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

Research output: Contribution to journalArticle

15 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)147-157
Number of pages11
JournalJournal of Structural and Functional Genomics
Volume5
Issue number1-2
DOIs
StatePublished - 2004
Externally publishedYes

Keywords

  • Cloning
  • Crystallization
  • Expression
  • High throughput
  • TB structural genomics

ASJC Scopus subject areas

  • Genetics
  • Structural Biology
  • Biochemistry

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    Segelke, B. W., Schafer, J., Coleman, M. A., Lekin, T. P., Toppani, D., Skowronek, K. J., Kantardjieff, K. A., & Rupp, B. (2004). Laboratory scale structural genomics. Journal of Structural and Functional Genomics, 5(1-2), 147-157. https://doi.org/10.1023/B:JSFG.0000029193.82120.d1