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© Research
Publication : Nucleic acids research

Towards systematic identification of Plasmodium essential genes by transposon shuttle mutagenesis

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Nucleic acids research - 10 Nov 2005

Sakamoto H, Thiberge S, Akerman S, Janse CJ, Carvalho TG, Ménard R

Link to Pubmed [PMID] – 16284199

Nucleic Acids Res. 2005;33(20):e174

After the deciphering of the genome sequences of several Plasmodium species, efforts must turn to elucidating gene function and identifying essential gene products. However, random approaches are lacking and gene targeting is inefficient in Plasmodium. Here, we established shuttle transposon mutagenesis in Plasmodium berghei. We constructed a mini-Tn5 derivative that can transpose into parasite genes cloned in Escherichia coli, providing an efficient means of generating knockout fragments. A 10(4)-fold increase in frequencies of double-crossover homologous recombination in the parasite using a new electroporation technology permits to reproducibly generate pools of distinct mutants after transfection with mini-Tn5-interrupted sequences. The procedure opens the way to the systematic identification of essential genes in Plasmodium.

http://www.ncbi.nlm.nih.gov/pubmed/16284199