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Screening Leishmania donovani Complex-Specific Genes Required for Visceral Disease

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Parasite Genomics Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1201))

Abstract

Leishmania protozoan parasites are the causing agent of leishmaniasis. Depending on the infecting species, Leishmania infection can causes a wide variety of diseases such as self-healing cutaneous lesions by L. major and fatal visceral leishmaniasis by L. donovani and L. infantum. Comparison of the visceral disease causing L. infantum genome with cutaneous disease causing L. major and L. braziliensis genomes has identified 25 L. infantum (L. donovani complex) species-specific genes that are absent or pseudogenes in L. major and L. braziliensis. To investigate whether these L. donovani complex species-specific genes are involved in visceral infection, we cloned these genes from L. donovani and introduced them into L. major and then determined whether the transgenic L. major had an increased ability to survive in liver and spleen of BALB/c mice. Several of these L. donovani complex specific genes were found to significantly increase L. major survival in visceral organs in BALB/c mice including the A2 and Ld2834 genes, while down regulation of these genes in L. donovani by either antisense RNA or gene knockout dramatically reduced L. donovani virulence in BALB/c mice. This demonstrated that L. donovani complex species-specific genes play important roles in visceral infection. In this chapter, we describe procedures to screen L. donovani complex specific genes required for visceral infection by cross species transgenic expression, gene deletion targeting and measuring infection levels in mice.

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Acknowledgments

We would like to thank the Canadian Institutes of Health Research for supporting this research.

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Authors and Affiliations

  1. Microbiology and Immunology, McGill University, 3775 University Street, Montreal, QC, Canada, H3A 2B4

    Wen-Wei Zhang & Greg Matlashewski

Authors
  1. Wen-Wei Zhang
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  2. Greg Matlashewski
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Corresponding author

Correspondence to Greg Matlashewski .

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Editors and Affiliations

  1. University of Western Australia, Nedlands, West Australia, Australia

    Christopher Peacock

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Zhang, WW., Matlashewski, G. (2015). Screening Leishmania donovani Complex-Specific Genes Required for Visceral Disease. In: Peacock, C. (eds) Parasite Genomics Protocols. Methods in Molecular Biology, vol 1201. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1438-8_20

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  • DOI: https://doi.org/10.1007/978-1-4939-1438-8_20

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1437-1

  • Online ISBN: 978-1-4939-1438-8

  • eBook Packages: Springer Protocols

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