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  • 1
    Electronic Resource
    Electronic Resource
    The daunting process of MIAME (2006)
    [s.l.] : Nature Publishing Group
    Nature 444 (2006), S. 31-31 
    Details
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Sir Michael Eisenstein, in his Technology Feature “Quality control” (〈weblink url="http://www.nature.com/nature/journal/v442/n7106/full/4421067a.html"〉Nature 442, 1067–1070; 2006), reports recent improvements in microarray technologies. He ...
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1038/444031b
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  • 2
    Electronic Resource
    Electronic Resource
    The effects of inhibitors of cell wall synthesis on tobacco protoplast development (1982)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 55 (1982), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The effect of 2,6-dichlorobenzonitrile (DB) and coumarin on tobacco protoplast development has been studied using a combination of flow microfluorimetry and quantitative fluorescence microscopy. The results indicated that, over the initial period of culture, DB largely inhibited cellulose production at the cell surface, but did not inhibit DNA synthesis or protein accumulation by the protoplasts. Continuous culture of protoplasts in DB resulted in the accumulation of an increased capacity for cellulose synthesis as expressed following the removal of inhibitor. The possible sites of the specific inhibitory action of DB are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1982.tb00279.x
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  • 3
    Electronic Resource
    Electronic Resource
    Flow sorting and culture of plant protoplasts (1984)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 60 (1984), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Conditions have been established for the rapid flow analysis of leaf protoplasts of Nicotiana tabacum L. cv. Xanthi using a flow cytometer-cell sorter. A procedure based upon chlorophyll autofluorescence was devised to permit the systematic evaluation of flow conditions in order to identify those under which protoplast damage was minimized. These conditions were employed for the flow sorting of protoplasts, following which it was possible to regenerate the sorted protoplasts into complete plants. The application of flow sorting is discussed for the rapid identification and selection of somatic hybrids produced by protoplast fusion.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1984.tb04247.x
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  • 4
    Electronic Resource
    Electronic Resource
    Microfluorimetric quantitation of cellulose biosynthesis by plant protoplasts using Calcofluor White (1981)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 53 (1981), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A microfluorimetric procedure, using Calcofluor White, has been developed for the measurement of cellulose biosynthesis by cultured protoplasts of tobacco (Nicotiana tabacum L. cv. Xanthi nc). The procedure was compared to a conventional method for cellulose estimation, that employing the anthrone reagent following exhaustive extraction of the developing cell walls. The results indicate that the amount of fluorescence emitted following Calcofluor White treatment is a specific measurement of cell wall cellulose levels. The procedure possesses the twin advantages of ease of manipulation and of greatly enhanced sensitivity (in the picogram range) as compared to other methods.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1981.tb04119.x
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  • 5
    Electronic Resource
    Electronic Resource
    Analysis of the initial stages of plant protoplast development using 33258 Hoechst: reactivation of the cell cycle (1981)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 51 (1981), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A microfluorimetric procedure, employing the fluorescent stain 33258 Hoechst, has been developed for the investigation of the process of DNA synthesis during the initial stages of culture of tobacco (N. tabacum cv. Xanthi) leaf protoplasts.In this system, the freshly-isolated protoplasts exhibited a unimodal distribution of nuclear DNA content characteristic of the diploid state. The almost immediate onset of DNA synthesis during culture resulted in a doubling of nuclear DNA levels prior to the first mitoses. Although the majority of the protoplasts subsequently entered into synchronous mitosis and cell division, a proportion of the remainder developed into large polyploid cells. Upon further culture, the polyploid cells became subdivided into clusters of small diploid cells. Measurement of total cell protein and cell volumes during culture indicated that a relationship existed between these parameters and the initiation of mitosis. The significance of these observations is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1981.tb05573.x
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  • 6
    Electronic Resource
    Electronic Resource
    Implementing microarrays for the study of plant genomics (1999)
    [s.l.] : Nature America, Inc.
    Nature genetics 23 (1999), S. 45-45 
    Details
    ISSN: 1546-1718
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Microarrays provide the means for the large-scale analysis of gene expression patterns in living organisms. My laboratory is part of three federally funded projects that are directed toward an understanding of the regulation of gene expression in higher plants. In the first project, funded by the ...
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1038/14307
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  • 7
    Electronic Resource
    Electronic Resource
    Selection of Somatic Hybrid Plants in Nicotiana Through Fluorescence-Activated Sorting of Protoplasts (1985)
    [s.l.] : Nature Publishing Company
    Nature biotechnology 3 (1985), S. 811-816 
    Details
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] We have developed general techniques for the successful selection of plant heterokaryons from parental populations of plant protoplasts subjected to somatic fusion treatments. These techniques involve a combination of artificial fluorescence labeling, natural chlorophyll autofluorescence and ...
