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2.
World J Biol Chem ; 5(2): 161-8, 2014 May 26.
Artigo em Inglês | MEDLINE | ID: mdl-24921006

RESUMO

Next-generation sequencing (NGS) technologies have made high-throughput sequencing available to medium- and small-size laboratories, culminating in a tidal wave of genomic information. The quantity of sequenced bacterial genomes has not only brought excitement to the field of genomics but also heightened expectations that NGS would boost antibacterial discovery and vaccine development. Although many possible drug and vaccine targets have been discovered, the success rate of genome-based analysis has remained below expectations. Furthermore, NGS has had consequences for genome quality, resulting in an exponential increase in draft (partial data) genome deposits in public databases. If no further interests are expressed for a particular bacterial genome, it is more likely that the sequencing of its genome will be limited to a draft stage, and the painstaking tasks of completing the sequencing of its genome and annotation will not be undertaken. It is important to know what is lost when we settle for a draft genome and to determine the "scientific value" of a newly sequenced genome. This review addresses the expected impact of newly sequenced genomes on antibacterial discovery and vaccinology. Also, it discusses the factors that could be leading to the increase in the number of draft deposits and the consequent loss of relevant biological information.

3.
PLoS One ; 8(1): e53818, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23342011

RESUMO

Corynebacterium pseudotuberculosis is a facultative intracellular pathogen and the causative agent of several infectious and contagious chronic diseases, including caseous lymphadenitis, ulcerative lymphangitis, mastitis, and edematous skin disease, in a broad spectrum of hosts. In addition, Corynebacterium pseudotuberculosis infections pose a rising worldwide economic problem in ruminants. The complete genome sequences of 15 C. pseudotuberculosis strains isolated from different hosts and countries were comparatively analyzed using a pan-genomic strategy. Phylogenomic, pan-genomic, core genomic, and singleton analyses revealed close relationships among pathogenic corynebacteria, the clonal-like behavior of C. pseudotuberculosis and slow increases in the sizes of pan-genomes. According to extrapolations based on the pan-genomes, core genomes and singletons, the C. pseudotuberculosis biovar ovis shows a more clonal-like behavior than the C. pseudotuberculosis biovar equi. Most of the variable genes of the biovar ovis strains were acquired in a block through horizontal gene transfer and are highly conserved, whereas the biovar equi strains contain great variability, both intra- and inter-biovar, in the 16 detected pathogenicity islands (PAIs). With respect to the gene content of the PAIs, the most interesting finding is the high similarity of the pilus genes in the biovar ovis strains compared with the great variability of these genes in the biovar equi strains. Concluding, the polymerization of complete pilus structures in biovar ovis could be responsible for a remarkable ability of these strains to spread throughout host tissues and penetrate cells to live intracellularly, in contrast with the biovar equi, which rarely attacks visceral organs. Intracellularly, the biovar ovis strains are expected to have less contact with other organisms than the biovar equi strains, thereby explaining the significant clonal-like behavior of the biovar ovis strains.


Assuntos
Corynebacterium/genética , Genoma Bacteriano/genética , Animais , Deleção de Genes , Genes Bacterianos/genética , Variação Genética , Ilhas Genômicas/genética , Família Multigênica/genética , Especificidade da Espécie , Fatores de Virulência/genética
4.
Microb Biotechnol ; 6(2): 150-6, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23199210

RESUMO

New sequencing platforms have enabled rapid decoding of complete prokaryotic genomes at relatively low cost. The Ion Torrent platform is an example of these technologies, characterized by lower coverage, generating challenges for the genome assembly. One particular problem is the lack of genomes that enable reference-based assembly, such as the one used in the present study, Corynebacterium pseudotuberculosis biovar equi, which causes high economic losses in the US equine industry. The quality treatment strategy incorporated into the assembly pipeline enabled a 16-fold greater use of the sequencing data obtained compared with traditional quality filter approaches. Data preprocessing prior to the de novo assembly enabled the use of known methodologies in the next-generation sequencing data assembly. Moreover, manual curation was proved to be essential for ensuring a quality assembly, which was validated by comparative genomics with other species of the genus Corynebacterium. The present study presents a modus operandi that enables a greater and better use of data obtained from semiconductor sequencing for obtaining the complete genome from a prokaryotic microorganism, C. pseudotuberculosis, which is not a traditional biological model such as Escherichia coli.


