| Expression profiling of the Leishmania life cycle: cDNA arrays identify developmentally regulated genes present but not annotated in the genome. | |
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MedLine Citation:
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PMID: 15138070 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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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:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Molecular and biochemical parasitology Volume: 136 ISSN: 0166-6851 ISO Abbreviation: Mol. Biochem. Parasitol. Publication Date: 2004 Jul |
Date Detail:
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Created Date: 2004-05-12 Completed Date: 2004-07-15 Revised Date: 2011-08-04 |
Medline Journal Info:
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Nlm Unique ID: 8006324 Medline TA: Mol Biochem Parasitol Country: Netherlands |
Other Details:
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Languages: eng Pagination: 87-100 Citation Subset: IM |
Affiliation:
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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.:
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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 |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals 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 | |
ID/Acronym/Agency:
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061343//Wellcome Trust |
| Chemical | |
Reg. No./Substance:
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0/Protozoan Proteins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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