Fine mapping of the pond snail left-right asymmetry (chirality) locus using RAD-Seq and fibre-FISH.

The left-right asymmetry of snails, including the direction of shell coiling, is determined by the delayed effect of a maternal gene on the chiral twist that takes place during early embryonic cell divisions. Yet, despite being a well-established classical problem, the identity of the gene and the m...

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Autores principales:	Mengning Maureen Liu, John W Davey, Ruby Banerjee, Jie Han, Fengtang Yang, Aziz Aboobaker, Mark L Blaxter, Angus Davison
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/858e1e88af104cc7abc2e0d653b8b83d
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spelling	oai:doaj.org-article:858e1e88af104cc7abc2e0d653b8b83d2021-11-18T09:00:13ZFine mapping of the pond snail left-right asymmetry (chirality) locus using RAD-Seq and fibre-FISH.1932-620310.1371/journal.pone.0071067https://doaj.org/article/858e1e88af104cc7abc2e0d653b8b83d2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23951082/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203The left-right asymmetry of snails, including the direction of shell coiling, is determined by the delayed effect of a maternal gene on the chiral twist that takes place during early embryonic cell divisions. Yet, despite being a well-established classical problem, the identity of the gene and the means by which left-right asymmetry is established in snails remain unknown. We here demonstrate the power of new genomic approaches for identification of the chirality gene, "D". First, heterozygous (Dd) pond snails Lymnaea stagnalis were self-fertilised or backcrossed, and the genotype of more than six thousand offspring inferred, either dextral (DD/Dd) or sinistral (dd). Then, twenty of the offspring were used for Restriction-site-Associated DNA Sequencing (RAD-Seq) to identify anonymous molecular markers that are linked to the chirality locus. A local genetic map was constructed by genotyping three flanking markers in over three thousand snails. The three markers lie either side of the chirality locus, with one very tightly linked (<0.1 cM). Finally, bacterial artificial chromosomes (BACs) were isolated that contained the three loci. Fluorescent in situ hybridization (FISH) of pachytene cells showed that the three BACs tightly cluster on the same bivalent chromosome. Fibre-FISH identified a region of greater that ∼0.4 Mb between two BAC clone markers that must contain D. This work therefore establishes the resources for molecular identification of the chirality gene and the variation that underpins sinistral and dextral coiling. More generally, the results also show that combining genomic technologies, such as RAD-Seq and high resolution FISH, is a robust approach for mapping key loci in non-model systems.Mengning Maureen LiuJohn W DaveyRuby BanerjeeJie HanFengtang YangAziz AboobakerMark L BlaxterAngus DavisonPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 8, p e71067 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Mengning Maureen Liu John W Davey Ruby Banerjee Jie Han Fengtang Yang Aziz Aboobaker Mark L Blaxter Angus Davison Fine mapping of the pond snail left-right asymmetry (chirality) locus using RAD-Seq and fibre-FISH.
description	The left-right asymmetry of snails, including the direction of shell coiling, is determined by the delayed effect of a maternal gene on the chiral twist that takes place during early embryonic cell divisions. Yet, despite being a well-established classical problem, the identity of the gene and the means by which left-right asymmetry is established in snails remain unknown. We here demonstrate the power of new genomic approaches for identification of the chirality gene, "D". First, heterozygous (Dd) pond snails Lymnaea stagnalis were self-fertilised or backcrossed, and the genotype of more than six thousand offspring inferred, either dextral (DD/Dd) or sinistral (dd). Then, twenty of the offspring were used for Restriction-site-Associated DNA Sequencing (RAD-Seq) to identify anonymous molecular markers that are linked to the chirality locus. A local genetic map was constructed by genotyping three flanking markers in over three thousand snails. The three markers lie either side of the chirality locus, with one very tightly linked (<0.1 cM). Finally, bacterial artificial chromosomes (BACs) were isolated that contained the three loci. Fluorescent in situ hybridization (FISH) of pachytene cells showed that the three BACs tightly cluster on the same bivalent chromosome. Fibre-FISH identified a region of greater that ∼0.4 Mb between two BAC clone markers that must contain D. This work therefore establishes the resources for molecular identification of the chirality gene and the variation that underpins sinistral and dextral coiling. More generally, the results also show that combining genomic technologies, such as RAD-Seq and high resolution FISH, is a robust approach for mapping key loci in non-model systems.
format	article
author	Mengning Maureen Liu John W Davey Ruby Banerjee Jie Han Fengtang Yang Aziz Aboobaker Mark L Blaxter Angus Davison
author_facet	Mengning Maureen Liu John W Davey Ruby Banerjee Jie Han Fengtang Yang Aziz Aboobaker Mark L Blaxter Angus Davison
author_sort	Mengning Maureen Liu
title	Fine mapping of the pond snail left-right asymmetry (chirality) locus using RAD-Seq and fibre-FISH.
title_short	Fine mapping of the pond snail left-right asymmetry (chirality) locus using RAD-Seq and fibre-FISH.
title_full	Fine mapping of the pond snail left-right asymmetry (chirality) locus using RAD-Seq and fibre-FISH.
title_fullStr	Fine mapping of the pond snail left-right asymmetry (chirality) locus using RAD-Seq and fibre-FISH.
title_full_unstemmed	Fine mapping of the pond snail left-right asymmetry (chirality) locus using RAD-Seq and fibre-FISH.
title_sort	fine mapping of the pond snail left-right asymmetry (chirality) locus using rad-seq and fibre-fish.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/858e1e88af104cc7abc2e0d653b8b83d
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Fine mapping of the pond snail left-right asymmetry (chirality) locus using RAD-Seq and fibre-FISH.

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