Random mutagenesis of the hyperthermophilic archaeon Pyrococcus furiosus using in vitro mariner transposition and natural transformation.

Abstract:

:Transposition mutagenesis is a powerful tool to identify the function of genes, reveal essential genes and generally to unravel the genetic basis of living organisms. However, transposon-mediated mutagenesis has only been successfully applied to a limited number of archaeal species and has never been reported in Thermococcales. Here, we report random insertion mutagenesis in the hyperthermophilic archaeon Pyrococcus furiosus. The strategy takes advantage of the natural transformability of derivatives of the P. furiosus COM1 strain and of in vitro Mariner-based transposition. A transposon bearing a genetic marker is randomly transposed in vitro in genomic DNA that is then used for natural transformation of P. furiosus. A small-scale transposition reaction routinely generates several hundred and up to two thousands transformants. Southern analysis and sequencing showed that the obtained mutants contain a single and random genomic insertion. Polyploidy has been reported in Thermococcales and P. furiosus is suspected of being polyploid. Yet, about half of the mutants obtained on the first selection are homozygous for the transposon insertion. Two rounds of isolation on selective medium were sufficient to obtain gene conversion in initially heterozygous mutants. This transposition mutagenesis strategy will greatly facilitate functional exploration of the Thermococcales genomes.

journal_name

Sci Rep

journal_title

Scientific reports

authors

Guschinskaya N,Brunel R,Tourte M,Lipscomb GL,Adams MWW,Oger P,Charpentier X

doi

10.1038/srep36711

subject

Has Abstract

pub_date

2016-11-08 00:00:00

pages

36711

issn

2045-2322

pii

srep36711

journal_volume

6

pub_type

杂志文章
  • Collision with duplex DNA renders Escherichia coli DNA polymerase III holoenzyme susceptible to DNA polymerase IV-mediated polymerase switching on the sliding clamp.

    abstract::Organisms possess multiple DNA polymerases (Pols) and use each for a different purpose. One of the five Pols in Escherichia coli, DNA polymerase IV (Pol IV), encoded by the dinB gene, is known to participate in lesion bypass at certain DNA adducts. To understand how cells choose Pols when the replication fork encounte...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-017-13080-1

    authors: Le TT,Furukohri A,Tatsumi-Akiyama M,Maki H

    更新日期:2017-10-16 00:00:00

  • Reliability in adolescent fMRI within two years - a comparison of three tasks.

    abstract::Longitudinal developmental fMRI studies just recently began to focus on within-subject reliability using the intraclass coefficient (ICC). It remains largely unclear which degree of reliability can be achieved in developmental studies and whether this depends on the type of task used. Therefore, we aimed to systematic...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-017-02334-7

    authors: Vetter NC,Steding J,Jurk S,Ripke S,Mennigen E,Smolka MN

    更新日期:2017-05-23 00:00:00

  • Generation of heritable germline mutations in the jewel wasp Nasonia vitripennis using CRISPR/Cas9.

    abstract::The revolutionary RNA-guided endonuclease CRISPR/Cas9 system has proven to be a powerful tool for gene editing in a plethora of organisms. Here, utilizing this system we developed an efficient protocol for the generation of heritable germline mutations in the parasitoid jewel wasp, Nasonia vitripennis, a rising insect...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-017-00990-3

    authors: Li M,Au LYC,Douglah D,Chong A,White BJ,Ferree PM,Akbari OS

    更新日期:2017-04-19 00:00:00

  • Functional spectroscopic imaging reveals specificity of glutamate response in mouse brain to peripheral sensory stimulation.

    abstract::Non-invasive investigation of physiological changes and metabolic events associated with brain activity in mice constitutes a major challenge. Conventionally, fMRI assesses neuronal activity by evaluating activity-evoked local changes in blood oxygenation levels (BOLD). In isoflurane-anaethetized mice, however, we fou...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-019-46477-1

    authors: Seuwen A,Schroeter A,Grandjean J,Schlegel F,Rudin M

    更新日期:2019-07-22 00:00:00

  • High Efficiency Inorganic/Inorganic Amorphous Silicon/Heterojunction Silicon Tandem Solar Cells.

