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Identification of differentially expressed cDNAs in Acanthamoeba culbertsoni after mouse brain passage

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Abstract

Free-living amoebae of the genus Acanthamoeba are causative agents of granulomatous amebicencephalitis and amebic keratitis. Because the virulence of Acanthamoeba culbertsoni cultured in the laboratory isrestored by consecutive brain passages, we examined the genes induced in mouse brain-passaged A. culbertsonibydifferential display reverse transcriptase polymerase chain reaction (DDRT-PCR). Enhanced A. culbertsonivirulencewas observed during the second mouse brain passage, i.e., infected mouse mortality increased from 5% to 70%. TencDNAs induced during mouse brain passage were identified by DDRT-PCR and this was confirmed by northern blotanalysis. BlastX searches of these cDNAs indicated the upregulations of genes encoding predictive NADH-dehydro-genase, proteasomal ATPase, and GDP-mannose pyrophosphorylase B, which have previously been reported to beassociated with A. culbertsonivirulence factors.

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