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1.
Sci Rep ; 10(1): 64, 2020 01 09.
Article in English | MEDLINE | ID: mdl-31919397

ABSTRACT

Chediak-Higashi Syndrome (CHS) is a well-characterized, autosomal recessively inherited lysosomal disease caused by mutations in lysosomal trafficking regulator (LYST). The feline model for CHS was originally maintained for ~20 years. However, the colonies were disbanded and the CHS cat model was lost to the research community before the causative mutation was identified. To resurrect the cat model, semen was collected and cryopreserved from a lone, fertile,  CHS carrier male. Using cryopreserved semen, laparoscopic oviductal artificial insemination was performed on three queens, two queens produced 11 viable kittens. To identify the causative mutation, a fibroblast cell line, derived from an affected cat from the original colony, was whole genome sequenced. Visual inspection of the sequence data identified a candidate causal variant as a ~20 kb tandem duplication within LYST, spanning exons 30 through to 38 (NM_001290242.1:c.8347-2422_9548 + 1749dup). PCR genotyping of the produced offspring demonstrated three individuals inherited the mutant allele from the CHS carrier male. This study demonstrated the successful use of cryopreservation and assisted reproduction to maintain and resurrect biomedical models and has defined the variant causing Chediak-Higashi syndrome in the domestic cat.


Subject(s)
Chediak-Higashi Syndrome/pathology , Vesicular Transport Proteins/genetics , Alleles , Animals , Cats , Cell Line , Chediak-Higashi Syndrome/genetics , Disease Models, Animal , Exons , Female , Fibroblasts/cytology , Fibroblasts/metabolism , Genotype , Male , Pedigree , Polymorphism, Genetic , Vesicular Transport Proteins/metabolism
2.
J Synchrotron Radiat ; 26(Pt 1): 263-271, 2019 Jan 01.
Article in English | MEDLINE | ID: mdl-30655494

ABSTRACT

GALAXIES is an in-vacuum undulator hard X-ray micro-focused beamline dedicated to the study of the electronic structure of materials with high energy resolution using both photoelectron spectroscopy and inelastic X-ray scattering and under both non-resonant (NR-IXS) and resonant (RIXS) conditions. Due to the penetrating power of hard X-rays and the `photon-in/photon-out' technique, the sample environment is not a limitation. Materials under extreme conditions, for example in diamond anvil cells or catalysis chambers, thus constitute a major research direction. Here, the design and performance of the inelastic X-ray scattering end-station that operates in the energy range from ∼4 keV up to 12 keV is reported, and its capabilities are highlighted using a selection of data taken from recently performed experiments. The ability to scan `on the fly' the incident and scattered/emitted X-ray energies, and the sample position enables fast data collection and high experimental throughput. A diamond X-ray transmission phase retarder, which can be used to generate circularly polarized light, will also be discussed in the light of the recent RIXS-MCD approach.

3.
J Synchrotron Radiat ; 22(1): 175-9, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25537606

ABSTRACT

The GALAXIES beamline at the SOLEIL synchrotron is dedicated to inelastic X-ray scattering (IXS) and photoelectron spectroscopy (HAXPES) in the 2.3-12 keV hard X-ray range. These two techniques offer powerful complementary methods of characterization of materials with bulk sensitivity, chemical and orbital selectivity, resonant enhancement and high resolving power. After a description of the beamline components and endstations, the beamline capabilities are demonstrated through a selection of recent works both in the solid and gas phases and using either IXS or HAXPES approaches. Prospects for studies on liquids are discussed.

