Description
General description
The pET System is the most powerful system yet developed for the cloning and expression of recombinant proteins in E. coli. Target genes are cloned in pET plasmids under control of strong bacteriophage T7 transcription and (optionally) translation signals; expression is induced by providing a source of T7 RNA polymerase in the host cell. T7 RNA polymerase is so selective and active that, when fully induced, almost all of the cell′s resources are converted to target gene expression; the desired product can comprise more than 50% of the total cell protein a few hours after induction. All of the pET vectors and companion products are available as kits designed for convenient cloning, expression, detection, and purification of target proteins. The pET-46 Ek/LIC Vector is a LIC-compatible derivative of pET-45b(+). The plasmid contains a strong T7lac promoter and an amino-terminal HIS TAG® coding sequence immediately followed by an Ek/LIC cloning site designed to allow the generation of fusion proteins with minimal vector-encoded sequence. An optional C-terminal S•Tag coding sequence is compatible with purification, detection, and quantification (1). This plasmid is compatible with the LIC Duet Adaptors for coexpression of two proteins as well as with pCDF-2 and pRSF-2 Ek/LIC vectors for coexpression of up to six proteins when using the LIC Duet Adaptors.
Other Notes
1 µg- Ek/LIC Vector
25 units- T4 DNA Polymerase, LIC-qualified
50 µl- 25 mM EDTA
40 µl- 25 mM dATP
8 µl- Ek/LIC β-gal Control Insert
100 µl- 100 mM DTT
50 µl- 10X T4 DNA Polymerase Buffer
1.5 ml- Nuclease-free Water
5 × 2 ml- SOC Medium
10 µl- Test Plasmid
0.2 ml- BL21(DE3)pLysS Competent Cells
0.2 ml- BL21(DE3) Competent Cells
22 × 50 µl- NovaBlue GigaSingles Competent Cells
Kim, J.S. and Raines, R.T. 1993. Protein Science 2, 348.
Legal Information
HIS TAG is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Commercial use of this Product 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 consideration; (3) use of the Product or its components (or any derivatives thereof) for therapeutic, diagnostic or prophylactic purposes (including as part of a device, chip, assay or other product); or (4) resale of the Product or its components, whether or not such Product or its components are resold for use in research. Nothing contained herein shall be deemed to represent or warrant that additional third party rights are not required for use of this Product.
This product contains genetically modified organisms (GMO). Within the EU GMOs are regulated by Directives 2001/18/EC and 2009/41/EC of the European Parliament and of the Council and their national implementation in the member States respectively. This legislation obliges MilliporeSigma to request certain information about you and the establishment where the GMOs are being handled. Clickhere for Enduser Declaration (EUD) Form.
Toxicity: Multiple Toxicity Values, refer to MSDS
