The Division of Structural Biology has a world leading reputation for structural biology research within the University of Oxford, UK
PlatformMammalian Expression systemThis facility is dedicated to protein expression in mammalian cells for structural biology. It includes (i) streamlined methodologies for transient expression in 293T cells, first for rapid small scale screening of multiple constructs, and then for scale up of selected constructs (Aricescu et al, 2006 Acta Cryst D), (ii) technologies for controlling glycosylation in secreted proteins (Chang et al, 2007, Structure) and SeMet labelling (e.g. Aricescu et al, 2006, EMBO J) and (iii) co-expression to produce heterodimeric protein complexes for structure determination. Development work includes membrane protein expression in mammalian cells.
Access to Instruct crystallisation provides full use of the Strubi-crystallisation facility for setting up crystallisation experiments and monitoring these experiments via a web based GUI http://www.oppf.ox.ac.uk/xtalpims/. The PX-Scanner is an integral part of the automated crystallisation screening process, allowing fast in-situ analysis of crystals, which can significantly shorten the path to diffraction quality crystals.
The thermal shift assay is designed to monitor the thermal stability of proteins and investigate factors affecting this stability. This is a rapid and simple technique used to screen buffer conditions, ligands, cofactors and drugs that may affect the thermal stability of purified proteins.
Producing a functional mammalian protein or protein complex often necessitates using a mammalian host to ensure appropriate post-translational modifications (e.g.,disulfide bonds, glycosylation, phosphorylation) are added to the protein. Expression in a variety of mammalian cells in Strubi utilizes constitutive and inducible expression. Transient expression is the most usual method, but targeted integration vectors for stable cell generation as well as lentiviral delivery vectors for difficult to transfect cells are also in use and available to the access visitor. Adherent and suspension cell lines are available for large-scale protein production. Expression in the mammalian cell system offered through Instruct also includes protein purification and can also manage post-translational modification (please enquire).
Nanolitre crystallization, using standard plate technology is the norm to produce diffraction quality crystals sufficient for structure determination. Miniaturization and automation has resulted in the growth of a wider variety of proteins, under a broad range of standardised conditions, that can be crystallized. In Strubi, crystallization can be set up using plate technologies or increasingly, microfluidic and capillary technologies, which allow in situ diffraction both in house and at the beamline, thereby accelerating data collection. Strubi has invested in a high degree of automation, for crystallization set-up, crystal storage and imaging. A range of crystallization screens are available for soluble and membrane proteins as well as crystallization at 5oC.
The Division of Structural Biology (STRUBI) at the University of Oxford is located about 1.5miles (2Km) from Oxford city centre, next to the Churchill Hospital site. Housed in the Wellcome Trust Centre for Human Genetics on the Old Road Campus, it has easy connections with bus and train transport from airports, London and Oxford Centre.
The Wellcome Trust Centre for Human Genetics has its own busy cafeteria as well as seminar and meeting rooms. Other academic buildings nearby have other catering facilities that may be available to you. There are shops in Headington (the nearest retail area) within walking distance (10-15 minutes from Strubi), and the centre of Oxford is a short bus or cycle ride away.
Oxford is a busy University town and many of the University colleges, museums and exhibitions are free and open to the public. More information about Oxford can be found here.