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A Comprehensive Reference Transcriptome Resource for the Common House Spider Parasteatoda tepidariorum
Parasteatoda tepidariorum is an increasingly popular model for the study of spider development and the evolution of development more broadly. However, fully understanding the regulation and evolution of P. tepidariorum development in comparison to other animals requires a genomic perspective. Althou...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4132015/ https://www.ncbi.nlm.nih.gov/pubmed/25118601 http://dx.doi.org/10.1371/journal.pone.0104885 |
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author | Posnien, Nico Zeng, Victor Schwager, Evelyn E. Pechmann, Matthias Hilbrant, Maarten Keefe, Joseph D. Damen, Wim G. M. Prpic, Nikola-Michael McGregor, Alistair P. Extavour, Cassandra G. |
author_facet | Posnien, Nico Zeng, Victor Schwager, Evelyn E. Pechmann, Matthias Hilbrant, Maarten Keefe, Joseph D. Damen, Wim G. M. Prpic, Nikola-Michael McGregor, Alistair P. Extavour, Cassandra G. |
author_sort | Posnien, Nico |
collection | PubMed |
description | Parasteatoda tepidariorum is an increasingly popular model for the study of spider development and the evolution of development more broadly. However, fully understanding the regulation and evolution of P. tepidariorum development in comparison to other animals requires a genomic perspective. Although research on P. tepidariorum has provided major new insights, gene analysis to date has been limited to candidate gene approaches. Furthermore, the few available EST collections are based on embryonic transcripts, which have not been systematically annotated and are unlikely to contain transcripts specific to post-embryonic stages of development. We therefore generated cDNA from pooled embryos representing all described embryonic stages, as well as post-embryonic stages including nymphs, larvae and adults, and using Illumina HiSeq technology obtained a total of 625,076,514 100-bp paired end reads. We combined these data with 24,360 ESTs available in GenBank, and 1,040,006 reads newly generated from 454 pyrosequencing of a mixed-stage embryo cDNA library. The combined sequence data were assembled using a custom de novo assembly strategy designed to optimize assembly product length, number of predicted transcripts, and proportion of raw reads incorporated into the assembly. The de novo assembly generated 446,427 contigs with an N50 of 1,875 bp. These sequences obtained 62,799 unique BLAST hits against the NCBI non-redundant protein data base, including putative orthologs to 8,917 Drosophila melanogaster genes based on best reciprocal BLAST hit identity compared with the D. melanogaster proteome. Finally, we explored the utility of the transcriptome for RNA-Seq studies, and showed that this resource can be used as a mapping scaffold to detect differential gene expression in different cDNA libraries. This resource will therefore provide a platform for future genomic, gene expression and functional approaches using P. tepidariorum. |
format | Online Article Text |
id | pubmed-4132015 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-41320152014-08-19 A Comprehensive Reference Transcriptome Resource for the Common House Spider Parasteatoda tepidariorum Posnien, Nico Zeng, Victor Schwager, Evelyn E. Pechmann, Matthias Hilbrant, Maarten Keefe, Joseph D. Damen, Wim G. M. Prpic, Nikola-Michael McGregor, Alistair P. Extavour, Cassandra G. PLoS One Research Article Parasteatoda tepidariorum is an increasingly popular model for the study of spider development and the evolution of development more broadly. However, fully understanding the regulation and evolution of P. tepidariorum development in comparison to other animals requires a genomic perspective. Although research on P. tepidariorum has provided major new insights, gene analysis to date has been limited to candidate gene approaches. Furthermore, the few available EST collections are based on embryonic transcripts, which have not been systematically annotated and are unlikely to contain transcripts specific to post-embryonic stages of development. We therefore generated cDNA from pooled embryos representing all described embryonic stages, as well as post-embryonic stages including nymphs, larvae and adults, and using Illumina HiSeq technology obtained a total of 625,076,514 100-bp paired end reads. We combined these data with 24,360 ESTs available in GenBank, and 1,040,006 reads newly generated from 454 pyrosequencing of a mixed-stage embryo cDNA library. The combined sequence data were assembled using a custom de novo assembly strategy designed to optimize assembly product length, number of predicted transcripts, and proportion of raw reads incorporated into the assembly. The de novo assembly generated 446,427 contigs with an N50 of 1,875 bp. These sequences obtained 62,799 unique BLAST hits against the NCBI non-redundant protein data base, including putative orthologs to 8,917 Drosophila melanogaster genes based on best reciprocal BLAST hit identity compared with the D. melanogaster proteome. Finally, we explored the utility of the transcriptome for RNA-Seq studies, and showed that this resource can be used as a mapping scaffold to detect differential gene expression in different cDNA libraries. This resource will therefore provide a platform for future genomic, gene expression and functional approaches using P. tepidariorum. Public Library of Science 2014-08-13 /pmc/articles/PMC4132015/ /pubmed/25118601 http://dx.doi.org/10.1371/journal.pone.0104885 Text en © 2014 Posnien et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Posnien, Nico Zeng, Victor Schwager, Evelyn E. Pechmann, Matthias Hilbrant, Maarten Keefe, Joseph D. Damen, Wim G. M. Prpic, Nikola-Michael McGregor, Alistair P. Extavour, Cassandra G. A Comprehensive Reference Transcriptome Resource for the Common House Spider Parasteatoda tepidariorum |
title | A Comprehensive Reference Transcriptome Resource for the Common House Spider Parasteatoda tepidariorum
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title_full | A Comprehensive Reference Transcriptome Resource for the Common House Spider Parasteatoda tepidariorum
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title_fullStr | A Comprehensive Reference Transcriptome Resource for the Common House Spider Parasteatoda tepidariorum
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title_full_unstemmed | A Comprehensive Reference Transcriptome Resource for the Common House Spider Parasteatoda tepidariorum
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title_short | A Comprehensive Reference Transcriptome Resource for the Common House Spider Parasteatoda tepidariorum
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title_sort | comprehensive reference transcriptome resource for the common house spider parasteatoda tepidariorum |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4132015/ https://www.ncbi.nlm.nih.gov/pubmed/25118601 http://dx.doi.org/10.1371/journal.pone.0104885 |
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