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1.
Nucleic Acids Res ; 48(4): e24, 2020 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-31925441

RESUMO

Recent years have witnessed increasing efforts to engineer artificial biological functions through recombination of modular-organized toolboxes of protein scaffolds and parts. A critical, yet frequently neglected aspect concerns the identity of peptide linkers or spacers connecting individual domains which remain poorly understood and challenging to assemble. Addressing these limitations, iFlinkC comprises a highly scalable DNA assembly process that facilitates the combinatorial recombination of functional domains with linkers of varying length and flexibility, thereby overcoming challenges with high GC-content and the repeat nature of linker elements. The capacity of iFLinkC is demonstrated in the construction of synthetic protease switches featuring PDZ-FN3-based affinity clamps and single-chain FKBP12-FRB receptors as allosteric inputs. Library screening experiments demonstrate that linker space is highly plastic as the induction of allosterically regulated protease switches can vary from >150-fold switch-ON to >13-fold switch-OFF solely depending on the identity of the connecting linkers and relative orientation of functional domains. In addition, Pro-rich linkers yield the most potent switches contradicting the conventional use of flexible Gly-Ser linkers. Given the ease and efficiency how functional domains can be readily recombined with any type of linker, iFLinkC is anticipated to be widely applicable to the assembly of any type of fusion protein.


Assuntos
Clonagem Molecular/métodos , DNA/genética , Proteínas/genética , Recombinação Genética/genética , Composição de Bases/genética , Peptídeos/genética , Domínios Proteicos/genética , Serina-Treonina Quinases TOR/genética
2.
Arch Virol ; 165(2): 419-424, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31823011

RESUMO

Bacteriophage TAC1 was isolated using a clinical isolate of Acinetobacter baumannii as the host. It showed stability over wide pH and temperature range and has exhibited in vitro antibacterial activity when applied at an MOI of 1. It demonstrated a broad intraspecies host range and infected 66% of the isolates tested. It has produced 454 virions from a single bacterium with a short latent period of 15 minutes. TAC1 has a linear dsDNA genome with a length of 101.77 kb and 37.5% GC content. The genome encodes 161 proteins and 13 putative tRNAs. Whole-genome sequence comparisons using BLASTn and phylogenetic analysis showed that TAC1 is related to unclassified bacteriophages of the family Myoviridae.


Assuntos
Acinetobacter baumannii/virologia , Bacteriófagos/genética , Myoviridae/genética , Composição de Bases/genética , DNA Viral/genética , Genoma Viral/genética , Especificidade de Hospedeiro/genética , Fases de Leitura Aberta/genética , Filogenia , RNA de Transferência/genética , Análise de Sequência de DNA/métodos , Vírion/genética
3.
Arch Virol ; 165(2): 387-396, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31865470

RESUMO

A pathogen of significance in the aquaculture sector, the Gram-negative marine bacterium Vibrio parahaemolyticus causes gastroenteritis associated with consumption of improperly prepared seafood. This bacterium can be controlled using lytic bacteriophages as an alternative to antibiotics. Ï•VP-1 is a lytic phage of V. parahaemolyticus that was isolated from an aquafarm water sample with the aim of assessing its potential as a bio-control agent and determining its physicochemical properties and genomic sequence. Morphological analysis by transmission electron microscopy and phylogenetic analysis based on the large terminase subunit gene showed that this phage belongs to the family Myoviridae. It could infect multiple-drug-resistant (MDR) V. parahaemolyticus and V. alginolyticus strains of mangrove and seafood origin. With a maximum adsorption time of 30 min, ϕVP-1 has a short latent period of 10 min with burst size of 44 particles/cell. Whole-genome sequencing was done using the Illumina platform, and annotation was done using GeneMarkS and Prodigal. The 150,764bp genome with an overall G+C content of 41.84% had 203 putative protein-encoding open reading frames, one tRNA gene, and 66 predicted promoters. A number of putative DNA replication and regulation, DNA packaging and structure, and host lysis genes were identified. Comparison of the ϕVP-1 genome sequence to those of known Vibrio phages indicated little discernible DNA sequence similarity, suggesting that ϕVP-1 is a novel Vibrio phage. Sequence analysis revealed the presence of 64 potential ORFs with a T4-like genomic organization. In silico analysis suggested an obligate lytic life cycle and showed the absence of lysogeny or virulence genes. The complete sequence of Ï•VP-1 was annotated and deposited in the GenBank database (accession no. MH363700). The genetic features of this novel phage suggest that it might be applicable for phage therapy against pathogenic strains of V. parahaemolyticus.


