Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:6.3.4.6 (urease)
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The nuclear DNA of 28 species (30 strains investigated) of yeasts classified currently or previously in the genus Trichosporon. was analysed for its molar percentage of guanine + cytosine (mol% G + C). This criterion, together with biochemical characteristics, suggested the separation of the organisms studied into two groups. The first group, which appears related to the Ascomycetes, includes thirteen species with a G + C content lower than 50 mol% (34.7-48.8), and lacks urease (except T. margaritiferum). The second group appears related to the Basidiomycetes and includes fifteen species with a G + C content higher than 50 mol% (57-64) and has the ability to hydrolyse urea. A DNA homology experiment with T. beigelii and twelve other species of the second group showed very low values of complementarity with T. beigelii-labeled DNA. All these species must be considered as taxa other than T. beigelii.
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PMID:DNA base composition and DNA relatedness among species of Trichosporon Behrend. 653 93

Quantitative transformation of streptomycin resistance marker was carried out with strains of Acinetobacter calcoaceticus. Standard recipient strain was the competent BD4-Ss. Transformation proceeded in a liquid system with a concentration of 20 micrograms/ml of streptomycin resistant DNA (Sr-DNA) from BD4, 17 reference strains of Acinetobacter, and 42 recent clinical isolates of the acid forming variant (anitratus) and 12 of the non-acid forming variant (lwoffi) as donors of Sr-DNA. The exposure time was 20 min before interruption with DNase and quantitated plating. Among the strains examined were differentiated 16 biotypes as based on oxidative acid production from glucose and lactose, haemolysis, urease, gelatinase, and growth with citrate as the sole carbon source. Autologous transformation gave a transformation of 2.94 (+/- 0.66)% of the recipient cells (quantitated by colony forming units). Sr-DNA from 50 of 63 strains were able to transform BD4-Ss. The transformation ratio was 0.06-6.4% of autologous BD4 transformation. Two further recent clinical isolates were weakly transformation competent. Competence was only found in autologous transformation. The major portion (50/63) of the strains belong to the BD4 genotype of A. calcoaceticus, but the taxonomic position of the 13 non-donors in respect to BD4 could not be evaluated by transformation because of lack of a competent recipient. The strains transforming BD4 belonged to both the glucose acidifyers and the non-acidifyers without genetic distinction. The results are consistent with recognition of both as belonging to the same species, but with their recognition as species variants: A calcoaceticus var, anitratum and A. calcoaceticus var. lwoffi.
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PMID:Taxonomic implications of quantitative transformation in Acinetobacter calcoaceticus. 658 37

We have cloned and sequenced the wild-type and suppressor alleles of the S. pombe sup8 tRNA gene. The wild-type allele has a leucine UAA anticodon and the suppressor (sup8-e) carries the opal suppressor anticodon UCA. The gene has a 16 base pair intervening sequence that, in the RNA, is predicted to form a secondary structure which involves base pairing to the 5', rather than the usual 3' side of the 5' splice site. When incubated in Saccharomyces cerevisiae cell-free extracts both alleles are efficiently transcribed, the 5' leader and 3' trailer sequences are removed and CCA is added to the 3' processed end; however, the intervening sequence is not excised. This finding implies that the structural requirements of the splicing endonucleases in the two yeasts have diverged. No other tRNA genes with related sequences were detected in S. pombe DNA by hybridization, suggesting that other UUA isoacceptors may be structurally dissimilar to sup8 or that the UUA codon may be decoded by a UUG leucine isoacceptor.
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PMID:The sup8 tRNALeu gene of Schizosaccharomyces pombe has an unusual intervening sequence and reduced pairing in the anticodon stem. 659 38

Urea-DNA glycosylase, an enzyme presumed to be active in the repair of DNA damage caused by oxidizing agents, has been identified previously in Escherichia coli. This enzyme has now been shown to be present in cell extracts of calf thymus and human fibroblasts. It catalyzes the release of free urea from a double-stranded polydeoxyribonucleotide containing thymine residues fragmented by KMnO4 and NaOH treatment. The calf thymus enzyme has been 400-fold purified and largely separated from previously identified mammalian DNA glycosylases. It has a molecular weight of about 25 000 and requires no cofactors. The identity of the enzymatically released product as unsubstituted urea has been verified by its susceptibility to urease.
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PMID:Urea--DNA glycosylase in mammalian cells. 662 1

