title: Improved negative selection protocol for Plasmodium berghei in the rodent malarial model.
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.
authors.affiliation: Wellcome Trust Centre for Molecular Parasitology, Institute of Infection, Immunity and Inflammation, University of Glasgow, UK.
authors.forename: Rachael Y
authors.initials: RY
authors.lastname: Orr
authors.forename: Nisha
authors.initials: N
authors.lastname: Philip
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Waters
mesh_terms: D000284:Administration, Oral; D000818:Animals; D043525:Cytosine Deaminase / Q000235:genetics / Q000378:metabolism; D004195:Disease Models, Animal; D005260:Female; D005437:Flucytosine / Q000008:administration & dosage; D008288:Malaria / Q000469:parasitology*; D051379:Mice; D030781:Organisms, Genetically Modified / Q000235:genetics; D010430:Pentosyltransferases / Q000235:genetics / Q000378:metabolism; D010962:Plasmodium berghei / Q000235:genetics / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D012641:Selection, Genetic; D014162:Transfection
chemical_list: D005437:Flucytosine; D010430:Pentosyltransferases; C023269:uracil phosphoribosyltransferase; D043525:Cytosine Deaminase