4.
Inorg Chem ; 53(1): 375-82, 2014 Jan 06.
Article in English | MEDLINE | ID: mdl-24350659

ABSTRACT

The valence state of uranium has been confirmed for the three sodium uranates NaU(V)O3/[Rn](5f(1)), Na4U(VI)O5/[Rn](5f(0)), and Na2U(VI)2O7/[Rn](5f(0)), using X-ray absorption near-edge structure (XANES) spectroscopy. Solid-state (23)Na magic angle spinning nuclear magnetic resonance (MAS NMR) measurements have been performed for the first time, yielding chemical shifts at -29.1 (NaUO3), 15.1 (Na4UO5), and -14.1 and -19 ppm (Na1 8-fold coordinated and Na2 7-fold coordinated in Na2U2O7), respectively. The [Rn]5f(1) electronic structure of uranium in NaUO3 causes a paramagnetic shift in comparison to Na4UO5 and Na2U2O7, where the electronic structure is [Rn]5f(0). A (23)Na multi quantum magic angle spinning (MQMAS) study on Na2U2O7 has confirmed a monoclinic rather than rhombohedral structure with evidence for two distinct Na sites. DFT calculations of the NMR parameters on the nonmagnetic compounds Na4UO5 and Na2U2O7 have permitted the differentiation between the two Na sites of the Na2U2O7 structure. The linear thermal expansion coefficients of all three compounds have been determined using high-temperature X-ray diffraction: αa = 22.7 × 10(-6) K(-1), αb = 12.9 × 10(-6) K(-1), αc = 16.2 × 10(-6) K(-1), and αvol = 52.8 × 10(-6) K(-1) for NaUO3 in the range 298-1273 K; αa = 37.1 × 10(-6) K(-1), αc = 6.2 × 10(-6) K(-1), and αvol = 81.8 × 10(-6) K(-1) for Na4UO5 in the range 298-1073 K; αa = 6.7 × 10(-6) K(-1), αb = 14.4 × 10(-6) K(-1), αc = 26.8 × 10(-6) K(-1), αß = -7.8 × 10(-6) K(-1), and αvol = -217.6 × 10(-6) K(-1) for Na2U2O7 in the range 298-573 K. The α to ß phase transition reported for the last compound above about 600 K was not observed in the present studies, either by high-temperature X-ray diffraction or by differential scanning calorimetry.

5.
Inorg Chem ; 52(19): 11669-76, 2013 Oct 07.
Article in English | MEDLINE | ID: mdl-24063301

ABSTRACT

A series of uranium carbide samples, prepared by arc melting with a C/U ratio ranging from 0.96 to 1.04, has been studied by X-ray diffraction (XRD), (13)C nuclear magnetic resonance (NMR), and extended X-ray absorption fine structure (EXAFS). XRD determines phase uniqueness and the increase of the lattice parameter versus the carbon content. In contrast, (13)C NMR detects the different carbon environments in the lattice and in this study, clearly identifies the presence of discrete peaks for carbon in the octahedral lattice site in UC and an additional peak associated with excess carbon in hyperstoichiometric samples. Two peaks associated with different levels of carbon deficiency are detected for all hypostoichiometric compositions. More than one carbon environment is always detected by (13)C NMR. This exemplifies the difficulty in obtaining a perfect stoichiometric uranium monocarbide UC(1.00). The (13)C MAS spectra of uranium carbides exhibit the effects resulting from the carbon content on both the broadening of the peaks and on the Knight shift. An abrupt spectral change occurs between hypo- and hyperstoichiometric samples. The results obtained by EXAFS highlight subtle differences between the different stoichiometries, and in the hyperstoichiometric samples, the EXAFS results are consistent with the excess carbon atoms being in the tetrahedral interstitial position.

6.
Inorg Chem ; 51(21): 11762-8, 2012 Nov 05.
Article in English | MEDLINE | ID: mdl-23072315

ABSTRACT

The electronical and structural properties of Th(0.80)Am(0.20)O(2-x) materials have been studied by the coupling of X-ray diffraction and X-ray absorption spectroscopy techniques. A substoichiometric fluorite Th(IV)(0.80)Am(III)(0.20)O(1.90) solid solution is found following sintering in moisturized Ar-H(2). In contrast, heating of this sample in air leads to a nondefective fluorite Th(IV)(0.80)Am(IV)(0.20)O(2.00) solid solution. The structures of these solid solution compounds were fully characterized by assessing the interatomic distances, the coordination numbers, and the structural disorder. The effect of the sintering atmosphere on these crystallographical parameters and on the cation valences has been determined and the capability of ThO(2) to accommodate tri- and tetravalent actinides in the fluorite structure assessed.

