Artificial polyploidy improves bacterial single cell genome recovery.

<h4>Background</h4>Single cell genomics (SCG) is a combination of methods whose goal is to decipher the complete genomic sequence from a single cell and has been applied mostly to organisms with smaller genomes, such as bacteria and archaea. Prior single cell studies showed that a signif...

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Autores principales:	Armand E K Dichosa, Michael S Fitzsimons, Chien-Chi Lo, Lea L Weston, Lara G Preteska, Jeremy P Snook, Xiaojing Zhang, Wei Gu, Kim McMurry, Lance D Green, Patrick S Chain, J Chris Detter, Cliff S Han
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/5ef31ebcf6a94a998d3b3ec3b3693318
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spelling	oai:doaj.org-article:5ef31ebcf6a94a998d3b3ec3b36933182021-11-18T07:17:46ZArtificial polyploidy improves bacterial single cell genome recovery.1932-620310.1371/journal.pone.0037387https://doaj.org/article/5ef31ebcf6a94a998d3b3ec3b36933182012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22666352/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Single cell genomics (SCG) is a combination of methods whose goal is to decipher the complete genomic sequence from a single cell and has been applied mostly to organisms with smaller genomes, such as bacteria and archaea. Prior single cell studies showed that a significant portion of a genome could be obtained. However, breakages of genomic DNA and amplification bias have made it very challenging to acquire a complete genome with single cells. We investigated an artificial method to induce polyploidy in Bacillus subtilis ATCC 6633 by blocking cell division and have shown that we can significantly improve the performance of genomic sequencing from a single cell.<h4>Methodology/principal findings</h4>We inhibited the bacterial cytoskeleton protein FtsZ in B.subtilis with an FtsZ-inhibiting compound, PC190723, resulting in larger undivided single cells with multiple copies of its genome. qPCR assays of these larger, sorted cells showed higher DNA content, have less amplification bias, and greater genomic recovery than untreated cells.<h4>Significance</h4>The method presented here shows the potential to obtain a nearly complete genome sequence from a single bacterial cell. With millions of uncultured bacterial species in nature, this method holds tremendous promise to provide insight into the genomic novelty of yet-to-be discovered species, and given the temporary effects of artificial polyploidy coupled with the ability to sort and distinguish differences in cell size and genomic DNA content, may allow recovery of specific organisms in addition to their genomes.Armand E K DichosaMichael S FitzsimonsChien-Chi LoLea L WestonLara G PreteskaJeremy P SnookXiaojing ZhangWei GuKim McMurryLance D GreenPatrick S ChainJ Chris DetterCliff S HanPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 5, p e37387 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Armand E K Dichosa Michael S Fitzsimons Chien-Chi Lo Lea L Weston Lara G Preteska Jeremy P Snook Xiaojing Zhang Wei Gu Kim McMurry Lance D Green Patrick S Chain J Chris Detter Cliff S Han Artificial polyploidy improves bacterial single cell genome recovery.
description	<h4>Background</h4>Single cell genomics (SCG) is a combination of methods whose goal is to decipher the complete genomic sequence from a single cell and has been applied mostly to organisms with smaller genomes, such as bacteria and archaea. Prior single cell studies showed that a significant portion of a genome could be obtained. However, breakages of genomic DNA and amplification bias have made it very challenging to acquire a complete genome with single cells. We investigated an artificial method to induce polyploidy in Bacillus subtilis ATCC 6633 by blocking cell division and have shown that we can significantly improve the performance of genomic sequencing from a single cell.<h4>Methodology/principal findings</h4>We inhibited the bacterial cytoskeleton protein FtsZ in B.subtilis with an FtsZ-inhibiting compound, PC190723, resulting in larger undivided single cells with multiple copies of its genome. qPCR assays of these larger, sorted cells showed higher DNA content, have less amplification bias, and greater genomic recovery than untreated cells.<h4>Significance</h4>The method presented here shows the potential to obtain a nearly complete genome sequence from a single bacterial cell. With millions of uncultured bacterial species in nature, this method holds tremendous promise to provide insight into the genomic novelty of yet-to-be discovered species, and given the temporary effects of artificial polyploidy coupled with the ability to sort and distinguish differences in cell size and genomic DNA content, may allow recovery of specific organisms in addition to their genomes.
format	article
author	Armand E K Dichosa Michael S Fitzsimons Chien-Chi Lo Lea L Weston Lara G Preteska Jeremy P Snook Xiaojing Zhang Wei Gu Kim McMurry Lance D Green Patrick S Chain J Chris Detter Cliff S Han
author_facet	Armand E K Dichosa Michael S Fitzsimons Chien-Chi Lo Lea L Weston Lara G Preteska Jeremy P Snook Xiaojing Zhang Wei Gu Kim McMurry Lance D Green Patrick S Chain J Chris Detter Cliff S Han
author_sort	Armand E K Dichosa
title	Artificial polyploidy improves bacterial single cell genome recovery.
title_short	Artificial polyploidy improves bacterial single cell genome recovery.
title_full	Artificial polyploidy improves bacterial single cell genome recovery.
title_fullStr	Artificial polyploidy improves bacterial single cell genome recovery.
title_full_unstemmed	Artificial polyploidy improves bacterial single cell genome recovery.
title_sort	artificial polyploidy improves bacterial single cell genome recovery.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/5ef31ebcf6a94a998d3b3ec3b3693318
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Artificial polyploidy improves bacterial single cell genome recovery.

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