Assuntos
Bacteriófagos/genética , Resistência a Múltiplos Medicamentos/genética , Genoma Viral/genética , Myoviridae/genética , Vibrio parahaemolyticus/virologia , Aquicultura/métodos , Composição de Bases/genética , Biofilmes , Genômica/métodos , Fases de Leitura Aberta/genética , Terapia por Fagos/métodos , Filogenia
4.
PLoS Genet ; 15(11): e1008493, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31703064

RESUMO

Genomic GC content varies widely among microbes for reasons unknown. While mutation bias partially explains this variation, prokaryotes near-universally have a higher GC content than predicted solely by this bias. Debate surrounds the relative importance of the remaining explanations of selection versus biased gene conversion favoring GC alleles. Some environments (e.g. soils) are associated with a high genomic GC content of their inhabitants, which implies that either high GC content is a selective adaptation to particular habitats, or that certain habitats favor increased rates of gene conversion. Here, we report a novel association between the presence of the non-homologous end joining DNA double-strand break repair pathway and GC content; this observation suggests that DNA damage may be a fundamental driver of GC content, leading in part to the many environmental patterns observed to-date. We discuss potential mechanisms accounting for the observed association, and provide preliminary evidence that sites experiencing higher rates of double-strand breaks are under selection for increased GC content relative to the genomic background.


Assuntos
Composição de Bases/genética , Evolução Molecular , Conversão Gênica/genética , Células Procarióticas , Quebras de DNA de Cadeia Dupla , Reparo do DNA por Junção de Extremidades/genética , Reparo do DNA/genética , Genoma/genética , Humanos
5.
Virus Genes ; 55(6): 834-842, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31420829

RESUMO

A novel Vibrio phage, P23, belonging to the family Siphoviridae was isolated from the surface water of the Yellow Sea, China. The complete genome of this phage was determined. A one-step growth curve showed that the latent period was approximately 30 min, the burst size was 24 PFU/cell, and the rise period was 20 min. The phage is host specific and is stable over a range of pH (5-10) and temperatures (4-65 °C). Transmission electron microscopy showed that phage P23 can be categorized into the Siphoviridae family, with an icosahedral head of 60 nm and a long noncontractile tail of 144 nm. The genome consisted of a linear, double-stranded 40.063 kb DNA molecule with 42.5% G+C content and 72 putative open reading frames (ORFs) without tRNA. The predicted ORFs were classified into six functional groups, including DNA replication, regulation and nucleotide metabolism, transcription, phage packaging, phage structure, lysis, and hypothetical proteins. The Vibrio phage P23 genome is a new marine Siphoviridae-family phage genome that provides basic information for further molecular research on interaction mechanisms between bacteriophages and their hosts.


Assuntos
Bacteriófagos/genética , Genoma Viral/genética , Filogenia , Sequenciamento Completo do Genoma , Bacteriófagos/classificação , Composição de Bases/genética , China , Genômica , Anotação de Sequência Molecular , Fases de Leitura Aberta/genética , Análise de Sequência de DNA , Siphoviridae/genética , Vibrio/genética , Vibrio/virologia
6.
Arch Virol ; 164(11): 2865-2871, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31401694

RESUMO

Phage Vp_R1 belongs to the family Podoviridae and has a C3 morphotype, with an elongated head with a diameter of 190 ± 1.1 nm and an ultrashort tail with a length of 9 ± 1.2 nm. The double-stranded DNA genome is 112.1 kb long, has a mol% G + C content of 40.3, contains 129 ORFs, and encodes four tRNAs. Phylogenetic analysis suggests that phage Vp_R1 is a novel member of the genus Kuravirus.


