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An Improved Method for the Isolation of Extrachromosomal DNA from the Pathogenic Free-Living Amoeba Naegleria fowleri

Colm P Roster1, James C Morris1

  • 1Eukaryotic Pathogens Innovation Center, Department of Genetics and Biochemistry, Clemson University, Clemson, SC 29634, USA.

Methods and Protocols
|July 24, 2026
PubMed

Insights

Researchers developed a new method to purify a unique DNA element from Naegleria fowleri, the amoeba causing deadly primary amebic meningoencephalitis (PAM). This breakthrough aids in studying the organism's pathobiology.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Parasitology

Background:

  • * *Naegleria fowleri* causes the lethal primary amebic meningoencephalitis (PAM).
  • * This amoeba possesses ~4000 copies of a unique nucleolar-localized closed circular extrachromosomal ribosomal DNA element (CERE).
  • * Historically, CERE purification has been difficult, hindering research into its structure and function.

Purpose of the Study:

  • * To develop an improved methodology for purifying CERE from *Naegleria fowleri*.
  • * To enhance CERE yield and preserve its integrity for further study.
  • * To facilitate research into the pathobiology of *Naegleria fowleri*.

Main Methods:

  • * Development of a novel purification protocol for CERE.
  • * Optimization of techniques to increase CERE yield.
  • * Implementation of methods to maintain CERE integrity during purification.

Main Results:

  • * A new CERE purification method was successfully established.
  • * The methodology significantly improved CERE yield compared to previous techniques.
  • * Processing times were reduced, and CERE integrity was maintained.

Conclusions:

  • * The developed method provides a reliable way to obtain substantial quantities of pure CERE.
  • * This advancement will enable detailed studies of CERE's unique DNA architecture.
  • * Understanding CERE will contribute to advancing the study of *Naegleria fowleri* pathobiology.
Keywords:
Naegleria fowlericircular extrachromosomal ribosomal DNA elementepisomefree-living pathogenic amoebae

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