Abstract
The technique of in situ RNA hybridization provides a means of examining RNA expression within individual parasites at many stages of development in both the vertebrate and mosquito hosts. The protocols described in this chapter have been developed with the aim of preserving the morphology of the parasites within the infected host cells or tissues, so that many examples of each parasite stage can be identified and observed in a single experiment. The subcellular morphology of the parasite is also preserved, and the localization of abundant RNA transcripts can be directly visualized (1,2; Fig. 1). The parasite samples are extremely simple to prepare and do not require specialized purification procedures; in situ hybridization has, for example, been used successfully to demonstrate the expression patterns of a number of sexual-stage specific genes within gametocytes present in smears of a few microliters of infected blood (1,3-5).
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References
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Thompson, J., van Spaendonk, R. M. L., Choudhuri, R., Sinden, R. E., Janse, C. J., and Waters, A. P. (1999) Heterogeneous ribosome populations are present in Plasmodium berghei during development in its vector. Mol. Microbiol. 31, 253ā260.
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Thompson, J. (2002). In Situ Detection of RNA in Blood- and Mosquito-Stage Malaria Parasites. In: Doolan, D.L. (eds) Malaria Methods and Protocols. Methods in Molecular Medicineā¢, vol 72. Humana Press. https://doi.org/10.1385/1-59259-271-6:225
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DOI: https://doi.org/10.1385/1-59259-271-6:225
Publisher Name: Humana Press
Print ISBN: 978-0-89603-823-3
Online ISBN: 978-1-59259-271-5
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