Expression of a bacterial aspartase gene in Aspergillus nidulans: an efficient system for selecting multicopy transformants.
The Escherichia coli aspartase gene aspA has been expressed in the fungus Aspergillus nidulans using the powerful constitutive gpdA promoter and trpC terminator, both from A. nidulans. Multiple, but not single, copies of aspA overcome nutritional deficiencies resulting from the loss of catabolic NAD-linked glutamate dehydrogenase. They also circumvent certain nutritional deficiencies resulting from loss of the positive-acting regulatory gene product mediating nitrogen metabolite repression. Both of these cases of physiological suppression involve the aspartase-catalyzed catabolism of aspartate to ammonium plus fumarate. No physiological evidence for the opposite reaction leading to aspartate synthesis was obtained as multiple copies of aspA did not affect the phenotype resulting from the loss of anabolic NADP-linked glutamate dehydrogenase. The use of vectors containing aspA and recipients lacking NAD-linked glutamate dehydrogenase is an efficient means of selecting multicopy transformants in A. nidulans and also offers the possibility to select strains having increased aspartase levels from original transformants.
Hunter
G D
GD
Department of Infectious Diseases and Bacteriology, Royal Postgraduate Medical School, London, England.
Bailey
C R
CR
Arst
H N
HN
D001220:Aspartate Ammonia-Lyase / Q000235:genetics* / Q000378:metabolism; D001233:Aspergillus nidulans / Q000235:genetics* / Q000302:isolation & purification; D015139:Blotting, Southern; D004271:DNA, Fungal; D004926:Escherichia coli / Q000201:enzymology*; D008040:Genetic Linkage; D005819:Genetic Markers; D005969:Glutamate Dehydrogenase / Q000235:genetics; D009243:NAD / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D013728:Terminator Regions, Genetic; D014170:Transformation, Genetic
D004271:DNA, Fungal; D005819:Genetic Markers; D009243:NAD; D005969:Glutamate Dehydrogenase; D001220:Aspartate Ammonia-Lyase