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Published on: July 31, 2012
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The pCri System: a vector collection for recombinant protein expression and purification.
Theodoros Goulas1, Anna Cuppari1, Raquel Garcia-Castellanos1
1Proteolysis Lab, Molecular Biology Institute of Barcelona, CSIC, Barcelona Science Park, Helix Building, Barcelona, Spain.
Plos One
|November 12, 2014
Summary
The pCri System enables efficient recombinant protein production across multiple expression hosts. This vector collection streamlines cloning and tag manipulation for high-throughput structural and biochemical studies.
Area of Science:
- Biochemistry
- Molecular Biology
- Structural Biology
Background:
- Recombinant protein overexpression is crucial for structural, biochemical, and biophysical studies.
- Obtaining large quantities of pure, homogenous protein remains a significant challenge.
Purpose of the Study:
- To introduce the pCri System, a novel vector collection for protein expression.
- To facilitate simultaneous assessment across prokaryotic and eukaryotic host cells.
Main Methods:
- The pCri System utilizes a single polymerase chain reaction product for directional cloning into multiple rare restriction sites.
- It allows incorporation of various fusion tags and signal peptides at N- and C-termini.
- Fusion tags can be cleaved using site-specific peptidases.
Main Results:
- The system supports expression in Escherichia coli, Bacillus subtilis, and Pichia pastoris.
- It facilitates protein expression, solubility, detection, and purification.
- Efficient removal of fusion tags leaves minimal additional residues.
Conclusions:
- The pCri System is a valuable tool for high-throughput protein studies.
- Its versatility in cloning and expression systems enhances protein research capabilities.
