New England Biolabs Canada

pMAL-c6T Vector

Catalog # Concentration Size
N0378S 10 µg
Catalog # Concentration Size
N0378S 10 µg

The pMAL-c6T Vector is designed to produce maltose-binding protein (MBP) fusions in the cytoplasm. The MBP has been engineered for tighter binding to amylose resin. The vector expresses an N-terminal his-tagged malE gene followed by a multiple cloning site containing a TEV protease recognition sequence, allowing MBP to be cleaved from the protein of interest after purification. 

  • View sequence details
  • pMAL-c6T is the recommended vector for use with the NEBExpress® MBP Fusion and Purification System (NEB #E8201)
  • Fusion to MBP significantly enhances the solubility and proper folding of proteins expressed in E. coli (1)
  • Vector includes His-tag and TEV Protease recognition site, allowing the protein of interest to be cleaved from MBP using TEV Protease (NEB #P8112)
  • Two-step purification: amylose resin elution followed by TEV Protease cleavage and Ni resin isolation results in a highly-pure, tag-free target protein
  • MBP has been engineered for tighter binding to amylose resin
  • Ampicillin resistant
 
The pMAL-c6T Vector provides a method for producing a protein expressed from a cloned gene or open reading frame. The cloned gene is inserted downstream from the malE gene of E. coli, which encodes maltose-binding protein (MBP), resulting in the expression of an MBP fusion protein in the cytoplasm (1,2). The MBP has been engineered for tighter binding to amylose resin. The method uses the strong “tac” promoter and the malE translation initiation signals to give high-level expression of the cloned sequences (3,4), and a one-step purification of the fusion protein using MBP’s affinity for maltose (5). The vector expresses an N-terminal hexahistidine tagged malE gene (lacking its secretory signal sequence) followed by a multiple cloning site containing a TEV protease recognition sequence and stop codons in all three frames. This allows MBP to be cleaved from the protein of interest by TEV Protease after purification. The vector also carries the lacIq gene, which encodes for the Lac repressor. This keeps expression from Ptac low in the absence of IPTG induction. 

Source: NEB-10 beta competent E. coli (pMAL-c6T)

Figure 1: pMAL-c6T Vector



The pMAL- c6T Vector has an exact deletion of the malE signal sequence. Arrows indicate the direction of transcription. Unique restriction sites in the multiple cloning site (MCS) are indicated.

Product Source
NEB-10 beta competent E. coli (pMAL-c6T)
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  pMAL-c6T Vector N0378SVIAL -20 1 x 10 µg 200 µg/ml
Features
  • The recommended vector for use with the NEBExpress® MBP Fusion and Purification System (NEB #E8201)
  • Fusion to MBP significantly enhances proper folding and solubility of target proteins
  • Vector includes His-tag and TEV Protease recognition site
  • Two-step purification: amylose resin elution followed by TEV Protease cleavage and Ni resin isolation results in a highly pure, tag-free target protein
An exception occurred during the operation, making the result invalid. Check InnerException for exception details.

Companion Products
Notes
  • A detailed product manual for the NEBExpress MBP Fusion and Purification System with protocols can be found here.
  • 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.
References
  • Guan, C. et al (1987). Gene. 67, 21–30.
  • Maina, C.V. et al (1988). Gene. 74, 365–373.
  • Amann, E. et al (1985). Gene. 40, 183–190..
  • Duplay, P. et al (1984). J. Biol. Chem. 259, 10606–10613.
  • Kellerman, O.K. et al (1982). Methods in Enzymology. 90, 459–463.
Quality Control Assay
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.
Certificate of Analysis
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]
Safety Data Sheets
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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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