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

BMC Genomics

Methodology Article

Hugues Parrinello¹  Christelle Dantec¹  Jérôme Boissier²  Christoph Grunau²  Julien Portela²  Céline Cosseau²  Sophie Beltran² 

[1] Plateforme MGX, Institut de Génomique Fonctionnelle, 41, rue de la Cardonille, Montpellier, France;UMR 5244 CNRS-EPHE-UPVD. Parasitologie Fonctionnelle et Evolutive, CBETM. Université de Perpignan, Perpignan, France;

关键词: Schistosomiasis;    Schistosoma Mansoni;    Massive Parallel Sequencing;    NCBI Sequence Read Archive;    Tandem Repeat Finder;   

DOI  :  10.1186/1471-2164-11-387

 received in 2010-02-22, accepted in 2010-06-21,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundEmerging 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 Schistosoma mansoni.ResultsGenomic DNA was extracted from male and female (heterogametic) S. mansoni adults and sequenced with a Genome Analyzer (Illumina). Sequences are available at the NCBI sequence read archive http://www.ncbi.nlm.nih.gov/Traces/sra/ 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 S. mansoni 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.ConclusionOur genome-to-genome comparison method that we call "whole-genome in-silico 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 (e.g. strains, pathovars) or even closely related species.

【 授权许可】

Unknown   
© Portela et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.