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  • pKLAC1-GLuc Control Plasmid

    Discontinued Date

    01/01/2013
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    Discontinued Products

    Description

    pKLAC1-GLuc is a yeast reporter plasmid that can be used with the K. lactis Expression Kit (NEB #E1000). The backbone integrative expression vector, pKLAC1, directs high-level secretion of Gaussia priniceps luciferase (GLuc) from the yeast Kluyveromyces lactis. The gene encoding GLuc is cloned in the NotI-StuI sites of pKLAC1, in-frame with the a-factor secretion leader domain. pKLAC1-GLuc can be used to test plasmid transformation, cell culture, and protein production procedures.

    Additionally, GLuc can be used as a carboxy-terminal fusion tag on a desired protein to track its secretion. GLuc activity is easily detected in K. lactis spent culture medium (Figure 1) using the Gaussia Luciferase Assay Kit (NEB #E3300).


    Figure 1. Detection of a secreted malE-Gluc fusion protein in spent culture medium. Medium (25 µl) from wild-type GG799 cells (lane 1) and GG799 cells contaning integrated pKLAC1-malE-Gluc was assyed using the Gaussia Luciferase Assay Kit (NEB #E3300S ).

    Advantages and Features

    Features

    • Expression of the fusion gene is under the control of the strong K. lactis PLAC4-PBI promoter.
    • K. lactis strain GG799 can be used as an expression host.
    • Gaussia luciferase generates over 1000-fold higher bioluminescent signal intensity, compared to Firefly and Renilla luciferases.
    • The secreted luciferase protein is very stable and can be stored for several days at 4°C without loss of activity.

    Properties and Usage

    Storage Temperature

    -20°C

    Notes

    1. NEB 5-alpha Competent E. coli (High Efficiency) (NEB #C2987), NEB 5-alpha Electrocompetent E. coli (NEB #C2989) and NEB 5-alpha Competent E. coli (Subcloning Efficiency) (NEB #C2988) are all recommended for propagation of this control plasmid.

    References

    1. Colussi, P.A. and Taron, C.H. (2005). Appl. Environ. Microbiol.. 71, 7092-7098.

    Supporting Documents

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

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