Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Transfection in Micromonospora spp.

J L Caso1, C Hardisson, J E Suárez

  • 1Departamento de Biología Funcional, Universidad de Oviedo, Spain.

Applied and Environmental Microbiology
|October 1, 1987
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Widespread use of <i>Lactobacillus</i> OppA, a surface located protein, as an adhesin that recognises epithelial cell surface glycosaminoglycans.

Beneficial microbes·2019
Same author

Two alkaline motifs in the Lactobacillus salivarius Lv72 OppA surface are important to its adhesin function.

Beneficial microbes·2018
Same author

A new contribution to the history of probiotics.

Beneficial microbes·2017
Same author

Lactococcin 972 : a homodimeric lactococcal bacteriocin whose primary target is not the plasma membrane.

Microbiology (Reading, England)·2011
Same author

Analysis of the morphogenetic cluster and genome of the temperate Lactobacillus casei bacteriophage A2.

Archives of virology·2003
Same author

Mycelial differentiation and spore formation by Streptomyces brasiliensis in submerged culture.

Canadian journal of microbiology·2002

Bacteriophage DNA transfection into Micromonospora protoplasts was optimized using specific polyethylene glycol (PEG) concentrations and liposomes. Protoplast properties, not DNA, are key for successful viral progeny production.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Micromonospora are important antibiotic-producing bacteria.
  • Bacteriophages are viruses that infect bacteria and can be used as tools in molecular biology.
  • Efficiently introducing foreign DNA into Micromonospora protoplasts is crucial for genetic manipulation and bacteriophage research.

Purpose of the Study:

  • To report the successful introduction of bacteriophage DNA into Micromonospora protoplasts.
  • To identify factors influencing the efficiency of this transfection process.
  • To understand the limitations and potential of this method for genetic studies.

Main Methods:

  • Protoplast isolation from Micromonospora mycelium.
  • Treatment with bacteriophage DNA in the presence of polyethylene glycol (PEG) and liposomes.

Related Experiment Videos

  • Optimization of PEG concentration and protoplast age.
  • Assessment of infective viral progeny production (PFU).

    • Main Results:

    • Transfection efficiency was maximized with 15-hour-old mycelium and 20% PEG.
    • Positively charged liposomes were essential for successful transfection.
    • Maximum efficiency reached 10(-3) to 10(-4) PFU per protoplast.
    • The method worked for multiple bacteriophage DNAs but only with a subset of Micromonospora strains.

    Conclusions:

    • Micromonospora protoplast competence, rather than DNA characteristics, is the primary determinant of transfection success.
    • A small subpopulation of protoplasts appears to be competent for DNA uptake.
    • This method provides a foundation for further genetic studies of Micromonospora and their bacteriophages.