Description:
pMYB5 is a control plasmid for the IMPACT™ Kit (NEB #E6901) (1,2). This plasmid carries the E. coli malE gene, encoding the maltose binding protein (MBP)(3), fused in-frame to the coding region of the Sce VMA intein-chitin binding domain (55 kDa)(1,4). pMYB5 can be used to test plasmid transformation, cell culture, induction and purification procedures. After induction with 0.3 mM IPTG at 30°C for 3 hours (or 15°C for 12-16 hours), 100 ml of cells should yield 2-3 mg of a 97 kDa fusion protein. After chitin column purification and cleavage, approximately 1.0-1.5 mg of the MBP (42 kDa) is usually obtained. This double stranded vector is 8602 bp in length.
Source:
pMYB5 is isolated from an E. coli strain (r-m-) by a standard plasmid purification procedure.
Advantages:
• Expression of the fusion gene is under the control of an IPTG -inducible T7 promoter (5).
• E. coli strains T7 Express Competent E. coli (High Efficiency) (NEB #C2566) or BL21(DE3) Competent E. coli (NEB #C2527) and derivatives can be used as an expression hosts.
• A pTYB1 derivative.
• Ampicillin resistance.
Storage Conditions:
10 mM Tris-HCl
1 mM EDTA
pH 8.0 @ 25°C
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