7.
Geobiology ; 7(4): 403-18, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19656214

ABSTRACT

Hydrothermal activity was common on the early Earth and associated micro-organisms would most likely have included thermophilic to hyperthermophilic species. 3.5-3.3 billion-year-old, hydrothermally influenced rocks contain silicified microbial mats and colonies that must have been bathed in warm to hot hydrothermal emanations. Could they represent thermophilic or hyperthermophilic micro-organisms and if so, how were they preserved? We present the results of an experiment to silicify anaerobic, hyperthermophilic micro-organisms from the Archaea Domain Pyrococcus abyssi and Methanocaldococcus jannaschii, that could have lived on the early Earth. The micro-organisms were placed in a silica-saturated medium for periods up to 1 year. Pyrococcus abyssi cells were fossilized but the M. jannaschii cells lysed naturally after the exponential growth phase, apart from a few cells and cell remains, and were not silicified although their extracellular polymeric substances were. In this first simulated fossilization of archaeal strains, our results suggest that differences between species have a strong influence on the potential for different micro-organisms to be preserved by fossilization and that those found in the fossil record represent probably only a part of the original diversity. Our results have important consequences for biosignatures in hydrothermal or hydrothermally influenced deposits on Earth, as well as on early Mars, as environmental conditions were similar on the young terrestrial planets and traces of early Martian life may have been similarly preserved as silicified microfossils.


Subject(s)
Fossils , Methanococcales/metabolism , Pyrococcus abyssi/metabolism , Silicon Dioxide/metabolism , Soil Microbiology
8.
J Bacteriol ; 189(12): 4510-9, 2007 Jun.
Article in English | MEDLINE | ID: mdl-17449623

ABSTRACT

Only one virus-like particle (VLP) has been reported from hyperthermophilic Euryarchaeotes. This VLP, named PAV1, is shaped like a lemon and was isolated from a strain of "Pyrococcus abyssi," a deep-sea isolate. Its genome consists of a double-stranded circular DNA of 18 kb which is also present at a high copy number (60 per chromosome) free within the host cytoplasm but is not integrated into the host chromosome. Here, we report the results of complete analysis of the PAV1 genome. All the 25 predicted genes, except 3, are located on one DNA strand. A transcription map has been made by using a reverse transcription-PCR assay. All the identified open reading frames (ORFs) are transcribed. The most significant similarities relate to four ORFs. ORF 180a shows 31% identity with ORF 181 of the pRT1 plasmid isolated from Pyrococcus sp. strain JT1. ORFs 676 and 678 present similarities with a concanavalin A-like lectin/glucanase domain, which could be involved in the process of host-virus recognition, and ORF 59 presents similarities with the transcriptional regulator CopG. The genome of PAV1 displays unique features at the nucleic and proteinic level, indicating that PAV1 should be attached at least to a novel genus or virus family.


Subject(s)
Archaeal Viruses/genetics , DNA, Viral/genetics , Genome, Viral/genetics , Pyrococcus abyssi/virology , Amino Acid Sequence , Archaeal Viruses/classification , DNA, Viral/chemistry , Genes, Regulator/genetics , Genes, Viral , Lectins/genetics , Molecular Sequence Data , Open Reading Frames , Plasmids/genetics , RNA, Messenger/genetics , RNA, Viral/genetics , Replication Origin , Reverse Transcriptase Polymerase Chain Reaction , Sequence Alignment , Sequence Analysis, DNA , Sequence Homology , Transcription, Genetic
9.
J Hered ; 96(4): 302-9, 2005.
Article in English | MEDLINE | ID: mdl-15817712

ABSTRACT

Type II tyrosinemia, designated Richner-Hanhart syndrome in humans, is a hereditary metabolic disorder with autosomal recessive inheritance characterized by a deficiency of tyrosine aminotransferase activity. Mutations occur in the human tyrosine aminotransferase gene, resulting in high levels of tyrosine and disease. Type II tyrosinemia occurs in mink, and our hypothesis was that it would also be associated with mutation(s) in the tyrosine aminotransferase gene. Therefore, the transcribed cDNA and the genomic tyrosine aminotransferase gene were sequenced from normal and affected mink. The gene extended over 11.9 kb and had 12 exons coding for a predicted 454-amino-acid protein with 93% homology with human tyrosine aminotransferase. FISH analysis mapped the gene to chromosome 8 using the Mandahl and Fredga (1975) nomenclature and chromosome 5 using the Christensen et al. (1996) nomenclature. The hypothesis was rejected because sequence analysis disclosed no mutations in either cDNA or introns that were associated with affected mink. This suggests that an unlinked gene regulatory mutation may be the cause of tyrosinemia in mink.


