Expression of tyrosinase in vegetative cultures of Neurospora crassa transformed with a metallothionein promoter/protyrosinase fusion gene.
Wild-type Neurospora crassa, strain Singapore, was transformed with a N. crassa metallothionein promoter/protyrosinase fusion gene. Transformants produced tyrosinase during vegetative growth, as determined by Western analyses and activity assays. This is in sharp contrast to wild-type strains, where this enzyme is only expressed in situations of starvation or sexual differentiation. Complete integration of a 400 bp metallothionein promoter-fragment leads to constitutive expression of protyrosinase, whereas a 3.6 kb promoter-fragment conferred copper inducibility on the reporter gene in four transformants. A transformant with high constitutive tyrosinase levels was able to produce melanin on complete medium agar plates supplemented with 1 mg/ml L-tyrosine.
Kupper
U
U
Biochemisches Institut der Universität Zürich, Switzerland.
Linden
M
M
Cao
K Z
KZ
Lerch
K
K
D003001:Cloning, Molecular; D003300:Copper; D004271:DNA, Fungal / Q000032:analysis; D004790:Enzyme Induction; D004792:Enzyme Precursors / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D008543:Melanins / Q000096:biosynthesis; D008668:Metallothionein / Q000235:genetics; D014442:Monophenol Monooxygenase / Q000235:genetics*; D009492:Neurospora crassa / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D014170:Transformation, Genetic
D004271:DNA, Fungal; D004792:Enzyme Precursors; D008543:Melanins; D011993:Recombinant Fusion Proteins; D003300:Copper; D008668:Metallothionein; D014442:Monophenol Monooxygenase