Cloning of the Aspergillus oryzae 5-aminolevulinate synthase gene and its use as a selectable marker.
The hemA gene encoding 5-aminolevulinate synthase, the first enzyme in heme biosynthesis, was cloned from Aspergillus oryzae and evaluated as a selectable marker for the transformation of filamentous fungi. Deletion of the hemA gene resulted in a lethal phenotype that could be rescued either by the supplementation of culture media with 5-aminolevulinic acid (ALA) or by transformation with the wild-type hemA gene, but not by growth on rich media, nor by the addition of exogenous heme. Transformation of a hemA deletion strain with the hemA gene linked to a lipase expression cassette yielded ALA prototrophs expressing lipase. The hemA gene can therefore be used as a selectable marker for the transformation of A. oryzae.
Elrod
S L
SL
Novo Nordisk Biotech Inc, Davis, CA 95616, USA. selrod@calpoly.edu
Jones
A
A
Berka
R M
RM
Cherry
J R
JR
D000624:5-Aminolevulinate Synthetase / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular; D017353:Gene Deletion; D005819:Genetic Markers; D006801:Humans; D008049:Lipase / Q000235:genetics; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010641:Phenotype; D010957:Plasmids; D011994:Recombinant Proteins / Q000737:chemistry; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D005819:Genetic Markers; D011994:Recombinant Proteins; D000624:5-Aminolevulinate Synthetase; D008049:Lipase