A rapid and robust selection procedure for generating drug-selectable marker-free recombinant malaria parasites
Citations Over TimeTop 10% of 2014 papers
Abstract
Experimental genetics have been widely used to explore the biology of the malaria parasites. The rodent parasites Plasmodium berghei and less frequently P. yoelii are commonly utilised, as their complete life cycle can be reproduced in the laboratory and because they are genetically tractable via homologous recombination. However, due to the limited number of drug-selectable markers, multiple modifications of the parasite genome are difficult to achieve and require large numbers of mice. Here we describe a novel strategy that combines positive-negative drug selection and flow cytometry-assisted sorting of fluorescent parasites for the rapid generation of drug-selectable marker-free P. berghei and P. yoelii mutant parasites expressing a GFP or a GFP-luciferase cassette, using minimal numbers of mice. We further illustrate how this new strategy facilitates phenotypic analysis of genetically modified parasites by fluorescence and bioluminescence imaging of P. berghei mutants arrested during liver stage development.
Related Papers
- → Role of host cellular response in differential susceptibility of nonimmunized BALB/c mice to Plasmodium berghei and Plasmodium yoelii sporozoites(1991)67 cited
- → Role of macrophages in malaria: O2 metabolite production and phagocytosis by splenic macrophages during lethal Plasmodium berghei and self-limiting Plasmodium yoelii infection in mice(1984)59 cited
- → The large difference in infectivity for mice of Plasmodium berghei and Plasmodium yoelii sporozoites cannot be correlated with their ability to enter into hepatocytes(1996)59 cited
- → Detection of Plasmodium berghei and Plasmodium yoelii Liver-Stage Parasite Burden by Quantitative Real-Time PCR(2015)13 cited
- → Induction of Hepatic Inflammatory Response by Plasmodium berghei Sporozoites Protects BALB/c Mice against Challenge with Plasmodium yoelii Sporozoites(1993)15 cited