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Transcripts of developmentally regulated Plasmodium falciparum genes quantified by real-time RT-PCR.

Blair, PL; Witney, A; Haynes, JD; Moch, JK; Carucci, DJ; Adams, JH (2002) Transcripts of developmentally regulated Plasmodium falciparum genes quantified by real-time RT-PCR. Nucleic Acids Res, 30 (10). 2224 - 2231. none
SGUL Authors: Witney, Adam Austin

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Abstract

Plasmodium falciparum intraerythrocytic development is a complex process. Development proceeds rapidly from the trophozoite phase of nutrient acquisition and growth through to the synthetic and reproductive schizont phase, which ends with production of new invasive merozoites. During this process, the malaria parasite must express a series of different gene products, depending on its metabolic and synthetic needs. We are particularly interested in the development of the merozoite's organelles in the apical complex, which form during the later schizont stages. We have used quantitative real-time RT-PCR fluorogenic 5' nuclease assays (TaqMan) for the first time on malaria parasites for analysis of erythrocytic stage-specific gene expression. We analyzed transcripts of the P.falciparum eba-175 and other erythrocyte binding-like (ebl) family genes in temperature-synchronized parasites and found ebl genes have tightly controlled, stage-specific transcription. As expected, eba-175 transcripts were abundant only at the end of schizont development in a pattern most common among ebl, including baebl, pebl and jesebl. The maebl transcript pattern was unique, peaking at mid-late trophozoite stage, but absent in late-stage schizonts. ebl-1 demonstrated another pattern of expression, which peaked during mid-schizont stage and then significantly diminished in late-stage schizonts. Our analysis demonstrates that using real-time RT-PCR fluorogenic 5' nuclease assays is a sensitive, quantitative method for analysis of Plasmodium transcripts.

Item Type: Article
Additional Information: PMCID: PMC115288
Keywords: Animals, Antigens, Protozoan, Blotting, Southern, Carrier Proteins, DNA, Protozoan, Gene Expression Regulation, Developmental, Genes, Protozoan, Plasmodium falciparum, Polymerase Chain Reaction, Protozoan Proteins, RNA, Protozoan, Receptors, Cell Surface, Reverse Transcriptase Polymerase Chain Reaction, Temperature, Time Factors, Transcription, Genetic
SGUL Research Institute / Research Centre: Academic Structure > Infection and Immunity Research Institute (INII)
Journal or Publication Title: Nucleic Acids Res
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Dates:
DateEvent
15 May 2002Published
PubMed ID: 12000842
Web of Science ID: 12000842
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URI: http://openaccess.sgul.ac.uk/id/eprint/103799
Publisher's version: none

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