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  • Genetics  (13)
  • Wiley-Blackwell  (12)
  • Springer  (1)
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  • 1990-1994  (13)
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  • 1945-1949
  • 1990  (13)
  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 319-330 
    ISSN: 0749-503X
    Keywords: δ-Aminolevulinate dehydratase purification ; Saccharomyces cerevisiae HEM2 transformants ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Saccharomyces cerevisiae transformed with a multicopy plasmid carrying the yeast structural gene HEM2, which codes for δ-aminolevulinate dehydratase, was enriched 20-fold in the enzyme. Beginning with cell-free extracts of transformed cells, the dehydratase was purified 193-fold to near-homogeneity. This represents a 3900-fold purification relative to the enzyme activity in normal, untransformed yeast cells. The specific activity of the purified enzyme was 16·2 μmol h-1 per mg protein at pH 9·4 and 37·5°C. In most respects the yeast enzyme resembles mammalian enzymes. It is a homo-octamer with an apparent Mr, of 275 000, as determined by centrifugation in glycerol density gradients, and under denaturing conditions behaved as a single subunit of Mr ≃ 37 000. The enzyme requires reduced thiol compounds to maintain full activity, and maximum activity was obtained in the presence of 1·0 mM-Zn2+. It is sensitive to inhibition by the heavy metal ions Pb2+ and Cu2+. The enzyme exhibits Michaelis-Menten kinetics and has an apparent Km of 0·359 mM. Like dehydratases from animal tissues, the yeast enzyme is rather thermostable. During the purification process an enhancement in total δ-aminolevulinate dehydratase activity suggested the possibility that removal of an inhibitor of the enzyme could be occurring.
    Additional Material: 5 Ill.
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  • 2
    ISSN: 1432-2242
    Keywords: Genetics ; Symbiosis ; Nitrogen fixation ; Coevolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary To determine the relationship between nodulation restriction by the Rj4 allele of soybean, rhizobitoxine-induced chlorosis, and taxonomic grouping of bradyrhizobia, 119 bradyrhizobial isolates were tested in Leonard jar culture for nodulation response and chlorosis induction. In addition to strain USDA 61, the strain originally reported as defining the Rj4 response, eight other isolates (i.e., USDA 62, 83, 94, 238, 252, 259, 260, and 340) were discovered to elicit the nodulation interdiction of the Rj4 allele. Only 16% of all the bradyrhizobial strains tested induced chlorosis, but seven of the nine strains (78%) interdicted by the Rj4 allele were chlorosis-inducing strains. Furthermore, in tests for antibiotic resistance profile, eight of the nine interdicted strains (89%) were classed in DNA homology group II. This evidence suggests that the Rj4 allele has a positive value to the host plant in shielding it from nodulation by certain chlorosis-inducing bradyrhizobia of a DNA homology group with impaired efficiency of nitrogen fixation with soybean.
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  • 3
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 318-325 
    ISSN: 0192-253X
    Keywords: Mice ; housekeeping genes ; liver ; tissue specificity ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In situ hybridization using nucleic acid probes was used to detect cell- and tissue-specific transcript(s) of embryonic genes during development and differentiation. This highly sensitive technique has the potential to provide valuable information on the regulation of low-abundance housekeeping genes during development. We have determined the experimental conditions required to detect the catalase message in adult mouse liver. Catalase effects the breakdown of H2O2 to O2 and H2O and offers protection against the toxic effects of oxygen radicals. We used a cloned 550 bp BamHI-Pstl fragment from a mouse catalase cDNA (pMCT-1) to generate 35S-labeled sense and antisense riboprobes. The experimental conditions used were sensitive enough to quantitate the abundance of silver grains generated by the antisense riboprobe on the adult liver, a tissue known to be positive for this message. The hybridization protocol was applied to serial sections of 13- and 18-day-old mouse embryos. The results suggest that the catalase expression in the liver and brain begins with somite formation and increases with development and differentiation. On the other hand, this message appears to be absent in mesenchyme, particularly in day 13 embryos. The message in positive tissues appears evenly distributed throughout the cell. The observed expression of the catalase message in the adult liver is approximately six times that in the embryonic liver. It is compatible with the enzyme activity results and emphasizes the sensitivity of the in situ hybridization method (over northern blot, etc.) used in this study.
    Additional Material: 5 Ill.
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  • 4
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 388-390 
    ISSN: 0192-253X
    Keywords: Transformation ; electroporotion ; gene targetting ; antisense RNA ; complementation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: DNA mediated transformation is a critical technique for uniting genetic, molecular genetic and reverse genetic approaches to a wide range of problems in cell and molecular biology. In Dictyostelium, there is now the capability not only to manipulate DNA sequences in vitro and put them back into the cell, but also to alter the sequences of endogenous chromosomal genes through high frequency homologous recombination. This means that the range of gene manipulation techniques that have made yeast such a useful system should now be applicable in Dictyostelium. For studying problems such as cellular motility, morphogenesis, and gene regulation, few organisms have the combination of features offered by Dictyostelium.
