Behaviour of a replicating mitochondrial DNA sequence from Aspergillus amstelodami in Saccharomyces cerevisiae and Aspergillus nidulans

Abstract

An amplified sequence of mitochondrial DNA from a ragged (rgd) mutant of Aspergillus amstelodami has been shown to exist in multimeric circular form, suggesting that it is excised from the genome and can exist independently of it. This sequence has replicative (ARS) activity in Saccharomyces cerevisiae, and a subfragment responsible for this activity has been identified and sequenced. A homologous sequence from Aspergillus nidulans mtDNA also has ARS activity in S. cerevisiae. Both A. amstelodami and A. nidulans ARS elements have been incorporated into the integrative transformation vector pDJB1 and the derived vectors used to transform A. nidulans. Inclusion of the A. nidulans ARS element enhanced the transformation frequency 5-fold relative to pDJB1. No increase in transformation frequency was evident with the ARS element from A. amstelodami. The stability of transformants was variable but in comparison to pDJB1, ARS-containing plasmids were mitotically unstable in A. nidulans. Although plasmid DNAs could be rescued in Escherichia coli from undigested transformant DNA, no freely replicating plasmids were detected by Southern hybridisation.