Mutations in maltose-binding protein that alter affinity and solubility properties.
Walker IH, Hsieh PC, Riggs PD
Appl Microbiol Biotechnol. 2010 Sep;88(1):187-97. doi: 10.1007/s00253-010-2696-y. Epub 2010 Jun 10.
(Link opens in a new window)
PubMed
(Link opens in a new window)
Article
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 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.
Learn more about MBP expression
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 75286 | pMAL-c2X | Expresses proteins in the cytoplasm as fusions to maltose-binding protein |
| 75287 | pMAL-p2X | Expresses proteins in the periplasm as fusions to maltose-binding protein |
| 75288 | pMAL-c4X | Expresses proteins in the cytoplasm as fusions to a high-binding mutant (A313V) maltose-binding protein |
| 75289 | pMAL-p4X | Expresses proteins in the periplasm as fusions to a high-binding mutant (A313V) maltose-binding protein |
| 75290 | pMAL-c2G | Expresses proteins in the cytoplasm as fusions to maltose-binding protein |
| 75291 | pMAL-c2E | Expresses proteins in the cytoplasm as fusions to maltose-binding protein |
| 75293 | pMAL-c4E | Expresses proteins in the cytoplasm as fusions to a high-binding mutant (A313V) maltose-binding protein |