Cloning and expression of a chitinase gene from the hyperparasitic fungus Aphanocladium album

Curr Genet. 1992 Jan;21(1):61-6. doi: 10.1007/BF00318656.

Abstract

Recombinant clones from a cDNA library of an Aphanocladium album chitinase-overproducing mutant strain were isolated by screening with antiserum against a 39 kDa chitinase purified from this hyperparasitic fungus. Analysis of the isolated positive clones indicated that most of them carried the same cDNA. A cDNA from this group was used as a hybridization probe to isolate an 8 kb DNA fragment from a genomic library of the wild-type strain. The chitinase 1 gene was mapped to this fragment by two independent approaches. Its partial DNA sequence was in perfect agreement with an amino-terminal peptide sequence obtained by sequencing 23 amino acids of the 39 kDa chitinase. Its transfer in Fusarium oxysporum resulted in a transformant producing both a protein of about 39 kDa that cross-reacted with the chitinase antiserum and a chitinase activity that was inhibited by the same antiserum. Northern blot analysis indicates that the cloned chitinase gene was subject to catabolite repression and appeared inducible by chitin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Chitin / pharmacology
  • Chitinases / genetics*
  • Cloning, Molecular
  • Fusarium / genetics
  • Gene Expression Regulation, Fungal
  • Gene Library
  • Genes, Fungal*
  • Glucose / pharmacology
  • Mitosporic Fungi / enzymology
  • Mitosporic Fungi / genetics*
  • Molecular Sequence Data
  • Transformation, Genetic

Substances

  • Chitin
  • Chitinases
  • Glucose

Associated data

  • GENBANK/M83092
  • GENBANK/S47168
  • GENBANK/S49998
  • GENBANK/S50001
  • GENBANK/S72771
  • GENBANK/S81303
  • GENBANK/X62697
  • GENBANK/X64094
  • GENBANK/X64095
  • GENBANK/X64096