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Related Experiment Videos

Pock forming plasmids from antibiotic-producing Streptomyces.

T Murakami, C Nojiri, H Toyama

    The Journal of Antibiotics
    |April 1, 1983
    PubMed
    Summary

    Four plasmids from antibiotic-producing Streptomyces showed pock-forming ability in Streptomyces lividans 66 after polyethylene glycol-mediated protoplast transformation. This study details their characteristics and enhanced transformation frequencies.

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    Area of Science:

    • Microbiology
    • Molecular Biology
    • Genetics

    Background:

    • Plasmids are crucial genetic elements in bacteria, often carrying genes for antibiotic resistance or production.
    • Streptomyces species are well-known for producing a wide range of antibiotics, making their genetic manipulation important for biotechnology.
    • Understanding plasmid behavior and transfer is key to harnessing antibiotic production capabilities.

    Purpose of the Study:

    • To investigate the pock-forming ability of plasmids isolated from antibiotic-producing Streptomyces.
    • To characterize the genetic stability and transformation efficiency of these plasmids in a heterologous host, Streptomyces lividans 66.
    • To construct expanded restriction enzyme maps for the identified pock-forming plasmids.

    Main Methods:

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  • Isolation and characterization of fifteen plasmids from antibiotic-producing Streptomyces strains.
  • Polyethylene glycol-mediated protoplast transformation of Streptomyces lividans 66.
  • Analysis of pock formation, plasmid integrity (restriction enzyme mapping), and transformation frequency.

    • Main Results:

    • Four plasmids (pSF765, pSF689, pSF674, and pSF601) exhibited pock-forming ability on S. lividans 66.
    • Transformants harbored plasmids with identical restriction enzyme cleavage sites as the originals, indicating genetic stability.
    • Transformation frequency of S. lividans 66 was significantly higher (up to 500-fold) using plasmids replicated in S. lividans 66 compared to those from the original strain.
    • Transformation frequency for pSF765 remained stable after replication in S. lividans 66.

    Conclusions:

    • The identified plasmids possess the ability to form pocks and can be stably maintained and efficiently transformed in Streptomyces lividans 66.
    • Replication in S. lividans 66 can enhance the transformation frequency of certain plasmids, like pSF689.
    • Expanded restriction maps provide valuable tools for further genetic manipulation and study of these antibiotic-related plasmids.