Assuntos
Genoma Viral/genética , Podoviridae/genética , Vibrio parahaemolyticus/virologia , Sequência de Aminoácidos , Composição de Bases/genética , DNA Viral/genética , Fases de Leitura Aberta/genética , Podoviridae/isolamento & purificação , Análise de Sequência de DNA , Proteínas Virais/genética
7.
Arch Virol ; 164(11): 2877-2880, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31451964

RESUMO

A temperate bacteriophage, IME1320_01, was induced by mitomycin C treatment from Corynebacterium striatum. This phage possesses a double-stranded DNA genome of 40,086 bp with a G+C content of 58%. A total of 53 putative open reading frames (ORFs) were identified in its genome. BLASTn analysis revealed that IME1320_01 had the highest sequence similarity to Corynebacterium striatum strain 216, with a genome homology coverage of 44% and highest sequence identity of 95%. The termini of the phage genome was non-redundant, with a 13-nt 3'-protruding cohesive end. To the best of our knowledge, phage IME1320_01 is the first inducible phage to be identified in Corynebacterium striatum.


Assuntos
Corynebacterium/virologia , Genoma Viral/genética , Siphoviridae/genética , Ativação Viral/efeitos dos fármacos , Composição de Bases/genética , DNA Viral/genética , Mitomicina/farmacologia , Fases de Leitura Aberta/genética , Análise de Sequência de DNA , Siphoviridae/classificação , Siphoviridae/isolamento & purificação
8.
World J Microbiol Biotechnol ; 35(8): 118, 2019 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-31332540

RESUMO

The fungal genus Ustilago consists of intimidating pathogens associated with disease manifestations in plants of agricultural importance and gravity. Rapid progress of genome sequencing has opened the floodgates for biological research. Availability of Ustilago genomes provides a scope to explore complex codon and amino acid usage patterns in the genus. An extensive scrutiny of the factors underlying the complex modalities of codon and amino acid usage in Ustilago has been executed in the present analysis. Multivariate statistical analysis revealed a dominant effect of natural selection pressure, aimed at translational accuracy, to be operative on the codon usage behavior. Subtle impact of GC compositional constraint was also evident on the codon usage patterns. Gene expressivity was inferred to be the most crucial determinant governing observed codon usage variations. Amino acid usage patterns were found to be significantly governed by aromatic and hydrophobic characters of the encoded proteins. GC content and length of protein coding sequences also had considerable influence on the amino acid usage signatures. Extensive analysis of codon context variations revealed that UpA dinucleotides were strictly avoided at the codon-codon junctions (cP3-cA1) which might be attributed to reduce the risk of nonsense mutations and subsequently, improve translational finesse. Identification of the optimal codons, employed preferentially among the genes with high expressivity, and estimation of preferred and avoided codon pairs in Ustilago promises to be useful pertaining to mutational experiments at the codonic level, targeted to thwart the growth of Ustilago and combat associated pathogenesis.


Assuntos
Perfilação da Expressão Gênica , Ustilago/genética , Aminoácidos/metabolismo , Composição de Bases/genética , Códon , Seleção Genética/genética , Ustilago/metabolismo , Sequenciamento Completo do Genoma
9.
BMC Evol Biol ; 19(1): 144, 2019 07 16.
Artigo em Inglês | MEDLINE | ID: mdl-31311498

RESUMO

BACKGROUND: Rapid accumulation of vertebrate genome sequences render comparative genomics a powerful approach to study macro-evolutionary events. The assessment of phylogenic relationships between species routinely depends on the analysis of sequence homology at the nucleotide or protein level. RESULTS: We analyzed mRNA GC content, codon usage and divergence of orthologous proteins in 55 vertebrate genomes. Data were visualized in genome-wide landscapes using a sliding window approach. Landscapes of GC content reveal both evolutionary conservation of clustered genes, and lineage-specific changes, so that it was possible to construct a phylogenetic tree that closely matched the classic "tree of life". Landscapes of GC content also strongly correlated to landscapes of amino acid usage: positive correlation with glycine, alanine, arginine and proline and negative correlation with phenylalanine, tyrosine, methionine, isoleucine, asparagine and lysine. Peaks of GC content correlated strongly with increased protein divergence. CONCLUSIONS: Landscapes of base- and amino acid composition of the coding genome opens a new approach in comparative genomics, allowing identification of discrete regions in which protein evolution accelerated over deep evolutionary time. Insight in the evolution of genome structure may spur novel studies assessing the evolutionary benefit of genes in particular genomic regions.


