Practical Course on rapid Cloning, Protein Expression and Purification, CPEP-2024 (3 ECTS), 18-27 March 2024

Course information

Date:  18 - 27 March 2024 (8 working days)

Start: 09:00, Monday, 18 March.

Place: Biochemistry teaching lab, KB.F4.04, Linnaeus väg 6, KBC-F building, 4th floor (2 stairs up).

Registration deadline: 8 March 2024.

Cost: 5.000 SEK per participant.

Registration form: see below.

Participants: max. 12 (PhD students, post-docs or technical staff)

Credits: 3 ECTS on a pass – no pass basis. Successful full-time participation is required to receive a pass certificate.

Leader: Uwe Sauer, E-mail: uwe.sauer@umu.se    

Instructor: Gunter Stier, Heidelberg Univ., Germany, E-mail: gunter.stier@bzh.uni-heidelberg.de  

The CPEP-24 course is organized in collaboration with the Protein Expertise Platform (PEP) at KBC and the Protein Production Sweden (PPS) national infrastructure.

This practical course is based on the bi-annual, one-week EMBO PEPC courses held at the EMBL-Hamburg outstation.

Aims

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
• Troubleshoot problems and apply strategies to solve them
• Apply special protein labelling techniques, such as ¹⁵N-labeling for quick NMR analysis
• Interpret simple NMR measurements
• Crystallize proteins
• Carry out electron microscopy sample preparation, e.g. negative staining techniques
• Participants will be guided to present short seminar talks on selected topics.

Special emphasis for the 2024 course

• Protein expression in eukaryotic cells
  • Expression in Human Embryonic Kidney (HEK) cells
    o Expression in Baculovirus infected Sf9 cells
    o Expression in plant cell suspensions

Additional demonstrations and seminars will include the use of:

• Protein purification with “Äkta” and BioRad machines
• Protein analysis using Mass-Photometry
• The Mosquito crystallization robot, crystal imaging facility
• NMR characterization of 15N labeled proteins
• EM facility – first approach to electron microscopy measurements

(*) If you bring your own project(s), please contact the instructors no later than 28 February 2024. You can also work on a project provided by the course leaders.