pET-49b(+) DNA - Novagen

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.

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.

NOVAGEN is a registered trademark of Merck KGaA, Darmstadt, Germany

Toxicity: Standard Handling (A)

This product is sold for internal research use only. Any commercial use of this product, its components, and/or any derivatives thereof (including but not limited to proteins produced using the product or its components) (together and hereinafter the “EMD Product”) requires signature of a written commercial use agreement with EMD Millipore Corporation or its successor-in-interest. Commercial use shall include but not be limited to: (1) use of the EMD Product to manufacture products for sale to third parties; (2) use of the EMD Product to provide services, information, or data to third parties in exchange for consideration; (3) use of the EMD Product for therapeutic, diagnostic or prophylactic purposes (including as part of a device, chip, assay or other product); or (4) resale of the EMD Product, whether or not such EMD Product is resold for research use. Nothing contained herein shall be deemed to represent or warrant that additional third party rights are not required for use of the EMD Product. Please direct any questions on these use restrictions to: licensing@milliporesigma.com.