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Structural Genomics of Eukaryotic Targets at a Laboratory Scale

  • Published:
Journal of Structural and Functional Genomics

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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Abbreviations

CCD:

charge-coupled device

DLS:

dynamic light scattering

FPLC:

fast performance liquid chromatography

GST:

Glutathione S-transferase

MBP:

Maltose binding protein

NusA:

N-utilizing substance A

PCR:

polymerase chain reaction

SDS-PAGE:

sodium dodecyl sulfate – polyacrylamide gel electrophoresis

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Authors and Affiliations

  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

    Didier Busso, Pierre Poussin-Courmontagne, David Rosé, Raymond Ripp, Alain Litt, Jean-Claude Thierry & Dino Moras

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  1. Didier Busso
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  2. Pierre Poussin-Courmontagne
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  3. David Rosé
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  4. Raymond Ripp
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  5. Alain Litt
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  6. Jean-Claude Thierry
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  7. Dino Moras
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Correspondence to Didier Busso.

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Busso, D., Poussin-Courmontagne, P., Rosé, D. et al. Structural Genomics of Eukaryotic Targets at a Laboratory Scale. J Struct Funct Genomics 6, 81–88 (2005). https://doi.org/10.1007/s10969-005-1909-6

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  • DOI: https://doi.org/10.1007/s10969-005-1909-6

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