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Vol. 12, Issue 10, 3114-3125, October 2001



and
*Department of Immunology and Infectious Diseases, Harvard School
of Public Health, Harvard University, Boston, Massachusetts 02115; and
Serial analysis of gene expression (SAGE) was applied to the
malarial parasite Plasmodium falciparum to characterize
the comprehensive transcriptional profile of erythrocytic stages. A
SAGE library of ~8335 tags representing 4866 different genes was
generated from 3D7 strain parasites. Basic local alignment search
tool analysis of high abundance SAGE tags revealed that a
majority (88%) corresponded to 3D7 sequence, and despite the low
complexity of the genome, 70% of these highly abundant tags matched
unique loci. Characterization of these suggested the major metabolic
pathways that are used by the organism under normal culture conditions.
Furthermore several tags expressed at high abundance (30% of tags
matching to unique loci of the 3D7 genome) were derived from previously
uncharacterized open reading frames, demonstrating the use of SAGE in
genome annotation. The open platform "profiling" nature of SAGE
also lead to the important discovery of a novel transcriptional
phenomenon in the malarial pathogen: a significant number of highly
abundant tags that were derived from annotated genes (17%)
corresponded to antisense transcripts. These SAGE data were validated
by two independent means, strand specific reverse
transcription-polymerase chain reaction and Northern analysis, where
antisense messages were detected in both asexual and sexual stages.
This finding has implications for transcriptional regulation of
Plasmodium gene expression.
The Institute for Genomic Research, Rockville, Maryland
20850
These authors contributed equally to this work.
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