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Sulzenbacher, G ; Gruez, A ; Roig-Zamboni, V ; Spinelli, S ; Valencia, C ; Pagot, F ; Vincentelli, R ; Bignon, C ; Salomoni, A ; Grisel, S ; Maurin, D ; Huyghe, C ; Johansson, K ; Grassick, A ; Roussel, A ; Bourne, Y ; Perrier, S ; Miallau, L ; Cantau, P ; Blanc, E ; Genevois, M ; Grossi, A ; Zenatti, A ; Campanacci, VR ; Cambillau, C

A medium-throughput crystallization approach

Acta Crystallographica Section D-Biological Crystallography 58 2109-2115 Part 12

par Administrateur - publié le

Abstract :

The first results of a medium-scale structural genomics program clearly demonstrate the value of using a medium-throughput crystallization approach based on a two-step procedure : a large screening step employing robotics, followed by manual or automated optimization of the crystallization conditions. The structural genomics program was based on cloning in the Gateway(TM) vectors pDEST17, introducing a long 21-residue tail at the N-terminus. So far, this tail has not appeared to hamper crystallization. In ten months, 25 proteins were subjected to crystallization ; 13 yielded crystals, of which ten led to usable data sets and five to structures. Furthermore, the results using a robot dispensing 50-200 nl drops indicate that smaller protein samples can be used for crystallization. These still partial results might indicate present and future directions for those who have to make crucial choices concerning their crystallization platform in structural genomics programs.