complexes using the baculovirus expression system.
The course is intended primarily for post?doctoral researchers, PhD students and researchers in structural biology. Selected participants will improve their theoretical background and gain experience in designing experimental strategies relevant for their projects.
1/ Engineered cell lines using the CRISPR/Cas9 genome editing technology. Strategies to generate human knock-in stable cell lines for isolation of difficult-to-express proteins and high resolution imaging.
2/ Production of recombinant multiprotein complexes using the baculovirus expression system. Construct and primer design, experimental setup and basic insect cell manipulations.
3/ Biophysical characterization and sample optimization using complementary methods including nanoDSF, AUC-SV, SEC-MALS and EM analysis of negatively stained particles.