This product is a replacement for NEB #N8108, pMAL™-c5X Vector
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.
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
Product Information
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)
This product is related to the following categories:
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
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.
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Certificate of Analysis
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