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71144 pET-44 Ek/LIC Vector Kit - Novagen

71144
  
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      This product has been discontinued.



      pET-44 Ek/LIC Vector Kit
      pET Expression Systems and pET Expression Systems plus Competent Cells provide core reagents needed for target gene cloning and expression.The pET-43.1 vectors incorporate the Nus•Tag fusion partner and are also compatible with the trxB/gor mutant Origami, Origami 2, Origami B, Rosetta-gami, Rosetta-gami 2, and Rosetta-gami B strains, which facilitatedisulfide bond formation in the cytoplasm (Davis 1998). Disulfide-bonded proteins maybe obtained by employing the combination of a pET-43.1 or pET-44 vector and these trxB/gor host strains. Unfused proteins can be produced by using the Nde I cloning site.

      Directional PCR cloning into the most powerful E. coli expression vectors

      The pET-44 vectors incorporate the Nus•Tag™ technology, which allows target proteins to be expressed as fusions to the highly soluble NusA protein. According to a modeling study, the NusA protein has the highest potential for solubility among more than 4000 E. coli proteins, and therefore increases the proportion of soluble protein when expressed in E. coli as a fusion partner (1, 2). Ek/LIC represents a highly flexible cloning strategy in which a single insert preparation can be simultaneously cloned into a wide variety of compatible Ligation Independent Cloning (LIC) vectors to allow high-throughput optimization of target protein expression (3). The Ek/LIC Nus•Tag vector, pET-43.1 Ek/LIC, carries an N-terminal Nus•Tag/His•Tag®/ S•Tag™ configuration, followed by protease cleavage sites, multiple cloning site region and optional C-terminal HSV•Tag® and His•Tag sequences. The new pET-44 Ek/LIC Vector encodes an additional His•Tag peptide at the N-terminus but is otherwise identical to pET-43.1 Ek/LIC. We have shown that the additional N-terminal His•Tag significantly enhances purification yields when utilizing Novagen's His•Mag™ Agarose Beads for high-throughput purification (4).

      The various pET Ek/LIC Vectors are shown below. Configurations of the pCDF-2 and pRSF-2 Ek/LIC Vectors, which are designed for coexpression, are also shown. For additional information about the LIC method and Ek/LIC primer design, refer to the previous section, The LIC Method. Ek/LIC vectors for insect and mammalian expression are also available.





      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.

      Catalogue Number71144
      Brand Family Novagen®
      References
      References1. Harrison, R. G. (2000) inNovations 11, 4–7.

      2. Davis, G. D., Elisee, C., Newham, D. M., and Harrison, R. G. (1998) Biotechnol. Bioeng. 65, 382–388.

      3. Novagen 2002–2003 Catalog, 44–47.

      4. Novy, R. and Drott, D. (2002) inNovations 14, 12–13.
      Product Information
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      Quality LevelMQ100
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      Ship Code Dry Ice Only
      Toxicity Multiple Toxicity Values, refer to MSDS
      Storage ≤ -70°C
      Do not freeze Ok to freeze
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      Global Trade Item Number
      Catalog Number GTIN
      71144 0

      Documentation

      pET-44 Ek/LIC Vector Kit - Novagen Certificates of Analysis

      TitleLot Number
      71144

      References

      Reference overview
      1. Harrison, R. G. (2000) inNovations 11, 4–7.

      2. Davis, G. D., Elisee, C., Newham, D. M., and Harrison, R. G. (1998) Biotechnol. Bioeng. 65, 382–388.

      3. Novagen 2002–2003 Catalog, 44–47.

      4. Novy, R. and Drott, D. (2002) inNovations 14, 12–13.

      Brochure

      Title
      Radiance! Multisystem Protein Expression

      User Protocols

      Title
      TB053 Academic and Non-profit Laboratory Assurance Letter
      TB055 pET System Manual
      TB163 Ek/LIC Cloning Kits
      TB184 Xa/LIC Cloning Kits

      Vector Map

      Title
      TB339

      Vector Sequence

      Title
      pET-44 Ek/LIC Vector Sequence

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      Categories

      Life Science Research > Genomic Analysis > Transfection and Protein Expression > Bacterial Expression > Bacterial Expression Vectors