Assuntos
Composição de Bases/genética , Evolução Molecular , Exoma/genética , Proteínas/genética , Vertebrados/genética , Animais , Códon/genética , Genoma , Humanos , Mamíferos/genética , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Répteis/genética
10.
Biomed Res Int ; 2019: 5857285, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31346520

RESUMO

Background: Flaviviridae viruses are single-stranded, positive-sense RNA viruses, which threat human constantly mediated by mosquitoes, ticks, and sandflies. Considering the recent increase in the prevalence of the family virus and its risk potential, we investigated the codon usage pattern to understand its evolutionary processes and provide some useful data to develop the medications for most of Flaviviridae viruses. Results: The overall extent of codon usage bias in 65 Flaviviridae viruses is low with the average value of GC contents being 50.5% and the highest value being 55.9%; the lowest value is 40.2%. ENC values of Flaviviridae virus genes vary from 48.75 to 57.83 with a mean value of 55.56. U- and A-ended codons are preferred in the Flaviviridae virus. Correlation analysis shows that the positive correlation between ENC value and GC content at the third nucleotide positions was significant in this family virus. The result of analysis of ENC, neutrality plot analysis, and correlation analysis revealed that codon usage bias of all the viruses was affected mainly by natural selection. Meanwhile, according to correspondence analysis (CoA) based on RSCU and phylogenetic analysis, the Flaviviridae viruses mainly are made up of two groups, Group I (Yellow fever virus, Apoi virus, Tembusu virus, Dengue virus 1, and others) and Group II (West Nile virus lineage 2, Japanese encephalitis virus, Usutu virus, Kedougou virus, and others). Conclusions: All in, the bias of codon usage pattern is affected not only by compositional constraints but also by natural selection. Phylogenetic analysis also illustrates that codon usage bias of virus can serve as an effective means of evolutionary classification in Flaviviridae virus.


Assuntos
/genética , Evolução Molecular , Infecções por Flaviviridae/genética , Flaviviridae/genética , Animais , Composição de Bases/genética , Códon/genética , Biologia Computacional , Flaviviridae/classificação , Flaviviridae/patogenicidade , Infecções por Flaviviridae/virologia , Genoma Viral/genética , Humanos , Nucleotídeos/genética , Filogenia , Seleção Genética/genética
11.
BMC Res Notes ; 12(1): 444, 2019 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-31331368

RESUMO

OBJECTIVE: Sulfodiicoccus acidiphilus HS-1T is the type species of the genus Sulfodiicoccus, a thermoacidophilic archaeon belonging to the order Sulfolobales (class Thermoprotei; phylum Crenarchaeota). While S. acidiphilus HS-1T shares many common physiological and phenotypic features with other Sulfolobales species, the similarities in their 16S rRNA gene sequences are less than 89%. In order to know the genomic features of S. acidiphilus HS-1T in the order Sulfolobales, we determined and characterized the genome of this strain. RESULTS: The circular genome of S. acidiphilus HS-1T is comprised of 2353,189 bp with a G+C content of 51.15 mol%. A total of 2459 genes were predicted, including 2411 protein coding and 48 RNA genes. The notable genomic features of S. acidiphilus HS-1T in Sulfolobales species are the absence of genes for polB3 and the autotrophic carbon fixation pathway, and the distribution pattern of essential genes and sequences related to genomic replication initiation. These insights contribute to an understanding of archaeal genomic diversity and evolution.


