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  • Fusion  (2)
  • Membrane  (2)
  • Springer  (3)
  • American Society of Hematology
  • Frontiers Media
  • Springer Nature
  • Wiley
  • ZBW - Deutsche Zentralbibliothek für Wirtschaftswissenschaften, Leibniz-Informationszentrum Wirtschaft Kiel, Hamburg
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  • Springer  (3)
  • American Society of Hematology
  • Frontiers Media
  • Springer Nature
  • Wiley
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 124 (1985), S. 65-70 
    ISSN: 1615-6102
    Keywords: Protoplast ; Fusion ; Fluorescence ; PEG
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two fluorescent compounds, scopoletin and carboxyfluorescein, have been used to label both tissue culture and leaf mesophyll cells and protoplasts. The compounds localized within the vacuoles of cells in approximately 15 hours. They remained in the vacuole during cell wall digestion, and fluorescence was observable for several hours after protoplast release. A one day pulse of these fluorescent labels had no deleterious effect on the growth of cells or protoplasts. When morphologically indistinguishable protoplasts were labeled and treated with polyethylene glycol, multicolored fluorescent fusion products were observable. These fluorescent labels provide a convenient method for selection of heterokaryon fusion products of whole plant and tissue culture cell protoplasts.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 120 (1984), S. 209-215 
    ISSN: 1615-6102
    Keywords: Fusion ; Calcium ; Protoplast ; Membrane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Rather than selecting for a chemical fusogen one can select for a fusogenic plant membrane (i.e., one that will fuse readily). Wild carrot suspension culture cells can be grown under conditions which cause the released protoplasts to have a high potential to fuse. Protoplast fusion is enhanced by calcium and inhibited by EGTA. When 10mM calcium (pH6.0) is added, fusion percentages of 60% are common. The mild fusion treatment appears to have no effect on callus regeneration and differentiation.
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  • 3
    ISSN: 1615-6102
    Keywords: Phosphatidylinositol ; Lysophosphatidylinositol ; Phosphoinositides ; Nuclei ; Membrane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Nuclei were isolated from carrot protoplasts and the distribution of [3H]inositol-labeled phospholipids was analyzed by thinlayer chromatography. Phosphatidylinositol (PI), lysophos-phatidylinositol (LPI), phosphatidylinositol monophosphate (PIP), lysophosphatidylinositol monophosphate (LPIP), and phosphatidylinositol bisphosphate (PIP2) were 55.7%, 12.3%, 5.0%, 11.5%, and 3.6% of the respective [3H]inositol-labeled lipids recovered from the nuclear fraction. While both the plasma membrane and nuclear fraction contained polyphosphoinositides, the distribution of the phosphoinositides and the amount of inositol-labeled lipid were distinct. For example, the nuclear fraction had a higher percentage of LPI and PIP2 and less PI and LPIP than the plasma membrane fraction. The amount of [3H]inositol-labeled lipid recovered from the nuclear fraction per mg protein was an order of magnitude lower than that recovered from either the plasma membrane of lower phase fraction isolated by aqueous two-phase partitioning, or from whole cells and protoplasts. In addition, when the ratio of the [3H]inositol-labeled lipid was compared to total [14C]myristate-labeled lipid recovered there was three to ten fold less [3H] relative to [14C] in the nuclear fraction. These data indicate that while the polyphosphoinositides are a relatively high percentage of the inositol lipid in the nuclear fraction, the inositol lipid was only a small portion of the total lipid in the nuclei. Despite this low concentration of inositol lipid, when [γ 32P]-ATP was added to the isolated nuclei,32P-labeled PIP and PIP2 were synthesized. Thus, the carrot nuclei contained PI and PIP kinase as well as the polyphosphoinositides.
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