Candidate phylum TM6 genome recovered from a hospital sink biofilm provides genomic insights into this uncultivated phylum
Jeffrey S. McLean,Jeffrey S. McLean,Mary-Jane Lombardo,Jonathan H. Badger,Anna Edlund,Mark Novotny,Joyclyn Yee-Greenbaum,Nikolay Vyahhi,Adam P. Hall,Youngik Yang,Christopher L. Dupont,Michael G. Ziegler,Hamidreza Chitsaz,Andrew E. Allen,Shibu Yooseph,Glenn Tesler,Pavel A. Pevzner,Pavel A. Pevzner,Robert Friedman,Kenneth H. Nealson,Kenneth H. Nealson,J. Craig Venter,Roger S. Lasken +22 more
TLDR
An approach for increasing throughput and effectively improving the likelihood of sampling rare events based on forming small random pools of single-flow–sorted cells, amplifying their DNA by multiple displacement amplification and sequencing all cells in the pool, creating a “mini-metagenome” is developed.Abstract:
The “dark matter of life” describes microbes and even entire divisions of bacterial phyla that have evaded cultivation and have yet to be sequenced. We present a genome from the globally distributed but elusive candidate phylum TM6 and uncover its metabolic potential. TM6 was detected in a biofilm from a sink drain within a hospital restroom by analyzing cells using a highly automated single-cell genomics platform. We developed an approach for increasing throughput and effectively improving the likelihood of sampling rare events based on forming small random pools of single-flow–sorted cells, amplifying their DNA by multiple displacement amplification and sequencing all cells in the pool, creating a “mini-metagenome.” A recently developed single-cell assembler, SPAdes, in combination with contig binning methods, allowed the reconstruction of genomes from these mini-metagenomes. A total of 1.07 Mb was recovered in seven contigs for this member of TM6 (JCVI TM6SC1), estimated to represent 90% of its genome. High nucleotide identity between a total of three TM6 genome drafts generated from pools that were independently captured, amplified, and assembled provided strong confirmation of a correct genomic sequence. TM6 is likely a Gram-negative organism and possibly a symbiont of an unknown host (nonfree living) in part based on its small genome, low-GC content, and lack of biosynthesis pathways for most amino acids and vitamins. Phylogenomic analysis of conserved single-copy genes confirms that TM6SC1 is a deeply branching phylum.read more
Citations
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SPAdes, a new genome assembly algorithm and its applications to single-cell sequencing ( 7th Annual SFAF Meeting, 2012)
TL;DR: SPAdes as mentioned in this paper is a new assembler for both single-cell and standard (multicell) assembly, and demonstrate that it improves on the recently released E+V-SC assembler and on popular assemblers Velvet and SoapDeNovo (for multicell data).
Journal ArticleDOI
MetaSPAdes: A new versatile metagenomic assembler
TL;DR: MetaSPAdes as mentioned in this paper addresses various challenges of metagenomic assembly by capitalizing on computational ideas that proved to be useful in assemblies of single cells and highly polymorphic diploid genomes.
Journal ArticleDOI
Assembling Single-Cell Genomes and Mini-Metagenomes From Chimeric MDA Products
Sergey Nurk,Anton Bankevich,Dmitry Antipov,Alexey Gurevich,Anton Korobeynikov,Alla Lapidus,Andrey D. Prjibelski,Alex Pyshkin,Alexander Sirotkin,Yakov Sirotkin,Ramunas Stepanauskas,Scott Clingenpeel,Tanja Woyke,Jeffrey S. McLean,Roger S. Lasken,Glenn Tesler,Max A. Alekseyev,Pavel A. Pevzner +17 more
TL;DR: Applications of the single-cell assembler SPAdes to a new approach for capturing and sequencing "microbial dark matter" that forms small pools of randomly selected single cells and further sequences all genomes from the mini-metagenome at once.
Journal ArticleDOI
Single-cell genome sequencing: current state of the science
TL;DR: An overview of the current state of the field of single-cell genome sequencing is provided, focusing on the technical challenges of making measurements that start from a single molecule of DNA, and how some of these recent methodological advancements have enabled the discovery of unexpected new biology.
Journal ArticleDOI
The First Microbial Colonizers of the Human Gut: Composition, Activities, and Health Implications of the Infant Gut Microbiota
Christian Milani,Sabrina Duranti,Francesca Bottacini,Eoghan Casey,Francesca Turroni,Jennifer Mahony,Clara Belzer,Susana Delgado Palacio,Silvia Arboleya Montes,Leonardo Mancabelli,Gabriele Andrea Lugli,Juan M. Rodríguez,Lars Bode,Willem M. de Vos,Willem M. de Vos,Miguel Gueimonde,Abelardo Margolles,Douwe van Sinderen,Marco Ventura +18 more
TL;DR: The infant microbiota, the mechanisms that drive its establishment and composition, and how microbial consortia may be molded by natural or artificial interventions are described and the relevance of key microbial players of the infant gut microbiota, in particular bifidobacteria, with respect to their role in health and disease are discussed.
References
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Journal ArticleDOI
SPAdes: A New Genome Assembly Algorithm and Its Applications to Single-Cell Sequencing
Anton Bankevich,Sergey Nurk,Dmitry Antipov,Alexey Gurevich,Mikhail Dvorkin,Alexander S. Kulikov,Valery M. Lesin,Sergey I. Nikolenko,Son Pham,Andrey D. Prjibelski,Alexey V. Pyshkin,Alexander Sirotkin,Nikolay Vyahhi,Glenn Tesler,Max A. Alekseyev,Pavel A. Pevzner +15 more
TL;DR: SPAdes generates single-cell assemblies, providing information about genomes of uncultivatable bacteria that vastly exceeds what may be obtained via traditional metagenomics studies.
Journal ArticleDOI
A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood.
Stéphane Guindon,Olivier Gascuel +1 more
TL;DR: This work has used extensive and realistic computer simulations to show that the topological accuracy of this new method is at least as high as that of the existing maximum-likelihood programs and much higher than the performance of distance-based and parsimony approaches.
SPAdes, a new genome assembly algorithm and its applications to single-cell sequencing ( 7th Annual SFAF Meeting, 2012)
TL;DR: SPAdes as mentioned in this paper is a new assembler for both single-cell and standard (multicell) assembly, and demonstrate that it improves on the recently released E+V-SC assembler and on popular assemblers Velvet and SoapDeNovo (for multicell data).
Journal ArticleDOI
The RAST Server: Rapid Annotations using Subsystems Technology
Ramy K. Aziz,Ramy K. Aziz,Daniela Bartels,Aaron A. Best,Matthew DeJongh,Terrence Disz,Terrence Disz,Robert Edwards,Kevin Formsma,Svetlana Gerdes,Elizabeth M. Glass,Michael Kubal,Folker Meyer,Folker Meyer,Gary J. Olsen,Gary J. Olsen,Robert Olson,Robert Olson,Andrei L. Osterman,Ross Overbeek,Leslie Klis McNeil,Daniel Paarmann,Tobias Paczian,Bruce Parrello,Gordon D. Pusch,Claudia I. Reich,Rick Stevens,Rick Stevens,Olga Vassieva,Veronika Vonstein,Andreas Wilke,Olga Zagnitko +31 more
TL;DR: A fully automated service for annotating bacterial and archaeal genomes that identifies protein-encoding, rRNA and tRNA genes, assigns functions to the genes, predicts which subsystems are represented in the genome, uses this information to reconstruct the metabolic network and makes the output easily downloadable for the user.
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