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  • 1
    ISSN: 1432-072X
    Keywords: Neurospora crassa ; “Slime” variant ; Cell coat ; Exocellular proteins ; Glycoproteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cells of the “slime” strain of Neurospora crassa synthesize a coherent extracellular material which remains attached to the cell surface, but is released into the liquid medium by shaking. The material was purified and studied by different criteria. By electron microscopy it appears as long wavy sheets which strongly bind concanavalin A, but not wheat germ agglutinin, and maintain their integrity in the absence of structural polysaccharides. Analysis of the purified material revealed that it was free of contaminating membranes; it contained more than 70% protein, 1% neutral sugars (glucose, mannose, fucose and galactose), less than 2% lipids and ca. 4% not-characterized hexosaminelike compounds. Its polypeptide pattern as determined by PAGE was complex. The significance of this material is discussed.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-072X
    Keywords: Neurospora crassa ‘Slime” variant ; Cell coat ; Glycoproteins ; Self-assembling
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The proteinaceous extracellular material (PEM) synthesized by the cells of the ‘slime” strain of Neurospora crassa (see Martinez et al. 1989) was solubilized by treatment with urea or guanidine. Removal of these chemicals by dialysis, caused reassembly of the solubilized proteins into material with the same microscopic appearance as the original PEM. Polypeptide patterns from both native and reassembled structures were identical. Dialysis-mediated reassembly of the solubilized proteins appeared to be dependent on both concentration of the soluble macromolecules and time. Gel chromatography of PEM solubilized with different agents revealed two discrete populations of complexes with molecular masses of 1,500 and 500 kDa respectively. These were able to reassemble into lamellar structures with a variable degree of efficiency.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-072X
    Keywords: Key words:Candida albicans– Dimorphism – Cell wall – Mannoprotein – EDTA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Hyphal development in Candida albicans was blocked by EDTA. This effect was not due to a general growth inhibition since the chelator did not affect protein and DNA synthesis. Recovery of mycelial growth was observed when EDTA-grown cells were incubated at 37 °C in EDTA-free medium. High-molecular-weight mannoproteins (HMWM) that are mycelium-specific wall components, and particularly a 260-kDa species (HMWM-260), were absent in the wall of cells grown under germination conditions in the presence of EDTA. Synthesis of the HMWM-260 species was not inhibited but its incorporation (secretion) into the wall structure seemed to be blocked in EDTA-treated cells.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-072X
    Keywords: “Slime” variant ; Neurospora ; Chitosomes ; Chitin synthetase ; Secretory vesicles ; Invertase ; Phosphatase ; Membranes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cells from the “slime” variant of Neurospora crassa were broken in isotonic conditions by use of triethanolamine buffer plus EDTA. After removal of large membranous structures by low-speed centrifugation, chitosomes and secretory vesicles were separated by means of gel filtration, precipitation of membranous contaminants with Concanavalin A, and centrifugation in sucrose or glycerol gradients. Polypeptidic composition of fractions enriched in secretory vesicles or chitosomes was found to be distinct. By these criteria we concluded that chitosomes and secretory vesicles represent different populations of microvesicles. Both microvesicular populations appeared free of endoplasmic reticulum and vacuolar contaminants as demonstrated by determination of appropriate enzymatic markers.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-0983
    Keywords: Key words Genetic linkage mapping ; Segregation distortion ; RAPD ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The inheritance of DNA markers was investigated in 27 F2 progeny from a single F1 hybrid derived from a wide cross in Uromyces appendiculatus. This cross was unusual because asexual spores were used to fertilize sexual fruiting structures. Sixty percent of the DNA markers failed to segregate according to simple Mendelian ratios. Segregation bias was evident, in that F2 progeny inherited on average 91% of maternal bands and 52% of paternal bands, which deviates significantly from the expected value for each of 75% for dominant markers. Because of these distortions, linkage mapping was not possible with this population. Evaluation of two F1s from a second wide cross, reciprocals obtained by normal fertilization, also showed non-Mendelian inheritance of one of three co-dominant RFLPs and five of six isozyme markers, indicating that the method of crossing was probably not responsible for the abnormal segregation patterns in the first cross. Either genetic incompatibility, similar to that of an interspecific cross, or selection of particular genotypes could explain the genetic anomalies reported here.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-0983