Subject(s)
DNA, Complementary/genetics , Mink/genetics , Tyrosine Transaminase/genetics , Tyrosinemias/genetics , Amino Acid Sequence , Animal Diseases/enzymology , Animal Diseases/genetics , Animals , Base Sequence , Chromosome Mapping , DNA Primers , In Situ Hybridization, Fluorescence , Introns/genetics , Molecular Sequence Data , Tyrosinemias/enzymology , Tyrosinemias/veterinary
10.
Virologie (Montrouge) ; 9(5): 357-366, 2005 Oct 01.
Article in French | MEDLINE | ID: mdl-34679282

ABSTRACT

Our knowledge of the diversity of the viruses infecting prokaryotic micro-organisms from extreme environments still remains very rudimentary. With about 5 150 viruses of prokaryotes described to date, only forty, were isolated from Archaea (Halophiles, methanogens, thermoacidophiles or hyperthermophiles). Nevertheless, the studies undertaken recently on hyperthermophilic Archaea from terrestrial or oceanic hydrothermal environments suggest the existence of an impressive morphological and genomic viral diversity. Among the different morphotypes observed, the lemon-shaped type prevailed but rigid rods, filaments and unique pleomorphic morphologies never yet observed were also detected. The majority of these new viruses was isolated from the phylum Crenarchaeota, mostly among representatives of the order Sulfolobales, whereas only one virus was described in the hyperthermophilic members of the phylum Euryarchaeota. Analysis of the genomes of these new viruses show that 90 to 100 % of the predicted proteins were not related to anything previously reported. The viruses of the hyperthermophiles thus represent an important reservoir of new proteinic structures and new biological functions.

11.
Biochem Soc Trans ; 32(Pt 2): 184-7, 2004 Apr.
Article in English | MEDLINE | ID: mdl-15046568

ABSTRACT

This minireview summarizes our current knowledge about archaeal genetic elements in the hyperthermophilic order Thermococcales in the phylum Euryarchaeota. This includes recent work on the first virus of Pyrococcus, PAV1, the discovery of novel unique virus morphotypes in hot deep-sea environments, and preliminary observations on novel cryptic plasmids. We also review the work accomplished over the last 5 years in the development of genetic tools for members of the Pyrococcus and Thermococcus genera, mainly in our laboratories.


Subject(s)
Genome, Archaeal , Pyrococcus/genetics , Thermococcales/genetics , Thermococcus/genetics , Antigens, Archaeal/chemistry , DNA, Archaeal/ultrastructure , Genetic Vectors , Hydrogen-Ion Concentration , Microscopy, Electron, Transmission , Organometallic Compounds/pharmacology , Plasmids/metabolism , Transgenes , Viruses/genetics
12.
J Bacteriol ; 185(13): 3888-94, 2003 Jul.
Article in English | MEDLINE | ID: mdl-12813083

ABSTRACT

We describe the first virus-like particle of a hyperthermophilic euryarchaeote which was discovered in a strain of "Pyrococcus abyssi" previously characterized in our laboratory. This particle, named PAV1, is lemon-shaped (120 nm x 80 nm), with a short tail terminated by fibers, and resembles the virus SSV1, the type member of the Fuselloviridae, isolated from Sulfolobus shibatae. Sensitivity of the virus-like particle to organic solvents and detergents suggested that the envelope of PAV1 may contain lipids in addition to proteins. It contains a double-stranded circular DNA of 18 kb which is also present in high copy number in a free form in the host cytoplasm. No integrated form of the PAV1 genome could be detected in the host chromosome. Under standard growth conditions, the host cells continuously release PAV1 particles into the culture supernatant without spontaneous lysis, with a maximum reached in the late stationary phase. UV, gamma irradiation, treatment with mitomycin C, and various physiological stresses had no effect on PAV1 production. Screening of a large number of Thermococcales isolates did not permit to find a sensitive host. These results suggest that PAV1 persists in the host strain in a stable carrier state rather than a prophage.


