CNRS - AIX MARSEILLE UNIV: UMR7257

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Structural Biology

High-throughput cloning, expression in bacteria, purification and interaction of recombinant proteins

Head Renaud VINCENTELLI

The IBiSA cloning, expression and protein purification-interaction high-throughput facility can perform all the steps from gene or multi-genes cloning to purified protein samples. This service uses innovative high-throughput strategies and methodologies that have been mostly developed in-house and validated in the lab using state-of-the-art equipment in partnerships with several structural genomics and big scale projects (More than ten ANR and ten EU grants including ; X-TB, MEPNET, SPINE, SPINE II, EMEP, P4EU, VENOMICS, PDZnet, ADDovenom and more). Since its launch, more than 10.000 proteins have been produced by this facility for customers or collaborators. A large portfolio of automated protocols is available, e.g. a HTP expression E. coli screen at a pace of up to 1152 purifications/analysis per week, a protein-DNA interaction assay (HTP SELEX 96) or a protein-protein interaction assay (HTP Pull-Down or HTP Hold-up in 96/384). New methods can be custom-developed for specific projects. This portfolio of new protocols is accessible once these methods are published. This service is closely linked to the biophysical and crystallography facilities of the lab.

Accessibility

Academics and industrials

How to access ?

Specific equipment

  • 1 liquid handling Tecan Freedon EVO 200 equipped with a 96 tips head, an 8-pipette arm, a plate gripper
  • 1 Pherastar 96/384/1536 UV-fluorescence reader
  • 2 Perkin Elmer GX II capillary electrophoresis system for 96/384 sample analysis
  • 1 dedicated minifors culture incubator for bacterial cultures with a maximum capacity of 48 DW24 or 48 DW96
  • 2 AKTA Xpress chromatography system at 4°C equipped with 8 autonomous modules

Cost

To be defined from the service requested

To go further

Publications

  • R. Vincentelli is the editor of the first "Methods in Molecular Biology" book on "High-Throughput Protein Production and Purification" (2019). https://urlz.fr/a6ZP
  • R. Vincentelli*, K. Luck*, J. Poirson, J. Polanowska, J. Abdat, M. Blémont, J. Turchetto, F. Iv, K. Ricquier, ML Straub, A. Forster, P. Cassonnet, JP. Borg, Y. Jacob, M. Masson, Y. Nominé, J. Reboul, N. Wolff, S. Charbonnier, G. Travé. Nature Methods Aug;12(8):787-93, * equal contribution
  • Saez NJ, Nozach H, Blemont M, Vincentelli R (2014) J Vis Exp. 89:e51464
  • Saez NJ, Vincentelli R (2014) Methods Mol Biol. 1091:33-53.
  • Jolma A, Yan J, Whitington T, Toivonen J, Nitta KR, Rastas P, Morgunova E, Enge M, Taipale M, Wei G, Palin K, Vaquerizas JM, Vincentelli R, Luscombe NM, Hughes TR, Lemaire P, Ukkonen E, Kivioja T, Taipale J (2013) Cell, 152:327-39
  • Gräslund S et al, (2008) Nat Methods 5:135-46
  • Vincentelli R, Bignon C, Gruez A, Canaan S, Sulzenbacher G, Tegoni M, Campanacci V, Cambillau C (2003) Acc Chem Res 36:165-17
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