Dataset Information


Synthesis vs. Abundance, Bradyzoite Development

ABSTRACT: Parasites grown as either tachyzoites or bradyzoites for 40 hours were pulsed with 2,4-dithiouracil (DTU) for 1 hour. Following the pulse, mRNA was extracted and either used directly in microarray experiments (Abundance Arrays) or biotinylated and purified to select DTU labeled RNAs that were then used in microarray experiments (Synthesis Arrays). Two independent tachyzoite and bradyzoite preparations were made and duplicate microarrays were performed for each sample. A development or differentiation experiment design type assays events associated with development or differentiation or moving through a life cycle. Development applies to organism(s) acquiring a mature state, and differentiation applies to cells acquiring specialized functions. User Defined

ORGANISM(S): Toxoplasma gondii  

SUBMITTER: unknown Stanford Microarray Database   Michael Cleary 

PROVIDER: E-GEOD-2946 | ArrayExpress | 2005-07-16



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Biosynthetic labeling of RNA with uracil phosphoribosyltransferase allows cell-specific microarray analysis of mRNA synthesis and decay.

Cleary Michael D MD   Meiering Christopher D CD   Jan Eric E   Guymon Rebecca R   Boothroyd John C JC  

Nature biotechnology 20050130 2

Standard microarrays measure mRNA abundance, not mRNA synthesis, and therefore cannot identify the mechanisms that regulate gene expression. We have developed a method to overcome this limitation by using the salvage enzyme uracil phosphoribosyltransferase (UPRT) from the protozoan Toxoplasma gondii. T. gondii UPRT has been well characterized because of its application in monitoring parasite growth: mammals lack this enzyme activity and thus only the parasite incorporates (3)H-uracil into its nu  ...[more]

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