Journal of Structural and Functional Genomics

, Volume 6, Issue 1, pp 13–20

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
Article

DOI: 10.1007/s10969-005-0462-7

Cite this article as:
Cowieson, N.P., Listwan, P., Kurz, M. et al. J Struct Funct Genomics (2005) 6: 13. doi:10.1007/s10969-005-0462-7

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.

Keywords

bacterial expressionmammalian proteinsmedium-throughputprotein expression screeningrecombinatorial cloning

Copyright information

© Springer 2005

Authors and Affiliations

  • Nathan P. Cowieson
    • 1
  • Pawel Listwan
    • 2
    • 3
    • 5
  • Mareike Kurz
    • 1
  • Anna Aagaard
    • 1
  • Timothy Ravasi
    • 1
    • 3
    • 5
  • Christine Wells
    • 1
    • 3
    • 5
  • Thomas Huber
    • 4
  • David A. Hume
    • 1
    • 3
    • 5
  • Bostjan Kobe
    • 1
    • 2
    • 5
  • Jennifer L. Martin
    • 1
    • 2
    • 5
  1. 1.Institute for Molecular BioscienceAustralia
  2. 2.School of Molecular and Microbial SciencesAustralia
  3. 3.Cooperative Research Centre for Chronic Inflammatory DiseasesAustralia
  4. 4.Department of MathematicsAustralia
  5. 5.ARC Special Research Centre for Functional and Applied Genomics, Queensland Bioscience PrecinctUniversity of QueenslandBrisbaneAustralia