The training will be primarily based on practical sessions in which challenging biological systems are produced and labelled. Systems such as large multi-domain proteins, large monomeric proteins, oligomeric protein assemblies, and large RNAs will be used as well as RNA and proteins of interest brought by the participants. Practical sessions will be linked to theoretical classes, and discussion sessions will be organized to promote exchange between participants and instructors. All labelled samples produced by the trainees will be characterized on the IBS high-field NMR platform.
The sessions will cover the following isotopic labeling methods:
• Adaptation, optimization and large scale production of protein in E. coli grown in perdeuterated media
(Theoretical)
• Cell-free expression, E. coli based extract, specific labelling and perdeuteration of proteins (Theoretical and Practical)
• Cell-free expression, Wheat Germ based extract, specific labelling of soluble and membrane proteins (Theoretical)
• Isotopic labeling of protein produced in eukaryotic cells (Theoretical)
• In-cell NMR (Theoretical and Practical)
• In vitro production of specifically labelled RNA (Theoretical and Practical)
• In vivo production of uniformally labelled RNA (Theoretical and Practical)
• Methyl specific labelling of large proteins (Theoretical)
• Segmental labelling of proteins & RNAs (Theoretical and Practical)