    abstract::We investigated high-efficiency two-terminal tandem photovoltaic (PV) devices consisting of a p/i/n thin film silicon top sub-cell (p/i/n-TFS) and a heterojunction with an intrinsic thin-layer (HIT) bottom sub-cell. We used computer simulations and experimentation. The short-circuit current density (Jsc) of the top su...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-018-33734-y

    authors: Park J,Dao VA,Kim S,Pham DP,Kim S,Le AHT,Kang J,Yi J

    更新日期:2018-10-18 00:00:00

  • Species identification by experts and non-experts: comparing images from field guides.

    abstract::Accurate species identification is fundamental when recording ecological data. However, the ability to correctly identify organisms visually is rarely questioned. We investigated how experts and non-experts compared in the identification of bumblebees, a group of insects of considerable conservation concern. Experts a...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/srep33634

    authors: Austen GE,Bindemann M,Griffiths RA,Roberts DL

    更新日期:2016-09-20 00:00:00

  • High levels of drug resistance in commensal E. coli in a cohort of children from rural central India.

    abstract::The world is experiencing crisis of antibiotic resistance not only in pathogenic but also in commensal bacteria. We determine the prevalence of antibiotic resistance in commensal Escherichia coli in young children in rural setting of central India and search for its correlations with demographic and behavioral factors...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-019-43227-1

    authors: Purohit MR,Lindahl LF,Diwan V,Marrone G,Lundborg CS

    更新日期:2019-04-30 00:00:00

  • Long-term passive acoustic recordings track the changing distribution of North Atlantic right whales (Eubalaena glacialis) from 2004 to 2014.

    abstract::Given new distribution patterns of the endangered North Atlantic right whale (NARW; Eubalaena glacialis) population in recent years, an improved understanding of spatio-temporal movements are imperative for the conservation of this species. While so far visual data have provided most information on NARW movements, pas...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-017-13359-3

    authors: Davis GE,Baumgartner MF,Bonnell JM,Bell J,Berchok C,Bort Thornton J,Brault S,Buchanan G,Charif RA,Cholewiak D,Clark CW,Corkeron P,Delarue J,Dudzinski K,Hatch L,Hildebrand J,Hodge L,Klinck H,Kraus S,Martin B,Mellin

    更新日期:2017-10-18 00:00:00

  • Computational Assessment of Thermokinetics and Associated Microstructural Evolution in Laser Powder Bed Fusion Manufacturing of Ti6Al4V Alloy.

    abstract::Although most of the near non-equilibrium microstructures of alloys produced by laser powder bed fusion (LPBF) additive manufacturing (AM) are being reported at a rapid rate, the accountable thermokinetics of the entire process have rarely been studied. In order to exploit the versatility of this AM process for the de...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-020-63281-4

    authors: Pantawane MV,Ho YH,Joshi SS,Dahotre NB

    更新日期:2020-05-05 00:00:00

  • Predicting Subtype Selectivity for Adenosine Receptor Ligands with Three-Dimensional Biologically Relevant Spectrum (BRS-3D).

    abstract::Adenosine receptors (ARs) are potential therapeutic targets for Parkinson's disease, diabetes, pain, stroke and cancers. Prediction of subtype selectivity is therefore important from both therapeutic and mechanistic perspectives. In this paper, we introduced a shape similarity profile as molecular descriptor, namely t...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/srep36595

    authors: He SB,Ben Hu,Kuang ZK,Wang D,Kong DX

    更新日期:2016-11-04 00:00:00

  • Genomic characterization of intrinsic and acquired resistance to cetuximab in colorectal cancer patients.

