The pMAL-c5X vector is designed to produce maltose-binding protein (MBP) fusions, where the protein of interest can be cleaved from MBP with Factor Xa protease (NEB #P8010). The vector creates MBP fusions expressed in the cytoplasm.
The vector pMAL-c5X 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 are expressed cytoplasmically. 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.
Visit DNA Sequences and Maps page for more information.
Source:
NEB 10-beta Competent E. coli (pMAL-c5X)
Concentration:
200 μg/ml
Storage Conditions:
10 mM Tris-HCl
1 mM EDTA
DNASU and Addgene are central repositories for plasmid clones and collections that may also be helpful.
NEB 10-beta Competent 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.
This vector conveys ampicillin resistance for propagation in E. coli.
Nagai, K. and Thogersen, H.C. (1987). Methods Enzymology. 153, 461-481.
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.), 16.6.1-16.6.12. New York: John Wiley & Sons, Inc.
Yanisch-Perron, C., Vieira, J. and Messing, J. (1985). Gene. 33, 103-119.
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 [email protected] or fill out the Technical Support Form for appropriate document.)
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
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 [email protected] or fill out the Technical Support Form for appropriate document.)
This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).
While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
Research Use Only
The buyer/user has a non-exclusive license to use the vector for Research Purposes Only. Commercial use of this vector requires a license from New England Biolabs, Inc.
U.S. Patent No. 5,643,758
Licenses
The Buyer/User has a non-exclusive license to use this vector for RESEARCH PURPOSES ONLY. Commercial use requires a license from New England Biolabs, Inc. This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
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