Analysis of Gene Expression by RT-PCR
To achieve a more complete understanding of malaria parasite biology and its interaction with its host, it is essential to be able to study the transcription and expression of parasite genes. Northern blot analysis and RNase protection assays are commonly used to study gene expression (1). However, both procedures require a significant amount of input RNA. In the case of the human malaria species, only Plasmodium falciparum can be grown in culture (2). This allows the production of a sufficient number of parasites to obtain the necessary amounts of RNA to use in these procedures. Unfortunately, when working with samples obtained from either animals or humans, the amount of RNA obtained is often insufficient to perform even one such experiment.
KeywordsPolymerase Chain Reaction Polymerase Chain Reaction Product Polymerase Chain Reaction Reaction Nest Polymerase Chain Reaction Reverse Transcription Step
- 1.Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Plainview, New York.Google Scholar
- 4.Snounou, G., Viriyakosol, S., Jarra, W., Thaithong, S., and Brown, K. N. (1993) Identification of the four human malaria parasite species in field samples by the polymerase chain reaction and detec tion of a high prevalence of mixed infections. Mol. Biochem. Parasitol. 58, 283–292.CrossRefPubMedGoogle Scholar
- 7.Smith, J. D., Chitnis, C. E., Craig, A. G., Roberts, D. J., Hudson-Taylor, D. E., Peterson, D. S., et al. (1995) Switches in expression of Plasmodium falciparum var genes correlate with changes in antigenic and cytoadherentphenotypes of infected erythrocytes. Cell 82, 101–110.CrossRefPubMedGoogle Scholar