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  • Disease resistance  (1)
  • β-Glucuronidase  (1)
  • 1990-1994  (2)
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  • 1990-1994  (2)
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  • 1
    Electronic Resource
    Electronic Resource
    Homologous recombination between plasmid DNA molecules in maize protoplasts (1991)
    Springer
    Molecular genetics and genomics 230 (1991), S. 209-218 
    Details
    ISSN: 1617-4623
    Keywords: Homologous recombination ; Protoplast transformation ; β-Glucuronidase ; Maize
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The requirements for homologous recombination between plasmid DNA molecules have been studied using the PEG (polyethylene glycol)-mediated transformation system of maize (Zea mays L.) protoplasts coupled with the transient expression assay for β-glucuronidase (GUS). Two plasmids were introduced into maize protoplasts; one plasmid (pB×26) contained a genomic clone of the Adh1 maize gene; the other plasmid (piGUS) was a promoterless construction containing part of intron A of the Adhl gene fused to the gusA coding sequence. Thus, the two vectors shared an effective homologous region consisting of a 459 by (Hindlll—PvuII) fragment of the yAdh1 intron A sequence. An active gusA fusion gene would result upon homologous recombination between the plasmids within the intron A sequence, and indeed GUS activity was observed in extracts following co-transformation of maize protoplasts with the two plasmids. The presence of recombinant DNA molecules in protoplast DNA isolated 1 day after co-transformation was verified using polymerase chain reactions (PCR) and Southern blots. For efficient homologous recombination, both plasmids had to be linearized. The recombination reaction was induced by restriction of the plasmid molecules either inside the effective homologous region or at the borders of the intron sequence. However, the presence of even small, terminal, nonhomologous sequences at the 3′ end of the pB×26 fragment inhibited the recombination reaction. Also, both ends of the linearized piGUS DNA molecules were involved in the recombination reaction. The results revealed some features of homologous recombination reactions occurring in plant cells which cannot be accommodated by mechanisms postulated for similar reactions in animal system and in lower eukaryotes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00290670
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  • 2
    Electronic Resource
    Electronic Resource
    Recombination at the Rp1 locus of maize (1991)
    Springer
    Molecular genetics and genomics 226 (1991), S. 377-382 
    Details
    ISSN: 1617-4623
    Keywords: Disease resistance ; Maize ; Recombination ; Unequal crossing-over ; Fine structure mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The Rp1 locus of maize determines resistance to races of the maize rust fungus (Puccinia sorghi). Restriction fragment length polymorphism markers that closely flank Rp1 were mapped and used to study the genetic fine structure and role of recombination in the instability of this locus. Susceptible progeny, lacking the resistance of either parent, were obtained from test cross progeny of several Rp1 heterozygotes. These susceptible progeny usually had non-parental genotypes at flanking marker loci, thereby verifying their recombinational origin. Seven of eight Rp1 alleles (or genes) studied were clustered within about 0.2 map units of each other. Rpl G, however, mapped from 1–3 map units distal to other Rp1 alleles. Rp5 also mapped distally to most Rp1 alleles. Other aspects of recombination at Rp1 suggested that some alleles carry duplicated sequences, that mispairing can occur, and that unequal crossing-over may be a common phenomenon in this region; susceptible progeny from an Rp1 A homozygote had recombinant flanking marker genotypes, and susceptible progeny from an Rp1 DlRp1 F heterozygote showed both possible nonparental flanking marker genotypes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00260649
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    Paper (German National Licenses)
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