Assuntos
Proteínas Arqueais/genética , DNA Arqueal/genética , Genoma Arqueal/genética , Sulfolobaceae/genética , Sequenciamento Completo do Genoma/métodos , Composição de Bases/genética , DNA Arqueal/química , Ordem dos Genes , Genes Arqueais/genética , Genômica/métodos , Fontes Termais/microbiologia , Japão , Filogenia , RNA Ribossômico 16S/genética , Especificidade da Espécie , Sulfolobaceae/classificação , Sulfolobaceae/isolamento & purificação
12.
Arch Microbiol ; 201(8): 1111-1118, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31154464

RESUMO

A novel Gram-stain-positive, motile, white color and endospore-forming bacterium, designated 18JY67-1T, was isolated from soil in Jeju Island, Korea. The strain grow at 15-42 °C (optimum 30 °C) in R2A medium at pH (6.0-9.5) (optimum 7.5). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain 18JY67-1T formed a distinct lineage within the family Paenibacillaceae (order Bacillales, class Bacilli), and was closely related to Paenibacillus rhizoryzae (KP675984; 96.9% 16S rRNA gene sequence similarity). The major cellular fatty acids of the strain 18JY67-1T were C16:0 and anteiso-C15:0. The predominant respiratory quinones were MK-7. The major polar lipid was identified as diphosphatidylglycerol. On the basis of phenotypic, chemotaxonomic and genotypic properties clearly indicated that isolate 18JY67-1T represents a novel species within the genus Paenibacillus, for which the name Paenibacillus flavus sp. nov. is proposed. The type strain of Paenibacillus flavus is 18JY67-1T (= KCTC 33959T = JCM 33184T).


Assuntos
Paenibacillus , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases/genética , DNA Bacteriano/genética , Ácidos Graxos/análise , Paenibacillus/genética , Paenibacillus/isolamento & purificação , Paenibacillus/efeitos da radiação , Filogenia , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Solo , Raios Ultravioleta
13.
Arch Microbiol ; 201(8): 1119-1127, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31165177

RESUMO

Strain KYPW7T, isolated from the Funglin Stream in Taiwan, was characterized using a polyphasic taxonomy approach. Cells of strain KYPW7T were Gram-stain-negative, aerobic, non-spore forming, non-motile rods and formed white colonies. Growth occurred at 15-30 °C (optimum 25 °C), at pH 6-8 (optimum pH 6.5) and with 0-1% NaCl (optimum 0%). Phylogenetic analyses based on 16S rRNA and coding sequences of 92 protein clusters showed that strain KYPW7T represents a novel genus in the family Flavobacteriaceae. The 16S rRNA gene sequence of strain KYPW7T was related to the species of the genera Chryseobacterium (91.8-96.0% sequence similarity), Bergeyella (95.1-95.8%), Cloacibacterium (94.5-95.7%), Daejeonia (95.6%) and Riemerella (94.0-95.0%). Strain KYPW7T showed less than 72% average nucleotide identity and less than 24% digital DNA-DNA hybridization identity compared to the type strains of related genera within the family Flavobacteriaceae. The predominant fatty acids were iso-C15:0 and iso-C17:0 3-OH. The major isoprenoid quinone was MK-6 and the DNA G + C content was 36.8 mol%. The polar lipids had phosphatidylethanolamine, three uncharacterized aminophospholipids and an uncharacterized phospholipid. The polyamines contained homospermidine, putrescine and spermidine. On the basis of the genotypic and phenotypic data, strain KYPW7T represents a novel species of a new genus in the family Flavobacteriaceae, for which the name Amniculibacterium aquaticum gen. nov., sp. nov. is proposed. The type strain is KYPW7T (= BCRC 81123T = LMG 30598T = KCTC 62512T).


Assuntos
Flavobacteriaceae , Rios/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases/genética , DNA Bacteriano/genética , Ácidos Graxos/análise , Flavobacteriaceae/classificação , Flavobacteriaceae/genética , Flavobacteriaceae/isolamento & purificação , Hibridização de Ácido Nucleico , Fosfatidiletanolaminas/análise , Fosfolipídeos/análise , Filogenia , Poliaminas/análise , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Taiwan
14.
Cells ; 8(6)2019 06 21.
Artigo em Inglês | MEDLINE | ID: mdl-31234415

