High-efficiency transformation of Plasmodium falciparum by the lepidopteran transposable element piggyBac

作者：Balu B; Shoue DA; Fraser MJ; Adams JH^*

来源：Proceedings of the National Academy of Sciences, 2005, 102(45): 16391-16396.

DOI：10.1073/pnas.0504679102

• malaria
• mutagenesis
• transposon

Functional analysis of the Plasmodium falciparum genome is restricted because of the limited ability to genetically manipulate this important human pathogen. We have developed an efficient transposon-mediated insertional mutagenesis method much needed for high-throughput functional genomics of malaria parasites. A drug-selectable marker, human dihydrofolate reductase, added to the lepidopteran transposon piggyBac, transformed parasites by integration into the A falciparum genome in the presence of a transposase-expressing helper plasmid. Multiple integrations occurred at the expected TTAA target sites throughout the genome of the parasite. We were able to transform A faliparum with this piggyBac element at high frequencies, in the range of 10(-3), and obtain stable clones of insertional mutants in a few weeks instead of 6-12 months. Our results show that the piggyBac transposition system can be used as an efficient, random integration tool needed for large-scale, whole-genome mutagenesis of malaria parasites. The availability of such an adaptable genetic tool opens the way for much needed forward genetic approaches to study this lethal human parasite.

• 出版日期2005-11-8