DUE TO EXTENSIVE INTEREST IN OUR MULTIBAC-EMBL PLATFORM, P-CUBE TEMPORARILY DOES NOT ACCEPT APPLICATIONS!!!
The facility in EMBL Grenoble (GRB) supports design and assembly of multigene expression vectors and production of multiprotein complexes for structural and functional studies. EMBL GRB has genetically engineered a novel baculovirus-based system, MultiBac-EMBL, for optimal production of eukaryotic multiprotein complexes in insect cells and have exploited advanced recombination methods to provide a flexible, automated platform for generation of multiprotein expression vectors. These advanced baculovirus technologies have yielded already many multiprotein assemblies suitable for structural studies, (e.g. Berger et al, 2004 Nature Biotech.; Fitzgerald et al, 2006 Nature Methods, Fitzgerald et al., 2007 Structure), and have been also used by many other research groups successfully in the meantime. EMBL GRB have streamlined baculovirus-based expression in insect cells so that it can be readily applied by a non-specialist user.
For further automation of multigene assembly, EMBL GRB is installing, independent from this project, a TECAN robot in addition to pre-existing equipment (second TECAN), and supporting instruments, computers and software. Once firmly established (projected for 2010), automated multigene assembly using this setup will also be offered to users (see Bieniossek et al., 2009 Nature Methods). The MultiBac-EMBL platform is located at the Partnership for Structural Biology (PSB) close to the ESRF site where there are also further technical platforms for protein quality control and crystallisation.
Modality of access and work
The user will perform molecular biology steps in the home laboratory then travel to EMBL GRB for a 7-10 working day stay. During this time, the user will use the MultiBac-EMBL platform for virus amplification and protein production screening as outlined above. The expected output is verified virus stocks including production virus that expresses the protein complex(es) of choice (typically production of 3-4 complexes is performed in parallel by users). Reagents, including the viruses, and results are returned to the home laboratory for scale-up and complex production.
Support offered
Advice for multigene vector assembly. Quality controlled starting reagents plus straightforward protocols for home laboratory preparatory work.
The PSB has significant experience in hosting scientific visitors (ESRF guesthouse or budget hotel accommodation could be used). |
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Users:
