pMAL™-p5X Vector

Description

The vector pMAL-p5X is designed to produce maltose-binding protein (MBP) fusions, where the protein of interest can be cleaved from MBP with the specific protease Factor Xa (NEB #P8010).

MBP fusions made with this vector include an N-terminal signal sequence, so the fusion protein is directed to the periplasm. The MBP has been engineered for tighter binding to amylose resin.

A gene or open reading frame is inserted into a restriction site of the vector polylinker, in the same translational reading frame as the malE gene (encoding maltose-binding protein). The fusion protein thus produced can be purified by amylose affinity chromatography. The sequence coding for the four amino acids Ile-Glu-Gly-Arg is present just upstream of the XmnI site. This allows the protein of interest to be cleaved from maltose-binding protein with the specific protease Factor Xa. Fragments inserted in the XmnI site (cleaves GAAGG↓ATTTC) will produce a fusion protein that, after Factor Xa cleavage, contains no vector-derived residues on the protein of interest.

VisitDNA Sequences and Maps page for more information.

DNASU and Addgene are central repositories for plasmid clones and collections that may also be helpful.

Properties and Usage

Affinity Tag

Maltose-Binding Protein (MBP)

Storage Temperature

-20°C

Sequence Files

Fasta  GenBank  

Notes

  1. NEB 10-beta E. coli (High Efficiency) (NEB #C3019) is recommended for propagation and subcloning. NEB Express Competent E.coli (High Efficiency) (NEB #C2523) is recommended for expression using this vector.
  2. This vector conveys ampicillin resistance for propagation in E. coli.

References

  1. Guan, C., Li, P., Riggs, P.D. and Inouye, H. (1987). Gene. 67, 21-30.
  2. Maina, C.V., Riggs, P.D., Grandea, A.G.III, Slatko, B.E., Moran, L.S., Tagliamonte, J.A., McReynolds, L.A. and Guan, C. (1988). Gene. 74, 365-373.
  3. Nagai, K. and Thogersen, H.C. (1987). Methods Enzymology. 153, 461-481.
  4. Riggs, P.D. (1990). Expression and Purification of Maltose-Binding Protein Fusions. In F.M. Ausebel, R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith and K. Struhl (Ed.), Methods Enzymology. 16.6.1-16.6.12. New York: John Wiley & Sons, Inc.
  5. Yanisch-Perron, C., Vieira, J. and Messing, J. (1985). Gene. 33, 103-119.

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

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Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.