Description
General description
The pET-49b(+) vector is designed for cloning and high-level expression of target proteins fused with the 220 aa schistosomal glutathione-S-transferase (GST•Tag) protein that is cleavable with the human rhinovirus (HRV) 3C protease (1). Schistosomal glutathione-S-transferase (GST) is commonly used as a fusion partner when expressing proteins in E. coli (2). The GST•Tag sequence has been reported to enhance the production and in some cases the solubility of its fusion partners. 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 glutathione-S-transferase activity. Cloning sites are available for producing fusion proteins also containing a cleavable N-terminal His•Tag sequence for detection and purification. The plasmid contains a strong T7lac promoter, optimized RBS, the coding sequence for the HRV 3C protease cleavage site (LeuGluValLeuPheGlnGlyPro) for N-terminal fusion tag removal and a multiple cloning site that contains restriction enzyme sites found in many other Novagen expression vectors to facilitate insert transfer. An optional C-terminal S•Tag coding sequence is compatible with purification, detection, and quantification (3).
The pET Vectors are supplied as purified plasmid DNA (10 µg). Each order of pET DNA also includes an Induction Control strain (supplied as a glycerol stock). Please contact technical service if you need additional information.
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.
Other Notes
1. Walker, P.A., Leong L. E.-C., Ng, P.W.P., Tan, S.H.,Waller, S., Murphy, D. and Porter, A.G. (1994) Bio/Technology12: 601-605.
2. Smith, D.B. and Johnson, K.S. (1988) Gene67, 31-40.
3. Kim, J.S. and Raines, R.T. (1993) Protein Science2, 348-356.
Legal Information
NOVAGEN is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Toxicity: Standard Handling (A)