    abstract::Anti-EGFR antibodies are effective in therapies for late-stage colorectal cancer (CRC); however, many tumours are unresponsive or develop resistance. We performed genomic analysis of intrinsic and acquired resistance to anti-EGFR therapy in prospectively collected tumour samples from 25 CRC patients receiving cetuxima...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-019-51981-5

    authors: Bray SM,Lee J,Kim ST,Hur JY,Ebert PJ,Calley JN,Wulur IH,Gopalappa T,Wong SS,Qian HR,Ting JC,Liu J,Willard MD,Novosiadly RD,Park YS,Park JO,Lim HY,Kang WK,Aggarwal A,Kim HC,Reinhard C

    更新日期:2019-10-25 00:00:00

  • Synthesis and characterization of water-soluble polythiophene derivatives for cell imaging.

    abstract::In this work, four water-soluble polythiophene derivatives (PT, PT-DDA, PT-ADA, and PT-ADA-PPR) with different pendant moieties were synthesized via oxidative copolymerization by FeCl3. By increasing the hydrophobic ability of side chain moieties, there is a gradually blue shift for the maximum absorption wavelength a...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/srep07617

    authors: Wang F,Li M,Wang B,Zhang J,Cheng Y,Liu L,Lv F,Wang S

    更新日期:2015-01-05 00:00:00

  • Polarized super-resolution structural imaging inside amyloid fibrils using Thioflavine T.

    abstract::Thioflavin T (ThT) is standardly used as a fluorescent marker to detect aggregation of amyloid fibrils by conventional fluorescence microscopy, including polarization resolved imaging that brings information on the orientational order of the fibrils. These techniques are however diffraction limited and cannot provide ...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-017-12864-9

    authors: Shaban HA,Valades-Cruz CA,Savatier J,Brasselet S

    更新日期:2017-10-02 00:00:00

  • Spatially- and vector-resolved momentum flux lost to a wall in a magnetic nozzle rf plasma thruster.

    abstract::Most of the artificial low-pressure plasmas contact with physical walls in laboratories; the plasma loss at the wall significantly affects the plasma device performance, e.g., an electric propulsion device. Near the surface of the wall, ions are spontaneously accelerated by a sheath and deliver their momentum and ener...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-020-58022-6

    authors: Takahashi K,Sugawara T,Ando A

    更新日期:2020-01-23 00:00:00

  • DeNeRD: high-throughput detection of neurons for brain-wide analysis with deep learning.

    abstract::Mapping the structure of the mammalian brain at cellular resolution is a challenging task and one that requires capturing key anatomical features at the appropriate level of analysis. Although neuroscientific methods have managed to provide significant insights at the micro and macro level, in order to obtain a whole-...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-019-50137-9

    authors: Iqbal A,Sheikh A,Karayannis T

    更新日期:2019-09-25 00:00:00

  • Chronic mercury exposure in Late Neolithic/Chalcolithic populations in Portugal from the cultural use of cinnabar.

    abstract::Cinnabar is a natural mercury sulfide (HgS) mineral of volcanic or hydrothermal origin that is found worldwide. It has been mined prehistorically and historically in China, Japan, Europe, and the Americas to extract metallic mercury (Hg(0)) for use in metallurgy, as a medicinal, a preservative, and as a red pigment fo...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/srep14679

    authors: Emslie SD,Brasso R,Patterson WP,Carlos Valera A,McKenzie A,Maria Silva A,Gleason JD,Blum JD

    更新日期:2015-10-01 00:00:00

  • Morphology-dependent Electrochemical Enhancements of Porous Carbon as Sensitive Determination Platform for Ascorbic Acid, Dopamine and Uric Acid.

    abstract::Using starch as the carbon precursor and different-sized ZnO naoparticles as the hard template, a series of porous carbon materials for electrochemical sensing were prepared. Experiments of scanning electron microscopy, transmission electron microscopy and Nitrogen adsorption-desorption isotherms reveal that the parti...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/srep22309

    authors: Cheng Q,Ji L,Wu K,Zhang W

    更新日期:2016-02-29 00:00:00

  • A quantitative analysis of 3D-cell distribution in regenerating muscle-skeletal system with synchrotron X-ray computed microtomography.