Subject(s)
Fuselloviridae/classification , Fuselloviridae/isolation & purification , Hot Temperature , Pyrococcus/virology , Virion/classification , Virion/isolation & purification , DNA/analysis , DNA, Circular/analysis , DNA, Viral/analysis , Electrophoresis/methods , Fuselloviridae/genetics , Fuselloviridae/ultrastructure , Genome, Viral , Microscopy, Electron , Seawater/microbiology , Thermococcales/virology , Viral Proteins/chemistry , Virion/genetics , Virion/ultrastructure
13.
FEMS Microbiol Lett ; 205(2): 329-35, 2001 Dec 18.
Article in English | MEDLINE | ID: mdl-11750823

ABSTRACT

The prokaryotic phylogenetic diversity was determined for a sample associated with an in situ growth chamber deployed for 5 days on a Mid-Atlantic Ridge hydrothermal vent (23 degrees 22'N, 44 degrees 57'W). The DNA was extracted from the sample and the 16S rDNA amplified by PCR. No Archaea were detected in the sample. Eighty-seven clones containing bacterial 16S rDNA inserts were selected. Based on restriction fragment length polymorphism analysis, 47 clones were unique, however, based on comparative sequence analysis some of these were very similar, and thus only 22 clones were selected for full sequence and phylogenetic analysis. The phylotypes were dominated by epsilon-Proteobacteria (66%). The remainder formed a novel lineage within the Proteobacteria (33%). One clone formed a distinct deeply branching lineage, and was a distant relative of the Aquificales. This report further expands the growing evidence that epsilon-Proteobacteria are important members in biogeochemical cycling at deep-sea hydrothermal ecosystems, participating as epibionts and free living bacteria.


Subject(s)
Epsilonproteobacteria/genetics , Seawater/microbiology , Water Microbiology , Ecosystem , Epsilonproteobacteria/classification , Epsilonproteobacteria/isolation & purification , Genetic Variation , Mid-Atlantic Region , Molecular Sequence Data , Phylogeny , Protein Structure, Secondary , RNA, Bacterial/chemistry , RNA, Ribosomal, 16S/chemistry , Temperature
14.
Mol Genet Genomics ; 266(1): 72-8, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11589580

ABSTRACT

Hyperthermophilic archaea of the genus Pyrococcus are resistant to gamma radiation, suggesting that efficient mechanisms for DNA repair exist in these organisms. To determine whether protective mechanisms might also be implicated in this radioresistance, we have estimated the linear density of DNA double-stranded breaks caused by gamma irradiation in the genomic DNA of two Pyrococcus species, using Escherichia coli and the radioresistant bacterium Deinococcus radiodurans as controls. The linear density of double-stranded breaks was essentially the same in all four microorganisms when irradiation was carried under similar anaerobic conditions, indicating that no specific DNA protection mechanisms exist in Pyrococcus species. Using one- and two-dimensional gel electrophoresis we compared the protein patterns from Pyrococcus abyssi and P. furiosus cells that had or had not been exposed to gamma rays. We did not detect any significant protein induction following DNA damage in either species.


Subject(s)
DNA, Archaeal/genetics , Pyrococcus/radiation effects , Radiation Tolerance/genetics , DNA Damage , Electrophoresis/methods , Escherichia coli/radiation effects , Gamma Rays , Pyrococcus/genetics , Species Specificity
15.
Article in English | MEDLINE | ID: mdl-11544095

ABSTRACT

The distribution of the enzyme tyrosine aminotransferase in tissues of mink, Mustela vison, was investigated. High levels of enzymatic activity were detected only in liver, documenting the hepatic-specific nature of this enzyme in this species. Further studies disclosed that tyrosine aminotransferase is not absent from non-hepatic tissues because of the lack of the use of a stabilized buffer, sensitivity to temperature, or due to the presence of an inhibitor. Collectively, these results suggest that the enzymatic assay of tyrosine aminotransferase will be unlikely to be an efficacious approach for identifying mink that are heterozygous for the autosomal recessive deficiency of this enzyme that is common in dark mink.


