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Rapid recombination among transfected plasmids, chimeric episome formation and trans gene expression in Plasmodium

M Kadekoppala1, P Cheresh, D Catron

  • 1Department of Pathology, Northwestern University Medical School, 303 E. Chicago Ave., Chicago, IL 60611-3008, USA.

Molecular and Biochemical Parasitology
|February 27, 2001
PubMed

Insights

Genetic manipulation of the malaria parasite Plasmodium falciparum is now feasible. Co-transfection with specific plasmids enables rapid homologous recombination in blood stages, facilitating genetic studies.

Area of Science:

  • Molecular Parasitology
  • Genetics of infectious diseases

Background:

  • Genetic manipulation of Plasmodium falciparum blood stages is challenging due to low transfection efficiency and slow DNA integration.
  • Recombination is crucial for generating diversity in P. falciparum.

Purpose of the Study:

  • To develop a method for efficient genetic manipulation of P. falciparum blood stages.
  • To investigate the mechanisms and timing of DNA recombination in P. falciparum.

Main Methods:

  • Co-transfection of P. falciparum with two plasmids: one for GFP reporter, another for drug resistance (Tgdhfr-ts M23).
  • Selection of GFP-producing parasites to identify successful transfections and recombination events.
  • Analysis of recombined episomes and their maintenance under drug pressure.

Main Results:

  • Successful selection of a P. falciparum population where ~30% of parasites produced GFP.
  • Formation of chimeric episomes (up to 20 kb) from transfected plasmids, maintained for at least 16 weeks.
  • Early chimera formation (7-14 days) mediated by homologous recombination, enhanced by shared P. falciparum 5'hrp3 UTR.

Conclusions:

  • High levels of homologous recombination activity exist in P. falciparum blood stages.
  • This method enables rapid DNA recombination for genetic studies and gene expression in P. falciparum.
  • Facilitates future research into malaria parasite genetics and the development of new interventions.

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