PEP logoPractical Course on rapid "Cloning, Protein Expression and Purification, CPEP-4" - 3 ECTS
Practical sessions are complemented with lectures, round-table discussions about techniques, problems and their solutions.

Organised and sponsored by the Protein Expertise Platform at KBC.

Date: Tuesday 04 to Thursday 13, November 2014
Time: about 8 hours laboratory work per day

Registration is closed now!


Start: Tuesday, November 04, 2014
Place: Chemistry teaching lab, KB4B111, KBC-building 4th floor (2 stairs up), Linnaeus väg 6
Time: about 8 hours laboratory work per day

Leader: Uwe Sauer, KBC, 786-5930, E-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.
Instructor: Gunter Stier, Heidelberg Univ., Germany, E-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.
Information: http://babel.ucmp.umu.se/cpep/
Participants: max. 12 (PhD students, post-docs or technical staff)
Credits: 3 ECTS and a certificate after successful participation. Full time participation is required!
The course is based on the one week EMBO PEPC5 course held at the EMBL-Hamburg Outstation.

Course description

Aims of the practical course:
• Guidelines in project planning: Work on your own project !!!
• Efficient use of bioinformatics tools for planning and analysis of a protein expression projects
• PCR primer design and gene optimization
• Mutagenesis techniques
• Insights in medium throughput cloning and protein expression in E. coli
• Fast parallel cloning of your gene(s) into vectors containing different tags and fusion proteins
• Test of different strains, growth conditions and fusion partners to optimize protein production
• Use of fusion proteins and associated purification strategies
• Co-expression and purification strategies
• The preparation and use of auto-inducing medium
• Protein expression mini-screens
• Scale-up of positive candidates to a protein midi-preparation using purification machines
• Introduction to cell-free protein expression
• Introduction to baculovirus expression vector system and protein expression in insect cells
• Insights in various trouble shooting problems and strategies to solve them
• Labeling techniques e.g. protein 15N-labeling for NMR analysis & simple NMR measurements
• Introduction to protein crystallization techniques and X-ray crystallography

Additional demonstrations will include the use of:
• The “Äkta” protein purification machines
• The Mosquito crystallization robot
• X-ray diffraction of crystals
• NMR characterization of N15 labeled proteins

Practical sessions are complemented with lectures, round-table discussions about techniques, problems and their solutions. Participants will present selected topics of interest in form of short seminars. We encourage the registered participants to bring their own projects*, in which case they should contact the instructor at least 3-4 weeks before the start of the course (no later than October 08, 2014).

(*) It is also possible to work on a project provided by the course leaders.