RESUMO

Single nucleotide polymorphisms located in 5' untranslated regions (5'UTRs) can regulate gene expression and have clinical impact. Recognition of functionally significant sequences within 5'UTRs is crucial in next-generation sequencing applications. Furthermore, information about the behavior of 5'UTRs during gene evolution is scarce. Using the example of the ATP-binding cassette transporter A1 (ABCA1) gene (Tangier disease), we describe our algorithm for functionally significant sequence finding. 5'UTR features (upstream start and stop codons, open reading frames (ORFs), GC content, motifs, and secondary structures) were studied using freely available bioinformatics tools in 55 vertebrate orthologous genes obtained from Ensembl and UCSC. The most conserved sequences were suggested as hot spots. Exon and intron enhancers and silencers (sc35, ighg2 cgamma2, ctnt, gh-1, and fibronectin eda exon), transcription factors (TFIIA, TATA, NFAT1, NFAT4, and HOXA13), some of them cancer related, and microRNA (hsa-miR-4474-3p) were localized to these regions. An upstream ORF, overlapping with the main ORF in primates and possibly coding for a small bioactive peptide, was also detected. Moreover, we showed several features of 5'UTRs, such as GC content variation, hairpin structure conservation or 5'UTR segmentation, which are interesting from a phylogenetic point of view and can stimulate further evolutionary oriented research.


Assuntos
Regiões 5' não Traduzidas/genética , Transportador 1 de Cassete de Ligação de ATP/genética , Vertebrados/genética , Transportador 1 de Cassete de Ligação de ATP/química , Sequência de Aminoácidos , Animais , Composição de Bases/genética , Sequência de Bases , Sequência Conservada/genética , Elementos Facilitadores Genéticos/genética , Humanos , Íntrons/genética , Mamíferos/genética , Anotação de Sequência Molecular , Conformação de Ácido Nucleico , Motivos de Nucleotídeos/genética , Fases de Leitura Aberta/genética , Filogenia , Processamento de RNA/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
15.
Gene ; 706: 146-153, 2019 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-31077734

RESUMO

Necrophagous Dermestes species have high forensic importance in relation to the estimation of elapsed time since death or death season. To further supplement the genome-level features for related species, the complete mitochondrial genome (mitogenome) of Dermestes species D. essellatocollis, D. frischii and D. coarctatus are amplified, sequenced, annotated, analyzed, and compared with other twelve species of the infraorder Bostrichoidea. The mitochondrial genomes were typical circular molecules with 16,218, 15,873 and 15,873 bp in length, respectively. They included 13 protein coding genes, two rRNAs, and 22 tRNAs, as well as the putative control region. The gene orders and orientations are identical to those of other recorded bostrichiformian species and had the ancestral insect gene composition. Furthermore, phylogenetic analyses based on all the mitochondrial protein coding genes for 13 Bostrichoidea and 16 outgroup taxa were performed using Bayesian and Maximum Likelihood analyses. The inferred trees indicate that the genus Dermestes is monophyletic. The monophyly of infraorder Bostrichiformia is not supported. This study provides genomic data for mitochondrial genome library of the genus Dermestes to investigate evolutionary and systematic studies.


Assuntos
Besouros/genética , Genoma Mitocondrial/genética , Mitocôndrias/genética , Animais , Composição de Bases/genética , Sequência de Bases , Ciências Forenses/métodos , Ordem dos Genes/genética , Filogenia , RNA Ribossômico/genética , RNA de Transferência/genética
16.
Arch Microbiol ; 201(8): 1019-1024, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31098768

RESUMO

A novel Gelidibacter strain, JCM 31967T, was isolated from seawater collected from the Inland Sea (Setonaikai) in Japan. It was characterized as a Gram-negative, halophilic, oxidase-negative, catalase-positive, aerobic, nonmotile, but gliding, rod-shaped bacterium without flagella. Based on 16S rDNA gene identity, strain JCM 31967T is closely related to Gelidibacter mesophilus (DSM 14095T, 96.6% identity), G. gilvus (IC158T, 96.4%), G. algens (DSM 12408T, 96.1%), G. sediminis (S11-41T, 94.7%), and G. salicanalis (IC162T, 94.7%). The G + C content of strain JCM 31967T DNA was found to be 39.1%. The average nucleotide identity (ANI) values of JCM 31967T against G. algens DSM 12408T and G. mesophilus DSM 14095T were 79.1% and 80.9%, respectively. Strain JCM 31967T phenotypically differed from the closest related Gelidibacter species in its utilization of methyl α-D-mannopyranoside, methyl α-D-glucopyranoside, and D-ribose and in its lack of utilization of L-arginine and D-arabinose. It was further differentiated based on its fatty acid composition, specifically properties of C18:0 and C20:2 ω6c, 9c, which were significantly different from those of G. algens, G. gilvus, G. mesophilus, G. salicanalis, and G. sediminis type strains. Overall, the results of DNA-DNA hybridization and physiological and biochemical analyses differentiated strain JCM 31967T from a previously described species of Gelidibacter. Based on these polyphasic taxonomic findings, it was concluded that strain JCM 31967T is a novel Gelidibacter species, for which the name Gelidibacter japonicus sp. nov. is proposed, with JCM 31967T (= LMG 30063T) as the type strain.


