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  • 1
    ISSN: 1432-1203
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Experimental conditions for detection of germline deletions of the von Hippel-Lindau (VHL) gene by means of long polymerase chain reaction have been established. Primers were designed to analyse the VHL gene in three overlapping fragments: 12.5 kb in length containing promoter and exons 1 and 2; 8.7 kb in length containing exons 2 and 3; and 16 kb in length containing exons 2 and 3 and the 3' untranslated region. Using the described procedure, it was possible to detect large deletions in four of five cases with such mutations previously detected by Southern blotting and in 5 of 11 unrelated Polish VHL patients in whom constitutional VHL gene mutations were not found by sequencing.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2242
    Keywords: Tissue culture mutant selection ; Herbicide tolerance ; Fatty acid biosynthesis ; Acetyl-CoA carboxylase ; maize
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The genetic relationship between acetyl-coenzyme A carboxylase (ACCase; EC 6.4.1.2.) activity and herbicide tolerance was determined for five maize (Zea mays L.) mutants regenerated from tissue cultures selected for tolerance to the ACCase-inhibiting herbicides, sethoxydim and haloxyfop. Herbicide tolerance in each mutant was inherited as a partially dominant, nuclear mutation. Allelism tests indicated that the five mutations were allelic. Three distinguishable herbicide tolerance phenotypes were differentiated among the five mutants. Seedling tolerance to herbicide treatments cosegregated with reduced inhibition of seedling leaf ACCase activity by sethoxydim and haloxyfop demonstrating that alterations of ACCase conferred herbicide tolerance. Therefore, we propose that at least three, and possible five, new alleles of the maize ACCase structural gene (Acc1) were identified based on their differential response to sethoxydim and haloxyfop. The group represented by Acc1-S1, Acc1-S2 and Acc1-S3 alleles, which had similar phenotypes, exhibited tolerance to high rates of sethoxydim and haloxyfop. The Acc1-H1 allele lacked sethoxydim tolerance but was tolerant to haloxyfop, whereas the Acc1-H2 allele had intermediate tolerance to sethoxydim but was tolerant to haloxyfop. Differences in tolerance to the two herbicides among mutants homozygous for different Acc1 alleles suggested that sites on ACCase that interact with the different herbicides do not completely overlap. These mutations in maize ACCase should prove useful in characterization of the regulatory role of ACCase in fatty acid biosynthesis and in development of herbicide-tolerant maize germplasm.
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  • 3
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Komparative genomische Hybridisierung ; Molekularpathologie ; Genomische Diagnostik ; Klassifikation von Tumoren ; Klassifikation von Malformationen ; Key words Comparative genomic hybridization ; Molecular pathology ; Genetic diagnostics ; Classification of tumors ; Classification of malformations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Comparative genomic hybridisation (CGH) is a new cytogenetic method, which is based on a combination of fluorescence microscopy and digital image analysis. The molecular genetic basis is the hybridization of a mixture of fluorescein labeled test-DNA and reference-DNA on normal metaphase chromosomes. Comparative analysis allows the identification of all unbalanced chromosomal aberrations of the test-DNA in a single experimental step. The resulting DNA gains or DNA losses on the chromosomal or subchromosomal level mirror possible amplifications of oncogenes or losses of suppress orgenes. As CGH can be performed with genomic DNA of formalin-fixed and fresh-frozen tissue or cells, this new method is a very effective tool for pathologists and cytologists in the extended genomic screening of tumors and genetically altered tissues. Despite CGH analysis at present is restricted to research applications; its widespread dissemination as a routine method in diagnostic pathology can be expected in the near future.
    Notes: Zusammenfassung Die komparative genomische Hybridisierung (CGH) ist eine neu entwickelte zytogenetische Methode, die auf einer Kombination von Fluoreszenzmikroskopie und digitaler Bildanalyse beruht. Grundlage stellt die Hybridisierung eines Gemisches aus zu testender und normaler DNA auf gesunde Metaphasechromosomen dar, deren vergleichende (komparative) Analyse die Identifizierung aller unbalancierten chromosomalen Aberrationen der Test-DNA in einem Versuchsansatz erlaubt. Die somit detektierten DNA-Gewinne oder DNA-Verluste auf chromosomaler und subchromosomaler Ebene stellen ein Spiegelbild für eine etwaige Amplifikation eines Onkogens oder den Verlust eines Tumorsuppressorgens dar. Da die CGH mit isolierter DNA sowohl aus formalinfixiertem als auch aus frischem Gewebe/Zellen durchgeführt werden kann, stellt sie für den Pathologen und Zytologen gleichermaßen ein effektives Werkzeug bei der erweiterten genomischen Analyse von Tumoren und anderen genetisch alterierten Geweben dar. Obwohl die CGH bislang ausschließlich in der Forschung eingesetzt wird, ist mit ihrer Anwendung als Routinemethode in der pathologisch-anatomischen Diagnostik bereits in naher Zukunft zu rechnen.
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