Six strains of a new species, Legionella sainthelensi, were isolated from freshwater in areas affected by the volcanic eruptions of Mt. St. Helens in the state of Washington. Strains of L. sainthelensi are culturally and biochemically similar to other legionellae. They grow on buffered charcoal yeast agar but not on media that lack cysteine. They are gram-negative, nonsporeforming, motile rods that are positive in reactions for catalase, oxidase, gelatin liquefaction, and beta-lactamase. They are negative in reactions for urease, hydrolysis of hippurate, reduction of nitrates, fermentation of glucose, and blue-white autofluorescence. Their cell wall fatty acid composition is qualitatively similar to those of other legionellae, with 50 to 62% branched-chain fatty acids. They contain the isobranched-chain 14- and 16-carbon acids and anteisobranched-chain 15- and 17-carbon acids and relatively large amounts of straight-chain 16-carbon acid. All strains of L. sainthelensi contain approximately equal amounts of ubiquinones Q9, Q10, Q11, and Q12, a pattern similar to those of Legionella bozemanii, Legionella dumoffi, and Legionella longbeachae. Serological cross-reactions were observed between L. sainthelensi, both serogroups of L. longbeachae, and Legionella oakridgensis. Three strains of L. sainthelensi were greater than 90% related by DNA hybridization. The type strain of L. sainthelensi, Mt. St. Helens 4, was 36% related to the type strain of L. longbeachae and 3 to 14% related to the other nine described Legionella species.
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PMID:Legionella sainthelensi: a new species of Legionella isolated from water near Mt. St. Helens. 671 10

The API STAPH-IDENT system was compared with conventional methods for the identification of 14 Staphylococcus species. Conventional methods included the Kloos and Schleifer simplified scheme and DNA-DNA hybridization. The API STAPH-IDENT strip utilizes a battery of 10 miniaturized biochemical tests, including alkaline phosphatase, urease, beta-glucosidase, beta-glucuronidase, and beta-galactosidase activity, aerobic acid formation from D-(+)-mannose, D-mannitol, D-(+)-trehalose, and salicin, and utilization of arginine. Reactions of cultures were determined after 5 h of incubation at 35 degrees C. Results indicated a high degree of congruence (greater than 90%) between the expedient API system and conventional methods for most species. The addition of a test for novobiocin susceptibility to the API system increased the accuracy of identification of S. saprophyticus, S. cohnii, and S. hominis, significantly. Several strains of S. hominis, S. haemolyticus, and S. warneri which were difficult to separate with the Kloos and Schleifer simplified scheme were accurately resolved by the API system.
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PMID:Identification of Staphylococcus species with the API STAPH-IDENT system. 675 90

The regions 5' proximal to many yeast tRNA genes exhibit a high frequency of DNA sequence polymorphisms. DNA sequence analysis of polymorphic variants of SUQ5, a tRNA Ser UCA gene, and SUP2, a tRNA Tyr gene, shows that in each case one sequence variant of the tRNA gene is 346 base pairs longer than the other. The longer variants appear to have arisen from the shorter ones by the insertion of nearly identical copies of a 341-base pair sigma element into a site 16 base pairs upstream from the 5' ends of the tRNA-coding regions. The sequences of the two copies of the sigma element differ at only five positions. The element has a number of properties that are typical of many transposable elements: (i) there is a perfect eight-base-pair inverted repeat at its ends, (ii) these ends are flanked by a five-base-pair direct repeat of a sequence that occurs only once in the target DNA, (iii) there are approximately 20 copies of the element in the yeast genome, and (iv) there is considerable strain-to-strain variation in the sizes of the restriction fragments on which these copies lie. The presence of the sigma element has no gross effect on the phenotype of a SUP2 ochre suppressor. Analysis of the SUQ5 and SUP2 sequences favors the hypothesis that sigma is a transposable element with a novel type of insertion specificity, which is primarily based on the presence of a tRNA-coding region a fixed distance from the insertion site, rather than on the immediate target sequences.
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PMID:Insertion of a repetitive element at the same position in the 5'-flanking regions of two dissimilar yeast tRNA genes. 676 Feb 1

