Some property of the nucleus determines the competence of Neurospora crassa for transformation.
In Neurospora, transformation of spheroplasts is quite efficient and usually occurs with the transforming DNA integrated at ectopic sites in the chromosome. However, only a small fraction of the spheroplasts is actually competent for transformation. To distinguish whether the limitation to competence is at the level of the plasma membrane or at the level of the nucleus, we performed experiments in which heterocaryotic spheroplasts were required to integrate two different plasmids in one transformation procedure. The cotransformants were then analyzed to determine into which nucleus or nuclei the separate plasmids had integrated. Results of such experiments confirm that successful ectopic transformation in Neurospora crassa requires a competent nucleus. The integration patterns of the two separate plasmids indicate that the availability of appropriate chromosomal sites for ectopic integration may be an aspect of nuclear competence.
Grotelueschen
J
J
Department of Biomolecular Chemistry, University of Wisconsin, Madison 53706, USA.
Metzenberg
R L
RL
D002467:Cell Nucleus / Q000378:metabolism; D015825:Chromosomes, Fungal; D004274:DNA, Recombinant; D009492:Neurospora crassa / Q000235:genetics*; D010957:Plasmids; D014170:Transformation, Genetic
D004274:DNA, Recombinant