TY - JOUR
T1 - Plasmodium falciparum
T2 - Differential display analysis of gene expression during gametocytogenesis
AU - Cui, Liwang
AU - Rzomp, Kimberly A.
AU - Fan, Qi
AU - Martin, Samuel K.
AU - Williams, Jackie
N1 - Funding Information:
We thank Drs. Kelli Hoover and Diana Cox-Foster for critically reviewing the manuscript. Malaria sequence data retrieved for reading frame analysis were obtained from the Malaria Genome Consortium: The Sanger Centre (chromosomes 1,3,4,5,6,7,8,9,13), The Institute for Genomic Research-TIGR (chromosomes 2,10,11,14), and The Stanford DNA Sequencing and Technology Center (chromosome 12). This work was supported by a grant from the National Institutes of Health (AI46472) to C.L.
PY - 2001
Y1 - 2001
N2 - With the Plasmodium falciparum genome sequencing near completion, functional analysis of individual parasite genes has become the major task of the postgenomic era. Understanding the expression patterns of individual genes is the initial step toward this goal. In this report, we have examined gene expression during gametocytogenesis of the malaria parasite, P. falciparum, using a modified differential display (DD) method. The modifications of this method include adjusting the dNTP mix, using upstream primers with higher AT contents, and reducing the extension temperature of the polymerase chain reaction (PCR). With a combination of 16 arbitrary upstream primers and 3 one-base-anchored oligo(dT) primers, we have successfully cloned 80 unique cDNA tags from stage IV-V gametocytes. Further analysis by dot blots and semiquantitative reverse transcriptase-PCR showed that at least 49 cDNAs had induced or elevated levels of expression in gametocytes. These results indicate that this modified DD procedure is suitable for large-scale identification of developmentally regulated genes in the AT-rich Plasmodium genome.
AB - With the Plasmodium falciparum genome sequencing near completion, functional analysis of individual parasite genes has become the major task of the postgenomic era. Understanding the expression patterns of individual genes is the initial step toward this goal. In this report, we have examined gene expression during gametocytogenesis of the malaria parasite, P. falciparum, using a modified differential display (DD) method. The modifications of this method include adjusting the dNTP mix, using upstream primers with higher AT contents, and reducing the extension temperature of the polymerase chain reaction (PCR). With a combination of 16 arbitrary upstream primers and 3 one-base-anchored oligo(dT) primers, we have successfully cloned 80 unique cDNA tags from stage IV-V gametocytes. Further analysis by dot blots and semiquantitative reverse transcriptase-PCR showed that at least 49 cDNAs had induced or elevated levels of expression in gametocytes. These results indicate that this modified DD procedure is suitable for large-scale identification of developmentally regulated genes in the AT-rich Plasmodium genome.
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U2 - 10.1006/expr.2001.4669
DO - 10.1006/expr.2001.4669
M3 - Article
C2 - 11888252
AN - SCOPUS:0035727588
SN - 0014-4894
VL - 99
SP - 244
EP - 254
JO - Experimental Parasitology
JF - Experimental Parasitology
IS - 4
ER -