Assuntos
Flavobacteriaceae , Água do Mar/microbiologia , Composição de Bases/genética , DNA Bacteriano/genética , DNA Ribossômico/genética , Ácidos Graxos/análise , Flavobacteriaceae/classificação , Flavobacteriaceae/genética , Flavobacteriaceae/isolamento & purificação , Japão , Hibridização de Ácido Nucleico , Fenótipo , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
17.
Arch Microbiol ; 201(8): 1053-1060, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31115641

RESUMO

Strain HPM-16T, isolated from seawater, was characterized using a polyphasic taxonomy approach. Phylogenetic analyses based on 16S rRNA gene sequences and coding sequences of an up-to-date bacterial core gene set (92 protein clusters) indicated that strain HPM-16T formed a phylogenetic lineage in the genus Neptunomonas. Strain HPM-16T was most closely related to Neptunomonas concharum LHW37T with 16S rRNA gene sequence similarity of 96.7%. Cells were Gram-stain negative, facultatively anaerobic, motile by means of a single polar flagellum, rod-shaped and formed white colonies. Optimal growth occurred at 30-35 °C, pH 6.5-8, and in the presence of 2-5% NaCl. C18:1ω7c and summed feature 3 (C16:1ω7c and/or C16:1ω6c) were the predominant fatty acids. The only isoprenoid quinone was Q-8. The polar lipid profile revealed the presence of phosphatidylethanolamine, phosphatidylglycerol and several uncharacterized lipids. The major polyamines were putrescine and spermidine. The draft genome was approximately 3.68 Mb in size with a G + C content of 50.5 mol%. Differential phenotypic properties, together with the phylogenetic inference, demonstrate that strain HPM-16T should be classified as a novel species of the genus Neptunomonas, for which the name Neptunomonas marina sp. nov. is presented. The type strain is HPM-16T (= BCRC 80980T = LMG 29560T = KCTC 52235T).


Assuntos
Oceanospirillaceae , Água do Mar/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases/genética , DNA Bacteriano/genética , Ácidos Graxos/análise , Oceanospirillaceae/classificação , Oceanospirillaceae/genética , Oceanospirillaceae/isolamento & purificação , Fosfatidiletanolaminas/análise , Fosfatidilgliceróis/análise , Fosfolipídeos/análise , Filogenia , Quinonas/análise , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
18.
Genome Res ; 29(5): 711-722, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30962178

RESUMO

The inclusion of exons during the splicing process depends on the binding of splicing factors to short low-complexity regulatory sequences. The relationship between exonic splicing regulatory sequences and coding sequences is still poorly understood. We demonstrate that exons that are coregulated by any given splicing factor share a similar nucleotide composition bias and preferentially code for amino acids with similar physicochemical properties because of the nonrandomness of the genetic code. Indeed, amino acids sharing similar physicochemical properties correspond to codons that have the same nucleotide composition bias. In particular, we uncover that the TRA2A and TRA2B splicing factors that bind to adenine-rich motifs promote the inclusion of adenine-rich exons coding preferentially for hydrophilic amino acids that correspond to adenine-rich codons. SRSF2 that binds guanine/cytosine-rich motifs promotes the inclusion of GC-rich exons coding preferentially for small amino acids, whereas SRSF3 that binds cytosine-rich motifs promotes the inclusion of exons coding preferentially for uncharged amino acids, like serine and threonine that can be phosphorylated. Finally, coregulated exons encoding amino acids with similar physicochemical properties correspond to specific protein features. In conclusion, the regulation of an exon by a splicing factor that relies on the affinity of this factor for specific nucleotide(s) is tightly interconnected with the exon-encoded physicochemical properties. We therefore uncover an unanticipated bidirectional interplay between the splicing regulatory process and its biological functional outcome.