Assuntos
Biologia Computacional/métodos , Corynebacterium pseudotuberculosis/genética , Genoma Bacteriano/genética , Semicondutores , Análise de Sequência de DNA , Animais , Infecções por Corynebacterium/microbiologia , Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/isolamento & purificação , DNA Bacteriano/análise , Técnicas Eletroquímicas/instrumentação , Técnicas Eletroquímicas/métodos , Desenho de Equipamento , Genômica/métodos , Doenças dos Cavalos/microbiologia , Cavalos , Análise de Sequência de DNA/instrumentação , Análise de Sequência de DNA/métodos , Software
5.
J Bacteriol ; 194(23): 6620-1, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23144380

RESUMO

The bacterium Corynebacterium pseudotuberculosis is of major veterinary importance because it affects livestock, particularly sheep, goats, and horses, in several countries, including Australia, Brazil, the United States, and Canada, resulting in significant economic losses. In the present study, we describe the complete genome of the Corynebacterium pseudotuberculosis Cp316 strain, biovar equi, isolated from the abscess of a North American horse.


Assuntos
Corynebacterium pseudotuberculosis/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Genoma Bacteriano , Análise de Sequência de DNA , Abscesso/microbiologia , Abscesso/veterinária , Animais , California , Infecções por Corynebacterium/microbiologia , Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/isolamento & purificação , Doenças dos Cavalos/microbiologia , Cavalos , Dados de Sequência Molecular
6.
J Bacteriol ; 194(23): 6663-4, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23144408

RESUMO

Corynebacterium pseudotuberculosis is of major veterinary importance because it affects many animal species, causing economically significant livestock diseases and losses. Therefore, the genomic sequencing of various lines of this organism, isolated from different hosts, will aid in the development of diagnostic methods and new prevention and treatment strategies and improve our knowledge of the biology of this microorganism. In this study, we present the genome of C. pseudotuberculosis Cp31, isolated from a buffalo in Egypt.


Assuntos
Corynebacterium pseudotuberculosis/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Genoma Bacteriano , Análise de Sequência de DNA , Animais , Búfalos/microbiologia , Corynebacterium pseudotuberculosis/isolamento & purificação , Egito , Dados de Sequência Molecular
7.
J Bacteriol ; 194(23): 6689-90, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23144424

RESUMO

Exiguobacterium antarcticum is a psychotropic bacterium isolated for the first time from microbial mats of Lake Fryxell in Antarctica. Many organisms of the genus Exiguobacterium are extremophiles and have properties of biotechnological interest, e.g., the capacity to adapt to cold, which make this genus a target for discovering new enzymes, such as lipases and proteases, in addition to improving our understanding of the mechanisms of adaptation and survival at low temperatures. This study presents the genome of E. antarcticum B7, isolated from a biofilm sample of Ginger Lake on King George Island, Antarctic peninsula.


Assuntos
Bacillales/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Genoma Bacteriano , Análise de Sequência de DNA , Regiões Antárticas , Bacillales/isolamento & purificação , Bacillales/fisiologia , Biofilmes/crescimento & desenvolvimento , Água Doce/microbiologia , Ilhas , Lagos , Dados de Sequência Molecular
8.
BMC Genomics ; 13 Suppl 5: S6, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23095951

