Cloning of an Aspergillus niger invertase gene by expression in Trichoderma reesei

Curr Genet. Jul-Aug 1993;24(1-2):53-9. doi: 10.1007/BF00324665.

Abstract

The filamentous fungus Aspergillus niger produces two glycosylated forms of the sucrose-hydrolysing enzyme, invertase. In contrast, some Trichoderma species lack invertase and are unable to utilise sucrose as a sole carbon source. Using an A. niger genomic library constructed in a cosmid vector containing the ura5 gene of Podospora anserina as a selectable marker, and the T. reesei ura5- strain as a sucrose-minus recipient strain, an A. niger invertase gene (suc1) has been cloned by a sib selection procedure. PAGE and enzyme analysis confirmed that transformants had acquired invertase activity. The cloned gene contained DNA sequences which were complementary to the amino-acid sequences of tryptic peptides found in invertase purified from A. niger. The suc1 invertase gene can be used as a dominant selectable marker for the transformation of Trichoderma strains.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aspergillus niger / enzymology
  • Aspergillus niger / genetics*
  • Cloning, Molecular
  • Cosmids
  • Gene Library
  • Genes, Fungal
  • Genetic Complementation Test
  • Genetic Markers
  • Glycoside Hydrolases / biosynthesis
  • Glycoside Hydrolases / genetics*
  • Restriction Mapping
  • Sucrose / metabolism
  • Transformation, Genetic
  • Trichoderma / genetics*
  • Trichoderma / growth & development
  • beta-Fructofuranosidase

Substances

  • Genetic Markers
  • Sucrose
  • Glycoside Hydrolases
  • beta-Fructofuranosidase