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pET GST Fusion System 41
Popular GST fusion tag in advanced pET vectors
pET Expression Systems and pET
pET Expression Systems and pET Expression Systems plus Competent Cells provide core reagents needed for target gene cloning and expression.
Components: pET Expression Systems
Components for pET Expression Systems are similar for all systems unless otherwise stated with the specific pET Expression System description. pET Expression Systems include:
• 10 µg pET vector DNA (for each indicated plasmid)
• 0.2 ml BL21 Glycerol Stock
• 0.2 ml BL21(DE3) Glycerol Stock
• 0.2 ml BL21(DE3)pLysS Glycerol Stock
• 0.2 ml Induction Control Glycerol Stock
Components: pET Expression Systems plus Competent Cells
pET Expression Systems plus Competent Cells contain all of the components of the specific pET Expression System, as well as the following additional components, unless otherwise stated with the specific pET Expression System description:
• 0.2 ml NovaBlue Competent Cells
• 0.2 ml BL21(DE3) Competent Cells
• 0.2 ml BL21(DE3)pLysS Competent Cells
• 2 × 0.2 ml SOC Medium
• 10 µl Test Plasmid
These components are sufficient for up to 10 transformations in each host.
Purification and Detection Reagents
Purification and detection reagents are available separately. For complete product descriptions and ordering information, refer to the Protein Purification and Antibodies, Conjugates & Detection Tools chapters.
pET Expression System 41
The pET-41a-c(+) and pET-42a-c(+) vectors incorporate the schistosomal glutathione-S-transferase (GST; GST•Tag) coding sequence as a fusion partner. The GST•Tag sequence has been reported to enhance the production and in some cases the solubility of its fusion partners (Smith 1998). When expressed in a soluble, properly folded form, GST•Tag fusion proteins can be purified with immobilized glutathione. Gentle elution is achieved with buffers containing reduced glutathione. Quantification of soluble GST fusions is also possible by assaying the transferase activity. The pET-41 and -42 series feature the powerful T7lac promoter, and encode the GST•Tag (220 aa) sequence, proteolytic sites, His•Tag (6 aa) sequence, and S•Tag (15 aa) sequence.
In contrast to other commercially available GST-fusion vectors, the Xa (IleGluGlyArg, pET-42 series) and enterokinase (AspAspAspAspLys, pET-41 series) proteolytic cleavage sites have been engineered to allow removal of 100% of the vector-encoded sequences and the generation of native proteins with their authentic N-terminal residues. Unfused proteins can be produced by using the Nde I cloning site. A version of pET-41 is available as a linearized vector prepared for ligation-independent cloning (LIC) of PCR products.
Another pET vector with the GST•Tag sequence is pET-49b(+). Please see the website description for more information. The His•Tag and S•Tag sequences enable alternative protein detection and purification procedures to be performed. For example, when enhancing purity with a separate purification method, or when purifying under denaturing conditions
The pET System allows for expression of target genes under control of the T7 promoter and is covered by US Patent 5,693,489. This patent is licensed by EMD Millipore Corporation from Brookhaven National Labs. Commercial entities need to obtain a license from Brookhaven National Labs prior to use of this Product. Ccommercial use of this Product also requires a commercial license from EMD Millipore Corporation. Commercial use shall include but not be limited to (1) use of the Product or its components in manufacturing; (2) use of the Product or its components to provide a service, information, or data to others in exchange for consid |