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  • 5
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 427-438 
    ISSN: 0192-253X
    Keywords: Cellular slime molds ; patterning ; development ; immunoblotting ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A series of monoclonal antibodies were previously raised against developing Polysphondylium pallidum cells. In this work, six of these antibodies have been used as probes to identify and characterize antigens regulated during development. Soluble and membrane fractions of P. pallidum cells at six stages of development or three stages of cyclic adenosine monophosphate (cAMP)-induced development were run in sodium dodecyl sulfate (SDS)-polyacrylamide gels and subjected to Western blot analysis. Three of the monoclonals, anti-Tp200, anti-Tp423, and antiPg 101, stain sorogen tips. Tp423 and Tp200 are membrane-associated antigens; both are stable to urea extraction, and Tp200 remains in the membrane after NaOH extraction. Tp423 is present in starved cells but is more prominent in sorogens and particularly in cAMP-developed cells. In contrast, Tp200 is first detected in early to mid-aggregation and is more abundant late in development. Pg101, which is expressed as a gradient with its highest concentration in tips, first appears in tight aggregates but is much more abundant in sorogens; unlike the Tp antigens, Pg101 is not greatly induced in cAMP-developed cells. All three of these antigens undergo changes in apparent molecular weight at the tight aggregate or sorogen stage: The gel mobilities of Tp200 and Pg101 increase, whereas that of Tp423 decreases. In addition to the tip-specific monoclonals, two monoclonals that stain all but the tips of sorogens have been used for analysis. One of these, anti-3D 10Pnk stains most cells within secondary tips, whereas anti-3D 10Dif does not. 3D 10Dif is membrane associated; it is present very early in development, increasing two- to threefold through the sorogen stage and diminishing in late cAMP-developed cells. 3D 10Pnk is a mostly soluble species first detected in late streaming. Anti-1c3, a sixth monoclonal, which stains nuclei uniformly throughout sorogens, is also developmentally expressed. 1c3 is mainly membrane associated and is expressed from late streaming through the sorogen stage.
    Additional Material: 6 Ill.
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  • 6
    ISSN: 0749-503X
    Keywords: Omnipotent suppressor ; gene structure ; evolutionary conservation ; codon bias analysis ; Pichia pinus ; yeast ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: SUP2(SUP35) is an omnipotent suppressor gene, coding for an EF-1α-like protein factor, intimately involved in the control of translational accuracy in yeast Saccharomyces cerevisiae.In the present study a SUP2 gene analogue from yeast Pichia pinus was isolated by complementation of the temperature-sensitive sup2 mutation of S. cerevisiae.The nucleotide sequence of the SUP2 gene of P. pinus codes for a protein of 82·4 kDa, exceeding the Sup2 protein of S. cerevisiae by 6 kDa. Like the SUP2 gene product of S. cerevisiae, the Sup2 protein of P. pinus represents a fusion of a unique N-terminal part of a region homologous to EF-1α. The comparison of amino acid sequences of the Sup2 proteins reveals high conservations (76%) of the C-terminal region and low conservation (36%) of the N-terminal part where, in addition, the homologous correspondence is ambiguous.Proteins related to the Sup2 of S. cerevisiae where found in P. pinus and some other yeast species by the immunoblotting technique.The relation between the evolutionary conservation of different regions of the Sup2 protein and their functional significance is discussed.
    Additional Material: 7 Ill.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 535-536 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 139-139 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Additional Material: 1 Ill.
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  • 9
    ISSN: 0749-503X
    Keywords: Protein sorting ; membranes ; phospholipid synthesis ; yeast ; Saccharomces cerevisiae ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The product of the yeast CHO1 gene, phosphatidylserine synthase (PSS), is an integral membrane protein that catalyses a central step in cellular phospholipid biosynthesis. A 1·2 kb fragment containing the regulatory and structural components of the CHO1 gene was sequenced. Transcription initiation in wild-type cells was found to occur between -1 and -15 relative to the first ATG of a large open reading frame capable of encoding a 30 804 molecular weight protein. This translation initiation site was active in vivo and in vivo in a hetrologous system. In both cases it supported production of a protein of approximately 30 000 molecular weight. A second potential translation initiation site was detected 225 or 228 bases dowstream from the first ATG. This second site was active in vitro where it supported production of a protein of 22 400 molecular weight. A subclone, lacking the 5′ regulatory region and the subsequence encoding the first 12 amino acids of the large open reading frame, allowed translation in vivo starting at the second ATG. The resulting protein was 22 000 moleculare weight, lacked the 74 N-terminal amino acids and was capable of complementing the choline auxotroy of a cho1 null-mutant. In transformants carrying this construct, PSS activity and 22 kDa protein was found to be associated with membrane fractions corresponding to mitochondria and endoplasmic reticulum. However, most of the truncated PSS protein accumulated in the cytosol in an inactive form. A hybrd-protein containing the 63 N-terminal amino acids of PSS protein accumulated in the cytosol in a active form. A hybrid-protein containing the 63 N-terminal amino acids of PSS fused to mouse dihydrofolate reductase was found exclusively in the cytosol when expressed in wild-type yeast. Thus, the hydrophilic, highly acidic N-terminus of PSS is required for efficient membrane insertion but does not appear to contain sequences required for a targeting to the membrane compartment.
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  • 10
    ISSN: 0749-503X
    Keywords: dsRNA ; yeast ; killer yeast ; neutral yeast ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: K2 neutral strain Saccharomyces cerevisiae USM12 was identified and characterized. This strain carried an M double-stranded RNA (dsRNA) genome encoding for resistance to K2 toxin. The M dsRNA was larger than the K2 killer yeast M dsRNA and homoduplex analysis of denatured and reannealed K2 neutral M dsRNA revealed an inverted duplication. Heteroduplex analysis showed that two thirds of the K2 M genome had homology with K2 neutral M genome. Hybridization showed that the USM12 M dsRNA has significant homology with the K2M dsRNA. Protein profiles of extracellular proteins from USMA12 and cured strain indicated that USM12 did not secrete any toxin. This is the first time that a K2 neutral yeast strain has been characterized.
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