Document Detail

Expression profiling using random genomic DNA microarrays identifies differentially expressed genes associated with three major developmental stages of the protozoan parasite Leishmania major.
MedLine Citation:
PMID:  15138069     Owner:  NLM     Status:  MEDLINE    
To complete its life cycle, protozoan parasites of the genus Leishmania undergo at least three major developmental transitions. However, previous efforts to identify genes showing stage regulated changes in transcript abundance have yielded relatively few. Here we used expression profiling to assess changes in transcript abundance in three stages: replicating promastigotes and infective non-replicating metacyclics, which occur in the sand fly vector, and in the amastigote stage residing with macrophage phagolysosomes in mammals. Microarrays were developed containing 11,484 PCR products that included a number of known genes and 10,464 random 1 kb genomic DNA fragments. Arrays were hybridized in triplicate and genes showing two-fold or greater changes in 2/3 experiments were scored as differentially expressed. Remarkably, only about one percent of the DNAs expression varied by this criteria, in either stage comparison. Northern blot analysis confirmed the predicted change in mRNA abundance for most of these (68%). This set of genes included most of those previously identified in the literature as differentially regulated as well as a number of novel genes. Notably, Leishmania maxicircle transcripts showed strong up-regulation in metacyclic and amastigote parasites, probably associated with changes in parasite energy metabolism. However, current data suggest that expression profiling using shotgun DNA libraries significantly underestimates the extent of regulated transcripts.
Natalia S Akopyants; Robin S Matlib; Elena N Bukanova; Matthew R Smeds; Bernard H Brownstein; Gary D Stormo; Stephen M Beverley
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Molecular and biochemical parasitology     Volume:  136     ISSN:  0166-6851     ISO Abbreviation:  Mol. Biochem. Parasitol.     Publication Date:  2004 Jul 
Date Detail:
Created Date:  2004-05-12     Completed Date:  2004-07-15     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  8006324     Medline TA:  Mol Biochem Parasitol     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  71-86     Citation Subset:  IM    
Department of Molecular Microbiology, Center for Infectious Disease Research, Washington University School of Medicine, Campus Box 8230, 660 South Euclid Avenue, Saint Louis, MO 63110, USA.
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MeSH Terms
Gene Expression Profiling*
Gene Expression Regulation*
Genome, Protozoan*
Leishmania major / genetics,  growth & development*,  metabolism
Leishmaniasis, Cutaneous / parasitology
Life Cycle Stages
Mice, Inbred BALB C
Oligonucleotide Array Sequence Analysis / methods*
Protozoan Proteins / genetics,  metabolism*
Transcription, Genetic
Grant Support
Reg. No./Substance:
0/Protozoan Proteins

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