Pilot studies on the parallel production of soluble mouse proteins in a bacterial expression system

Nathan P Cowieson, Pawel Listwan, Mareike Kurz, Anna Aagaard, Timothy Ravasi, Christine Wells, Thomas Huber, David A Hume, Bostjan Kobe, Jennifer L Martin

Research output: Contribution to journalArticlepeer-review

Abstract

We investigated the parallel production in medium throughput of mouse proteins, using protocols that involved recombinatorial cloning, protein expression screening and batch purification. The methods were scaled up to allow the simultaneous processing of tens or hundreds of protein samples. Scale-up was achieved in two stages. In an initial study, 30 targets were processed manually but with common protocols for all targets. In the second study, these protocols were applied to 96 target proteins that were processed in an automated manner. The success rates at each stage of the study were similar for both the manual and automated approaches. Overall, 15 of the selected 126 target mouse genes (12%) yielded soluble protein products in a bacterial expression system. This success rate compares favourably with other protein screening projects, particularly for eukaryotic proteins, and could be further improved by modifications at the cloning step.
Original languageEnglish
Pages (from-to)13-20
Number of pages8
JournalJournal of Structural and Functional Genomics
Volume6
Issue number1
DOIs
Publication statusPublished - 2005

Keywords

  • Animals
  • Cloning, Molecular
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli
  • Mice
  • Pilot Projects
  • Plasmids
  • Recombinant Fusion Proteins

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