Multiprotein expression strategy for structural biology of eukaryotic complexes

Daniel J Fitzgerald, Christiane Schaffitzel, Philipp Berger, Ralf Wellinger, Christoph Bieniossek, Timothy J Richmond, Imre Berger

Research output: Contribution to journalArticle (Academic Journal)peer-review

43 Citations (Scopus)

Abstract

The concept of the cell as a collection of multisubunit protein machines is emerging as a cornerstone of modern biology, and molecular-level study of these machines in most cases will require recombinant production. Here, we present and validate a strategy to rapidly produce, permutate, and posttranslationally modify large, eukaryotic multiprotein complexes by using DNA recombination in a process that is fully automatable. Parallel production of 12 protein complex variants within a period of weeks resulted in specimens of sufficient quantity and homogeneity for structural biology applications.

Original languageEnglish
Pages (from-to)275-9
Number of pages5
JournalStructure
Volume15
Issue number3
DOIs
Publication statusPublished - Mar 2007

Keywords

  • Animals
  • Baculoviridae
  • Cell Line
  • Escherichia coli
  • Genetic Vectors
  • Humans
  • Multiprotein Complexes
  • Spodoptera

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