"Understanding Protein Expression PECF" course
This short course will describe many aspects of heterologous expression in E.coli, from choice of construct design through to methods to improve levels of soluble expression and co-expression. In addition we will also describe expression in Eukaryotic hosts, including secreted proteins and ECDs, and options for co-expression. The course will finish with a discussion and problem solving session using examples brought along by the attendees.
Date: Tuesday, 20th June 2017
Time: 09:30 – 13:00
Place: Fèlix Serratosa, Parc Cientific de Barcelona (PCB)
The workshop is offered exclusively to IRB members and places reserved for IRB Barcelona alumni. To register, please send an email to firstname.lastname@example.org
If anyone has an expression problem that they would like to discuss in the final session please submit a one-slide powerpoint summary to Nick before the course.
09.30 – 09.45 – Introduction to the PECF
09.45 - 11.00 – Construct design and optimization, E.coli the expression "wor-horse"
Topics will include: A look at cloning methodologies, the effect of sequence on expression levels, controlling expression, choosing a fusion protein for stability and solubility, strain and media selection, basic co-expression.
11.00 – 11.20 – COFFEE BREAK
11.20 – 12.00 –Protein expression in Eukaryotic hosts and expression of complexes.
Topics will include: Expression/co-expression in Mammalian cells, Extra-Cellular Domains (ECDs), membrane and secreted proteins, ‘HSP-free’ expression of cytoplasmic proteins, introduction to the baculoviral (BEVs) expression system, co-expression of multi-subunit complexes in insect cells.
12.00 – 12.40– Discussion and problem solving. Please bring along a single slide summary of your protein expression or purification problems for discussion.
12.00 – 12.40– Lab visit for interested attendees.