Genetic complementation in apicomplexan parasites

Boris Striepen; Michael W White; Catherine Li; Michael N Guerini; S.-Banoo Malik; John M Logsdon; Chang Liu; Mitchell S Abrahamsen

doi:10.1073/pnas.092525699

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Genetic complementation in apicomplexan parasites

Journal article

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Peer reviewed

Genetic complementation in apicomplexan parasites

Boris Striepen, Michael W White, Catherine Li, Michael N Guerini, S.-Banoo Malik, John M Logsdon, Chang Liu and Mitchell S Abrahamsen

Proceedings of the National Academy of Sciences - PNAS, Vol.99(9), pp.6304-6309

04/30/2002

DOI: 10.1073/pnas.092525699

PMCID: PMC122944

PMID: 11959921

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Abstract

A robust forward genetic model for Apicomplexa could greatly enhance functional analysis of genes in these important protozoan pathogens. We have developed and successfully tested a genetic complementation strategy based on genomic insertion in Toxoplasma gondii . Adapting recombination cloning to genomic DNA, we show that complementing sequences can be shuttled between parasite genome and bacterial plasmid, providing an efficient tool for the recovery and functional assessment of candidate genes. We show complementation, gene cloning, and biological verification with a mutant parasite lacking hypoxanthine-xanthine-guanine phosphoribosyltransferase and a T. gondii cDNA library. We also explored the utility of this approach to clone genes based on function from other apicomplexan parasites using Toxoplasma as a surrogate. A heterologous library containing Cryptosporidium parvum genomic DNA was generated, and we identified a C. parvum gene coding for inosine 5-monophosphate-dehydrogenase (IMPDH). Interestingly, phylogenetic analysis demonstrates a clear eubacterial origin of this gene and strongly suggests its lateral transfer from ɛ-proteobacteria. The prokaryotic origin of this enzyme might make it a promising target for therapeutics directed against Cryptosporidium .

Biological Sciences

purine salvage‖lateral gene transfer‖Cryptosporidium parvum‖ Toxoplasma gondii

Details

Title: Subtitle: Genetic complementation in apicomplexan parasites
Creators: Boris Striepen - Department of Cellular Biology and Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA 30602; Department of Veterinary Molecular Biology, Montana State University, Bozeman, MT 59717; Department of Biology, Emory University, Atlanta, GA 30322; and Department of Veterinary Pathobiology, University of Minnesota, St. Paul, MN 55108
Michael W White - Department of Cellular Biology and Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA 30602; Department of Veterinary Molecular Biology, Montana State University, Bozeman, MT 59717; Department of Biology, Emory University, Atlanta, GA 30322; and Department of Veterinary Pathobiology, University of Minnesota, St. Paul, MN 55108
Catherine Li - Department of Cellular Biology and Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA 30602; Department of Veterinary Molecular Biology, Montana State University, Bozeman, MT 59717; Department of Biology, Emory University, Atlanta, GA 30322; and Department of Veterinary Pathobiology, University of Minnesota, St. Paul, MN 55108
Michael N Guerini - Department of Cellular Biology and Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA 30602; Department of Veterinary Molecular Biology, Montana State University, Bozeman, MT 59717; Department of Biology, Emory University, Atlanta, GA 30322; and Department of Veterinary Pathobiology, University of Minnesota, St. Paul, MN 55108
S.-Banoo Malik - Department of Cellular Biology and Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA 30602; Department of Veterinary Molecular Biology, Montana State University, Bozeman, MT 59717; Department of Biology, Emory University, Atlanta, GA 30322; and Department of Veterinary Pathobiology, University of Minnesota, St. Paul, MN 55108
John M Logsdon - Department of Cellular Biology and Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA 30602; Department of Veterinary Molecular Biology, Montana State University, Bozeman, MT 59717; Department of Biology, Emory University, Atlanta, GA 30322; and Department of Veterinary Pathobiology, University of Minnesota, St. Paul, MN 55108
Chang Liu - Department of Cellular Biology and Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA 30602; Department of Veterinary Molecular Biology, Montana State University, Bozeman, MT 59717; Department of Biology, Emory University, Atlanta, GA 30322; and Department of Veterinary Pathobiology, University of Minnesota, St. Paul, MN 55108
Mitchell S Abrahamsen - Department of Cellular Biology and Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA 30602; Department of Veterinary Molecular Biology, Montana State University, Bozeman, MT 59717; Department of Biology, Emory University, Atlanta, GA 30322; and Department of Veterinary Pathobiology, University of Minnesota, St. Paul, MN 55108
Resource Type: Journal article
Publication Details: Proceedings of the National Academy of Sciences - PNAS, Vol.99(9), pp.6304-6309
DOI: 10.1073/pnas.092525699
PMID: 11959921
PMCID: PMC122944
NLM abbreviation: Proc Natl Acad Sci U S A
ISSN: 0027-8424
eISSN: 1091-6490
Publisher: National Academy of Sciences
Language: English
Date published: 04/30/2002
Academic Unit: Biology
Record Identifier: 9984217424802771

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