Presentation
This short workshop will describe many aspects of heterologous expression in E.coli, from choice of construct design through to methods to improve the levels of soluble expression and options for co-expression. In addition, it will also address expression in eukaryotic hosts, including secreted proteins and ECDs, and options for co-expression.
The workshop will finish with a session on protein purification and protein quality and a discussion and problem-solving session using examples brought along by the attendees*.
Speaker:
Nick Berrow - IRB Barcelona Facility Manager
Date: Monday, 17 May 2021
Time: 09:30 – 13:30
Place: Online (Zoom)
Audience: All IRB Barcelona members are welcome (previous registration requested).
Registration: Fill in the registration form here before 4 May. A confirmation email will be sent to all participants. Participants will receive access details prior to the start of the workshop.
If you are registered to participate and finally cannot attend, please send an e-mail to phd@irbbarcelona.org in order to give others the opportunity.
Programme
9.30-10.00 Introduction to PECF
Genes to proteins: Low or High Through-Put approaches, overview of construct design, fusion proteins and purification tags, different expression hosts and strains.
10.00-11.00 E.coli-the expression ‘work-horse’
Including basics and background theory, protein engineering, cloning and expression methods, purification approaches and co-expression methods for complexes.
11.00-11.30 Break
11.30-12.15 Eukaryotic hosts
Including overviews of current Mammalian and Baculoviral systems, secreted proteins, membrane proteins, co-expression methods for complexes.
12.15-13.30 Purifying proteins and Protein Quality issues including Discussion and problem solving session*
*If you have a protein-related problem that you would like to troubleshoot within the group please send a one-slide explanation of the problem two days in advance of the course.