Transformation of Neurospora crassa by an integrative transforming plasmid is not enhanced by ribosomal DNA sequences

Russell, Peter; Welsch, Jo Anne; Wagner, Sheryl

doi:10.1016/0167-4781(80)90015-9

Cited by 4 publications

(2 citation statements)

References 22 publications

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“…This, however, is at variance with the observation that DNA sequences present once and about 40 times (the genes for ribosomal RNA) in the genome gave the same transformation frequencies. Similar results were obtained in N. crassa which contains about 200 copies of the genes for ribosomal RNA in its genome (Russell et al 1989). The competence of protoplasts, i.e, the degree of access of exogenous DNA to the nucleus, is likely to be the limiting factor for the transformation of F. fujikuroi.…”

Section: Discussionsupporting

confidence: 75%

Homologous recombination and allele replacement in transformants of Fusarium fujikuroi

Fernández-Martı́n¹,

Cerdá‐Olmedo²,

Ávalos³

2000

Mol Gen Genet

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The ascomycete Fusarium fujikuroi could be transformed stably to hygromycin resistance only when the transforming plasmid contained a fragment of DNA from the fungus. The transformation frequencies were roughly independent of the sequence of the particular fungal DNA fragment used, of its size (1.8 or 6 kb), and of whether this DNA was present only once in the fungal genome or about forty times (the genes for ribosomal RNA). The plasmid was integrated into the fungal genome by homologous recombination in the eighteen transformants tested; ectopic integration was never observed. The carB gene of F. fujikuroi was cloned and shown to complement unpigmented mutants deficient in phytoene dehydrogenase. A mutant carB allele was prepared in vitro and used to transform wild-type protoplasts; the transformants contained a genomic duplication and were heterozygous for carB; the mutant allele replaced the original wild-type allele when this was spontaneously lost in the transformants. This loss was due to gene conversion in some cases and to recombination between repeated sequences in others.

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Section: Discussionsupporting

confidence: 75%

Homologous recombination and allele replacement in transformants of Fusarium fujikuroi

Fernández-Martı́n¹,

Cerdá‐Olmedo²,

Ávalos³

2000

Mol Gen Genet

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“…The aim of these studies was to increase the copy number of an integrative plasmid by targeting it to a DNA sequence present in multiple copies in the fungus genome. This group of fungi included S. proli®cans (Ruiz-Dõ Âez and Martõ Ânez-Suarez 1999), Alternaria alternata (Tsuge et al 1990;Shiotani and Tsuge 1995), Cephalosporium acremonium (Skatrud et al 1987) and N. crassa (Russell et al 1989). Generally, the transformation ef®ciencies increased, and the occurrence of homologous integration of the exogenous DNA into the chromosome is mediated by the rDNA sequences.…”

Section: Selection Of Transforming Dnamentioning

confidence: 99%