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

Abstract

Two integrative transforming plasmids of Neurospora crassa that differed only by the presence of almost all of a ribosomal DNA repeat unit on one plasmid were constructed. The plasmids were used to test the target concentration hypothesis which states that the transformation frequency is proportional to the number of genomic copies of a homologous sequence located on the transforming plasmid. Since there are approx. 200 copies of the rDNA sequences in the genome, the target concentration hypothesis would have been proved if the transformation frequency was 200-fold higher for the rDNA-containing plasmid compared with the plasmid without rDNA. The results indicated no difference in the transformation for the two plasmids, thereby providing no support for the hypothesis. The target concentration hypothesis has been proved for yeast, and thus mechanisms different from that responsible for integrative transformation in yeast must operate in N. crassa, perhaps including non-homologous recombination events.