Whole-genome <it>in-silico </it>subtractive hybridization (WISH) - using massive sequencing for the identification of unique and repetitive sex-specific sequences: the example of <it>Schistosoma mansoni</it>

<p>Abstract</p> <p>Background</p> <p>Emerging methods of massive sequencing that allow for rapid re-sequencing of entire genomes at comparably low cost are changing the way biological questions are addressed in many domains. Here we propose a novel method to compare two...

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Main Authors: Parrinello Hugues, Dantec Christelle, Beltran Sophie, Cosseau Céline, Grunau Christoph, Portela Julien, Boissier Jérôme
Format: Article
Language:English
Published: BMC 2010-06-01
Series:BMC Genomics
Online Access:http://www.biomedcentral.com/1471-2164/11/387
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author Parrinello Hugues
Dantec Christelle
Beltran Sophie
Cosseau Céline
Grunau Christoph
Portela Julien
Boissier Jérôme
author_facet Parrinello Hugues
Dantec Christelle
Beltran Sophie
Cosseau Céline
Grunau Christoph
Portela Julien
Boissier Jérôme
author_sort Parrinello Hugues
collection DOAJ
description <p>Abstract</p> <p>Background</p> <p>Emerging methods of massive sequencing that allow for rapid re-sequencing of entire genomes at comparably low cost are changing the way biological questions are addressed in many domains. Here we propose a novel method to compare two genomes (genome-to-genome comparison). We used this method to identify sex-specific sequences of the human blood fluke <it>Schistosoma mansoni</it>.</p> <p>Results</p> <p>Genomic DNA was extracted from male and female (heterogametic) <it>S. mansoni </it>adults and sequenced with a Genome Analyzer (Illumina). Sequences are available at the NCBI sequence read archive <url>http://www.ncbi.nlm.nih.gov/Traces/sra/</url> under study accession number SRA012151.6. Sequencing reads were aligned to the genome, and a pseudogenome composed of known repeats. Straightforward comparative bioinformatics analysis was performed to compare male and female schistosome genomes and identify female-specific sequences. We found that the <it>S. mansoni </it>female W chromosome contains only few specific unique sequences (950 Kb i.e. about 0.2% of the genome). The majority of W-specific sequences are repeats (10.5 Mb i.e. about 2.5% of the genome). Arbitrarily selected W-specific sequences were confirmed by PCR. Primers designed for unique and repetitive sequences allowed to reliably identify the sex of both larval and adult stages of the parasite.</p> <p>Conclusion</p> <p>Our genome-to-genome comparison method that we call "whole-genome <it>in-silico </it>subtractive hybridization" (WISH) allows for rapid identification of sequences that are specific for a certain genotype (e.g. the heterogametic sex). It can in principle be used for the detection of any sequence differences between isolates (<it>e.g</it>. strains, pathovars) or even closely related species.</p>
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spelling doaj.art-1dea0745c9d3453eb0153ad36c553fd12022-12-21T19:11:41ZengBMCBMC Genomics1471-21642010-06-0111138710.1186/1471-2164-11-387Whole-genome <it>in-silico </it>subtractive hybridization (WISH) - using massive sequencing for the identification of unique and repetitive sex-specific sequences: the example of <it>Schistosoma mansoni</it>Parrinello HuguesDantec ChristelleBeltran SophieCosseau CélineGrunau ChristophPortela JulienBoissier Jérôme<p>Abstract</p> <p>Background</p> <p>Emerging methods of massive sequencing that allow for rapid re-sequencing of entire genomes at comparably low cost are changing the way biological questions are addressed in many domains. Here we propose a novel method to compare two genomes (genome-to-genome comparison). We used this method to identify sex-specific sequences of the human blood fluke <it>Schistosoma mansoni</it>.</p> <p>Results</p> <p>Genomic DNA was extracted from male and female (heterogametic) <it>S. mansoni </it>adults and sequenced with a Genome Analyzer (Illumina). Sequences are available at the NCBI sequence read archive <url>http://www.ncbi.nlm.nih.gov/Traces/sra/</url> under study accession number SRA012151.6. Sequencing reads were aligned to the genome, and a pseudogenome composed of known repeats. Straightforward comparative bioinformatics analysis was performed to compare male and female schistosome genomes and identify female-specific sequences. We found that the <it>S. mansoni </it>female W chromosome contains only few specific unique sequences (950 Kb i.e. about 0.2% of the genome). The majority of W-specific sequences are repeats (10.5 Mb i.e. about 2.5% of the genome). Arbitrarily selected W-specific sequences were confirmed by PCR. Primers designed for unique and repetitive sequences allowed to reliably identify the sex of both larval and adult stages of the parasite.</p> <p>Conclusion</p> <p>Our genome-to-genome comparison method that we call "whole-genome <it>in-silico </it>subtractive hybridization" (WISH) allows for rapid identification of sequences that are specific for a certain genotype (e.g. the heterogametic sex). It can in principle be used for the detection of any sequence differences between isolates (<it>e.g</it>. strains, pathovars) or even closely related species.</p>http://www.biomedcentral.com/1471-2164/11/387
spellingShingle Parrinello Hugues
Dantec Christelle
Beltran Sophie
Cosseau Céline
Grunau Christoph
Portela Julien
Boissier Jérôme
Whole-genome <it>in-silico </it>subtractive hybridization (WISH) - using massive sequencing for the identification of unique and repetitive sex-specific sequences: the example of <it>Schistosoma mansoni</it>
BMC Genomics
title Whole-genome <it>in-silico </it>subtractive hybridization (WISH) - using massive sequencing for the identification of unique and repetitive sex-specific sequences: the example of <it>Schistosoma mansoni</it>
title_full Whole-genome <it>in-silico </it>subtractive hybridization (WISH) - using massive sequencing for the identification of unique and repetitive sex-specific sequences: the example of <it>Schistosoma mansoni</it>
title_fullStr Whole-genome <it>in-silico </it>subtractive hybridization (WISH) - using massive sequencing for the identification of unique and repetitive sex-specific sequences: the example of <it>Schistosoma mansoni</it>
title_full_unstemmed Whole-genome <it>in-silico </it>subtractive hybridization (WISH) - using massive sequencing for the identification of unique and repetitive sex-specific sequences: the example of <it>Schistosoma mansoni</it>
title_short Whole-genome <it>in-silico </it>subtractive hybridization (WISH) - using massive sequencing for the identification of unique and repetitive sex-specific sequences: the example of <it>Schistosoma mansoni</it>
title_sort whole genome it in silico it subtractive hybridization wish using massive sequencing for the identification of unique and repetitive sex specific sequences the example of it schistosoma mansoni it
url http://www.biomedcentral.com/1471-2164/11/387
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