Analysis of promoter activity by transformation of Acremonium chrysogenum.
Promoter activity was examined in the beta-lactam-producing fungus, Acremonium chrysogenum, by assessment of the properties of transformant isolates. Transformation was achieved using plasmid constructs specifying hygromycin B resistance (HyR) linked to the promoter elements of gpdA (the glucose-6-phosphate dehydrogenase-encoding gene of Aspergillus nidulans), and pcbC [the gene encoding the isopenicillin N synthetase (IPNS) enzyme of A. chrysogenum]. Transformation frequency, HyR levels, and Hy phosphotransferase (HPT) levels suggested that the transformants of constructs using the gpdA promoter showed a higher level of expression of the HyR gene than in transformants obtained using the pcbC promoter. The patterns of integration of the transforming DNA also differed in that pcbC promoter construct transformants appeared to have tandem repeats. All integrations of plasmid DNA occurred on a single chromosome which was different in four out of five transformants studied. Multiple copy transformants of constructs using the pcbC promoter did not show the regulated pattern of expression of HPT activity observed with IPNS in untransformed strains.
Smith
A W
AW
Department of Life Sciences, University of Nottingham, UK.
Ramsden
M
M
Peberdy
J F
JF
D000164:Acremonium / Q000235:genetics*; D015139:Blotting, Southern; D002934:Cinnamates; D004351:Drug Resistance / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000494:pharmacology; D010088:Oxidoreductases / Q000235:genetics; D010770:Phosphotransferases / Q000235:genetics / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor); D011401:Promoter Regions, Genetic; D014170:Transformation, Genetic
D002934:Cinnamates; D006921:Hygromycin B; C026273:hygromycin A; D010088:Oxidoreductases; D005954:Glucosephosphate Dehydrogenase; C038370:isopenicillin N synthetase; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase