CNRS - AIX MARSEILLE UNIV: UMR7257

Home page > en > Facilities > Structural Biology > High-throughput cloning, expression in bacteria, purification of (...)

Structural Biology

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

Head Renaud VINCENTELLI

The cloning, expression and protein purification 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 developed and validated in the lab using state-of-the-art equipment in partnerships with several structural genomics projects (ASG, X-TB, SPINE, SPINE II, EMEP, VIZIER, VENOMICS). Since its launch, several thousands of protein samples have been produced by this facility for customers or collaborators. A large portfolio of automated protocols is available, e.g. 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 liquid handling Tecan Genesis 200 equipped with an 8-pipette arm, a plate gripper and a 96/384 UV-fluorescence reader
  • 1 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
  • 1 AKTA Xpress chromatography system at 4°C equipped with 4 autonomous modules

Cost

To be defined from the service requested

To go further

Publications

  • 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
  • Structural Genomics Consortium; China Structural Genomics Consortium; Northeast Structural Genomics Consortium, Gräslund S, Nordlund P, Weigelt J, Hallberg BM, Bray J, Gileadi O, Knapp S, Oppermann U, Arrowsmith C, Hui R, Ming J, dhe-Paganon S, Park HW, Savchenko A, Yee A, Edwards A, Vincentelli R, Cambillau C, Kim R, Kim SH, Rao Z, Shi Y, Terwilliger TC, Kim CY, Hung LW, Waldo GS, Peleg Y, Albeck S, Unger T, Dym O, Prilusky J, Sussman JL, Stevens RC, Lesley SA, Wilson IA, Joachimiak A, Collart F, Dementieva I, Donnelly MI, Eschenfeldt WH, Kim Y, Stols L, Wu R, Zhou M, Burley SK, Emtage JS, Sauder JM, Thompson D, Bain K, Luz J, Gheyi T, Zhang F, Atwell S, Almo SC, Bonanno JB, Fiser A, Swaminathan S, Studier FW, Chance MR, Sali A, Acton TB, Xiao R, Zhao L, Ma LC, Hunt JF, Tong L, Cunningham K, Inouye M, Anderson S, Janjua H, Shastry R, Ho CK, Wang D, Wang H, Jiang M, Montelione GT, Stuart DI, Owens RJ, Daenke S, Schütz A, Heinemann U, Yokoyama S, Büssow K, Gunsalus KC (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
© AFMB UMR7257  W3C validation