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Related Experiment Videos

Versatile mercury-resistant cloning and expression vectors.

B D Gambill, A O Summers

    Gene
    |January 1, 1985
    PubMed
    Summary
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    New cloning vectors offer broad or narrow host ranges and resistance to kanamycin and mercury. One vector enables mercury-inducible gene expression, enhancing cloning applications.

    Area of Science:

    • Molecular Biology
    • Microbiology
    • Biotechnology

    Background:

    • Development of novel cloning vectors is crucial for genetic engineering.
    • Existing vectors may have limitations in host range, copy number, or selectable markers.

    Purpose of the Study:

    • To construct and characterize two new cloning vectors, pDG105 and pDG106, based on IncQ and P15A replicons.
    • To evaluate their utility in various bacterial species and as regulated expression systems.

    Main Methods:

    • Construction of plasmids pDG105 (IncQ) and pDG106 (P15A) with resistance to kanamycin and mercuric ions.
    • Transformation of Escherichia coli, Acinetobacter calcoaceticus, and Pseudomonas putida.
    • Analysis of cloning sites within the kanamycin-resistance gene and mer operon.

    Related Experiment Videos

  • Assessment of mercury-inducible gene expression using the galK gene.

    • Main Results:

    • Both vectors confer resistance to kanamycin (Km) and mercuric ions (Hg2+).
    • pDG105 demonstrates broad-host-range transformation capabilities.
    • pDG106 is a narrow-host-range vector compatible with pBR322.
    • Cloning into the mer operon's EcoRI site creates a mercury-supersensitive phenotype.
    • Insertion of galK results in Hg2+-inducible galactokinase activity.

    Conclusions:

    • The novel IncQ and P15A-based vectors provide versatile tools for molecular cloning.
    • The mercury-inducible expression system offers a controllable method for gene regulation.
    • These vectors expand options for genetic manipulation in diverse bacterial hosts.