Document Detail

Expression profiling of the Leishmania life cycle: cDNA arrays identify developmentally regulated genes present but not annotated in the genome.
MedLine Citation:
PMID:  15138070     Owner:  NLM     Status:  MEDLINE    
As genomic sequencing of Leishmania nears completion, functional analyses that provide a global genetic perspective on biological processes are important. Despite polycistronic transcription, RNA transcript abundance can be measured using microarrays. To provide a resource to evaluate cDNA arrays, we undertook 5' expressed sequence tag analysis of 2183 full-length randomly selected cDNAs from Leishmania major promastigote (days 3, 7, 10 of culture in vitro), and lesion-derived amastigote libraries. PCR-amplified inserts from 1830 of these cDNA representing 1001 unique genes were spotted onto microarrays, and compared internally with PCR-amplified open reading frames (ORFs) from 904 genes representing 842 unique genes annotated in the L. major genome. Microarrays were screened with RNA from procyclic, metacyclic and amastigote populations of L. major. Redundant clones on the array gave highly reproducible results, providing confidence in identification of stage-specific gene expression. Four hundred and thirty unique (i.e. non-redundant) stage-specific genes were identified. A higher percentage of stage-specific gene expression was observed in amastigotes ( approximately 35%) compared to metacyclics ( approximately 12%) for both cDNAs and ORFs, but cDNAs provided a richer source of regulated genes than currently annotated ORFs from the Leishmania genome. In mapping cDNAs onto the Leishmania genome, we noted that approximately 42% aligned to regions not recognised as genes using current predictive annotation tools. These genes are highly represented in our stage-specific genes, and therefore represent important drug targets and vaccine candidates. Careful annotation of cDNAs onto the Leishmania genome will be important before producing the next generation of oligonucleotide arrays based on annotated genes of the genomic sequencing project.
Renata Almeida; Brian J Gilmartin; Sharon H McCann; Alan Norrish; Alasdair C Ivens; Danial Lawson; Mark P Levick; Deborah F Smith; Sabrina D Dyall; David Vetrie; Tom C Freeman; Richard M Coulson; Iracilda Sampaio; Horacio Schneider; Jenefer M Blackwell
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
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:  2011-08-04    
Medline Journal Info:
Nlm Unique ID:  8006324     Medline TA:  Mol Biochem Parasitol     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  87-100     Citation Subset:  IM    
Cambridge Institute for Medical Research, Wellcome Trust/MRC Building, Addenbrooke's Hospital, Hills Road, Cambridge CB2 2XY, UK.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/AA060734;  AA060735;  AA060736;  AA060737;  AA060738;  AA060739;  AA060740;  AA060741;  AA060742;  AA060743;  AA060744;  AA060745;  AA060746;  AA060747;  AA060748;  AA060749;  AA060750;  AA060751;  AA060752;  AA060753;  AA060754;  AA060755;  AA060756;  AA060757;  AA060758;  AA060759;  AA060760;  AA060761;  AA060762;  AA060763;  AA060764;  AA060765;  AA060766;  AA060767;  AA060768;  AA060769;  AA060770;  AA060771;  AA060772;  AA060773;  AA060774;  AA060775;  AA060776;  AA060777;  AA060778;  AA060779;  AA060780;  AA060781;  AA060782;  AA060783;  AA060784
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MeSH Terms
Expressed Sequence Tags
Gene Expression Profiling
Gene Expression Regulation
Leishmania / genetics,  growth & development*,  metabolism
Leishmania major / genetics,  growth & development,  metabolism
Life Cycle Stages
Molecular Sequence Data
Oligonucleotide Array Sequence Analysis
Protozoan Proteins / genetics,  metabolism
Sequence Analysis, DNA
Grant Support
061343//Wellcome Trust
Reg. No./Substance:
0/Protozoan Proteins

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