Assuntos
Processamento Alternativo , Éxons/genética , Sítios de Splice de RNA/genética , Fatores de Processamento de RNA/metabolismo , Aminoácidos/química , Composição de Bases/genética , Linhagem Celular , Código Genético , Ribonucleoproteínas Nucleares Heterogêneas/metabolismo , Humanos , Íntrons/genética , Motivos de Nucleotídeos/genética , Análise de Sequência de Proteína , Análise de Sequência de RNA , Fatores de Processamento de Serina-Arginina/metabolismo
19.
Biomed Res Int ; 2019: 5046958, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31016191

RESUMO

Sorghum comprises 31 species that exhibit considerable morphological and ecological diversity. The phylogenetic relationships among Sorghum species still remain unresolved due to lower information on the traditional DNA markers, which provides a limited resolution for identifying Sorghum species. In this study, we sequenced the complete chloroplast genomes of Sorghum sudanense and S. propinquum and analyzed the published chloroplast genomes of S. bicolor and S. timorense to retrieve valuable chloroplast molecular resources for Sorghum. The chloroplast genomes ranged in length from 140,629 to 140,755 bp, and their gene contents, gene orders, and GC contents were similar to those for other Poaceae species but were slightly different in the number of SSRs. Comparative analyses among the four chloroplast genomes revealed 651 variable sites, 137 indels, and nine small inversions. Four highly divergent DNA regions (rps16-trnQ, trnG-trnM, rbcL-psaI, and rps15-ndhF), which were suitable for phylogenetic and species identification, were detected in the Sorghum chloroplast genomes. A phylogenetic analysis strongly supported that Sorghum is a monophyletic group in the tribe Andropogoneae. Overall, the genomic resources in this study could provide potential molecular markers for phylogeny and species identification in Sorghum.


Assuntos
Cloroplastos/genética , Genoma de Cloroplastos/genética , Sorghum/genética , Composição de Bases/genética , DNA de Cloroplastos/genética , Ordem dos Genes/genética , Marcadores Genéticos/genética , Genômica/métodos , Filogenia , Poaceae/genética , Análise de Sequência de DNA/métodos
20.
Mol Genet Genomics ; 294(3): 715-727, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30848356

RESUMO

Among invertebrates, only a few groups still have uncertain phylogenetic position, Orthonectida, a small group of rare multi-cellular parasites of marine invertebrates, being one of them. Recent molecular and morphological findings suggest that orthonectids belong to Lophotrochozoa and are close to Annelida. Nevertheless, phylogenetic relationships between orthonectids and annelids are unclear, and the phylogeny within the group itself has never been studied. Sequencing of mitochondrial genomes is used here to clarify this issue. Complete mt genomes of the orthonectids Intoshia variabili and Rhopalura litoralis were characterized and compared with Intoshia linei mt genome. Our results show that Orthonectida mt genomes have undergone reduction and gene loss, and that they have complicated organization revealed in strand asymmetry in nucleotide composition, in some features of intergenic non-coding regions, tRNA duplication and folding. Moreover, all species of Orthonectida have a unique gene order with complicated rearrangement landscape. Significant differences in mitochondrial genomes in the three orthonectid species could be explained by the fact that their host species belong to different taxa (flat worms, nemertines and gastropods). Among the analyzed mt genomes of Orthonectida, I. linei possesses the closest gene order to the ancestral genome. All Orthonectida species are monophyletic, and in the phylogenetic tree are close to Pleistoannelida, and specifically, to Clitellata.


Assuntos
Evolução Molecular , Ordem dos Genes , Genes Mitocondriais/genética , Genoma Mitocondrial/genética , Invertebrados/genética , Animais , Composição de Bases/genética , DNA Mitocondrial/química , DNA Mitocondrial/genética , Gastrópodes/parasitologia , Helmintos/parasitologia , Interações Hospedeiro-Parasita , Invertebrados/classificação , Invertebrados/fisiologia , Filogenia , Análise de Sequência de DNA , Especificidade da Espécie
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