Improved negative selection protocol for Plasmodium berghei in the rodent malarial model

Malar J. 2012 Mar 31;11:103. doi: 10.1186/1475-2875-11-103.

Abstract

An improved methodology is presented here for transgenic Plasmodium berghei lines that express the negative selectable marker yFCU (a bifunctional protein that combines yeast cytosine deaminase and uridyl phosphoribosyl transferase (UPRT)) and substitutes delivery of selection drug 5-fluorocytosine (5FC) by intraperitoneal injection for administration via the drinking water of the mice. The improved methodology is shown to be as effective, less labour-intensive, reduces animal handling and animal numbers required for successful selection thereby contributing to two of the "three Rs" of animal experimentation, namely refinement and reduction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Administration, Oral
  • Animals
  • Cytosine Deaminase / genetics
  • Cytosine Deaminase / metabolism
  • Disease Models, Animal
  • Female
  • Flucytosine / administration & dosage
  • Malaria / parasitology*
  • Mice
  • Organisms, Genetically Modified / genetics
  • Pentosyltransferases / genetics
  • Pentosyltransferases / metabolism
  • Plasmodium berghei / genetics
  • Plasmodium berghei / isolation & purification*
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae / genetics
  • Selection, Genetic*
  • Transfection

Substances

  • Flucytosine
  • Pentosyltransferases
  • uracil phosphoribosyltransferase
  • Cytosine Deaminase