Expression of tyrosinase in vegetative cultures of Neurospora crassa transformed with a metallothionein promoter/protyrosinase fusion gene

Curr Genet. 1990 Nov;18(4):331-5. doi: 10.1007/BF00318214.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular
  • Copper
  • DNA, Fungal / analysis
  • Enzyme Induction
  • Enzyme Precursors / genetics
  • Gene Expression Regulation, Fungal*
  • Genes, Fungal*
  • Melanins / biosynthesis
  • Metallothionein / genetics
  • Monophenol Monooxygenase / genetics*
  • Neurospora crassa / genetics*
  • Nucleic Acid Hybridization
  • Plasmids
  • Promoter Regions, Genetic
  • Recombinant Fusion Proteins / genetics*
  • Transformation, Genetic

Substances

  • DNA, Fungal
  • Enzyme Precursors
  • Melanins
  • Recombinant Fusion Proteins
  • Copper
  • Metallothionein
  • Monophenol Monooxygenase