Structural Genomics of Eukaryotic Targets at a Laboratory Scale |
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Authors: | Didier Busso Pierre Poussin-Courmontagne David Rosé Raymond Ripp Alain Litt Jean-Claude Thierry Dino Moras |
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Institution: | (1) Département de Biologie et de Génomique Structurales, IGBMC, CNRS/INSERM/Université Louis Pasteur, Parc d’Innovation, 1 rue Laurent Fries, BP10142, 67404, Illkirch, cedex, France |
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Abstract: | Structural genomics programs are distributed worldwide and funded by large institutions such as the NIH in United-States,
the RIKEN in Japan or the European Commission through the SPINE network in Europe. Such initiatives, essentially managed by
large consortia, led to technology and method developments at the different steps required to produce biological samples compatible
with structural studies. Besides specific applications, method developments resulted mainly upon miniaturization and parallelization.
The challenge that academic laboratories faces to pursue structural genomics programs is to produce, at a higher rate, protein
samples. The Structural Biology and Genomics Department (IGBMC – Illkirch – France) is implicated in a structural genomics
program of high eukaryotes whose goal is solving crystal structures of proteins and their complexes (including large complexes)
related to human health and biotechnology. To achieve such a challenging goal, the Department has established a medium-throughput
pipeline for producing protein samples suitable for structural biology studies. Here, we describe the setting up of our initiative
from cloning to crystallization and we demonstrate that structural genomics may be manageable by academic laboratories by
strategic investments in robotic and by adapting classical bench protocols and new developments, in particular in the field
of protein expression, to parallelization. |
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Keywords: | cloning crystallization expression TM" target="_blank">Gateway TM high throughput screening recombinant protein structural genomics |
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