RESUMO

BACKGROUND: Pan-genomic studies aim, for instance, at defining the core, dispensable and unique genes within a species. A pan-genomics study for vaccine design tries to assess the best candidates for a vaccine against a specific pathogen. In this context, rather than studying genes predicted to be exported in a single genome, with pan-genomics it is possible to study genes present in different strains within the same species, such as virulence factors. The target organism of this pan-genomic work here presented is Corynebacterium pseudotuberculosis, the etiologic agent of caseous lymphadenitis (CLA) in goat and sheep, which causes significant economic losses in those herds around the world. Currently, only a few antigens against CLA are known as being the basis of commercial and still ineffective vaccines. In this regard, the here presented work analyses, in silico, five C. pseudotuberculosis genomes and gathers data to predict common exported proteins in all five genomes. These candidates were also compared to two recent C. pseudotuberculosis in vitro exoproteome results. RESULTS: The complete genome of five C. pseudotuberculosis strains (1002, C231, I19, FRC41 and PAT10) were submitted to pan-genomics analysis, yielding 306, 59 and 12 gene sets, respectively, representing the core, dispensable and unique in silico predicted exported pan-genomes. These sets bear 150 genes classified as secreted (SEC) and 227 as potentially surface exposed (PSE). Our findings suggest that the main C. pseudotuberculosis in vitro exoproteome could be greater, appended by a fraction of the 35 proteins formerly predicted as making part of the variant in vitro exoproteome. These genomes were manually curated for correct methionine initiation and redeposited with a total of 1885 homogenized genes. CONCLUSIONS: The in silico prediction of exported proteins has allowed to define a list of putative vaccine candidate genes present in all five complete C. pseudotuberculosis genomes. Moreover, it has also been possible to define the in silico predicted dispensable and unique C. pseudotuberculosis exported proteins. These results provide in silico evidence to further guide experiments in the areas of vaccines, diagnosis and drugs. The work here presented is the first whole C. pseudotuberculosis in silico predicted pan-exoproteome completed till today.


Assuntos
Corynebacterium pseudotuberculosis/genética , Genes/genética , Genoma Bacteriano/genética , Genômica/métodos , Proteoma/genética , Vacinas Bacterianas/genética , Proteínas de Membrana/genética , Software
9.
J Bacteriol ; 194(20): 5718-9, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23012291

RESUMO

Corynebacterium pseudotuberculosis is a pathogen of great veterinary and economic importance, since it affects livestock, mainly sheep and goats, worldwide, together with reports of its presence in camels in several Arabic, Asiatic, and East and West African countries, as well as Australia. In this article, we report the genome sequence of Corynebacterium pseudotuberculosis strain Cp162, collected from the external neck abscess of a camel in the United Kingdom.


Assuntos
Corynebacterium pseudotuberculosis/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Genoma Bacteriano , Análise de Sequência de DNA , Abscesso/microbiologia , Abscesso/veterinária , Animais , Camelus , Infecções por Corynebacterium/microbiologia , Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/isolamento & purificação , Dados de Sequência Molecular , Reino Unido
10.
Gene ; 508(2): 145-56, 2012 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-22890137

RESUMO

The genus Campylobacter contains pathogens causing a wide range of diseases, targeting both humans and animals. Among them, the Campylobacter fetus subspecies fetus and venerealis deserve special attention, as they are the etiological agents of human bacterial gastroenteritis and bovine genital campylobacteriosis, respectively. We compare the whole genomes of both subspecies to get insights into genomic architecture, phylogenetic relationships, genome conservation and core virulence factors. Pan-genomic approach was applied to identify the core- and pan-genome for both C. fetus subspecies and members of the genus. The C. fetus subspecies conserved (76%) proteome were then analyzed for their subcellular localization and protein functions in biological processes. Furthermore, with pathogenomic strategies, unique candidate regions in the genomes and several potential core-virulence factors were identified. The potential candidate factors identified for attenuation and/or subunit vaccine development against C. fetus subspecies contain: nucleoside diphosphate kinase (Ndk), type IV secretion systems (T4SS), outer membrane proteins (OMP), substrate binding proteins CjaA and CjaC, surface array proteins, sap gene, and cytolethal distending toxin (CDT). Significantly, many of those genes were found in genomic regions with signals of horizontal gene transfer and, therefore, predicted as putative pathogenicity islands. We found CRISPR loci and dam genes in an island specific for C. fetus subsp. fetus, and T4SS and sap genes in an island specific for C. fetus subsp. venerealis. The genomic variations and potential core and unique virulence factors characterized in this study would lead to better insight into the species virulence and to more efficient use of the candidates for antibiotic, drug and vaccine development.


