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Laboratory scale structural genomics

  • Brent W. Segelke
  • Johana Schafer
  • Matthew A. Coleman
  • Tim P. Lekin
  • Dominique Toppani
  • Krzysztof J. Skowronek
  • Katherine A. Kantardjieff
  • Bernhard Rupp
Article

Abstract

At Lawrence Livermore National Laboratory, the development of the TB structural genomics consortium crystallization facility has paralleled several local proteomics research efforts that have grown out of gene expression microarray and comparative genomics studies. Collective experience gathered from TB consortium labs and other centers involved in the NIH-NIGMS protein structure initiative allows us to explore the possibilities and challenges of pursuing structural genomics on an academic laboratory scale. We discuss our procedures and protocols for genomic targeting approaches, primer design, cloning, small scale expression screening, scale-up and purification, through to automated crystallization screening and data collection. The procedures are carried out by a small group using a combination of traditional approaches, innovative molecular biochemistry approaches, software automation, and a modest investment in robotic equipment.

cloning crystallization expression high throughput TB structural genomics 

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Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Brent W. Segelke
    • 1
  • Johana Schafer
    • 1
  • Matthew A. Coleman
    • 2
  • Tim P. Lekin
    • 1
  • Dominique Toppani
    • 1
  • Krzysztof J. Skowronek
    • 1
  • Katherine A. Kantardjieff
    • 3
  • Bernhard Rupp
    • 1
  1. 1.Macromolecular Crystallography and Structural Genomics Group, Biology and Biotechnology Research ProgramLivermoreUSA
  2. 2.Health Effects Genetics Division, Biology and Biotechnology Research ProgramLivermoreUSA
  3. 3.W.M. Keck Foundation Center for Molecular Structure, Department of Chemistry and BiochemistryCalifornia State University FullertonFullertonUSA

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