Practical Course on rapid Cloning, Protein Expression and Purification, CPEP-2020 (3 ECTS), 29 January - 7 February 2020
Graduate course run in collaboration with the Protein Expertise Platform (PEP). Practical sessions are complemented with lectures, round-table discussions about techniques, problems and their solutions.
Registration deadline: Friday January 17, 2020. Admission on a first-come first-served basis.
Registeration form: see below
Participants: max. 16 (PhD students, post-docs or technical staff)
Credits: 3 ECTS on a pass – no pass basis. Full time participation is required to obtain a pass certificate.
After the CPEP course you should know how to:
Plan and successfully execute your project. Bring your own project(s)*!!!
Make efficient use of bioinformatics tools for planning and analysis of protein expression projects
Design PCR primers and codon optimize genes
Apply various fast cloning and mutagenesis techniques
Use medium-throughput approaches for cloning and protein expression in E. coli
Carry out fast parallel cloning of your gene(s) into vectors containing different tags and fusion proteins
Use of fusion proteins and associated purification strategies
Test for optimal protein production: different strains, growth conditions and fusion partners
Co-express components of protein complexes and their purification strategies
Prepare and use of auto-inducing medium
Do protein expression mini-screens
Scale-up positive screening candidates to a protein midi-preparation using purification machines
Trouble shoot problems and apply strategies to solve them
Apply special protein labeling techniques, such as 15N-labeling for quick NMR analysis
Interpret simple NMR measurements
Carry out electron microscopy sample preparation e.g. negative staining techniques
Participants will be guided to present short seminar talks of selected topics.
Special emphasis for the 2020 course
Protein expression in eukaryotic cells
Expression in Human Embryonic Kidney (HEK) cells (Alexej Schmid)
Expression in Baculovirus infected Sf9 cells (Gunter Stier)
Expression in plant cell suspensions (Laszlo Bako)
Additional demonstrations and seminars will include the use of:
Protein purification with “Äkta” and BioRad machines
Protein stability measurement using Thermofluor assay (Melanie Oelker)
The Mosquito crystallization robot, crystal imaging facility (Uwe Sauer)
NMR characterization of 15N labeled proteins (Anders Öhman)
EM facility – first approach to electron microscopy measurements (Linda Sandblad)
(*) If you bring your own project(s), please contact the instructors at least 2 weeks before the start of the course, no later than January 15, 2020). It is also possible to work on a project provided by the course leaders.