Assuntos
Campylobacter fetus/classificação , Campylobacter fetus/genética , Genes Bacterianos , Genoma Bacteriano , Ilhas Genômicas/genética , Fatores de Virulência/genética , Virulência/genética , Animais , Infecções por Campylobacter/microbiologia , Campylobacter fetus/patogenicidade , Bovinos , DNA Bacteriano/genética , Humanos , Filogenia , Análise de Sequência de DNA , Especificidade da Espécie
12.
Stand Genomic Sci ; 7(2): 189-99, 2012 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-23408795

RESUMO

The Actinobacteria, Corynebacterium pseudotuberculosis strain P54B96, a nonmotile, non-sporulating and a mesophile bacterium, was isolated from liver, lung and mediastinal lymph node lesions in an antelope from South Africa. This strain is interesting in the sense that it has been found together with non-tuberculous mycobacteria (NTMs) which could nevertheless play a role in the lesion formation. In this work, we describe a set of features of C. pseudotuberculosis P54B96, together with the details of the complete genome sequence and annotation. The genome comprises of 2.34 Mbp long, single circular genome with 2,084 protein-coding genes, 12 rRNA, 49 tRNA and 62 pseudogenes and a G+C content of 52.19%. The analysis of the genome sequence provides means to better understanding the molecular and genetic basis of virulence of this bacterium, enabling a detailed investigation of its pathogenesis.

13.
J Bacteriol ; 193(22): 6420-1, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22038974

RESUMO

In this work, we report the complete genome sequence of a Corynebacterium pseudotuberculosis PAT10 isolate, collected from a lung abscess in an Argentine sheep in Patagonia, whose pathogen also required an investigation of its pathogenesis. Thus, the analysis of the genome sequence offers a means to better understanding of the molecular and genetic basis of virulence of this bacterium.


Assuntos
Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/genética , Genoma Bacteriano , Abscesso Pulmonar/microbiologia , Doenças dos Ovinos/microbiologia , Animais , Argentina , Sequência de Bases , Corynebacterium pseudotuberculosis/isolamento & purificação , Corynebacterium pseudotuberculosis/patogenicidade , Dados de Sequência Molecular , Ovinos , Virulência
14.
J Bacteriol ; 193(24): 7025-6, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22123771

RESUMO

In this work, we report the whole-genome sequence of Corynebacterium pseudotuberculosis bv. equi strain CIP 52.97 (Collection Institut Pasteur), isolated in 1952 from a case of ulcerative lymphangitis in a Kenyan horse, which has evidently caused significant losses to agribusiness. Therefore, obtaining this genome will allow the detection of important targets for postgenomic studies, with the aim of minimizing problems caused by this microorganism.


Assuntos
Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/genética , Genoma Bacteriano , Doenças dos Cavalos/microbiologia , Animais , Sequência de Bases , Infecções por Corynebacterium/microbiologia , Corynebacterium pseudotuberculosis/isolamento & purificação , Cavalos , Quênia , Dados de Sequência Molecular
15.
J Bacteriol ; 193(20): 5871-2, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21952544

RESUMO

Campylobacter fetus subsp. venerealis is the etiologic agent of bovine genital campylobacteriosis, a sexually transmitted disease of cattle that is of worldwide importance. The complete sequencing and annotation of the genome of the type strain C. fetus subsp. venerealis NCTC 10354(T) are reported.


Assuntos
Infecções por Campylobacter/veterinária , Campylobacter fetus/genética , Doenças dos Bovinos/microbiologia , Genoma Bacteriano , Animais , Sequência de Bases , Infecções por Campylobacter/microbiologia , Campylobacter fetus/isolamento & purificação , Bovinos , Feminino , Masculino , Dados de Sequência Molecular
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