Amid Biosciences| Protein Expression and Purification Products
Maltose-binding Protein Fusion Vector: pTrham-6XHis-MBP Expression Vector | Amid Biosciences
E. coli maltose-binding protein (MBP) enhances the solubility and promotes the proper folding of its fusion partners.
Amid Biosciences has engineered and successfully tested pTrham-6XHis-MBP vector based on the rhamnose-inducible pTrham series of expression vectors for the production of dual His-MBP-tagged fusion proteins in the cytoplasm of E. coli. The MBP moiety improves the yield and solubility of its fusion partners while the His-tag facilitates and simplifies their purification with Immobilized Metal Ion Affinity Chromatography (IMAC) resins. Furthermore, the MBP tag allows for affinity purification of the fusion product on the amylose resin.
pTrham-6xHis-MBP vector encodes an N-terminal 6xHis-MBP tag followed by a TEV (Tobacco Etch Virus) protease cleavage site. The TEV site enables removal of the MBP tag from the protein following production. TEV is reported to have better specificity for its recognition site compared to EKT Thrombin or Factor Xa proteases.
pTrham-6xHis-MBP has the same features as Amid Biosciences’ pTrham vector to ensure highly efficient transcription and translation of the cloned gene, the rhaBAD promoter is complemented with the rrnG anti-termination region, the bacteriophage T7 gene 10 translation enhancer, and ribosome binding site. The multiple cloning site (MCS) provides directional cloning options using a variety of restriction enzymes.
The fusion proteins are expressed under control of L-rhamnose-inducible rhaBAD promoter of Escherichia coli which ensures a tight control of expression of cloned genes. PrhaBAD promoter is capable of high expression levels but at same time displays low baseline gene expression in the absence of inducer. Combination of the tight control of expression with MBP fusion will often enable polypeptides that normally accumulate as inclusion bodies in E. coli to fold in to their biologically active conformations.
- High level expression of proteins with cleavable N-terminal 6xHis MBP tag
- MBP tag improves protein solubility and yield
- L-Rhamnose-inducible E. coli promoter ensures minimal basal expression in the absence of inducer
- Directional cloning with a choice of restriction enzymes
- The 6xHis affinity tag is for efficient purification.
- TEV protease cleavage site for MBP removal
- ampicillin resistance
Figure 1. The nucleotide sequence encoding the polyhistidine tag, and the first few N-terminal and the last few C-terminal residues of MBP, along with the corresponding amino acid sequences in single letter code, are shown. The Multiple Cloning Site sequence is underlined.
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This product is intended for laboratory use only.