Related Experiment Video
Updated: Jun 23, 2026
13:51
The MultiBac Protein Complex Production Platform at the EMBL
Published on: July 11, 2013
Automated unrestricted multigene recombineering for multiprotein complex production.
Christoph Bieniossek1, Yan Nie, Daniel Frey
1European Molecular Biology Laboratory, Grenoble Outstation, Grenoble, France.
Nature Methods
|May 5, 2009
Summary
Automated protein complex expression is vital for research. The Acembl system uses recombineering for efficient multigene assembly and diversification in E. coli, enabling complex protein production.
Area of Science:
- Molecular Biology
- Biochemistry
- Protein Engineering
Background:
- Recombinant protein overexpression is essential for studying multiprotein complexes.
- Efficient revision and diversification of expression experiments are critical for project success.
- Automation is increasingly indispensable in protein expression research.
Purpose of the Study:
- To introduce Acembl, a versatile and automatable system for protein-complex expression.
- To facilitate multigene assembly and diversification using recombineering.
- To demonstrate the system's capability in producing complex protein structures.
Main Methods:
- Development of the Acembl system for automated protein expression.
- Utilization of recombineering for multigene assembly and diversification.
- Application in Escherichia coli expression systems.
Main Results:
- Successful demonstration of protein-complex expression using Acembl.
- Production of the complete prokaryotic holotranslocon complex.
- Validation of Acembl's versatility and automatable nature.
Conclusions:
- Acembl provides an efficient and automatable solution for recombinant protein-complex expression.
- The system facilitates rapid experimental revision and diversification of protein complexes.
- Acembl is a valuable tool for advancing structural and functional studies of multiprotein complexes.
Related Concept Videos
02:53Conservative Site-specific Recombination and Phase Variation
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
01:09Recombinant DNA
Overview
01:09Recombinant DNA
Overview
01:57Protein Complexes with Interchangeable Parts
Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order to...
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order to...
02:31Homologous Recombination
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...