Analysis Note

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Application

Thirteen pGEX vectors are available (see Figure). Nine of the vectors have an expanded multiple cloning site (MCS) that contains six restriction sites. The expanded MCS facilitates the unidirectional cloning of cDNA inserts obtained from libraries constructed using many available lambda vectors. pGEX-6P-1, pGEX-6P-2, and pGEX-6P-3 each encode the recognition sequence for site-specific cleavage by PreScission^™ Protease, (see PreScission^™ Protease) between the GST domain and the multiple cloning site. pGEX-4T-1, pGEX-4T-2, and pGEX-4T-3 are derived from pGEX-2T and contain a Thrombin recognition site. pGEX-5X-1, pGEX-5X-2, and pGEX5X-3 are derivatives of pGEX-3X and possess a Factor Xa recognition site.pGEX-2TK is designed to allow the detection of expressed Proteins by directly labeling the fusion products in vitro (1). This vector contains the recognition sequence for the catalytic subunit of cAMP-dependent Protein kinase obtained from heart muscle. The Protein kinase site is located between the GST domain and the MCS. Expressed Proteins can be directly labeled using Protein kinase and [gamma-32P]ATP and readily detected using standard radiometric or autoradiographic techniques. pGEX-2TK is a derivative of pGEX-2T; its fusion Proteins can be cleaved with Thrombin.Cleavage of pGEX-6P GST fusion Proteins occurs between the Gln and Gly residues of the recognition sequence Leu-Glu-Val-Leu-Phe-Gln-Gly-Pro (2). Low temperature (5°C) digestion minimizes the degradation of the Protein of interest. Because PreScission^™ Protease has been engineered with a GST tag, it can also be removed from the cleavage mixture simultaneously with the GST portion of the fusion Protein. The pGEX-6P Expression Vectors permit convenient site-specific cleavage and simultaneous purification on Glutathione Sepharose. The pGEX-6P series provides all three translational reading frames linked between the GST coding region and the multiple cloning site.

Features and Benefits

A tac promoter for chemically inducible, high-level expression of GST-tagged recombinant Proteins. An internal lacIq gene for use in any E. coli host. Very mild elution conditions for release of fusion Proteins from the affinity matrix, thus minimizing effects on antigenicity and functional activity. PreScission^™ Protease, Thrombin, or Factor Xa recognition sites for cleaving the desired Protein from the fusion product.

General description

Simplified package with only the pGEX vector in the pack (E. coli BL21 will continue to be available separately, code no 27-1542-01. Updated Product Specification sheets.

Legal Information

pGEX VectorspGEX Vectors are to be used for scientific investigation and research and for no other purpose whatsoever and a license for commercial use of the licensed products and the processes claimed in US patent 5,654,176 and equivalent patents and patent applications in other countries must be negotiated directly with Millipore Corp (formerly Chemicon International Inc) by the purchaser prior to such use.

PreScission is a trademark of Cytiva