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Exploring the malarial transcriptome...

Munasinghe, Anusha Dharshini.

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Exploring the malarial transcriptome by the application of serial analysis of gene expression to Plasmodium falciparum.

紀錄類型:	書目-語言資料,印刷品 : Monograph/item
正題名/作者:	Exploring the malarial transcriptome by the application of serial analysis of gene expression to Plasmodium falciparum./
作者:	Munasinghe, Anusha Dharshini.
面頁冊數:	167 p.
附註:	Adviser: Dyann F. Wirth.
Contained By:	Dissertation Abstracts International63-04B.
標題:	Biology, Molecular. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3051246
ISBN:	0493657789

Exploring the malarial transcriptome by the application of serial analysis of gene expression to Plasmodium falciparum.
Munasinghe, Anusha Dharshini.

Exploring the malarial transcriptome by the application of serial analysis of gene expression to Plasmodium falciparum. - 167 p.

Adviser: Dyann F. Wirth.

Thesis (Ph.D.)--Harvard University, 2002.

This work demonstrates the successful adaptation of the genome-wide profiling technique, SAGE in <italic>P. falciparum</italic>. More importantly, it has provided insights into the global modulation of transcript levels in asexual forms. This study comprises one of the first aimed at comprehensive characterization of gene expression in the malarial parasite. Firstly, the feasibility of applying the SAGE methodology to the 3D7 strain was demonstrated, following several modifications to the original protocol; these were required to bypass specific problems presented by the AT richness of its genome. Secondly, BLAST analysis of the 187 abundant tags from a SAGE library of mixed blood stages revealed that 88% of these tags corresponded to the 3D7 genome, with 70% of them matching single loci. The latter tags identified genes involved in various metabolic and parasite-specific functions, transcripts encoding unknown and novel ORFs, and antisense RNAs. SAGE data for both sense and antisense transcription was confirmed by alternate methods, demonstrating that antisense tags reflect <italic> bona fide</italic> transcription off of the minus strand. Thirdly, comprehensive annotation of SAGE tags extending over a 350-fold expression range, defined both a sense and antisense transcriptome in mixed erythrocytic stages. A relational database integrating SAGE expression information with genome annotation data in PlasmoDB was constructed for this purpose. Major contributors to the sense transcriptional profile included ORFs encoding membrane-associated proteins, carbohydrate metabolism genes, mitochondrial metabolism genes and signal transduction factors. These results likely reflect the unique contribution of each pathway to cellular life in the parasite. Global characterization of antisense transcription revealed that this phenomenon was largely restricted to the nucleus, where it was detected from 20% of expressed loci. Furthermore the inverse relationship between antisense and sense transcript abundance across the transcriptome suggests a role for the former in modulating gene expression. Finally we observed a differential regulation of over one hundred unique genes in a chloroquine-sensitive strain following exposure to the drug. A few, such as <italic>PfMDR1</italic>, have been previously implicated in the mechanism of chloroquine resistance. In conclusion, we hope that the current body of work will contribute to the expansion of the field of malarial genomics.

ISBN: 0493657789Subjects--Topical Terms:

1017719
Biology, Molecular.

Exploring the malarial transcriptome by the application of serial analysis of gene expression to Plasmodium falciparum.
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