    abstract::One of the greatest enigmas of modern biology is how the geometry of muscular and skeletal structures are created and how their development is controlled during growth and regeneration. Scaling and shaping of vertebrate muscles and skeletal elements has always been enigmatic and required an advanced technical level in...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-018-32459-2

    authors: Tesařová M,Mancini L,Simon A,Adameyko I,Kaucká M,Elewa A,Lanzafame G,Zhang Y,Kalasová D,Szarowská B,Zikmund T,Novotná M,Kaiser J

    更新日期:2018-09-20 00:00:00

  • Dietary flavonoids, lignans and colorectal cancer prognosis.

    abstract::Flavonoids and lignans are polyphenol classes with anticarcinogenic activities against colorectal cancer (CRC). However, very limited epidemiological evidence exists on their effects on CRC prognosis. This study aimed to evaluate the association between flavonoid and lignan intakes with the risk of CRC recurrence and ...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/srep14148

    authors: Zamora-Ros R,Guinó E,Alonso MH,Vidal C,Barenys M,Soriano A,Moreno V

    更新日期:2015-09-15 00:00:00

  • Distillation of photon entanglement using a plasmonic metamaterial.

    abstract::Plasmonics is a rapidly emerging platform for quantum state engineering with the potential for building ultra-compact and hybrid optoelectronic devices. Recent experiments have shown that despite the presence of decoherence and loss, photon statistics and entanglement can be preserved in single plasmonic systems. This...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/srep18313

    authors: Asano M,Bechu M,Tame M,Kaya Özdemir Ş,Ikuta R,Güney DÖ,Yamamoto T,Yang L,Wegener M,Imoto N

    更新日期:2015-12-16 00:00:00

  • Publisher Correction: A humanized mouse model identifies key amino acids for low immunogenicity of H7N9 vaccines.

    abstract::An amendment to this paper has been published and can be accessed via a link at the top of the paper. ...

    journal_title:Scientific reports

    pub_type: 已发布勘误

    doi:10.1038/s41598-019-50828-3

    authors: Wada Y,Nithichanon A,Nobusawa E,Moise L,Martin WD,Yamamoto N,Terahara K,Hagiwara H,Odagiri T,Tashiro M,Lertmemongkolchai G,Takeyama H,De Groot AS,Ato M,Takahashi Y

    更新日期:2019-10-08 00:00:00

  • Macromolecular Characterization of Swordfish Oocytes by FTIR Imaging Spectroscopy.

    abstract::During folliculogenesis, primary oocytes of teleosts grow by several orders of magnitude by-self synthesizing proteins and mRNA, or sequestering from blood specific macromolecular components, such as fatty acids and vitellogenin. All these materials are stored into cortical alveoli, yolk globules or oil droplets durin...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-019-45065-7

    authors: Carnevali O,Candelma M,Sagrati A,Pignalosa P,Giorgini E,Gioacchini G

    更新日期:2019-06-20 00:00:00

  • Earth tectonics as seen by GOCE - Enhanced satellite gravity gradient imaging.

    abstract::Curvature components derived from satellite gravity gradients provide new global views of Earth's structure. The satellite gravity gradients are based on the GOCE satellite mission and we illustrate by curvature images how the Earth is seen differently compared to seismic imaging. Tectonic domains with similar seismic...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-018-34733-9

    authors: Ebbing J,Haas P,Ferraccioli F,Pappa F,Szwillus W,Bouman J

    更新日期:2018-11-05 00:00:00

  • LEGO® Block Structures as a Sub-Kelvin Thermal Insulator.

    abstract::We report measurements of the thermal conductance of a structure made from commercial Acrylonitrile Butadiene Styrene (ABS) modules, known as LEGO® blocks, in the temperature range from 70 mK to 1.8 K. A power law for the sample's thermal conductivity κ = (8.7 ± 0.3) × 10-5 T 1.75±0.02 WK-1 m-1 was determined. We conc...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-019-55616-7

    authors: Chawner JMA,Jones AT,Noble MT,Pickett GR,Tsepelin V,Zmeev DE

    更新日期:2019-12-23 00:00:00

  • Advanced P Wave Detection in Ecg Signals During Pathology: Evaluation in Different Arrhythmia Contexts.

