Transformation of Acremonium chrysogenum and Saccharomyces cerevisiae using an antibiotic resistance marker.

Abstract

Plasmid pCYG97 was constructed, which carried the kanamycin (G418) resistance gene of Tn903 and a DNA fragment from Acremonium chrysogenum. The DNA fragment from A. chrysogenum functions as an autonomously replicating sequence (ARS) in Saccharomyces cerevisiae. The transformation of A. chrysogenum and S. cerevisiae with pCYG97 DNA was attempted. Transformants resistant to the antibiotic, G418, were obtained from both strains after treatment of protoplasts with polyethylene glycol. The transformation efficiency of A. chrysogenum was lower than that of S. cerevisiae. The DNA sequence of the A. chrysogenum DNA fragment of pCYG97 was determined. The fragment contains multiple copies of sequences that have at least 10 out of 11 bases of homology to a previously reported core consensus sequence of S. cerevisiae, (A/T)TTTAT(A/G)TTT(A/T).