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  • 1
    Electronic Resource
    Electronic Resource
    Construction of a 550 kb BAC contig spanning the genomic region containing the apple scab resistance gene Vf (1999)
    Springer
    Molecular genetics and genomics 262 (1999), S. 884-891 
    Details
    ISSN: 1617-4623
    Keywords: Key wordsVf ; Malus×domestica ; Resistance gene ; Chromosome walking ; Inverse PCR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A positional cloning project was started in apple with the aim of isolating the Vf resistance gene of Malus floribunda 821. Vf confers resistance against apple scab, the most important disease in apple orchards. A chromosome walk starting from two molecular markers (M18-CAPS and AM19-SCAR) flanking Vf was performed, using a bacterial artificial chromosome (BAC) library containing inserts of the cultivar Florina, which is heterozygous for Vf. Thirteen BAC clones spanning the region between the two markers were identified in nine chromosome walking steps. The size of the resulting contig is approximately 550 kb. In order to map the Vf region in more detail, we analyzed over 2000 plants from different populations segregating for Vf with markers produced from BAC end sequences. In this way, we were able to restrict the possible location of the Vf gene to a minimum of five clones spanning an interval of approximately 350 kb.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004380051154
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    Construction and characterization of a bacterial artificial chromosome library of apple (1998)
    Springer
    Theoretical and applied genetics 97 (1998), S. 1183-1190 
    Details
    ISSN: 1432-2242
    Keywords: Key words Bacterial artificial chromosome (BAC) ; High-molecular-weight (HMW) DNA ; Malus×domestica ; Venturia inaequalis ; Resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A bacterial artificial chromosome (BAC) library has been constructed from apple (Malus×domestica Borkh.) using the variety “Florina”, which is resistant to scab (Venturia inaequalis) by virtue of the Vf gene. Since apple leaves are rich in polyphenols, high-molecular-weight DNA was extracted from leaf nuclei with a protocol adapted to apple. The nuclei were then embedded in agarose microbeads and partially digested by varying ratios of EcoRI to EcoRI methylase. The resulting DNA fragments were size-selected by pulsed-field gel electrophoresis, ligated to the BAC cloning vector pECBAC1 and transformed into Escherichia coli cells by electroporation. A total of 36 864 recombinant clones (BACs) were obtained. The library has an average insert size of 120 kb and represents approximately 5×apple haploid-genome equivalents. It was screened with six cDNA probes using the chemiluminescent DIG system. An average of 4.4 clones was detected for each locus. The apple BAC library will be used to isolate the Vf scab resistance gene through map-based cloning. In this connection the library was screened with a marker closely linked to the Vf gene and six positive clones have been isolated. This library should thus be well suited for map-based gene cloning, in particular for the isolation of the Vf gene and for the construction of a physical map of the apple genome.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s001220051008
    Library Location Call Number Volume/Issue/Year Availability
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    Paper (German National Licenses)
    Fulltext
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