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Home Discontinued Products K. lactis Protease Deficient Competent Cell Sampler

K. lactis Protease Deficient Competent Cell Sampler

  • Catalog # C1007 was discontinued on September 15, 2014 ;
  • Product Information
    Chemically competent Kluyveromyces lactis cells are suitable hosts for expressing proteins that have exhibited proteolysis in the parental GG799 background (NEB #C1001S). Proteolysis of secreted heterologous proteins in K. lactis is most often casued by aspartyl poroteases that are present in the secretory pathway. In many cases, a single aspartyl protease causes the majority of the detrimental protein degredation. The Protease Deficient Competent Cell Sampler contains a wild-type GG799 strain, as well as four protease deficient K. lactis strains, each having selectable marker-free deletion of a specific secretory pathway aspartyl protease gene. Each strain can be transformed with any linearized pKLAC-series expression vector, permitting a comparison of heterologous proein quality and yield in each background. 

    The Sampler contains one transformation each of five K.lactis strains. K. lactis GG799 is a wild-type expression strain. K. lactis YCT389 contains a selectable marker-free deletion of the KLLA0E0398g locus encoding an extracellular aspartyl protease (KlYps1p) that is homologous to cerevisiae Yps1p. K. lactis YCT390 carries a selectable marker-free deletion of the KLLA0F2208g locus encoding an extracellular aspartyl protease (Klyps7p) that is homologous to S.cervisaiae Yps7p. K. lactis strain YCT 569 carries a selectable marker-free deletion of the KLLA0D01507g locus encoding an extracellular aspartyl protease belonging to pfam00026. K. lactis strain YCT598 carries a selectable marker-free deletion of the KLLA0D15917g locus encoding an extracellular aspartyl protease that is homologous to S.cervisaiae Bar1p. 

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C) Concentration
    NEB® Yeast Transformation Reagent (5 ml) 4 1 X
    Product Categories:
    Discontinued Products
    • Properties & Usage

      Storage Temperature

      -80°C
    • Product Notes
      1. Due to the high transformation efficiency of K. lactis Competent Cells, plating multiple dilutions of the cell mixture is necessary to ensure formation of plates with distinct single colonies. Growth time should not exceed 5 days as small colonies that lack an integrated expression fragment may form. 

        Plates containing colonies can be stored at 4°C for up to two weeks. 

        Growth of K. lactis Competent Cells carrying an integrated expression vector for protein expression should be performed in YPGal medium (1% yeast extract, 2% peptone, 2% galactose) supplemented with a final concentration of 38 mM (NH4)2SO4. In the absence of 38 mM (NH4)2SO4, YCT569, YCT389, YCT390 and YCT598 cells will grow slower than the GG799 parent strain (#C1001).
      2. Storage: Store Competent Cells at -80°C. Once Thawed, Do Not Re-freeze.
    • References
      1. Colussi, P.A. et al. (2005). Appl.Environ. Microbiol. 71, 2862-2869.
  • Protocols, Manuals & Usage
  • FAQs & Troubleshooting
    • Tech Tips

      Always store immediately at -80 C.  Yeast competent cells will quickly lose competency if handled improperly.

  • Quality, Safety & Legal
    • Quality Control Assays
      Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
    • Safety Data Sheets
      The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
    • Datacards
      The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the Technical Support Form for appropriate document.)
    • Legal and Disclaimers
      This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

      While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

      For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].

      This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

      New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

      Licenses

      K. lactis Competent Cells are a component of an expression system that was developed from basic research at New England Biolabs and DSM Biologics Company B.V. The buyer and user has a non-exclusive sublicense to use this system or any component thereof, including the K. lactis  Protease Deficient Competent Cell Sampler, for RESEARCH PURPOSES ONLY. A license to use this system for manufacture of clinical grade material or commercial purposes is available from New England Biolabs, Inc., or DSM Biologics Company B.V.

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