Subject(s)
Liver/enzymology , Mink/metabolism , Tyrosine Transaminase/metabolism , Animals , Enzyme Stability , Fasting , Gene Dosage , Kidney/enzymology , Liver/physiology , Sonication , Spleen/enzymology , Telencephalon/enzymology , Temperature , Tissue Distribution
16.
Res Microbiol ; 152(10): 901-5, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11766965

ABSTRACT

Superoxide dismutases (SODs) are vital components that defend against oxidative stress through decomposition of superoxide radical. Escherichia coli contains two highly homologous SODs, a manganese- and an iron-containing enzyme (Mn-SOD and Fe-SOD, respectively). In contrast, a single Mn-SOD is present in Bacillus subtilis. In E. coli, the absence of SODs was found to be associated with an increased sensitivity to cadmium, nickel and cobalt ions. Mutants lacking either sodA or sodB exhibited metal resistance to levels comparable to that of the wild-type strain. Although sod-deficient mutant cells were more resistant to zinc than their wild-type counterpart, no differences between the strains were observed in the presence of copper. In B. subtilis, the sodA mutation had no effect on cadmium and copper resistance. These results suggest that intracellular generation of superoxide by cadmium, nickel and cobalt is toxic in E. coli. They support the participation of sod genes in its protection against metal stress.


Subject(s)
Bacterial Proteins/physiology , Escherichia coli/drug effects , Metals, Heavy/pharmacology , Superoxide Dismutase/physiology , Bacterial Proteins/genetics , Cadmium/pharmacology , Cations , Cobalt/pharmacology , Escherichia coli/enzymology , Microbial Sensitivity Tests , Mutation , Nickel/pharmacology , Superoxide Dismutase/genetics
17.
Curr Microbiol ; 41(3): 201-5, 2000 Sep.
Article in English | MEDLINE | ID: mdl-10915208

ABSTRACT

Thirty thermophilic strains isolated from heavy metal-rich hydrothermal vent sites at Lau Basin were tested for their susceptibility to cadmium, zinc, cobalt, and nickel. The 14 aerobic spore formers belonging to the genus Bacillus, 6 anaerobic fermenters from the order Thermotogales, and 10 anaerobic sulfur reducers from the order Thermococcales could be clearly distinguished according to their metal susceptibilities. The Thermococcales were found to exhibit the highest resistance to cadmium and zinc, whereas Thermotogales were highly sensitive to these metals. In contrast, the Thermotogales displayed the highest resistance to cobalt ions. No clear distinction could be established between the metal susceptibilities of these strains and seven reference organisms used for comparative studies. Cadmium resistance, slightly inducible in some cadmium-resistant bacilli, was not plasmid mediated. The amount of cadmium immobilized by the Thermotogales was related to their level of resistance to this metal.


Subject(s)
Bacillus/drug effects , Bacteria, Anaerobic/drug effects , Metals, Heavy/pharmacology , Thermococcales/drug effects , Water Microbiology , Bacillus/growth & development , Bacteria, Anaerobic/metabolism , Cadmium/analysis , Cadmium/pharmacology , Cobalt/pharmacology , Culture Media , Drug Resistance, Microbial , Hydrogen-Ion Concentration , Metals, Heavy/analysis , Microbial Sensitivity Tests , Pacific Ocean , Time Factors , Zinc/pharmacology
18.
Can J Microbiol ; 46(5): 465-73, 2000 May.
Article in English | MEDLINE | ID: mdl-10872082

