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  • 1
    Electronic Resource
    Electronic Resource
    Liposome-mediated delivery of DNA to carrot protoplasts (1981)
    Springer
    Planta 153 (1981), S. 90-94 
    Details
    ISSN: 1432-2048
    Keywords: Daucus ; DNA transfer ; Lipid vesicle transfer ; Liposome transfer ; Protoplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The encapsulation of DNA within liposomes and subsequent fusion of the liposomes with carrot (Daucus carota L.) protoplasts were examined to determine optimum conditions for effective liposome-mediated delivery of DNA to protoplasts. Escherichia coli [3H]DNA could be encapsulated with 50% efficiency using encapsulation volumes as low as 0.5 ml. Incorporation of liposome-encapsulated [3H]DNA by carrot protoplasts increased linearly for 2.5 h, and increasing the ratio of protoplasts to liposomes increased the total amount of radioactive label incorporated within the protoplasts. Liposome-mediated incorporation of [3H]DNA by protoplasts increased over a range of polyethylene glycol concentrations up to 20%, but Ca2+ did not increase liposome-mediated incorporation when present in the liposome-protoplast incubation mixture. Optimum incorporation was observed when the pH of the liposome-protoplast incubation medium was decreased to 4.8. Encapsulation experiments using DNA of the plasmid pBR322 indicated that an average of 200–1,000 intact copies of pBR322 were sequestered within each nucleus after liposome delivery.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00385322
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    Osmotic stress inhibits thymidine incorporation into soybean protoplast DNA (1982)
    Springer
    Plant cell reports 1 (1982), S. 186-188 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract DNA synthesis in protoplasts isolated from soybean cell suspension cultures has been investigated by [3H] thymidine uptake and incorporation kinetics. Initial rates of incorporation in exponential and 5-fluorodeoxyuridine synchronized protoplasts are inhibited by increased osmolarities of the medium. The inhibition was not readily reversible during 3 h culture in low osmotic medium. Velocity sedimentation analyses of replicating DNA from such protoplasts shows a complex pattern of inhibition. The inhibition probably effects replicon initiation as well as strand elongation and ligation of replication intermediates.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00270230
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    Site-specific mutagenesis of potato spindle tuber viroid cDNA: (1986)
    Springer
    Plant molecular biology 6 (1986), S. 179-192 
    Details
    ISSN: 1573-5028
    Keywords: Agrobacterium tumefaciens ; infectious cDNAs ; potato spindle tuber viroid ; site-specific mutagenesis ; Ti plasmid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The infectivity of cloned viroid cDNAs permits investigation of structure/function relationships in these unusual pathogenic RNAs by systematic site-specific mutagenesis of the cDNAs and subsequent bioassay. We have used three different strategies to create nucleotide substitutions within premelting region 2, a region of potato spindle tuber viroid (PSTV) believed to be important in viroid replication: sodium bisulfitecatalyzed deamination of deoxycytosine residues, oligonucleotide-directed mutagenesis, and construction of chimeric viroid cDNAs from fragments of infectious PSTV and tomato apical stunt viroid cDNAs. Although their effects upon the rod-like native structure of PSTV should be minimal, C → U transitions at positions 92 or 284 appeared to be lethal. When inoculation with PSTV cDNA containing a single nucleotide substitution was mediated by the Ti plasmid of Agrobacterium tumefaciens, PSTV progeny with an unaltered ‘wild type’ sequence was obtained. Two factors, the high error frequency characteristic of RNA synthesis and the use of a systemic bioassay for PSTV replication, may explain such sequence reversion and emphasize the importance of an appropriate bioassay system for screening mutant viroid cDNAs.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00021487
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  • 4
    Electronic Resource
    Electronic Resource
    Genetic analysis of somatic embryogenesis in carrot cell culture: Initial characterization of six classes of temperature-sensitive variants (1988)
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 9 (1988), S. 49-67 
    Details
    ISSN: 0192-253X
    Keywords: Daucus carota ; auxin ; gene expression ; mutant ; filtration-enrichment procedure ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Cell cultures of the carrot Daucus carota are a useful experimental system for studying the genetic regulation of plant embryogenesis. A modified filtration-enrichment procedure was used to isolate 21 temperature-sensitive variants in somatic embryogenesis; the variants display normal embryo development at the permissive temperature (24°C) and altered development at the restrictive temperature (33°C). Temperature-shift experiments were performed on these variants to determine the timing of gene action for the putative temperature-sensitive alleles. According to their phenotypes at the restrictive temperature, these variants can be divided into six classes: No Growth, Callus Proliferation, Globularstage Block, Oblong-stage Block, Lateral Growth, and Root Formation. Although many variants exhibit lengthy temperature-sensitive periods, the temperature sensitivity of some variants is restricted to one or two embryonic stages. These results plus those in the literature are incorporated into a preliminary model concerning the genetic regulation of carrot embryogenesis.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/dvg.1020090106
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