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Related Experiment Videos

Characterization of conjugative plasmid EDP208.

B B Finlay, W Paranchych, S Falkow

    Journal of Bacteriology
    |October 1, 1983
    PubMed
    Summary

    Researchers mapped the 90kb conjugative plasmid EDP208, identifying key regions for replication, pilus formation, and transfer. A 16.5kb segment is crucial for pilus synthesis, elucidated through transposon mutagenesis.

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    Area of Science:

    • Molecular Biology
    • Genetics
    • Microbiology

    Background:

    • Conjugative plasmids are crucial for horizontal gene transfer in bacteria.
    • Understanding plasmid structure and function is vital for controlling bacterial dissemination.

    Purpose of the Study:

    • To construct a restriction map of the IncF0 lac conjugative plasmid EDP208.
    • To identify and localize genes essential for plasmid replication, pilus formation, and transfer.

    Main Methods:

    • Restriction endonuclease mapping using BamHI, HindIII, PvuI, SstI, and XhoI.
    • Construction and analysis of plasmid clones and fragments.
    • Transposon mutagenesis (Tn1 and Tn5) and phenotypic analysis (f1 phage resistance).

    Main Results:

    • A 90kb restriction map of plasmid EDP208 was established.
    • The lac genes and origin of vegetative replication (oriV) were localized to the HindIII-20.5 fragment.
    • Genes for pilus synthesis and the origin of transfer (oriT) were mapped, with a 16.5kb region identified as essential for pilus biosynthesis.

    Conclusions:

    • The study provides a detailed map of the EDP208 plasmid, delineating functional regions.
    • Identified genetic elements are critical for conjugative plasmid replication, transfer, and pilus formation.

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