Gene expression from replicating plasmids in Aspergillus nidulans.

Abstract

Plasmids bearing the AMA1 replicator from Aspergillus nidulans are capable of extrachromosomal replication in this fungus as well as in other species. Synthetic plasmids bearing the moderately expressed argB gene and the highly expressed, inducible beta-galactosidase gene (bgaS) were introduced into fungal cells. Expression of both genes was monitored by Northern hybridization. It was demonstrated that transcription of bgaS is induced and repressed normally, irrespective of whether the gene is integrated into the chromosome or maintained on an extrachromosomal supercoiled plasmid. Transcription of the strongly expressed bgaS gene stimulates transcription of the argB gene located on the same replicating plasmid irrespective of orientation. This effect also occurs with chromosomally integrated vectors, but to a lesser extent. Episomal vectors are present in 10 copies per nucleus, and the expression level of the argB gene is approximately proportional to copy number. However, the amount of mRNA transcribed from the highly expressed bgaS gene on the multi-copy replicating plasmid does not exceed that from single-copy integrants. High levels of expression of the plasmid-borne gene do not affect plasmid mitotic stability or copy number.