    abstract::Reliable P wave detection is necessary for accurate and automatic electrocardiogram (ECG) analysis. Currently, methods for P wave detection in physiological conditions are well-described and efficient. However, methods for P wave detection during pathology are not generally found in the literature, or their performanc...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-019-55323-3

    authors: Maršánová L,Němcová A,Smíšek R,Vítek M,Smital L

    更新日期:2019-12-13 00:00:00

  • A map of the PGC-1α- and NT-PGC-1α-regulated transcriptional network in brown adipose tissue.

    abstract::Transcriptional coactivator PGC-1α and its splice variant NT-PGC-1α play crucial roles in regulating cold-induced thermogenesis in brown adipose tissue (BAT). PGC-1α and NT-PGC-1α are highly induced by cold in BAT and subsequently bind to and coactivate many transcription factors to regulate expression of genes involv...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-018-26244-4

    authors: Chang JS,Ghosh S,Newman S,Salbaum JM

    更新日期:2018-05-18 00:00:00

  • Complementary Role of Fibroblast Growth Factor 21 and Cytokeratin 18 in Monitoring the Different Stages of Nonalcoholic Fatty Liver Disease.

    abstract::Fibroblast growth factor 21 (FGF21) and cytokeratin 18 (CK18) were previously reported to be elevated in nonalcoholic fatty liver disease (NAFLD). We aim to analyze the differential roles of FGF21, cell apoptosis marker CK18 fragment M30 and total cell death marker CK18 M65ED in monitoring the different stages of NAFL...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-017-05257-5

    authors: Wu G,Li H,Fang Q,Zhang J,Zhang M,Zhang L,Wu L,Hou X,Lu J,Bao Y,Jia W

    更新日期:2017-07-11 00:00:00

  • Structural modulation of lithium metal-electrolyte interface with three-dimensional metallic interlayer for high-performance lithium metal batteries.

    abstract::The use of lithium (Li) metal anodes has been reconsidered because of the necessity for a higher energy density in secondary batteries. However, Li metal anodes suffer from 'dead' Li formation and surface deactivation which consequently form a porous layer of redundant Li aggregates. In this work, a fibrous metal felt...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/srep30830

    authors: Lee H,Song J,Kim YJ,Park JK,Kim HT

    更新日期:2016-08-03 00:00:00

  • Assembly, growth and conductive properties of tellurium nanorods produced by Rhodococcus aetherivorans BCP1.

    abstract::Tellurite (TeO32-) is a hazardous and toxic oxyanion for living organisms. However, several microorganisms can bioconvert TeO32- into the less toxic form of elemental tellurium (Te0). Here, Rhodococcus aetherivorans BCP1 resting (non-growing) cells showed the proficiency to produce tellurium-based nanoparticles (NPs) ...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-018-22320-x

    authors: Presentato A,Piacenza E,Darbandi A,Anikovskiy M,Cappelletti M,Zannoni D,Turner RJ

    更新日期:2018-03-02 00:00:00

  • Phylogenetic evidence of the intercontinental circulation of a Canine distemper virus lineage in the Americas.

    abstract::Canine distemper virus (CDV) is the cause of a multisystem disease in domestic dogs and wild animals, infecting more than 20 carnivore and non-carnivore families and even infecting human cell lines in in vitro conditions. Phylogenetic classification based on the hemagglutinin gene shows 17 lineages with a phylogeograp...

    journal_title:Scientific reports

    pub_type: 杂志文章

    doi:10.1038/s41598-019-52345-9

    authors: Duque-Valencia J,Forero-Muñoz NR,Díaz FJ,Martins E,Barato P,Ruiz-Saenz J

    更新日期:2019-10-31 00:00:00