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Related Experiment Videos

Genetic differences between nitrate-negative and nitrate-positive C. pseudotuberculosis strains using restriction

S S Sutherland1, R A Hart, N B Buller

  • 1Animal Industries, Department of Agriculture, Australia.

Veterinary Microbiology
|March 1, 1996
PubMed
Summary

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Restriction fragment length polymorphism (RFLP) differentiated nitrate-positive and nitrate-negative strains of Corynebacterium pseudotuberculosis. Genetic analysis confirmed phenotypic differences, aiding bacterial classification.

Area of Science:

  • Microbiology
  • Bacteriology
  • Genetics

Background:

  • Corynebacterium pseudotuberculosis biotypes are defined by nitrate breakdown.
  • Restriction enzyme analysis (REA) shows differentiation but is complex.
  • Restriction fragment length polymorphism (RFLP) is a viable genetic tool for bacterial differentiation.

Purpose of the Study:

  • To genetically differentiate Corynebacterium pseudotuberculosis biotypes using RFLP.
  • To investigate genetic variations correlating with phenotypic traits like nitrate reduction.
  • To assess RFLP uniqueness for C. pseudotuberculosis compared to related species.

Main Methods:

  • Phenotypic characterization of 13 C. pseudotuberculosis isolates and related bacteria.
  • Genetic analysis using non-radioactive ribosomal RNA (rRNA) probes.

Related Experiment Videos

  • Southern blotting of restriction digests (ApaI, PstI, SstI).

    • Main Results:

    • Distinct genetic differences were identified between nitrate-positive and nitrate-negative C. pseudotuberculosis strains.
    • RFLP analysis revealed clear distinctions correlating with nitrate reduction ability.
    • No genetic variations were observed related to carbohydrate fermentation differences.
    • No genetic differentiation was found between strains from different animal species or geographic locations.
    • RFLP bands were unique to C. pseudotuberculosis compared to Corynebacterium ulcerans, Actinomyces pyogenes, and Rhodococcus equi.

    Conclusions:

    • RFLP effectively differentiates C. pseudotuberculosis into nitrate-reducing groups.
    • Genetic markers correlate with key phenotypic traits, enhancing bacterial classification.
    • RFLP provides a simpler alternative to REA for C. pseudotuberculosis strain differentiation.