MBP tag properties and uses:
The MBP tag is derived from the E. coli maltose/maltodextrin system and is widely used as both a solubility tag (notably for recombinant proteins grown in E. coli) and also as an affinity tag, since it binds to amylose columns. MBP has a molecular mass of 42.5 kDa.
Oxford Genetics provides a range of plasmid vectors containing MBP tags for use in mammalian, bacterial and yeast cells. We provide MBP in a range of orientations, to meet all your cloning requirements. Please browse through our products using the ‘Browse Plasmids’ menu. For example we provide MBP at both N and C sides of the MCS, allowing versatile positioning of your gene, we sell it with and without enzyme cleavage sites (for simple removal of the tag, if required, after protein production), and we sell it in a range of configurations with other tags (such as hexahistidine) to give you maximum flexibility.
Purification of proteins using MBP tags: the principle
A typical MBP purification is as follows:
- Harvest and lyse host cells with expressed MBP-tagged protein, and clarify by centrifugation
- Pass the lysate slowly through an amylose column and wash carefully with buffer.
- Elute the bound MBP-tagged protein and its binding partner(s) with a buffer containing high levels of free maltose
- Purify by dialysis.
Our product range
Oxford Genetics provides a broad range of plasmids containing N-terminal and C-terminal MBP tags for mammalian, bacterial and yeast systems. In some plasmids the tags flank reporter genes, while in others they are positioned adjacent the MCS enabling you to insert your own gene in frame as required.
The plasmid structure shown includes an MBP tag positioned upstream of the MCS, for inclusion at the N terminus of your gene of interest, and an EKT cleavage site allowing simple removal of the MBP, if required, after protein production. Expression is regulated by the TEF1 promoter in yeast cells. The structure also contains URA3 for selection of transfected yeast cells in uracil-deficient media, and kanamycin resistance for selection in bacteria.
Our Plasmid Builder facility also provides a simple means for you to design an efficient strategy for any modifications or cloning manipulations to these plasmids. Finally, as always, although we have designed these plasmids for simple and efficient cloning, we are happy to undertake any cloning steps that you prefer to outsource. You can access all these options through the Plasmid Builder button below.