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  • Indirect immunofluorescence  (2)
  • interphase kinetochore association  (1)
  • 1995-1999  (3)
  • 1
    Electronic Resource
    Electronic Resource
    Molecular-cytogenetic characterization of a higher plant centromere/kinetochore complex (1996)
    Springer
    Theoretical and applied genetics 93 (1996), S. 477-484 
    Details
    ISSN: 1432-2242
    Keywords: Key words Microdissection of plant centromeres ; Subtractive hybridization ; Fluorescence in situ hybridization ; Indirect immunofluorescence ; Immunoadsorption
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The centromeric region of a telocentric field bean chromosome that resulted from centric fission of the metacentric satellite chromosome was microdissected. The DNA of this region was amplified and biotinylated by degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR)/linker-adapter PCR. After fluorescence in situ hybridization (FISH) the entire chromosome complement of Vicia faba was labelled by these probes except for the nucleolus organizing region (NOR) and the interstitial heterochromatin, the chromosomes of V. sativa and V. narbonensis were only slightly labelled by the same probes. Dense uniform labelling was also observed when a probe amplified from a clearly delimited microdissected centromeric region of a mutant of Tradescantia paludosa was hybridized to T. paludosa chromosomes. Even after six cycles of subtractive hybridization between DNA fragments amplified from centromeric and acentric regions no sequences specifically located at the field bean centromeres were found among the remaining DNA. A mouse antiserum was produced which detected nuclear proteins of 33 kDa and 68 kDa; these were predominantly located at V. faba kinetochores during mitotic metaphase. DNA amplified from the chromatin fraction adsorbed by this serum out of the sonicated total mitotic chromatin also did not cause specific labelling of primary constrictions. From these results we conclude: (1) either centromere-specific DNA sequences are not very conserved among higher plants and are – at least in species with large genomes – intermingled with complex dispersed repetitive sequences that prevent the purification of the former, or (2) (some of) the dispersed repeats themselves specify the primary constrictions by stereophysical parameters rather than by their base sequence.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s001220050305
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    Molecular-cytogenetic characterization of a higher plant centromere/kinetochore complex (1996)
    Springer
    Theoretical and applied genetics 93 (1996), S. 477-484 
    Details
    ISSN: 1432-2242
    Keywords: Microdissection of plant centromeres ; Subtractive hybridization ; Fluorescence in situ hybridization ; Indirect immunofluorescence ; Immunoadsorption
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The centromeric region of a telocentric field bean chromosome that resulted from centric fission of the metacentric satellite chromosome was microdissected. The DNA of this region was amplified and biotinylated by degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR)/linker-adapter PCR. After fluorescence in situ hybridization (FISH) the entire chromosome complement of Vicia faba was labelled by these probes except for the nucleolus organizing region (NOR) and the interstitial heterochromatin, the chromosomes of V. sativa and V. narbonensis were only slightly labelled by the same probes. Dense uniform labelling was also observed when a probe amplified from a clearly delimited microdissected centromeric region of a mutant of Tradescantia paludosa was hybridized to T. paludosa chromosomes. Even after six cycles of subtractive hybridization between DNA fragments amplified from centromeric and acentric regions no sequences specifically located at the field bean centromeres were found among the remaining DNA. A mouse antiserum was produced which detected nuclear proteins of 33 kDa and 68 kDa; these were predominantly located at V. faba kinetochores during mitotic metaphase. DNA amplified from the chromatin fraction adsorbed by this serum out of the sonicated total mitotic chromatin also did not cause specific labelling of primary constrictions. From these results we conclude: (1) either centromere-specific DNA sequences are not very conserved among higher plants and are — at least in species with large genomes — intermingled with complex dispersed repetitive sequences that prevent the purification of the former, or (2) (some of) the dispersed repeats themselves specify the primary constrictions by stereophysical parameters rather than by their base sequence.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00417938
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  • 3
    Electronic Resource
    Electronic Resource
    Immunostaining and interphase arrangement of field bean kinetochores (1995)
    Springer
    Chromosome research 3 (1995), S. 27-31 
    Details
    ISSN: 1573-6849
    Keywords: CREST sera ; interphase kinetochore association ; kinetochore ; nuclear proteins ; Vicia faba
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract More than 100 sera from patients with scleroderma CREST (calcinosis, Raynaud phenomenon, esophageal dismotility, sclerodactyly, telangiectasia) were tested in order to detect antigenic nuclear components of the field beanVicia faba (2n=12). Kinetochores of mitotic chromosomes and prekinetochores of interphase cells from root-tip meristems were specifically labelled via an indirect immunofluorescence procedure by antibodies of one of these sera. In 44% of interphase nuclei in which centromeres could be identified, only half (6) of the number of expected prekinetochores (12) was detected, circumstantially indicating at least transient association of homologous centromeres. Some nuclei showed clustering of centromeres at one pole (Rabl configuration). In metaphase chromosomes, each sister kinetochore contained a fluorescent spot. Western blotting of field bean nuclear proteins revealed four antigenic proteins of 28, 30, 64 and 68 kDa.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00711158
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    Paper (German National Licenses)
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