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1038/nbt0985-811
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  • 8
    Electronic Resource
    Electronic Resource
    Biosynthesis, processing and targeting of the G-protein of vesicular stomatitis virus in tobacco protoplasts (1992)
    Springer
    Planta 186 (1992), S. 324-336 
    Details
    ISSN: 1432-2048
    Keywords: Nicotiana (transfection with VSVG) ; Protoplast (transfection) ; Plasma membrane ; Rhabdoviruses ; Protein (secretion, targeting) ; Vesicular stomatitis virus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Leaf protoplasts of tobacco (Nicotlana tabacum L.) were employed for transfection of chimeric transcriptional gene fusions comprising the 35S promoter from cauliflower mosaic virus, the coding sequence of the G-protein from vesicular stomatitis virus (VSVG) and the transcriptional terminator from the Agrobacterium tumefaciens nopaline-synthetase gene. Transient expression of the chimeric gene was monitored through Northern analysis of total protoplast RNA using a labeled VSV cDNA probe, and through Western-blot analysis of protoplast proteins using a polyclonal and-VSV antiserum. Although a single species of mRNA was detected in the transfected protoplasts, two glycoproteins differing in mass by approx. 9 kDa were detected by the antiserum. Biosynthesis of the VSVG isoforms was not impeded by chemical inhibitors of cell-wall production or of proline hydroxylation. Transfection using mutant forms of the VSVG coding sequence in which either one or both consensus glycosylation sites were removed resulted in the production of progressively smaller VSVG proteins. Those proteins produced from the double mutant had mobilities on sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were very similar to those produced from the wild-type construct in the presence of tunicamycin. Analysis of protoplast homogenates by differential centrifugation showed that the two VSVG isoforms were exclusively associated with cellular membranes. The larger protein co-localized with the plasma membrane and with the organelles of the endomembrane-secretory pathway leading to the plasma membrane. The smaller protein was associated with membranes of lower isopycnic densities which were not identical to the endoplasmic reticulum. The larger protein displayed greater sensitivity than did the smaller to degradation in vivo by exogenously added protease. Immunofluorescence microscopy demonstrated that the VSVG isoforms were present both within the protoplasts and at the surface of the plasma membrane. The intracellular distribution was either punctate or reticulate. These results are consistent with the progressive and accurate glycosylation of the newly synthesized VSVG polypeptide during its passage through the endomembrane-secretory pathway, the access of the larger isoform to the cell surface, and the conversion of the larger to the small isoform by selective proteolysis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00195312
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  • 9
    Electronic Resource
    Electronic Resource
    Isolation and characterization of an Arabidopsis thaliana C-8,7 sterol isomerase: functional and structural similarities to mammalian C-8,7 sterol isomerase/emopamil-binding protein (1998)
    Springer
    Plant molecular biology 38 (1998), S. 807-815 
    Details
    ISSN: 1573-5028
    Keywords: cloning ; isomerase ; phytosterol ; yeast complementation ; Arabidopsis thaliana ; sigma receptor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The yeast C-8,7 sterol isomerase contains a polyvalent high-affinity drug binding site similar to mammalian sigma receptors. Exogenously supplied sigma ligands inhibit sterol biosynthesis in yeast, demonstrating a pharmacological relationship between sigma ligand-binding and C-8,7 sterol isomerase activity. We report the isolation of an Arabidopsis thaliana C-8,7 sterol isomerase by functional complementation of the corresponding sterol mutant in yeast and its characterization by exposure to sigma ligands. The yeast erg2 mutant, which lacks the C-8,7 sterol isomerase gene and activity, was transformed with an Arabidopsis cDNA yeast expression library. Transformed colonies were selected for restoration of C-8,7 sterol isomerase activity (i.e. wild-type ergosterol production) by enhanced resistance to the antibiotic cycloheximide. Sterols produced in complemented lines were characterized by gas chromatography-mass spectroscopy (GC-MS). The full-length A. thaliana cDNA (pA.t.SI1) that complemented the erg2 mutation contains an open reading frame encoding a 21 kDa protein that shares 68% similarity and 35% amino acid identity to the recently isolated mouse C-8,7 sterol isomerase. The sigma ligands, haloperidol, ifenprodil and verapamil inhibited the production of ergosterol in wild-type Saccharomyces cerevisiae and in the erg2 mutant complemented with pA.t.SI1. Structural and biochemical similarities between the A. thaliana C-8,7 sterol isomerase and the mammalian emopamil-binding protein (EBP) are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006028623875
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  • 10
    Electronic Resource
    Electronic Resource
    Characterization of Zea mays endosperm C-24 sterol methyltransferase: one of two types of sterol methyltransferase in higher plants (1997)
    Springer
    Plant molecular biology 34 (1997), S. 891-896 
    Details
    ISSN: 1573-5028
    Keywords: cloning ; methyltransferase ; phytosterol ; yeast complementation ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We report the characterization of a higher-plant C-24 sterol methyltransferase by yeast complementation. A Zea mays endosperm expressed sequence tag (EST) was identified which, upon complete sequencing, showed 46% identity to the yeast C-24 methyltransferase gene (ERG6) and 75% and 37% amino acid identity to recently isolated higher-plant sterol methyltransferases from soybean and Arabidopsis, respectively. When placed under GAL4 regulation, the Z. mays cDNA functionally complemented the erg6 mutation, restoring ergosterol production and conferring resistance to cycloheximide. Complementation was both plasmid-dependent and galactose-inducible. The Z. mays cDNA clone contains an open reading frame encoding a 40 kDa protein containing motifs common to a large number of S-adenosyl-L-methionine methyltransferases (SMTs). Sequence comparisons and functional studies of the maize, soybean and Arabidopsis cDNAs indicates two types of C-24 SMTs exist in higher plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1005818210641
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