ABSTRACT

The nitrification activity of a thermophilic heterotrophic bacterium, Bacillus MS30 isolated from a deep-sea hydrothermal vent, was studied under various growth conditions. Nitrification was estimated from the nitrogen balance calculations in the culture media. The results showed that this isolate actively nitrified in culture conditions similar to those prevailing in hydrothermal sites. Therefore, its ecological significance was considered. In standard aerobic conditions, MS30 produced nitrite from ammonia and acetate (1.13 mumol NO2-.mg-1 dry wt), but nitrate was never produced, and a low nitrite reduction was often observed. Higher nitrification activities were observed in defined optimal conditions (simple carbon substrate, 65 degrees C, pH 7.5, and 15 g sea salts.L-1). In addition, discrepancies between the optima for growth and nitrification were observed, showing the ability of MS30 to adapt to changing environmental conditions typical of hydrothermal sites.


Subject(s)
Bacillus/metabolism , Nitrates/metabolism , Nitrites/metabolism , Seawater/microbiology , Ammonia/metabolism , Bacillus/growth & development , Culture Media , Hydrogen-Ion Concentration , Salts/pharmacology , Temperature
19.
Extremophiles ; 3(4): 239-45, 1999 Nov.
Article in English | MEDLINE | ID: mdl-10591013

ABSTRACT

Fourteen strains of hyperthermophilic organotrophic anaerobic marine Archaea were isolated from shallow water and deep-sea hot vents, and four of them were characterized. These isolates, eight previously published strains, and six type strains of species of the order Thermococcales were selected for the study of cell wall components by means of thin sectioning or freeze-etching electron microscopy. The cell envelopes of most isolates were shown to consist of regularly arrayed surface protein layers, either single or double, with hexagonal lattice (p6) symmetry, as the exclusive constituents outside the cytoplasmic membrane. The S-layers studied differed in center-to-center spacing and molecular mass of the constituent protein subunits. Polyclonal antisera raised against the cells of 10 species were found to be species-specific and allowed 12 new isolates from shallow water hot vents to be identified as representatives of the species Thermococcus litoralis, Thermococcus stetteri, Thermococcus chitonophagus, and Thermococcus pacificus. Of the 7 deep-sea isolates, only 1 was identified as a T. litoralis strain. Thus, hyperthermophilic marine organotrophic isolates obtained from deep-sea hot vents showed greater diversity with regard to their S-layer proteins than shallow water isolates.


Subject(s)
Antigens, Archaeal/analysis , Thermococcales/classification , Thermococcus/classification , Cell Membrane/immunology , Cell Membrane/ultrastructure , Seawater/microbiology , Thermococcales/immunology , Thermococcales/ultrastructure , Thermococcus/immunology , Thermococcus/ultrastructure
20.
Extremophiles ; 3(4): 247-51, 1999 Nov.
Article in English | MEDLINE | ID: mdl-10591014

ABSTRACT

We examined a single, non-spore-forming, aerobic, thermophilic strain that was isolated from a deep-sea hydrothermal vent in the Guaymas Basin at a depth of 2000 m and initially placed in a phenetic group with Thermus scotoductus (X-1). We identified this deep-sea isolate as a new strain belonging to Thermus thermophilus using several parameters. DNA-DNA hybridization under stringent conditions showed 74% similarity between the deep-sea isolate and T. thermophilus HB-8T (T = type strain). Phenotypic characteristics, such as the utilization of carbon sources, hydrolysis of different compounds, and antibiotic sensitivity were identical in the two strains. The polar lipids composition showed that strain Gy1211 belonged to the genus Thermus. The fatty acids composition indicated that this strain was related to the marine T. thermophilus strain isolated from the Azores. The new isolate T. thermophilus strain Gy1211 grew optimally at 75 degrees C, pH 8.0; and 2% NaCl. A hydrostatic pressure of 20 MPa, similar to the in situ hydrostatic pressure of the deep-sea vent from which the strain was isolated, had no effect on growth. Strain HB-8T, however, showed slower growth under these conditions.


Subject(s)
Seawater/microbiology , Thermus thermophilus/classification , Thermus thermophilus/physiology , Cell Division , DNA, Bacterial/genetics , Microbial Sensitivity Tests , Pressure , Spores, Bacterial , Thermus/classification , Thermus thermophilus/isolation & purification
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