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  • Terminal inverted repeats  (1)
  • Transformation  (1)
  • 1
    ISSN: 1432-0983
    Keywords: Fusarium solani ; Mitochondrial plasmids ; Terminal inverted repeats ; Terminal proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The mitochondria of isolate FS37 from Nectria haematococca mating population I (Fusarium solani f. sp. cucurbitae) contain two linear plasmids, pFSCI and pFSC2, of 9.2 and 8.3 kbp, respectively. Evidence for a protein blocking the 5′ termini of these plasmids was obtained from exonuclease digestion experiments. A single protein band with an apparent Mr of 80 K was labeled when the DNA-protein complex of either plasmid was reacted with [125I] Bolton-Hunter reagent and then digested with DNase I. DNA sequence analysis of the termini of both plasmids revealed long inverted repeats of 1,211 by (pFSC1) and 1,027bp (pFSC2). No sequence similarity was found between the terminal inverted repeats (TIRs) of pFSC1 and pFSC2, nor was any similarity identified between the TIRs of the these plasmids and sequences of TIRs from other linear DNAs. A restriction fragment containing the TIR of pFSCI conferred autonomous replication when incorporated into an integrative transformation vector of Ustilago maydis.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0983
    Keywords: Transformation ; Stability ; Hygromycin B resistance ; Rice blast fungus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Magnaporthe grisea was transformed with cosmid pAN7-2 encoding hygromycin B resistance but containing no homology with the M. grisea genome. Rearrangement of the integrated DNA was detected in several hygromycin B-resistant progeny from cross Guy11-T10-1 (a single-copy integration site transformant)x2539 (sensitive wild-type parent), but not in hygromycin B-resistant progeny from four other crosses. Transformants produced typical lesions when inoculated onto host plants. Southern hybridization revealed rearrangements of integrated DNA in single conidial isolates of highcopy transformant 2539-T1-1 re-isolated from host plants, characterized by excision of one or more copies of the transforming plasmid. Plasmid loss and rearrangement were also observed within single conidial isolates derived from transformant 2539-T1-1 following ten asexual generations on non-selective agar medium. These examples of instability of integrated DNA in M. grisea transformants suggest that caution should be exercised in the use of transformation for assessing the phenotypic effects of specific introduced genes.
    Type of Medium: Electronic Resource
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