    Keywords: Genetic linkage mapping ; Segregation distortion ; RAPD ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The inheritance of DNA markers was investigated in 27 F2 progeny from a single F1 hybrid derived from a wide cross inUromyces appendiculatus. This cross was unusual because asexual spores were used to fertilize sexual fruiting structures. Sixty percent of the DNA markers failed to segregate according to simple Mendelian ratios. Segregation bias was evident, in that F2 progeny inherited on average 91 % of maternal bands and 52% of paternal bands, which deviates significantly from the expected value for each of 75% for dominant markers. Because of these distortions, linkage mapping was not possible with this population. Evaluation of two F1s from a second wide cross, reciprocals obtained by normal fertilization, also showed non-Mendelian inheritance of one of three co-dominant RFLPs and five of six isozyme markers, indicating that the method of crossing was probably not responsible for the abnormal segregation patterns in the first cross. Either genetic incompatibility, similar to that of an interspecific cross, or selection of particular genotypes could explain the genetic anomalies reported here.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 17 (1990), S. 133-138 
    ISSN: 1432-0983
    Keywords: RFLPs ; Septoria tritici ; DNA fingerprinting ; Genetic variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A set of probes that detect restriction fragment length polymorphisms (RFLPs) in nuclear DNA has been developed for genetic studies of the phytopathogenic fungus Septoria tritici. Two plasmid libraries containing 0.5–1.3 or 1.3–2.4 kb fragments of S. tritici nuclear DNA were constructed. Seventeen random clones from each library were used as probes to screen for RFLP variation among a geographically-diverse group of six S. tritici isolates. Among the 196 probe-enzyme combinations tested, 145 detected RFLPs among the six isolates. The restriction enzymes EcoRV and PstI detected RFLPs most efficiently. Three probes detected deletions. A ribosomal DNA probe from yeast did not detect a significant amount of variation. These probes will be useful for studying genetic variation, population genetics, and genome organization of S. tritici.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 19 (1991), S. 265-271 
    ISSN: 1432-0983
    Keywords: Electrophoretic karyotype ; Transverse alternating field electrophoresis ; Septoria tritici ; Chromosome length polymorphism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Transverse alternating field electrophoresis (TAFE) was used to compare the electrophoretic karyotypes of seven Septoria tritici isolates sampled from a single population. Isolates were selected based on differences at 12 DNA restriction fragment length polymorphism (RFLP) loci. Significant differences in electrophoretic karyotype existed among the isolates. Isolates had 14–16 bands, believed to correspond to chromosomes, ranging in size from approximately 330 to 3500 kb. Homologous chromosomes were identified by hybridization of anonymous single-locus and multiplelocus nuclear DNA sequences to Southern blots of TAFE gels. Length differences of up to 20% existed among homologous chromosomes. Densitometry and probe hybridization data showed that several bands contained two chromosomes. Probe pSTS192 hybridized to two chromosomes in each isolate, supporting previous data which suggested that this probe hybridized to two unlinked RFLP loci. Hybridization with probe pSTL53 provided additional evidence of partial diploidy at an RFLP locus in one isolate.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1432-2242
    Keywords: Key words Chromosome walking ; Gene mapping ; Glycine max ; Heterodera glycines ; High-molecular-weight DNA ; Positional cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  We constructed a bacterial artificial chromosome (BAC) library for soybean (Glycine max) consisting of approximately 30 000 clones with an average insert size of 120 kilobase pairs. The library was successfully screened with restriction fragment length polymorphism (RFLP) and microsatellite markers tightly linked to a major resistance gene for the cyst nematode, Heterodera glycines. Since many soybean RFLPs hybridize to duplicate loci, BACs homologous to duplicate RFLP loci were distinguished by digestion with the restriction enzyme originally used to map the RFLP, followed by a comparison of the hybridizing fragments. Linkage mapping of BAC clones identified with markers linked to the cyst nematode resistance gene demonstrated that these clones were located at the expected chromosomal positions and that there were no indications of chimeras within the genomic inserts.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1435-4373
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Diagnosis of systemicCandida infections was attempted by the use of an enzyme-linked immunosorbent assay (EIA) to detect IgG antibodies towards cell wall-bound and cytoplasmic candidal antigens. Cell wall antigens were sequentially solubilized by treatment of germinated blastoconidia ofCandida albicans (ATCC 26555 strain) with β-mercaptoethanol (βME extract) and digestion with Zymolyase 20T, a β-glucanase preparation (Zymolyase extract). Protoplasts obtained after treatment with Zymolyase were osmotically lysed (cytoplasmic antigens). Sera were obtained from patients with systemic (n=28) and superficial (n=46) candidiasis. Control sera were obtained from normal healthy individuals (n=31) and from hospitalized patients at low (n=36) and at high (n=13) risk of developing systemic candidiasis yet showing no symptoms of candidal infection. Detection of antibodies in crude sera samples by EIA using all of these antigenic extracts was highly specific (98–100 %), but sensitivity of the method was low (3.5–17.8 %). However, adsorption of sera with latex microspheres coated with purifiedCandida mannan in order to selectively remove antimannan antibodies prior to EIA improved the diagnostic efficiency of this test. Improvement was particularly noticeable when the βME extract was used as antigenic preparation, yielding a sensitivity of 89.2 % and a specificity of 98.6 %.
    Type of Medium: Electronic Resource
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