A simplified translation system coupled to DNA transcription that involves assaying the synthesis of the first dipeptide of a gene product has been described recently [Robakis, N., Meza-Basso, L., Brot, N. & Weissbach, H. (1981) Proc. Natl. Acad. Sci. USA 78, 4261--4264]. Using this dipeptide system, we have investigated the expression of genes carried on plasmids coding for beta-lactamase, ribosomal protein L12, and the chloroplast large subunit (LS) of ribulosebisphosphate carboxylase (RbuBPCase). Although all three nascent gene products begin with the sequence fMet-Ser, the formation of fMet-Ser can be used to distinguish between the synthesis of beta-lactamase and either L12 or the LS of RbuBPCase by using different serine isoacceptor tRNA species. In beta-lactamase, the serine codon is AGU, which utilizes the serine isoacceptor species tRNASer3; in L12 and the LS of RbuBPCase, the serine codewords are UCU and UCA, respectively, both of which are recognized by the serine isoacceptor species tRNASer1. By using either pure tRNASer1 or pure tRNASer3, the expression of each gene can be quantitated. In this system, guanosine-5'-diphosphate-3'-diphosphate inhibits the expression of the beta-lactamase and L12 genes but stimulates the synthesis of the LS. In addition, the ratio of fMet-Ser/fMet-Ala (L12/L10) synthesized was about 1 as compared with the ratio of 4 that has been obtained previously in vivo or in vitro protein-synthesizing systems in which the entire gene product was measured.
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PMID:Use of different tRNASer isoacceptor species in vitro to discriminate between the expression of plasmid genes. 680 42

During the summer of 1981, 3,887 sucrose-negative vibrios were isolated from seawater, sediment, plankton, and animal samples taken from 80 sites from Miami, Fla., to Portland, Maine. Of these, 4.2% were able to ferment lactose. The lactose-positive strains isolated from the various samples correlated positively with pH and turbidity of the water, vibrios in the sediment and oysters, and total bacterial counts in oysters. Negative correlations were obtained for water salinity. Numerical taxonomy was performed on 95 of the lactose-fermenting environmental isolates and 23 reference strains. Five clusters resulted, with the major cluster containing 33 of the environmental isolates and all of the Vibrio vulnificus reference strains. The 33 isolates, which produced an acid reaction in lactose broth within hours of initial inoculation, represented 20% of all lactose-fermenting vibrios studied. These isolates were nearly identical phenotypically to clinical strains of V. vulnificus studied by the Centers for Disease Control, Atlanta, Ga., and by our laboratory, and their identification was confirmed by DNA-DNA hybridization studies. V. vulnificus was isolated from all sample types and from Miami to Cape Cod, Mass., and comparison of the environmental parameters of the eight subsites yielding this species with those of all 80 subsites revealed no significant differences. The majority of the isolates were obtained from animals, with clams providing most (84%) of these. On injection into mice, 82% of the V. vulnificus isolates resulted in death. Members of the remaining four clusters contained strains which differed from V. vulnificus in such phenotypic traits as luminescence and in urease or H(2)S production. None of the other reference cultures, including nine other Vibrio species, were contained in the remaining clusters, and these isolates could not be identified. Most of these were also lethal for mice. Phenotypic differences, potential pathogenicity, and geographic distribution of the five clusters were examined. It is concluded that V. vulnificus is a ubiquitous organism, both geographically and in a variety of environmental sources, although it occurs in relatively low numbers. The public health significance of this organism and of the other unidentified lactose-fermenting Vibrio species is discussed.
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PMID:Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment. 684 90

1. The Rumen Simulation Technique (Rusitec) was used in a series of long-term experiments to study the distribution and changes of urease (EC 3.5.1.5) activity in a heterogeneous fermentation system. 2. It was shown that in Rusitec the high urease activity from the inoculum decreased to low values, that the rate of decrease was consistent with simple dilution of ureolytic micro-organisms and that the urease activity could be restored to original values by infusion of urea into the reaction vessels. The magnitude of this urease activity was a direct function of the amounts of urea infused. Single daily additions of the same or greater amounts of urea in food or as solid failed to increase the urease activity significantly. 3. In general, urease activity increased 2-6 h after feeding and the increases were greater with roughage diets. 4. The ureolytic activity per unit volume was always higher in compartment 2(space occupied by micro-organisms that are loosely associated with the solid) than in compartment 1 (strained rumen contents) or compartment 3 (space occupied by microbial population that cannot be washed out of the solid matrix). 5. The distribution of urease activity between the compartments was different from the distribution of certain other enzymes (e.g. protease and alkaline phosphatase (EC 3.1.3.1)). 6. Apart from the boundary region, the concentrations of urease, ammonia and volatile fatty acids in compartment 2 were constant, while the concentrations of protein, DNA and another enzyme (alkaline phosphatase) increased with the depth of the compartment. Specific urease activity (per unit weight of protein or DNA) was much higher in compartment 1 than in compartment 2 and it decreased markedly with depth of compartment. 7. The concentrations of ammonia were always much higher in the solid matrix (compartments 2 and 3) than in the free suspension of micro-organisms (compartment 1). There was a linear relation between these two quantities. 8. The results are discussed in relation to published work on the entry and metabolism of urea in the rumen.
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PMID:Distribution and changes in urease (EC 3.5.1.5) activity in Rumen Simulation Technique (Rusitec). 703 70


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