Maltose binding protein (MBP) is a common protein expression tag. Originally developed by NEB in the late 1980’s, it is one of the most well-known and accomplished means of tagging proteins expressed in microbes. Fusion of a target protein to MBP permits its one-step purification using amylose resin. Additionally, in E. coli, MBP is known to have significantly enhanced the solubility of many proteins it has been fused to.
The pMAL™ Protein Fusion and Purification System (NEB #E8200) allows researchers to use E. coli to create a fusion between an expressed target protein and MBP, and purify the fusion protein in a single step. A gene of interest is cloned into a pMAL vector downstream of the malE gene that encodes MBP. The strong Ptac promoter and MBP translation initiation signals control expression of the gene. Upon induction, this system fuses the target protein sequence with a portion of MBP to create a fusion protein that is isolated using amylose affinity chromatography. Different pMAL vectors permit fusion proteins to be secreted into the periplasm or produced in the cytoplasm, and each introduce a protease cleavage site between MBP and the target protein to facilitate tag removal.
MBP fusion proteins can also be expressed in yeast. The pKLMF-series vectors permit intracellular expression of MBP fusions in the yeast Kluyvermyces lactis. These vectors stably integrate into the host chromosome and utilize the strong yeast LAC4 promoter for expression. Both vectors also introduce a protease cleavage site between MBP and the target protein.
pMAL™ is a trademark of New England Biolabs, Inc.
FAQs for Maltose Binding Protein Expression
Protocols for Maltose Binding Protein Expression
- 5 Minute Transformation (C2523)
- 5 Minute Transformation Protocol (C2530)
- 5 Minute Transformation Protocol (C3037)
- Affinity Purification and On-column Cleavage (E6901)
- Cleavage of the Fusion Protein Generated Using The pMAL Protein Fusion and Purification System (E8200)
- Cloning a PCR Fragment Into a pMAL Expression Vector (E8200)
- Construction of the Fusion Plasmid (E6901)
- Cross-linking of IgG to Protein A or G Beads
- Expression Using NEB Express (C2523)
- Expression Using SHuffle (C3029)
- Fusion Constructs (E6901)
- Fusion Protein Expression (E6901)
- Goat Anti-Mouse IgG Magnetic Beads Protocol
- Goat Anti-Rabbit IgG Magnetic Beads Protocol
- Goat Anti-Rat IgG Magnetic Beads (S1433)
- High Efficiency Transformation (C2523)
- High Efficiency Transformation Protocol (C3037)
- Immunoprecipitation using Protein A/G Magnetic Beads
- Isolation of MBP-fusion protein using Amylose Magnetic Beads
- mRNA Isolation using Streptavidin Magnetic Beads (NEB #S1420 and NEB #S1421)
- Phage Display: Solution-phase Panning with Affinity Bead Capture
- Preparation of Media and Solutions (E6901)
- Primer Design for Restriction Enzyme Cloning (E6901)
- Protein Expression with T7 Strains
- Protocol for Expression Using NEB Express Iq (C3037)
- Protocol for Protein Expression Using BL21 (C2530)
- Purification of a Fusion Protein generated by The pMAL Protein Fusion and Purification System (E8200)
- Purification of IgG using Protein A/G Magnetic Beads
- Quick Start Guide (E8200)
- Separating the Protein of Interest from MBP after Protease Cleavage Using The pMAL Protein Fusion and Purification System (E8200)
- Simplified Expression and Purification Protocol (E6901)
- Transformation Protocol (C2530)
- Use SNAP-Capture Magnetic Beads (S9145)
Application Notes Maltose Binding Protein Expression
Bypassing Common Obstacles in Protein Expression
Competent Cells Brochure
The Competent Cells brochure provides information on the different competent cell strains for cloning and protein expression available from NEB.
Protein Expression & Purification Brochure
The Protein Expression and Purification brochure provides information on the advanced tools for protein expression and purification offered by NEB.
- Competent Cell Product Comparison
- Protein Expression and Purification Selection Chart
- Convenient Formats of Competent Cells