Identification of differentially expressed cDNAs in Acanthamoeba culbertsoni after mouse brain passage.
Free-living amoebae of the genus Acanthamoeba are causative agents of granulomatous amebic encephalitis and amebic keratitis. Because the virulence of Acanthamoeba culbertsoni cultured in the laboratory is restored by consecutive brain passages, we examined the genes induced in mouse brain-passaged A. culbertsoni by differential display reverse transcriptase polymerase chain reaction (DDRT-PCR). Enhanced A. culbertsoni virulence was observed during the second mouse brain passage, i.e., infected mouse mortality increased from 5% to 70%. Ten cDNAs induced during mouse brain passage were identified by DDRT-PCR and this was confirmed by northern blot analysis. BlastX searches of these cDNAs indicated the upregulations of genes encoding predictive NADH-dehydrogenase, proteasomal ATPase, and GDP-mannose pyrophosphorylase B, which have previously been reported to be associated with A. culbertsoni virulence factors.
Han
Kyu-Lee
KL
Department of Parasitology and Institute of Tropical Medicine, Yonsei University College of Medicine, Seoul, Korea.
Lee
Jongweon
J
Kim
Don-Soo
DS
Park
Soon-Jung
SJ
Im
Kyung-il
KI
Yong
Tai-Soon
TS
D000048:Acanthamoeba / Q000235:genetics* / Q000472:pathogenicity*; D000562:Amebiasis / Q000401:mortality / Q000469:parasitology*; D000818:Animals; D015152:Blotting, Northern / Q000379:methods; D001921:Brain / Q000469:parasitology; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000096:biosynthesis; D016054:DNA, Protozoan / Q000096:biosynthesis / Q000502:physiology*; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation; D017125:Genes, Protozoan / Q000235:genetics; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012692:Serial Passage; D015854:Up-Regulation; D014774:Virulence / Q000235:genetics
D018076:DNA, Complementary; D016054:DNA, Protozoan