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PMID: 16260745 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

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

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 102 ·No. 45 ·2005-11-08 ·Pages 16391-6

Balu B, Shoue DA, Fraser MJ, Adams JH

Abstract

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 P. 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 P. falciparum 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.

MeSH Terms
Animals DNA Transposable Elements Genome, Protozoan Lepidoptera/genetics Plasmids Plasmodium falciparum/genetics Transformation, Genetic
Chemicals
DNA Transposable Elements
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Balu Bharath
Department of Biological Sciences, Center for Tropical Disease Research and Training, University of Notre Dame, Notre Dame, IN 46556, USA.
Shoue Douglas A
Fraser Malcolm J
Adams John H
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0504679102
Published
2005-11-08
Epub
2005-00-31
Pages
16391-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC1275597
Subset
IM
Grants
NIAID NIH HHS · R01 AI033656-13 · United States
NIAID NIH HHS · R01 AI033656 · United States
NIAID NIH HHS · R01AI33656 · United States
NIAID NIH HHS · R01 AI048561 · United States
NIAID NIH HHS · R01AI48561 · United States
External Links
PubMed source DOI: 10.1073/pnas.0504679102 PMC Full Text View journal details
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