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FRENCH INFRASTRUCTURE FOR INTEGRATED STRUCTURAL BIOLOGY

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INSECT CELLS EXPRESSION

This facility is dedicated to protein and multi-protein complex production in insect cells using the baculovirus expression system for structural biology as well as for functional studies. It includes streamlined procedures for (i) the generation of recombinant baculoviruses (transfections, co-transfections) and amplification of a high titer viral stock, (ii) evaluation of recombinant baculoviruses and expression optimization using adherent cells or suspensions and (iii) protein expression in various scales and formats including production of multi-protein complexes.

IGBMC baculovirus expression platform offers two modes of access with typical duration of stays ranging from a single to several weeks, depending on the project:

1/ The platform provides the transfer vectors to users who generate constructs in their home institutions and come to IGBMC to produce viruses and perform the expression assays.

2/ The external users perform cloning and expression assays at the IGBMC platform.

Support includes training and supervision in all aspects of the production run, including construct design and choice of purification strategies for reconstitution and purification of multi-protein complexes.

https://instruct-eric.eu/platform/protein-production-strasbourg-france/

BIBLIOGRAPHY

In TFIIH the Arch Domainof XPD is mechanistically essential for transcription and DNA repair. Peissert S., Sauer F., Grabarczyk D.B , Braun C., Sander G., Poterszman A., Egly J-M.,  Kuper J., Kisker C. (2020) Nat Commun 11: 1667

Identification and characterization of the Onchocerca volvulus Excretory Secretory Product Ov28CRP, a putative GM2 activator protein.Njume FN, Ghogomu SM, Shey RA, Gainkam LOT, Poelvoorde P, Humblet P, Kamgno J, Robert A, Mutesa L, Lelubre , Edelweiss E, Poterszman A, Anheuser S, Vanhamme L, Souopgui J. (2019) PloS Negl Trop Dis 13:e0007591 <o:p></o:p>

The production of multiprotein complexes in insect cells using the baculovirus expression system. Abdulrahman, W., Radu, L., Garzoni, F., Kolesnikova, O., Gupta, K., Osz-Papai, J., Berger, I. & Poterszman, A. (2015). Methods in molecular biology1261, 91-114.

Insect cells-baculovirus system for the production of difficult to express proteins. Osz-Papai J, Radu L, Abdulrahman W, Kolb-Cheynel I, Troffer-Charlier N, Birck C, Poterszman A. Methods Mol Biol. 2015;1258:181-205. doi: 10.1007/978-1-4939-2205-5_10.

In TFIIH, XPD Helicase Is Exclusively Devoted to DNA Repair. Kuper J, Braun C, Elias A, Michels G, Sauer F, Schmitt DR, Poterszman A, Egly JM, Kisker C. PLoS Biol. 2014 Sep; 12(9): e1001954.